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Objective: To study the changes of morphology and function in rat hippocampus induced by high power microwave (HPM) radiation., Methods: Fifty male Wistar rats were radiated by HPM. Then their learning and memory abilities were tested with Y maze and were sacrificed 6 h, 1 d, 3 d and 7 d after radiation. The hippocampus was taken out to study the basic pathologic changes, apoptosis and the expressions of neuron-specific enolase (NSE) and glial fibrillary acidic protein (GFAP) by means of HE staining, Nissel body staining, in situ terminal end labeling and immunohistochemistry., Results: The learning and memory abilities of rats reduced significantly after HPM radiation. HPM also resulted in rarefaction, edema and hemangiectasia of hippocampus, nervous cells degeneration and necrosis, decrease or disappearance of Nissel bodies. The injuries were more serious in field CA4 and dentate gyrus, which showed dose-effect relationship, and were progressively aggravated within 7 days. The apoptosis cells were significantly increased. NSE was increased in neurons. The NSE positive areas were also seen in the interstitial matrix and blood vessels. GFAP was increased in astrocytes, which became shorter and thicker., Conclusion: HPM can damage the abilities of learning and memory and results in morphologic changes in hippocampus. The major pathologic changes are degeneration, apoptosis and necrosis of neurons and edema in interstitium. NSE and GFAP play an important role in the pathologic process.

Purpose: This retrospective study was performed to analyze the causes, 3D spiral CT examination and the treatment of ZMC fractures., Methods: Rigid internal fixation was used in 25 patients with ZMC fractures after 3D spiral CT examination and anatomical reduction. The patients were followed up for 3 to 12 months., Results: In the 25 cases, 13 cases had traffic accident injury, 5 had industrial accident injury. Local collapsed teratism were found in 21 cases,malocclusion was noted in 19 cases, limitation of mouth opening was found in 15 cases, diplopia was in 17 cases, infraorbital numbness in 11 cases, 5 cases had sagittal maxillary fracture. Spiral 3D CT images showed the steric anatomical images and relationship to the adjacent structures clearly in 25 patients. All patients were treated via rigid internal fixation with titanium plates and pins. 22 patients had satisfied maxillofacial appearance., Conclusion: Using topical, coronal and vestibular incisions accordingly, and rigid internal fixation with titanium plates, satisfied outcomes can be achieved in patients with ZMC fractures.

Tetrandrine (TET) is a well known naturally occurred nonspecific Ca(2+) channel blocker. It has long been used for the treatment of arrhythmia, hypertension, and occlusive cardiovascular disorders. The objective of the present study was to investigate the effect of TET on the proliferation of primary cultured rat aortic smooth muscle cells (RASMCs). TET significantly inhibited both 10% fetal bovine serum (FBS) and 50 ng/ml platelet-derived growth factor (PDGF)-BB-induced proliferation, [(3) H] ]thymidine incorporation into DNA, and p42/p44 mitogen-activated protein kinase (ERK1/2) phosphorylation at the concentration of 1.0 and 5.0 microM. Flow cytometry analysis of DNA content in synchronized cells revealed blocking of the FBS-inducible cell cycle progression by TET. In accordance with these findings, TET 5 microM caused a 48% decrease in the early elevation of c-fos expression induced after 10% FBS addition. Furthermore, in contrast to its distinguishable higher potency of Ca(2+) antagonistic activity, verapamil showed lower potent antiproliferative activities than TET. These results suggest that TET can exert antiproliferative effects against mitogenic stimuli for RASMCs in vitro by a mechanism that involves the MAPK pathway, altering cell cycle progression, and the inhibitory action cannot be limited to its Ca(2+) modulation.

Objective: To evaluate the clinical effect of abdominal multiorgan transplantation in patients with multiorgan failure., Methods: Simultaneous kidney-pancreas transplantation (SKPT) with enteric drainage of pancreatic exocrine secretions was performed in 2 patients with type 1 diabetes and end-stage renal disease. A combined liver-kidney transplantation (CLKT) was done in a 66-year-old patient with alcoholic liver cirrhosis and uremia. Simultaneous orthotopic liver and heterotopic pancreas-duodenum transplantation (SLPT) was performed in a patient with hepatitis B, hepatocirrhosis, hepatic cellular cancer, and type 1 diabetes., Results: The function of kidney grafts became normal 5 days postoperatively and insulin-independent after treatment with low dose insulin for 10 days in the 2 SKPT patients. For the CLKT patient, both transplanted organs rapidly achieved normal functions after operation but suffered-acute liver graft rejection on postoperation day 10 and the rejection was controlled after methylprednisolone pulse therapy. In the SLPT patient, insulin was withdrawn 5 days after operation, liver allograft function recovered well. All the patients are alive with stable allograft function after following-up for 29, 26, 9 and 6 months, respectively., Conclusions: Abdominal multi organ transplantation was effective therapy to patients with multiple organ failure. SLPT can reduce acute pancreas rejection and promote the recovery of liver allograft.

The regeneration of motor and sensory neurons and the morphological changes of the target muscle after phrenic nerve transfer were investigated in adult rats. Six months following nerve transfer, 326.0 +/- 16.31 phrenic motoneurons regenerated into musculocutaneous nerve, which is not different from the normal number of phrenic motoneurons. The regenerated motoneurons exhibited a 14% nonsignificant hypertrophy. Of the dorsal root ganglia (DRG) neurons, 255.8 +/- 45.26 regenerated, which was significantly lower than the number of normal phrenic DRG neurons. The regenerated phrenic DRG neurons showed a 24% close-to-significant atrophy. The target muscle fiber morphology changed considerably after reinnervation. The present results suggest that the phrenic nerve has very good regenerative ability in terms of its motoneurons and a relatively insufficient sensory neuronal regeneration., (Copyright 2004 Wiley-Liss, Inc.)

Objective: To study the effect of combined transplantation of the liver and the pancreas in diabetic patients with end-stage liver disease, and explore the optimal surgical procedure., Methods: Simultaneous orthotopic liver and heterotopic pancreas-duodenum transplantations were performed in a patient diagnosed as having chronic hepatitis B, hepatocirrhosis, hepatic cellular cancer, and insulin-dependent diabetes. Immunosuppression therapy utilized prednisone, tacrolimus (FK506), mycophenolate mofetil (MMF), and simulect. The function of the liver graft, serum amylase and lipase, blood glucose, and C-peptide were monitored after transplantation., Results: Insulin was withdrawn at the 6th day after operation, good liver allograft functional recovery was achieved, without such complications as pancreatitis, thrombosis, and localized infections., Conclusion: End-stage liver disease with concomitant insulin-dependent diabetes is the indication for combined liver-pancreas transplantations, for which simultaneous orthotopic liver and heterotopic pancreas-duodenum transplantations may constitute the optimal surgical approaches as the primary choice.

Objective: To compare the efficacy and safety of tacrolimus (FK506)- versus cyclosporine (CsA)-based immunosuppression protocol in the treatment of patients with renal transplantation for diabetic end-stage renal disease., Methods: A total of 64 patients with end-stage renal disease were randomized into FK506 (n=33) and CsA (n=31) group after cadaveric renal transplantation, the former group adopting small-dose FK506 therapy with low trough concentration, and the latter receiving CsA therapy. The dose of insulin was adjusted according to blood glucose level of the patients after the operation. The incidence of acute rejections, blood glucose level, the dose of insulin and changes in blood pressure, lipid metabolism and liver function were compared between the two groups., Results: The 1-year patient/graft survival rates were 96.97%/93.94% in FK506 group and 96.77%/90.32% in CsA group. Four patients (12.12%) developed acute rejection in the FK506 group, while 11 (35.48%) did in the CsA group during the first year after the operation (P<0.05). The dose of the insulin of the FK506 group (34.35 U/d, 14.09 U/d) was not significantly different from that in the CsA group (28.15 U/d, 13.05 U/d) at the first month and 1 year after the operation (P>0.05). One year after the operation, 21 patients (63.63%) required anti-hypertension treatment, 5 (15.15%) needed anti-hyperlipidemia treatment and 3 (9.09%) had abnormal liver function in FK506 group, while in the CsA group, 28 patients (90.32%) needed anti-hypertension therapy, 13 (41.94%) received anti-hyperlipidemia treatment and 11 (35.48%) showed signs of abnormal liver function, with significant differences between the two groups., Conclusions: Small-dose FK506 therapy with low concentrations is effective in the postoperative management of patients with renal transplantation for diabetic end-stage renal disease, with only mild adverse effect. FK506 is comparable with CsA in terms of the effect on glucose metabolism of the patients.

Objective: To study the surgical techniques, perioperative management, management of infections and graft rejection in patients with combined liver-kidney transplantation (CLKT)., Methods: CLKT was performed in a 66-year-old patient with alcoholic liver cirrhosis and uremia. Lavage in situ with University of Wisconsin (UW) solution of the donor organs and en hoc resection was performed. Orthotopic liver transplantation (OLT) and routine kidney transplantation were respectively carried out. Immunosuppression therapy consisted of tacrolimus (FK506), antithymocyte globulin (ATG), mycophenolate mofetil (CellCept, MMF) and corticosteroid., Results: Both of the transplanted organs rapidly recovered normal functions after operation, and acute rejection of the liver graft occurred on day 10 after operation but was controlled after methylprednisolone pulse therapy. The patient fully recovered and was discharged from hospital on day 29 after operation., Conclusions: CLKT is effective against both liver and renal function failure. Well-matched HLA tissue typing, proficient surgical skills, adequate application of immunosuppressants and effective management of postoperative complications are crucial for successful CLKT.

Objective: To investigate the effect of anti-tumor necrosis factor-alpha monoclonal antibody (anti-TNF-alpha mAb) in alleviating renal ischemia-reperfusion injury., Methods: Fifty normal male Sprague-Dawley rats were randomly divided into 3 groups, namely group A that was subjected to ischemia-reperfusion injury with intravenous administration of anti-TNF-alpha mAb (0.1mg/kg.b.w.) 5 min before reperfusion (treatment group), group B with the same injury followed by saline administration in the same manner (control group), and group C with only anesthetization and leparotomy but not ischemia (sham operation group). Routine assays were performed for testing the levels of blood creatine (Cr), blood urea nitrogen (BUN), plasma TNF-alpha and the cell apoptosis. Ultrastructure of the kidney was also observed., Results: Renal ischemia-reperfusion resulted in significant increase of the levels of Cr, BUN and TNF-alpha in the plasma (P<0.01), but these effects were offset by administration of anti-TNF-alpha mAb (P<0.01). In group B, widespread pathological changes and cell apoptosis were observed in the renal tissue following renal ischemia-reperfusion injury, while similar changes were scarcely visible in group A due to the protective effect of intravenous administration of anti-TNF-alpha mAb 5 min before reperfusion., Conclusion: Renal ischemia-reperfusion injury can be alleviated by anti-TNF-alpha mAb treatment.

Objective: To evaluate the experiences, attitudes, and perceptions of the caregivers of children with cataracts who were visually rehabilitated with contact lenses., Participants: One hundred twenty-three caregivers of children <8.1 years old treated for unilateral and bilateral cataracts at one pediatric hospital., Design: Survey by questionnaire., Intervention: Primary caregivers were asked to complete an anonymous questionnaire., Main Outcome Measures: Caregiver responses to questions assessing background and demographic and clinical information, as well as perceptions, attitudes, levels of compliance, and anxiety with respect to treatment, were reviewed. Caregivers were also asked to choose between aphakic rehabilitation with contact lenses, aphakic glasses, or intraocular lenses, given various hypothetical scenarios differing in regard to their final visual prognosis, risks of treatment complications, and cost., Results: The response rate was 82.9%. Absolute average stress levels for contact lens use were 1.36 +/- 1.79 and 0.79 +/- 1.48 (scale, 0-5) for insertion and removal, respectively, compared with 4.03 +/- 1.64 and 2.40 +/- 1.92 for cataract surgery and patching therapy, respectively. Although average paired initial resistance to treatment (RT) levels for contact lens insertion and removal on a scale of 0 to 3 were high (2.09 +/- 1.15) and moderate (1.63 +/- 1.20), respectively, final RT levels were significantly lower (1.09 +/- 1.14 and 0.66 +/- 1.07, respectively; P < 0.0001). The vast majority of caregivers chose contact lens use in hypothetical scenarios that depicted realistic expectations for other forms of aphakic rehabilitation., Conclusions: In our study, contact lenses seemed to be well tolerated by most patients, as assessed by caregivers. Although initial resistance to contact lens use is high, this decreases with time. Relative to other events in the treatment of pediatric cataracts, contact lens use is not a major stressor for most caregivers and patients. This study supports the notion that contact lenses should continue to receive serious consideration as a treatment option for pediatric cataracts.

Objective: To evaluate the role of panel reactive antibody (PRA) screening and human leukocyte antigen (HLA) typing in renal transplantation., Methods: PRA screening and HLA typing were performed in 1 700 patients eligible for the first group of renal transplantation who had 3 to 6 HLA matches in HLA-A, B and DR with the donor, and in cases positive for PRA, plasma exchange was conducted. Another 423 patients who did not receive PRA screening or HLA typing constituted the second group. The changes of immune variables, incidences of acute rejection and the effect of HLA-A, B, DR matching on long-term graft survival were observed., Results: In 1 700 cases of group 1, post-transplantation CsA dose was reduced to 5 to 7 mg*kg(-1)*d(-1) and the graft function recovery time ranged from 2 to 16 d, averaging 5 d. Acute graft rejection occurred in 252 (14.8%) cases, but no hyper-acute rejection was observed. The 1-, 3- and 5-year patient/graft survival rates were 98.6%/96.7%, 93.1%/87.3% and 88.1%/83.6% respectively. In group 2, CsA dose ranged from 8 to 12 mg*kg(-1)*d(-1) and the graft function recovery time was 4 to 30 d, averaging 13 d. Hyper-acute rejection occurred in 9 (2.1%) and acute rejection in 198 (46.8%) cases, and the 1-, 3- and 5-year patient/graft survival rates were 86.7%/76.3%, 72.5%/67.9% and 69.5%/59.3% respectively., Conclusions: Negative PRA and good HLA matching can eliminate the incidences of hyper-acute rejection, decrease the rate of acute rejection and improve both patient and graft survival rates.

Corneal ulceration is a significant cause of visual morbidity. Although this discussion has primarily focused on the local factors involved in non-immune related sterile corneal ulcerations, an understanding of these mechanisms is important since a final common pathway that is conceivably relevant to all forms of corneal ulceration exists. With respect to the treatment of chronic sterile ulcerations, our contemporary armamentarium is extensive; an acknowledgement to the inadequacies of any single strategy. A sound appreciation of the subtleties involved in the pathogenesis of corneal ulceration and the orchestration of wound repair will lead us towards more effective strategies to decrease inflammation associated with ulceration, promote corneal wound healing, and ultimately provide better care for our patients.

Objective: The purpose of this article is to define and illustrate the radiologic findings in patients with enterohemorrhagic Escherichia coli colitis., Conclusion: Although not definitive, imaging studies in conjunction with an appropriate clinical history can aid in the early diagnosis of E. coli colitis and exclude surgical conditions. CT is more sensitive than conventional radiography for detection. Contiguous involvement, including the transverse colon, was seen in all patients. Because CT is becoming routine in the initial workup of patients with acute abdominal pain, it is important for the radiologist to suggest E. coli colitis in the proper setting.

Human amniotic membrane (AM) is composed of three layers: a single epithelial layer, a thick basement membrane, and an avascular stroma. Amniotic membrane has anti-adhesive properties and is felt to promote epithelialization and decrease inflammation, neovascularization, and fibrosis. Amniotic membrane transplantation (AMT) is currently being used for a continuously widening spectrum of ophthalmic indications. Amniotic membrane transplantation has been shown to be effective in the reconstruction of the corneal surface in the setting of persistent epithelial defects, sterile corneal ulcerations, and partial limbal stem cell (LSC) deficiency states, including those secondary to chemical or thermal burns. Amniotic membrane transplantation also has been used in conjunction with limbal stem cell transplantation (LSCT) both in a concurrent fashion as well as in preparation for LSCT. Amniotic membrane transplantation also has been used in place of conjunctival autografting after pterygium excision and to reconstruct the conjunctival surface after removal of conjunctival lesions. Most recently, ex vivo cultivation and expansion of limbal epithelial cells has been performed utilizing AM as a matrix. However, the superiority of AMT over other treatment modalities in many of these settings needs to be substantiated by controlled clinical trials.

Production of nitric oxide (NO) by inducible nitric oxide synthase (iNOS) has been proposed as a pathogenic factor in acute pancreatitis, but its role has still not been fully examined. The present study explored the role of iNOS in cerulein-induced acute pancreatitis using iNOS-deficient mice. Twelve- to 14-week-old male mice (C57B1/6 and iNOS-deficient) were administered cerulein by intraperitoneal (i.p.) injection at hourly intervals for 7 hours and killed 24 hours later after the first dose. Pancreatic wet weight, pancreatic myeloperoxidase (MPO) activity, and levels of plasma nitrite and serum amylase were measured. In another experiment isosorbide dinitrate (an NO donor) was given by oral gavage every 6 hours for 24 hours beginning simultaneously with cerulein injections in iNOS-deficient mice. Cerulein administration dose-dependently increased pancreatic wet weight, myeloperoxidase activity, and levels of nitrite and amylase in C57B1/6 mice. These parameters (except nitrite levels) were significantly intensified in iNOS-deficient mice. At the dose employed, cerulein failed to increase nitrite levels in iNOS-deficient mice. The susceptibility to cerulein toxicity in iNOS-deficient mice was abolished by NO donor treatment. NO release from an iNOS source appears to play a protective role in cerulein-induced pancreatitis. At least in part, NO may prevent neutrophil accumulation after cerulein administration.

Choroidal neovascularization in age-related macular degeneration is a frequent and poorly treatable cause of vision loss in elderly Caucasians. This choroidal neovascularization has been associated with the expression of vascular endothelial growth factor (VEGF). In current animal models choroidal neovascularization is induced by subretinal injection of growth factors or vectors encoding growth factors such as VEGF, or by disruption of the Bruch's membrane/retinal pigment epithelium complex with laser treatment. We wished to establish a transgenic murine model of age-related macular degeneration, in which the overexpression of VEGF by the retinal pigment epithelium induces choroidal neovascularization. A construct consisting of a tissue-specific murine retinal pigment epithelium promoter (RPE(65) promoter) coupled to murine VEGF(164) cDNA with a rabbit beta-globin-3' UTR was introduced into the genome of albino mice. Transgene mRNA was expressed in the retinal pigment epithelium at all ages peaking at 4 months. The expression of VEGF protein was increased in both the retinal pigment epithelium and choroid. An increase of intravascular adherent leukocytes and vessel leakage was observed. Histopathology revealed intrachoroidal neovascularization that did not penetrate through an intact Bruch's membrane. These results support the hypothesis that additional insults to the integrity of Bruch's membrane are required to induce growth of choroidal vessels into the subretinal space as seen in age-related macular degeneration. This model may be useful to screen for inhibitors of choroidal vessel growth.

Nicotine intensifies experimental gastric ulceration by reducing gastric mucosal blood flow (GMBF) and mucus. As both these parameters can be improved by nitric oxide (NO), we evaluated the impact of a NO donor in ethanol-induced gastric mucosal injury in rats administered nicotine. A nicotine solution or water was administered for 20 days to Sprague-Dawley rats. NO donor (isosorbide dinitrate) was given 60 and 10 min before preparation of ex vivo gastric chambers and exposure to ethanol. Chronic nicotine intake significantly reduced GMBF and gastric mucus content. Nicotine intensifies ethanol-induced gastric injury and short-term administration of NO donor failed to antagonize the ulcerogenic action from either nicotine or alcohol. In another study, rats drank nicotine solution for 20 days, after which the nicotine was withdrawn and replaced by water for 10 additional days. NO donor was provided during these last 10 days. The gastric effects of nicotine persisted for at least 10 days after nicotine was withdrawn but then these effects could be abolished by prolonged NO treatment. Nicotine reduces plasma nitrite level, but gastric mucosal MPO activity remained unchanged. Our data suggest that nicotine cessation plus a longer period of NO donor administration can completely abolish the gastric effects of nicotine., (Copyright 2001 S. Karger AG, Basel)

Invasive aspergillosis is a rare but important cause of morbidity and mortality in patients with AIDS. In the majority of patients, invasive aspergillosis involves the lungs. Extrapulmonary involvement with total splenic infarction has not been previously described. We illustrate a patient who developed total splenic infarction and septic emboli to the brain from Aspergillus.

Invasive aspergillosis is a rare but important cause of morbidity and mortality in patients with AIDS. In the majority of patients, invasive aspergillosis involves the lungs. Extrapulmonary involvement with total splenic infarction has not been previously described. We illustrate a patient who developed total splenic infarction and septic emboli to the brain from Aspergillus.

We studied genetic polymorphisms in the tumor necrosis factor (TNF) region as well as HLA-DRB1 of 42 patients with Western-type multiple sclerosis (MS) and 38 with Asian-type MS in Japan. The sex ratio (Female:Male) was significantly higher in Asian than in Western type MS (3.8 vs. 1.3, P = 0.038). The frequency of HLA-DRB1 * 1501 allele in the Western-type MS group increased significantly compared with the control group, while Asian-type MS and control groups showed similar distribution in the frequencies of HLA-DRB1 alleles. No significant differences existed in the TNF region, however, including TNF-a microsatellite alleles. The results suggest that MS in Asians may present two different clinical and immunogenetic manifestations.

We studied genetic polymorphisms in the tumor necrosis factor (TNF) region in 81 Japanese patients with Guillain-Barré syndrome (GBS) and 85 controls. A significantly higher frequency of the 100-base pair (bp) (TNFa2) allele of the TNFa microsatellite marker, which is associated with high TNF alpha production, existed in Campylobacter jejuni-positive (Cj+) GBS patients than in controls, suggesting the involvement of a genetic predisposition to high TNF alpha secretion in the development of C. jejuni-related GBS.

Objective: We examined a possible involvement of genetic factors influencing the development of Guillain-Barré syndrome (GBS)., Methods: We studied T-cell receptor (TCR), alpha-chain constant (AC), and beta-chain variable (BV) gene polymorphisms using microsatellite markers and serologic HLA class I antigens, HLA-DRB1, and HLA-DQB1 alleles in 81 Japanese patients with GBS and 87 controls., Results: There were no significant differences in these genetic markers between GBS patients and controls. Subgrouping of GBS patients according to recent Campylobacter jejuni infection, the presence of anti-GM1 antibody in the sera, or their combinations also failed to reveal significant associations with these genetic markers. There was, however, a tendency for an increased frequency of HLA-DRB1*0803 in the C. jejuni + GM1 + GBS group, when compared with controls., Conclusions: The data suggest that the roles of TCRAC, T-cell receptor beta-chain variable (TCRBV), HLA class I or class II in the development of GBS are not critical, and further research is necessary to clarify other genes encoded within the HLA region for genetic susceptibility to GBS.

The effects of 5-hydroxytryptamine (5-HT) on ethanol-induced gastric mucosal damage and on epithelial and vascular integrity were investigated. Male Sprague-Dawley rats were administered with 5-HT (5 or 10 mg/kg, IP) 30 min prior to the challenge with ethanol (40% v/v, 10 ml/kg, PO). 5-HT dose dependently aggravated ethanol-induced injury in the gastric mucosa. Both xanthine oxidase (XO) and myeloperoxidase (MPO) activities in the mucosa were significantly increased with the high dose of 5-HT, which also potentiated the elevation of these enzyme activities by ethanol. However, the mucosal superoxide dismutase activity was left unaltered. In neutropenic (antineutrophil serum-treated) animals, the ethanol-induced gastric mucosal injury was significantly ameliorated, with or without the pretreatment of 5-HT (10 mg/kg). In addition, the effect of 5-HT on the activity of MPO, but not of XO, was also attenuated in these animals. In the ex vivo gastric chamber study on pentobarbital-anesthetized animals, volume of gastric secretion was significantly decreased in the 5-HT-treated groups, with further reduction after ethanol incubation. Transmucosal potential difference (PD) was significantly reduced in 5-HT-treated rats, which also potentiated the ethanol-induced drop in PD. Nevertheless, 5-HT dose dependently increased mucosal vascular permeability and further enhanced during ethanol incubation. These findings suggest that 5-HT adversely affects the defense mechanisms of the gastric mucosa by reducing the secretory function of the mucosal cells and to weaken the epithelial and vascular integrity. Neutrophil activation appears to be responsible for the detrimental effects of 5-HT partly through the elevation in MPO activity. The increase in mucosal XO activity by 5-HT may induce free radical production and possibly modulate the ulcerogenic processes.

Forgetting during recall may be one limit on memory span. Output time and accuracy of immediate serial recall using spoken and keypress responses were measured for digit, letter, and word sets approximately matched in phonemic discriminability and in immediate recognition memory. Nonetheless, the materials differed from one another in recall span, in output time during recall, and in pronunciation time (speech rate). Recall output times accounted precisely and completely for the measured memory span for these matched materials. Pronunciation times are correlated with recall output times, but output time gives a slightly better account of recall accuracy. The output time equivalent to the rule that short-term memory span corresponds to the number of items that can be said in about 1.5-2 s is that span corresponds to the number of items that can be recalled in about 4-6 s. Additional variations in span reflect differential item interference.

Campylobacter jejuni is a major pathogen preceding Guillain-Barré syndrome (GBS), and most C. jejuni isolates from GBS patients belong to Penner serotype 19 (heat-stable; HS-19). We analyzed sixteen independent clinical isolates from GBS patients, twelve of which belonged to HS-19, three to HS-2, and one to HS-4, using PCR-based RFLP analysis of a flagellin-A (flaA) gene. Two isolates from patients with Miller Fisher syndrome (MFS), and 27 from patients with uncomplicated enteritis were also examined. All HS-19 isolates, regardless of GBS, showed an identical pattern (Cj-1) by RFLP typing and were distinguishable from those of the other Penner serogroups. In contrast, HS-2 and HS-4 isolates were divided into several different RFLP groups, suggesting HS-19 strains are genetically distinctive among C. jejuni isolates. A DNA fingerprinting method also failed to detect any specific band pattern for GBS-related C. jejuni isolates. We examined relationships among anti-GM1 antibody titres in the sera of GBS patients, clinical forms of GBS, serotype of C. jejuni, and the presence of GM1-like structures in lipopolysaccharide (LPS) components from C. jejuni isolates by immunoblotting. HS-19 related GBS was significantly associated with elevated anti-GM1 antibody titers in the sera of the patients, but not associated with any clinical pattern of GBS. No significant correlations were found between anti-GM1 antibody and the pattern of disease, or between GBS-related C. jejuni strains and the presence of GM1-like structures. HS-19 strains seem to be unique among C. jejuni isolates, and HS-19-related GBS may provide an excellent model for clarification of the pathogenesis of GBS.

There are now six recognized neuropeptide Y (NPY) receptor subtypes (Y1-Y4 and two recently cloned distinct receptors labeled Y5), of which Y1 and one of the Y5's have been suggested could mediate the effect of NPY on feeding. The fragments NPY(2-36) and NPY(3-36), which bind Y1 only poorly, were injected intracerebroventricularly (icv) and found to have similar dose-response relationships to NPY in the stimulation of feeding. However NPY (13-36), which stimulates both Y2 and Y5, caused no increase in food intake, even at high doses. Maximal stimulation with the classical Y1 agonist [Pro34]-NPY produced only 50% of the maximum effect of NPY itself despite fully inhibiting adenylyl cyclase activity in vitro in a Y1 system. The novel fragment [Pro34]-NPY(3-36) is as effective at stimulating food intake as the classical Y1 analogue [Pro34]-NPY but bound to the Y1 receptor with only 1/20th of the affinity of NPY and failed to inhibit adenylyl cyclase through this receptor. [Pro34]-NPY(3-36) is therefore a relatively appetite-selective ligand. Coadministration of high dose NPY(13-36) and [Pro34]NPY did not enhance feeding compared with [Pro34]-NPY alone. In addition, the NPY Y1 receptor antagonist BIBP-3226, which does not bind Y2, Y4, or Y5 receptors, significantly reduced NPY induced feeding. These results indicate that the feeding effect of icv NPY involves a novel receptor and that it is functionally distinct from the recognized receptor subtypes.

IL-6 is a multifunctional cytokine involved in differentiation and proliferation of immune cells. Moreover, it has diverse effects on the proliferation of tumor cells in vivo and in vitro. Although stimulating cell growth of multiple myeloma cells, it inhibits the proliferation of B16 melanoma cells and lung cancer cells. B9.55 cells, B-cell lymphoma, are IL-6-dependent cells, definitely requiring exogenous IL-6 for growth. When the cDNA for IL-6 was transfected into B9.55 cells, they began growing in an autocrine pattern without exogenous IL-6. To investigate the effects of IL-6 on B9.55 lymphoma in vivo, IL-6-transfected B9.55 cells (B9.G7) or neotransfected B9.55 cells (B9.vec) were injected subcutaneously into syngeneic mice. Initially, B9.G7 outgrew B9.vec, but after 3 weeks, B9.G7 grew slower than B9.vec. In addition, 5 micro g of recombinant human IL-6 was injected daily into the tumor site. Reduced tumor sizes of IL-6-treated rats, similar to those observed in mice which received B9.G7, indicated that IL-6 itself is the mediator of tumor regression. When B9.G7 cells were injected into the irradiated normal mice, tumor regression was released compared with the untreated normal control, suggesting that radiosensitive host components were involved in the regression of B9.G7 cell growth. However, the tumor regression of B9.G7 cells was not released in SCID mice. Histologically, B9.G7 tumor demonstrated severe necrosis and apoptotic cells with infiltration of host inflammatory cells. Above data indicate that IL-6 functions as an autocrine growth factor for B9.G7 cells in vitro, but behaves as an autocrine inhibiting factor in vivo. These contrasting effects of IL-6 on tumor cells in vitro and in vivo will be facilitative in understanding the interaction of cytokines and host immune systems.

A molecular typing approach for Campylobacter jejuni was applied with restriction fragment length polymorphism (RFLP) analysis of a 702-bp PCR-amplified portion of the flagellin-A (flaA) gene. We analyzed a total of 179 strains, including 69 independent clinical isolates from diarrheic patients in Japan, 85 isolates in China, and 25 heat-stable (HS) serotype strains by Penner and Hennessy (1980) J. Clin. Microbiol. 12, 732-737). Six AfaI, seven MboI, and five HaeIII RFLPs were found in the 702-bp flaA segment from the 179 strains. Using a combination of these three enzymes, 25 separate RFLP groups were recognized. While 59 of 154 (38.3%) strains obtained in Japan and China were nontypeable by the HS antigenic scheme, all but two of 154 (98.7%) could be typed by RFLP typing. All 11 isolates of HS-19 strains, which are frequently isolated from Guillain-Barré syndrome (GBS) patients, showed an identical RFLP pattern (Cj-1), and Cj-1 consisted only of HS-19 strains. This suggests that the HS-19:Cj-1 strain is distinct among C. jejuni strains. This molecular typing method provides a rapid and reliable typing scheme for epidemiological studies of C. jejuni, and may also be useful for the analysis of C. jejuni subtypes from GBS patients.

Objective: To estimate the incidence of Wernicke-Korsakoff syndrome (WKS) before and after the introduction of thiamine enrichment of bread flour in 1991., Design: Retrospective survey of hospital records. Patient records with the diagnostic codes for Wernicke's encephalopathy (WE) or Korsakoff's psychosis (KP) were reviewed and details of acceptable cases were entered onto a data form., Setting: All 17 major public general hospitals in the Sydney area (New South Wales), between 1978 and 1993., Outcome Measures: Numbers of confirmed or probable diagnoses of WE, KP or WKS and associated deaths, patient demographic and social characteristics and alcohol intake., Results: 1,267 patients with WKS were found, with 1,012 acute cases. Although numbers of acute cases may have started to fall before 1991, numbers for the last two years were the lowest of all the 16 years (P = 0.004). Cases of KP outnumbered those of WE by about 3:1 and men outnumbered women 4:1. The peak age was 60-64 years (17%) and beer was the most commonly cited alcoholic drink (71%). The red-cell transketolase test was seldom used for diagnosis (3% of acute cases)., Conclusions: The lower number of "acute" cases in 1992 and 1993 is consistent with a preventive effect of mandatory enrichment of bread with thiamine, but is not conclusive evidence. Longer follow-up of Sydney hospitals, results of postmortem examinations and follow-up in other areas of Australia are required.

The time-course effects of portal hypertension on gastric secretory function, mucosal blood flow, vascular permeability, and ethanol-induced gastric mucosal damage were examined in anesthetized rats. Partial ligation of the portal vein effectively produced portal hypertension one to three days later but the raised pressure returned to normal on the sixth day after ligation. This time-course effect coincided with reduced pepsin secretion and mucosal blood flow and also with potentiated ethanol-induced mucosal damage during the first to third days. These effects started to tail off on the sixth day. However, gastric acid output was significantly reduced on the third day, and this was strongest on the sixth day after operation. Portal vein ligation also reduced basal vascular permeability, which was markedly potentiated after ethanol treatment. It is concluded that: (1) portal vein blood pressure changes are a time-dependent process following ligation; (2) changes in gastric mucosal blood flow (GMBF) and lesion formation are closely related to portal hypertension; (3) gastric mucosal injury is associated with vascular damage, as evidenced by increased in vascular permeability; and (4) pepsin but not acid secretion is closely related to the state of the GMBF.

The yeast Saccharomyces cerevisiae SOC8 DNA fragment was isolated as a wildtype dominant suppressor of the cdc8 mutation. The SOC8 has previously been proved to be allelic with URA6, encoding the UMP kinase in yeast cells. The protein has been purified to homogeneity. In this report, we describe the characteristics of the UMP kinase from yeast. The yeast enzyme requires a divalent cation and is active over the entire range of pH from 6 to 9.5. The enzyme can use UMP and dUMP as phosphate acceptors with high activity; to a lesser extent, it can also use IMP, GMP, dGMP, 5-iodo-dUMP, XMP, and dTMP as substrates. ATP and dATP are the best phosphate donors; the enzyme could use GTP, dGTP, dCTP, and dTTP to some degree (30-50%). CTP and UTP were poor phosphate donors for the UMP kinase reaction (10-14%). Like other monophosphate kinases, UMP kinase contains a conserved nucleotide-binding site at its N-terminus following a cysteine residue, and its enzymatic activity is inhibited by sulfhydryl inhibitors such as 5,5'-dithio-bis(2-nitrobenzoic acid) and N-ethylmaleimide. Subcellular localization studies indicate that the UMP kinase locates primarily in the cytoplasm (approximately 80%) and also in the nucleus (approximately 20%), but not in the mitochondria. These results suggest that it may exert its function in the nucleus, such as in RNA synthesis, as well as in the cytoplasm, but not in the mitochondria. The presence of UMP kinase in the nucleus might facilitate its suppression of cdc8 mutant cells, which are defective in nuclear DNA synthesis.

Twelve healthy adults and 12 epileptics had quantitative EEG tests while taking valproic acid. The results indicated that the total power of dominant frequency band and the frequency of dominant occipital rhythm were relatively stable in healthy awake adults. A single 400 mg oral dose of sodium valproate had similar influence on the total power of dominant frequency band in both healthy adults and epileptics. The total power of dominant frequency band tended to rise at 2 and 6 hours after medication in subjects with serum VPA concentrations reaching a peak after 2 hours. There was a tendency to rise at 4 and 8 hours in subjects with serum VPA concentrations which peaked after 4 hours. The curve of the total power with time had two peaks. A single oral dose of sodium valproate did not have a marked influence on the frequency of dominant rhythm in the occipital areas in healthy adults or in epileptics.

The effects of moxibustion on experimental hyperbilirubinemia and cholangitis were studied in male rats (n = 33). The experimental group were treated with moxibustion, 18, 12 and 6 hours prior to and 6, 12 and 18 hours after challenge with oral alpha-naphthylisothiocyanate (ANIT, 100 mg per kg). Rats were sacrificed 48 hours after challenge. Serum and liver samples were taken for biochemical and histological analysis, respectively. The results revealed that rats treated with ANIT exhibited elevations in bilirubin, SGOT and SGPT as well as cholangitis. In rats receiving moxibustion and ANIT, biochemical and morphological parameters of liver injury were significantly reduced. Thus, this study shows that moxibustion therapy may be able to prevent ANIT-induced hyperbilirubinemia and cholangitis.

Nucleoside-diphosphate kinase is an enzyme which catalyzes the phosphorylation of nucleoside diphosphates into the corresponding triphosphates for nucleic acid biosynthesis. In this communication, we describe the purification and characterization of nucleoside-diphosphate kinase from yeast. The purified protein appears to be homogeneous by sodium dodecyl sulfate-polyacrylamide gel analysis, with a molecular weight of about 17,000-18,000. An estimate from the fast protein liquid chromatography Superose 12 gel filtration shows a native molecular weight of about 68,000 to 70,000. The results suggest that yeast nucleoside-diphosphate kinase is composed of four subunits. Substrate specificity studies show that the relative activity of nucleoside diphosphates (NDP) as phosphate acceptors is in the order of dTDP greater than CDP greater than UDP greater than dUDP greater than GDP greater than or equal to dGDP greater than dCDP greater than dADP greater than ADP; and the relative activity of triphosphate donors is in the order of UTP greater than dTTP greater than CTP greater than dCTP greater than dATP greater than ATP greater than or equal to dGTP greater than GTP. The Km and Vm of dTDP, dGDP, dCDP, dUDP, CDP, and UDP have been determined. The rate constant studies indicate that the purified NDP kinase prefers using, to a slight extent, dTDP (approximately 800 min-1) as the substrate rather than other tested deoxyribo- and ribonucleotides (350-450 min-1). The broad substrate specificity and kinetic data suggest that the enzyme is involved in both DNA and RNA metabolism.

The mechanism of transmission in skeletal muscle EC coupling is still an open question. There is some indirect evidence in favour of the mechanical coupling hypothesis, deriving mostly from consideration of the structure of the Ca2+ release channel protein. A new functional approach is proposed, that consists in comparing the properties of the complete system--EC coupling in a skeletal muscle fibre--with those of the EC coupling molecules in bilayers. In this approach, those properties of the whole system that are not traceable to its constitutive molecules, are ascribed to the physiological interaction, and are expected to yield new information on the nature of this interaction.

We isolated and sequenced the gene for the chlamydial heat shock protein 60 (HSP-60) from a Chlamydia trachomatis genomic library by molecular genetic methods. The DNA sequence derived revealed an operon-like gene structure with two open reading frames encoding an 11,122- and a 57,956-Da protein. The translated amino acid sequence of the larger open reading frame showed a high degree of homology with known sequences for HSP-60 from several bacterial species as well as with plant and human sequences. By using the determined nucleotide sequence, fragments of the gene were cloned into the plasmid vector pGEX for expression as fusion proteins consisting of glutathione S-transferase and peptide portions of the chlamydial HSP-60. HSP-60 antigenic identity was confirmed by an immunoblot with anti-HSP-60 rabbit serum. Sera from patients that exhibited both high antichlamydial titers and reactivity to chlamydial HSP-60 showed reactivity on immunoblots to two fusion proteins that represented portions of the carboxyl-terminal half of the molecule, whereas fusion proteins defining the amino-terminal half were nonreactive. No reactivity with the fusion proteins was seen with sera from patients that had been previously screened as nonreactive to native chlamydial HSP-60 but which had high antichlamydial titers. Sera from noninfected control subjects also exhibited no reactivity. Definition of recognized HSP-60 epitopes may provide a predictive screen for those patients with C. trachomatis infections who may develop damaging sequelae, as well as providing tools for the study of immunopathogenic mechanisms of Chlamydia-induced disease.

The SOC8 gene was isolated as an extragenic suppressor of cdc8 mutant cells. It has been suggested that SOC8 is allelic with the URA6 gene which was originally identified as a uridine monophosphate kinase. In this article, we describe the purification of the uridine monophosphate kinase from a yeast Saccharomyces cerevisae strain that overproduces the activity 8-fold. The protein was purified through Fast-Flow Q-Separose, phosphocellulose, blue-agarose, and fast protein liquid chromatography Superose 12 columns, and appears homogeneous by sodium dodecyl sulfate-polyacrylamide gel analysis. The uridine monophosphate kinase contains a single polypeptide with a molecular weight of 25,000, as evidence by both sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration analysis. The amino acid composition has also been determined. Substrate specificity studies show that the relative activity of nucleoside monophosphates is in order of UMP greater than dUMP, and to a lesser extent, dTMP, GMP, and dGMP. The Km and Vm of UMP, dUMP, and dTMP have been determined.

Previous studies have demonstrated a defect associated with the calcium release mechanism of sarcoplasmic reticulum (SR) from individuals susceptible to malignant hyperthermia (MH). To examine whether SR calcium release channels were indeed altered in MH, SR vesicles were purified from normal and MH susceptible (MHS) porcine muscle. The Ca2+ dependence of calcium efflux rates from 45Ca2(+)-filled SR vesicles was then compared with the Ca2+ dependence of single-channel recordings of SR vesicles incorporated into planar lipid bilayers. The rate constants of 45Ca2+ efflux from MHS SR were two to threefold larger than from normal SR over a wide range of myoplasmic Ca2+. Normal and MHS single channels were progressively activated in a similar fashion by cis Ca2+ from pCa 7 to 4. However, below pCa 4, normal channels were inactivated by cis Ca2+, whereas MHS channels remained open for significantly longer times. The altered Ca2+ dependence of channel inactivation in MHS SR was also evident when Ca2+ was increased on the trans side while cis Ca2+ was held constant. We propose that a defect in a low-affinity Ca2+ binding site is responsible for the altered gating of MHS SR channels. Such a defect could logically result from a mutation in the gene encoding the calcium release channel, providing a testable hypothesis for the molecular basis of this inherited disorder.

Proteolytic peptides containing the serological epitopes present on the major outer membrane protein of Chlamydia trachomatis were studied by immunoblots. Monoclonal antibodies which have been defined by micro-immunofluorescence typing as serovar-, subspecies and species-specific were utilized. The reactivity of either serovar-specific or species-specific monoclonal antibodies in the immunoblots was similar to that in the micro-immunofluorescence test. However, monoclonal antibodies which demonstrated subspecies-specific reactivity by the micro-immunofluorescence test expressed species-specific reactivity by the immunoblot method. The serovar-specific epitope which was identified on the major outer membrane protein was lost after proteolytic digestion with Staphylococcus aureus V8 protease. Immunoblots of the proteolytic fragments identified the species-specific epitope on several peptides and the subspecies-specific epitope on fewer peptides than those in which the species-specific epitope was identified. In addition, both the species and subspecies epitopes were present on the same fragments of different serovars. The data indicate that the species epitope is on conserved regions of the major outer membrane protein and is structurally clustered with a subspecies-specific epitope.

A purified dihydropyridine-receptor complex (DHPR) of skeletal muscle consisting of a major polypeptide of Mr 150K under reducing conditions induces divalent cation selective channels when incorporated into planar lipid bilayers. Channels were inserted into preformed planar bilayers by two techniques: (i) direct dilution of detergent-solubilized DHPR into the aqueous chambers adjacent to the bilayer membrane or (ii) reconstitution of DHPR into phospholipid vesicles followed by fusion of the preformed vesicles to the planar bilayer membrane. Unlike native membrane preparations of t-tubules, which only have one major Ca channel type of slope conductance of 12 pS in symmetrical 100 mM Ba, the purified DHPR complex induced at least two channel types with conductances of 12-14 and 22 pS. Some recordings suggest that these two channels are statistically coupled in time, i.e., that they may correspond to substrates of the same DHPR channel. Activity was found to occur spontaneously in the absence of the Ca channel agonist Bay k 8644. The 12-14-pS channel from DHPR exhibits voltage-dependent kinetics, is highly selective for barium ions, and was inhibited by micromolar nitrendipine. The 12-14-pS DHPR channel appears to be identical with functional Ca channels previously described in native t-tubules.