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In this study, 15 Acanthamoeba isolates from AK patients and 10 environmental samples (water, soil and animal-origin samples) were classified at the genotype level based on the sequence analysis of the Diagnostic Fragment 3 (DF3) of Acanthamoeba small subunit rRNA gene. The obtained results revealed that most of these Acanthamoeba strains belonged to genotype T4 both in clinical and environmental samples. The presence T11 genotype in clinical samples was also revealed after the genotyping analysis and to our knowledge this is the first report of T11 genotype in Iran. Moreover, the isolation of T4 genotype from cow faeces in this study highlights a possible transmission of Acanthamoeba through animal faeces in Iran. Overall, the widespread distribution of pathogenic Acanthamoeba T4 across the environmental sources and the increasing number of contact lens wearers in Iran, demands more awareness within the public and health professionals as this pathogen is emerging as a risk for human health in Iran and worldwide.

Aims: This study aimed to analyse the diversity of the vanB gene cluster in enterococcal species isolated from sewage treatment plants (STP) in Tehran, Iran., Methods and Results: The enterococcal isolates were collected from three sewage treatment plants in Tehran, Iran, during 2005. A total of 203 enterococcal isolates, collected over six rounds of sampling from three STPs were tested for the presence of vanB gene. Long-PCR showed that amongst the isolates, three Enterococcus faecium, one Enterococcus gallinarum and one Enterococcus casseliflavus harboured the complete vanB gene cluster. Restriction fragment length polymorphism (RFLP) of the vanB1 gene cluster (5900 bp) from the isolates showed an identical pattern to a standard strain of Enterococcus faecalis (V583). None of the isolates were able to transfer the vanB gene in conjugation experiments. Different pulsed-field gel electrophoresis patterns were obtained for the three E. faecium isolates with vanB gene clusters., Conclusions: Our results indicated that the dissemination of vanB is not widespread in Tehran. Although only a few vanB positive isolates were detected, vanB was found in several enterococcal species., Significance and Impact of the Study: In view of the lack of information on vanB resistance genes and their diversity in Iran, knowledge of the global dissemination of vanB genes in Enterococcus spp. is noteworthy.

Objective: A phase II study was performed to assess the activity of oxaliplatin plus 5-fluorouracil (5-FU) modulated by leucovorin, as second-line treatment in locally advanced or metastatic pancreas adenocarcinoma pretreated with gemcitabine-containing schedule., Methods: Patients received weekly intravenous infusions of oxaliplatin 40 mg/m, 5-FU 500 mg/m, and leucovorin 250 mg/m (3 weeks on, 1 week off)., Results: Twenty-three patients affected with metastatic (16) or locally advanced (7) pancreas adenocarcinoma were involved in this study. A total of 148 weeks of chemotherapy was delivered (median 2 courses each patient). Among 17 assessable patients, no objective response was registered and 4 patients had stable disease, whereas 13 had tumor progression. Median duration of stable disease was 14 weeks. Median time to progression of disease (TTP) was 11.6 weeks [95% confidence interval (CI), 7.6-5.6]. Kaplan-Meier estimated median overall survival (OS) was 17.1 week (95% CI, 4.0-30.1) and 3 months survival rate was 69.6%. Seven patients experienced grade 3 to 4 toxicity. The regimen was associated with 36% clinical benefit., Conclusions: The median TTP and median OS in this population with poor prognosis suggests some activity, however, only further investigations will be able to establish the clinical value of this combination.

The assembly of amyloidogenic proteins into toxic oligomers is a seminal event in the pathogenesis of protein misfolding diseases, including Alzheimer's, Parkinson's, and Huntington's diseases, hereditary amyotrophic lateral sclerosis, and type 2 diabetes. Owing to the metastable nature of these protein assemblies, it is difficult to assess their oligomer size distribution quantitatively using classical methods, such as electrophoresis, chromatography, fluorescence, or dynamic light scattering. Oligomers of amyloidogenic proteins exist as metastable mixtures, in which the oligomers dissociate into monomers and associate into larger assemblies simultaneously. PICUP stabilizes oligomer populations by covalent cross-linking and when combined with fractionation methods, such as sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) or size-exclusion chromatography (SEC), PICUP provides snapshots of the oligomer size distributions that existed before cross-linking. Hence, PICUP enables visualization and quantitative analysis of metastable protein populations and can be used to monitor assembly and decipher relationships between sequence modifications and oligomerization(1). Mechanistically, PICUP involves photo-oxidation of Ru(2+) in a tris(bipyridyl)Ru(II) complex (RuBpy) to Ru(3+) by irradiation with visible light in the presence of an electron acceptor. Ru(3+) is a strong one-electron oxidizer capable of abstracting an electron from a neighboring protein molecule, generating a protein radical(1,2). Radicals are unstable, highly-reactive species and therefore disappear rapidly through a variety of intra- and intermolecular reactions. A radical may utilize the high energy of an unpaired electron to react with another protein monomer forming a dimeric radical, which subsequently loses a hydrogen atom and forms a stable, covalently-linked dimer. The dimer may then react further through a similar mechanism with monomers or other dimers to form higher-order oligomers. Advantages of PICUP relative to other photo- or chemical cross-linking methods(3,4) include short (

Biofeedback signals have been frequently used for rehabilitation purposes, and in design and calibration of orthotic and prosthetic devices. Whenever one or a couple of muscles of a joint are chosen for rehabilitation or control of a device, it's assumed that a specific load sharing or activation pattern exists among them for each individual and for each specific joint demand. Indeterminacy or a load sharing problem arises from having more muscles crossing a joint than needed to perform all possible movements. It's proven that muscle activation patterns depend on fatigue, the task (isometric/isokinetic, concentric/eccentric), load type, mental demands, etc. The most used biofeedbacks are electromyogram of one of the muscles or the joint torque signal. An important question is if they can be used interchangeably. This study investigated if the choice of biofeedback can also change the activation pattern in the two main elbow flexors. The results of this experiment on six healthy subjects and seven activation levels, indicated that change in biofeedback type had a significant effect on the activation ratio of these two muscles.

While the majority of tremor-afflicted Parkinso-nian (PD) patients suffer from rest tremors, which is not considered highly disabling, a portion of these PD patients also demonstrate action tremors that interfere with their daily lives. Two main considerations in designing an orthosis that aims at suppressing the tremor, are the frequency bands of the tremor and the joints tremor affects. Nine subjects, which included six healthy people, two PD patients with typical tremor afflictions, and a PD patient with severe tremor of not only in her fingers and wrist, but also in her elbow, participated in this study. The highly afflicted patient displayed the need for tremor suppression in action as well as when in rest. The study focuses on uncommon elbow tremors and demonstrates that, for typically afflicted patients, tremor amplitudes are comparable to healthy subjects, but the frequency distribution of the tremors are different at high levels of elbow torque. For the highly afflicted patient, both tremor amplitude and its frequency distribution are different at all levels of elbow torque. The study further investigates the tremors in two bands of frequency on both hands of the highly troubled patient before, and after medication. The two bands are those of classical Parkinsonian tremor (4-6 Hz) and physiological (or enhanced physiological) tremor (8-12 Hz). Power spectrum and tremor amplitude comparisons reveal that, for part of tremulous PD patients, both tremors coexist and, depending on the level of affliction, the designed orthosis needs to suppress tremors in both bands, even at more proximal joints, such as the elbow.

Background: This study was conducted to determine the treatment outcome of incompletely removed, histopathologically documented ocular surface squamous neoplasia with mitomycin-C., Methods: Through an interventional case series, 17 eyes of 17 patients presented with incompletely removed ocular surface squamous neoplasia were treated according to a protocol using two to three alternate seven-day courses of 0.04% mitomycin-C. Clinical recurrence was re-treated with the same protocol. All patients had weekly follow-up visits to the end of the treatment course, then biweekly visits for three months, and monthly visits, thereafter., Results: The mean+/-SD follow-up period was 30.76+/-4.4 (range: 24.5 - 41) months. Five patients (29.4%) experienced recurrence after the initial treatment; four of them responded to retreatment and were disease-free till the end of follow-up. Survival analysis with Kaplan-Meier method was performed. Taking into account four recurrences, the 41-month nonrecurrence rate was 70.6%. However, including four of five patients with recurrence who responded to retreatment, the final outcome in survival analysis was 94.1% nonrecurrence for 41 months of follow-up. All patients reported mild to moderate redness and irritation which were controlled with lubricants and mild corticosteroid eye drops. No serious ocular or systemic side effects were observed., Conclusion: Point zero four percent (0.04%) mitomycin-C drop used as two to three alternate seven-day courses seems to be a safe and effective treatment for ocular surface squamous neoplasia.

Purpose: This study was designed to evaluate whether probiotics improve symptoms in patients with irritable bowel syndrome., Methods: PubMed, Embase, Scopus, Web of Science, and Cochrane Central Register of Controlled Trials were searched for studies that investigated the efficacy of probiotics in the management of irritable bowel syndrome. Clinical improvement was the key outcome of interest. Data were searched within the time period of 1966 through September 2007., Results: Eight randomized, placebo-controlled, clinical trials met our criteria and were included in the analysis. Pooling of eight trials for the outcome of clinical improvement yielded a significant relative risk of 1.22 (95 percent confidence interval, 1.07-1.4; P = 0.0042)., Conclusions: Probiotics may improve symptoms of irritable bowel syndrome and can be used as supplement to standard therapy.

Objective: To evaluate whether probiotics maintain remission in patients with Crohn's disease (CD)., Design: A meta-analysis of controlled clinical trials., Methods: PUBMED and Cochrane Central Register of Controlled Trials were searched for clinical trial studies investigated the efficacy of probiotics for the maintenance of remission in Crohn's disease. Clinical relapse and endoscopic relapse were the key outcomes of interest. Data were searched within the time period of 1966 through May 2007., Result: Eight randomized placebo-controlled clinical trials met our criteria and were included in the analysis. Seven determined clinical relapse and three evaluated endoscopic relapse among patients with CD received probiotics for maintenance of remission. Pooling of seven trials for the outcome of clinical relapse yielded an odds ratio of 0.92 (95% confidence interval of 0.52-1.62, P = 0.8853), a nonsignificant odds ratio. The odds ratio for three studies for the outcome of endoscopic relapse was 0.97 (95% confidence interval of 0.54-1.78, P = 0.93), a nonsignificant odds ratio., Conclusion: This meta-analysis fails to demonstrate the efficacy of probiotics in maintaining remission and preventing clinical and endoscopic recurrence in CD. It is suggested to use probiotic preparations containing a mixture of lactobacillus with E. coli or Saccharomyces.

Several neurodegenerative diseases, including Alzheimer's, Parkinson's, Huntington's and prion diseases, are characterized pathognomonically by the presence of intra- and/or extracellular lesions containing proteinaceous aggregates, and by extensive neuronal loss in selective brain regions. Related non-neuropathic systemic diseases, e.g., light-chain and senile systemic amyloidoses, and other organ-specific diseases, such as dialysis-related amyloidosis and type-2 diabetes mellitus, also are characterized by deposition of aberrantly folded, insoluble proteins. It is debated whether the hallmark pathologic lesions are causative. Substantial evidence suggests that these aggregates are the end state of aberrant protein folding whereas the actual culprits likely are transient, pre-fibrillar assemblies preceding the aggregates. In the context of neurodegenerative amyloidoses, the proteinaceous aggregates may eventuate as potentially neuroprotective sinks for the neurotoxic, oligomeric protein assemblies. The pre-fibrillar, oligomeric assemblies are believed to initiate the pathogenic mechanisms that lead to synaptic dysfunction, neuronal loss, and disease-specific regional brain atrophy. The amyloid beta-protein (Abeta), which is believed to cause Alzheimer's disease (AD), is considered an archetypal amyloidogenic protein. Intense studies have led to nominal, functional, and structural descriptions of oligomeric Abeta assemblies. However, the dynamic and metastable nature of Abeta oligomers renders their study difficult. Different results generated using different methodologies under different experimental settings further complicate this complex area of research and identification of the exact pathogenic assemblies in vivo seems daunting. Here we review structural, functional, and biological experiments used to produce and study pre-fibrillar Abeta assemblies, and highlight similar studies of proteins involved in related diseases. We discuss challenges that contemporary researchers are facing and future research prospects in this demanding yet highly important field.

The antimicrobial activity of the methanol extracts of the dried flowering aerial parts of Stachys byzantina, S. inflata, S. lavandulifolia and S. laxa (Labiatae) were studied using the disc diffusion method and determination of minimum inhibitory concentration (MIC) values against Staphylococcus aureus, Streptococcus sanguis, Escherichia coli, Pseudomonas aeroginosa, Klebsiella pneumoniae, Aspergilus niger and Candida albicans. The extracts of plants exhibited concentration-dependent antibacterial activity against the bacteria tested. The extracts were more active against Gram-positive microorganisms. The extracts, however, did not show any antifungal activity.

We investigated the prevalence of vancomycin-resistant enterococci (VRE) isolated from wastewater (n = 593) and clinical (n = 450) samples, and the genetic linkage between the isolates was compared. Out of the total samples, 38 Enterococcus faecium (3.6%) from sewage (n = 19) and clinical (n = 19) isolates were found to be highly resistant to vancomycin. The majority of the VRE isolates from the two sources showed distinct phenotyping and genotyping patterns. At the same time, one common pulsed-field gel electrophoresis pattern was found among the VRE obtained from wastewater and human clinical isolates, suggestive of an epidemiological link.

Background: Enterococci are important because of their role as the leading cause of nosocomial infections which have a significant role in the dissemination and persistence of antimicrobial resistance genes., Methods: In this study, we determined the distribution of enterococcal species in the sewage treatment plants in Iran. Furthermore, we improved a rapid and specific PCR method using primers (sodA and ddl genes) for identification of enterococci spp., Results and Conclusion: A total number of 712 enterococci spp. were isolated and the results showed that 56%, 24%, 12%, 4%, 2%, 1% and 1% isolates were E. faecium, E. hirae, E. faecalis, E. gallinarum, E. casseliflavus, E. mundtii and other enterococcal spp., respectively. The use of species-specific PCR was in agreement with the biochemical tests. Furthermore, multiplex PCR was developed to study the presence of vancomycin resistant genes in E. faecium or E. faecalis. The multiplex PCR appeared to be a useful, rapid and specific method for detecting and discriminating genotypes for vancomycin-resistant Enterococcus.

Neurilemmoma rarely develops in the biliary tree. Here, we report a 39-year-old Iranian woman with neurilemmoma in the extrahepatic bile duct presenting with progressively deepening jaundice. On the basis of clinical and radiological features, this tumor was initially suspected as Klatskin tumor. Histologically, the tumor was a typical neurilemmoma. Immunostaining showed that tumor cells were strongly and diffusely positive for S-100 protein, which supported the diagnosis of neurilemmoma. Neurilemmoma should be considered in the differential diagnosis of obstructive jaundice.

Purpose: To evaluate the outcome of microkeratome-assisted posterior lamellar keratoplasty for replacing diseased endothelium in pseudophakic and aphakic corneal edema., Methods: Microkeratome-assisted posterior lamellar keratoplasty was performed on 10 eyes with pseudophakic or aphakic corneal edema. A nasal hinged 130- to 250-microm-thick flap was created with an automated microkeratome. The underlying 7.0-mm trephined button of deep stroma and endothelium was substituted with the same size donor button prepared by removing a same-sized flap and trephination of the remaining donor bed. The donor lenticule was fixated without sutures in position; the retracted flap was repositioned and sutured to the peripheral recipient tissue. The viscoelastic content of the anterior chamber was replaced by air to hold the graft in position., Results: All surgeries were uneventful except for one case of donor button posterior dislocation forming a secondary anterior chamber, which was corrected through donor button exchange 1 month postoperatively. All uncomplicated cases had stable refraction and corneal topography as early as 1 month after surgery, which was maintained in follow-up examinations. In two eyes, corneal astigmatism > 5.00 diopters was recorded. One case of prolonged re-epithelialization and two cases of epithelial interface ingrowth occurred., Conclusions: Microkeratome-assisted posterior lamellar keratoplasty is an alternative to conventional penetrating keratoplasty for patients with diseased corneal endothelium, significantly reducing the time of visual rehabilitation. Determination of the relative advantages of this technique over penetrating keratoplasty and other recent alternative endothelial graft procedures relies on controlled prospective studies.

Background: Breast cancer is the leading cause of carcinoma death in women. Proper treatment depends on the consideration of molecular biology status of tumor cells, which may determine the patient's treatment and prognosis. To determine the prognostic models for this disease, we evaluated the role of cell proliferation-related antigens including ki-67 (a nuclear antigen, expressed in G1, G2, and M phases of cell cycle) and repp86 (an 86-kDa nuclear protein expressed in S, G2, and M phases of cell cycle) for detection of biologic behavior of breast cancer., Methods: We studied 60 women with grade I and II lymph node-negative and 27 with grade III lymph node-positive breast cancers. The mean follow-up periods for these two groups were 60 and 72 months, respectively. Tumor cell proliferation was measured by immunohistochemical methods with monoclonal antibodies directed against the nuclear antigens ki-67 and repp86., Results: The ki-67, repp86 labeling indices (percentage of antibody-stained tumor cell nuclei) were not statistically different between the cases and controls of lymph node-negative patients (ki-67 with P = 0.33; repp86 with P = 0.40). The odds ratio (the mean chance of ki-67 labeling index > 10%, repp86 labeling index >10%) in patients with recurrence was 4 (CI = 0.2 - 76.5) for ki-67 and 3.6 (CI = 0.4 - 32.5) for repp86. Both indices were statistically different in lymph node-positive cases and controls (P < 0.0001). The odds ratio in patients with recurrence was 87 (CI = 4 - 18.71) for ki-67 and 71.5 (CI = 5.7 - 899.2) for repp86., Conclusion: The present study confirms the importance of cell proliferation as a determinant of biologic behavior of breast cancer. Measurement of ki-67 and repp86 labeling indices may be very helpful for physicians to detect high-risk patients and to adopt appropriate procedure such as adjuvant therapy.

In this study, alginate microspheres containing BCG were prepared at a diameter of approximately 10 microm by emulsification-internal gelation of an alginate-BCG solution dispersed in olive oil using a high rate speed stirrer. The stability of BCG was assayed at 4 degrees C showing that the encapsulated BCG was more stable than free BCG at least for 5 weeks; however, BCG in sodium alginate solution was not stable at all. On the other hand, the studies using media with different pH (1.2, 4.4, 6.2, 6.8 and 7.5) have demonstrated that the alginate microspheres are stable in acidic medium for upto 1.5 h without any sign of disintegration. Moreover, BCG incorporated in alginate microspheres demonstrated an almost 9-fold increase in viable bacilli in simulated gastric fluid (SGF) after 1.5 h in comparison with free BCG.

Inflammation, insoluble protein deposition and neuronal cell loss are important features of the Alzheimer's disease (AD) brain. S100B is associated with the neuropathological hallmarks of AD where it is thought to play a role in neuritic pathology. S100A8, S100A9 and S100A12 comprise a new group of inflammation-associated proteins that are constitutively expressed by neutrophils and inducible in numerous inflammatory cells. We investigated expression of S100B, S100A8, S100A9 and S100A12 in brain samples from sporadic and familial (PS-1) AD cases and controls using immunohistochemistry and Western blot analysis. S100B, S100A9 and S100A12, but not S100A8, were consistently associated with the neuropathological hallmarks of AD. Western blot analysis confirmed significant increases in soluble S100A9 in PS-1 AD compared to controls. S100A9 complexes that were resistant to reduction were also evident in brain extracts. A reactive component of a size consistent with hexameric S100A12 was seen in all cases. This study indicates a potential role for pro-inflammatory S100A9 and S100A12 in pathogenesis caused by inflammation and protein complex formation in AD.

Aim: To determine whether insulin resistance occurs in patients with chronic hepatitis B (CHB) and chronic hepatitis C (CHC) and its relationship with the presence of liver fibrosis and steatosis., Methods: Untreated patients with CHC (n=60) or CHB (n=40), similar in age, gender, body mass index and waist-hip ratio, were studied. Relationship between anthropometric, biochemical (fasting serum insulin, C-peptide, ferritin, iron, TNF-alpha, cholesterol, triglyceride, bilirubin, hemoglobin and platelet concentrations) and liver biopsy (43 CHC and 20 CHB patients) findings was investigated by insulin resistance determined via the homeostasis model assessment (HOMA-IR)., Results: The mean fasting serum insulin was 14.9 (11.9) mU/mL in CHC and 21.4 (17.4) in the CHB group (normal range 0.7-9; p=0.049) and mean HOMA-IR was 3.1 (2.6) in CHC versus 4.7 (4.1) in the CHB group (normal range 0.12-4.61; p=0.036). HOMA-IR was significantly associated with fibrosis stage in the CHC group (p=0.015), but not in the CHB group., Conclusion: Hyperinsulinemia occurs in chronic viral hepatitis B and hepatitis C; insulin resistance is associated with stage of fibrosis in hepatitis C.

Formalin is a routine fixative facilitating tissue preservation and histopathology. Proteomic techniques require freshly frozen specimens, which are often difficult to procure, and methods facilitating proteomic analysis of archival formalin-fixed brain tissue are lacking. We employed antigen-epitope-retrieval principles to facilitate proteomic analysis of brain tissue that had been fixed and stored in formalin for 3-7 years. Twenty-micrometer-thick cryopreserved OCT-embedded sections from inferior temporal cortex of human (7 years in formalin) or mouse brain specimens (3 years in formalin) were hematoxylin-/eosin-stained. Approximately 16-64-mm2 areas of the tissue sections were manually scraped off slides, or approximately 2 mm2 of human brain cortex was captured off membrane-coated slides using laser microdissection. Tissue was treated using various pH and temperature conditions prior to trypsin digestion and nano-LC-MS/MS. The largest number of proteins were retrieved by solubilization at pH 9 at 95 degrees C for 1 h; treatments at pH 4 or 6 at 25 or 65 degrees C were generally ineffective. Three-year formalin-fixed murine tissue did not yield more proteins compared to human tissue. Use of formalin-fixed tissue for proteomics is an invaluable tool for medical research. The combination of proteomics and microdissection enables selective enrichment and identification of novel, unique, or abundant proteins that may be important in pathogenesis.

Background: The present study is aimed to assess the usefulness of silver nitrate staining of nucleolar organizer regions (NORs) as a quantitative criterion for the diagnosis of selected head and neck tumors., Materials and Methods: The silver nitrate staining technique was used on 195 paraffin blocks collected from 85 patients. The samples consisted of 21 squamous cell carcinoma (SCC) of larynx, 28 SCC of oral mucosa and 36 samples of most common salivary gland tumors. Mann-Whitney U-Test was used for data analysis., Results: A significant difference was seen in the number of AgNOR dots between oral and laryngeal SCC with surrounding dysplastic and normal tissues (P < 0.001) and also between mucoepidermoid carcinoma and adenoid cystic carcinoma with pleomorphic adenoma and normal salivary gland tissue (P < 0.001)., Conclusion: The silver nitrate staining for NORs is a useful method for aiding the diagnosis of malignant and dysplastic mucosal lesions and also malignant and benign salivary gland tumors.

Atherogenesis is a complex process involving inflammation. S100A8 and S100A9, the Ca2+-binding neutrophil cytosolic proteins, are associated with innate immunity and regulate processes leading to leukocyte adhesion and transmigration. In neutrophils and monocytes the S100A8-S100A9 complex regulates phosphorylation, NADPH-oxidase activity, and fatty acid transport. The proteins have anti-microbial properties, and S100A8 may play a role in oxidant defense in inflammation. Murine S100A8 is regulated by inflammatory mediators and recruits macrophages with a proatherogenic phenotype. S100A9 but not S100A8 was found in macrophages in ApoE-/- murine atherosclerotic lesions, whereas both proteins are expressed in human giant cell arteritis. Here we demonstrate S100A8 and S100A9 protein and mRNA in macrophages, foam cells, and neovessels in human atheroma. Monomeric and complexed forms were detected in plaque extracts. S100A9 was strongly expressed in calcifying areas and the surrounding extracellular matrix. Vascular matrix vesicles contain high levels of Ca2+-binding proteins and phospholipids that regulate calcification. Matrix vesicles characterized by electron microscopy, x-ray microanalysis, nucleoside triphosphate pyrophosphohydrolase assay and cholesterol/phospholipid analysis contained predominantly S100A9. We propose that S100A9 associated with lipid structures in matrix vesicles may influence phospholipid-Ca2+ binding properties to promote dystrophic calcification. S100A8 and S100A9 were more sensitive to hypochlorite oxidation than albumin or low density lipoprotein and immunoaffinity confirmed S100A8-S100A9 complexes; some were resistant to reduction, suggesting that hypochlorite may contribute to protein cross-linking. S100A8 and S100A9 in atherosclerotic plaque and calcifying matrix vesicles may significantly influence redox- and Ca2+-dependent processes during atherogenesis and its chronic complications, particularly dystrophic calcification.

Growth factors, including fibroblast growth factor-2 (FGF-2) and transforming growth factor-beta (TGF-beta) regulate fibroblast function, differentiation and proliferation. S100A8 and S100A9 are members of the S100 family of Ca2+-binding proteins and are now accepted as markers of inflammation. They are expressed by keratinocytes and inflammatory cells in human/murine wounds and by appropriately activated macrophages, endothelial cells, epithelial cells and keratinocytes in vitro. In this study, regulation and expression of S100A8 and S100A9 were examined in fibroblasts. Endotoxin (LPS), interferon gamma (IFNgamma), tumour-necrosis factor (TNF) and TGF-beta did not induce the S100A8 gene in murine fibroblasts whereas FGF-2 induced mRNA maximally after 12 h. The FGF-2 response was strongly enhanced and prolonged by heparin. Interleukin-1beta (IL-1beta) alone, or in synergy with FGF-2/heparin strongly induced the gene in 3T3 fibroblasts. S100A9 mRNA was not induced under any condition. Induction of S100A8 in the absence of S100A9 was confirmed in primary fibroblasts. S100A8 mRNA induction by FGF-2 and IL-1beta was partially dependent on the mitogen-activated-protein-kinase pathway and dependent on new protein synthesis. FGF-2-responsive elements were distinct from the IL-1beta-responsive elements in the S100A8 gene promoter. FGF-2-/heparin-induced, but not IL-1beta-induced responses were significantly suppressed by TGF-beta, possibly mediated by decreased mRNA stability. S100A8 in activated fibroblasts was mainly intracytoplasmic. Rat dermal wounds contained numerous S100A8-positive fibroblast-like cells 2 and 4 days post injury; numbers declined by 7 days. Up-regulation of S100A8 by FGF-2/IL-1beta, down-regulation by TGF-beta, and its time-dependent expression in wound fibroblasts suggest a role in fibroblast differentiation at sites of inflammation and repair.

S100A8 (A8) has roles in inflammation, differentiation and development and is associated with oxidative defense. Murine A8 (mA8) is up-regulated in macrophages, fibroblasts, and microvascular endothelial cells by LPS. Glucocorticoids (GCs) amplified LPS-induced mA8 in these cells. Relative to stimulation by LPS, GCs increased mA8 gene transcription and mRNA half-life. Enhancement required new protein synthesis, IL-10 and products of the cyclooxygenase-2 pathway, and both ERK1/2 and p38 MAPK. Protein kinase A positively and protein kinase C negatively regulated this process. Promoter analysis indicated element(s) essential for LPS and dexamethasone enhancement colocated within the region -178 to 0 bp. In the absence of glucocorticoid response elements, NF1 motif at -58 is a candidate for mediation of enhancement. Gel shift analysis detected no differences between LPS- and LPS/dexamethasone-treated complexes within this region. GCs increased constitutive levels of A8 and S100A9 (A9) mRNA in human monocytes. The synovial membrane of rheumatoid patients treated with high dose i.v. methylprednisolone contained higher numbers of A8/A9-positive macrophages than pre- or posttreatment samples. Results support the proposal that A8 has anti-inflammatory properties that may be independent of hetero-complex formation with A9 and may also enable localized defense in the absence of overriding deleterious host responses.

Presenilin-1 (PS-1) mutations can cause Pick's disease without evidence of Alzheimer's disease (AD). We describe a family with a PS-1 M146L mutation and both Pick bodies and AD. Sarkosyl-insoluble hyperphosphorylated tau showed three bands consistent with AD, although dephosphorylation showed primarily three-repeat isoforms. M146L mutant PS-1 may predispose to both Pick's disease and AD by affecting multiple intracellular pathways involving tau phosphorylation and amyloid metabolism.

The EF-hand superfamily of calcium binding proteins includes the S100, calcium binding protein, and troponin subfamilies. This study represents a genome, structure, and expression analysis of the S100 protein family, in mouse, human, and rat. We confirm the high level of conservation between mammalian sequences but show that four members, including S100A12, are present only in the human genome. We describe three new members of the S100 family in the three species and their locations within the S100 genomic clusters and propose a revised nomenclature and phylogenetic relationship between members of the EF-hand superfamily. Two of the three new genes were induced in bone-marrow-derived macrophages activated with bacterial lipopolysaccharide, suggesting a role in inflammation. Normal human and murine tissue distribution profiles indicate that some members of the family are expressed in a specific manner, whereas others are more ubiquitous. Structure-function analysis of the chemotactic properties of murine S100A8 and human S100A12, particularly within the active hinge domain, suggests that the human protein is the functional homolog of the murine protein. Strong similarities between the promoter regions of human S100A12 and murine S100A8 support this possibility. This study provides insights into the possible processes of evolution of the EF-hand protein superfamily. Evolution of the S100 proteins appears to have occurred in a modular fashion, also seen in other protein families such as the C2H2-type zinc-finger family., (Copyright 2004 Elsevier Inc.)

We report a 5.5 year-old girl with a benign adrenocortical adenoma who presented with virilization and rapid growth. She did not have any clinical features of isosexual precocity or, except for hypertension, Cushing's syndrome. Measurement of hormones in adrenal vein blood obtained at surgery showed high concentrations of testosterone, cortisol, estradiol and intermediary substrates. Elevated levels of hormones detected in the peripheral blood were released directly from the tumor and were not the result of peripheral interconversion. Hyperandrogenism can obscure the clinical features of Cushing's syndrome and estrogen hypersecretion in patients with functional adrenal tumors.

Dactylolysis spontanea is an idiopathic condition affecting the fifth toe, and sometimes other toes, that is frequently bilateral, with lesions in different stages. Between 1977 and 1999, a total 6000 radiographic studies of the feet were reviewed in a mainly African American population in Chicago, Illinois. After an initial screening based on the Cole criteria, 581 patients were selected and re-examined, and amplification techniques were performed. After reviewing the complementary exams, 102 patients were diagnosed with dactylolysis spontanea or ainhum. Soft-tissue constriction was the most frequently presented radiological sign on the initial screening. Kurtosis at the digit plantar fold and marked rotation of the fifth toe were normal findings in asymptomatic patients. Demographics, comorbidities, and radiological findings were analyzed in the selected population. Associated diseases occurring in these patients appeared to have no specific etiologic correlation with ainhum. African Americans and the dark-skinned population are affected exclusively by this condition, presumably due to the fibrogenic tendency of these individuals. Early diagnosis and accurate staging of ainhum are facilitated by radiological examination of the feet. The findings suggest that this condition is underdiagnosed and overlooked because its low prevalence and variable clinical presentations that might mimic more common etiologies, including localized trauma.

Despite decades of study, the exact mechanism of action of lead, a potent neurotoxic agent, have not been fully elucidated. One of the suggested mechanism of lead neurotoxicity is apoptotic cell death. The present study sought to examine the effect of acute lead poisoning on apoptosis in rat hippocampus. Two to four and 12-14 week old rats were treated for 7 days with 15 mg/kg daily dose of lead acetate intraperitoneally. Control animals received distilled water. In treated groups, the blood lead levels was increased by about 17-19-folds. Histological study of hippocampus revealed apoptotic cells, using light and electron microscopy. In Western blot analysis, the ratio of Bax/Bcl-2 protein expression in hippocampus was significantly increased compared to controls. In conclusion, the lead induced cell death in hippocampus in vivo may partly be due to apoptosis., (Copyright 2002 Elsevier Science Ireland Ltd.)

We report a prepubertal boy who developed true precocious puberty following unilateral orchidectomy for the treatment of a Leydig cell tumor.

Plain-film radiography is commonly used in the assessment of joints affected by osteoarthritis. It has been shown that the condition of articular cartilage in the knee, when grossly assessed, does not always correlate well to the radiographic evaluation of the joint. The aim of this study was to determine whether a relationship existed between the radiographic appearance of the subchondral bone and the morphological condition of the articular cartilage surfaces of the first metatarsophalangeal joint. Forty-nine pairs of cadaveric metatarsals and their respective proximal phalanges were studied in order to determine the correlation between articular cartilage degeneration and radiographic grade using a modified Kellgren-Lawrence scale. The relationship between cartilage degeneration and age, gender, osteophytes, metatarsal length, and measured angles (proximal articular set angle, distal articular set angle, intermetatarsal angle) was also investigated. It was found that there was a significant correlation between radiographic grade and cartilage degeneration on both the metatarsal head and the base of the proximal phalange (r2 = .2038 and .1733, respectively). However, while useful in the clinical setting, bony changes of the first metatarsophalangeal joint, as seen radiographically, fail to provide a full representation of the level of cartilage degeneration on the articular surfaces of this joint.

The use of coralline hydroxyapatite has become a viable bone grafting alternative. Its efficacy has been well established through multiple human and animal studies. Coralline hydroxyapatite enhances osteogenesis by providing a biocompatible lattice for the passage and assembly of vascular, fibroblastic, and osteoblastic tissues. It also provides support for surrounding osseous structures. The uses of this material are expanding into the realm of foot and ankle surgery. Its consideration as an appropriate bone graft substitute as well as multiple case studies demonstrating its surgical applicability are discussed. The implants utilized at Thorek Hospital and Medical Center over the past eight years, with an average follow-up of three and one-half years, have proven to be a valuable resource for augmentation where an osseous defect has occurred.

A 14 year old boy is presented who was admitted to the psychiatric ward because of delirium. He also had gynaecomastia, facial oedema, striae, and hypertension. He was finally proved to have a mixed oestrogen and cortisol producing adrenal tumour which was removed surgically with complete improvement and no relapse.

The inheritance of the sesquiterpene zingiberene was analyzed in segregating progeny of interspecific crosses among Lycopersicon hirsutum f. hirsutum Humb. and Bonpl. (hir), L. esculentum Mill. cv 'Nova' (esc), and L. hirsutum f. glabratum C.H. Mull (gla). The presence of zingiberene was inherited as a single dominant gene from hir in F2 and BC progeny of esc x hir, and as a single recessive gene in F2 and BC progeny of gla x hir. The segregation of esc x hir, gla x hir, and esc x gla progeny supported a single locus allelomorphic model in which the presence of zingiberene is controlled at a single locus, Z, where the allele from hir confers presence of zingiberene and is dominant to the allele from esc but recessive to that from gla. The presence of zingiberene in esc x hir progeny was not linked to the ability to set fruit or to several fruit characters, and progeny with zingiberene levels comparable to hir were recovered.

Heterorhabditis bacteriophora is an entomopathogenic nematode that has potential as a biological control agent. As a first step in the genetic analysis of this nematode, we induced and isolated a short nematode (dumpy) from the F2 progeny of an EMS mutagenized nematode. Phenotypically, the dumpy adult is 60% shorter than the wild type. There is almost no longitudinal growth after the infective juvenile stage. Dumpy is completely recessive with 100% penetrance and uniform expressivity. When F1 progeny were allowed to self, F2 progeny segregated wild: dumpy in a 15:1 ratio (chi 2 = 0.634, .5 > P > .2). This dumpy mutation (Rhdpy) complements another dumpy mutation isolated in Israel and is the second gene known for this species.

Stump neuromas and recurrent neuromas are common problems following amputation and neuroma excision, respectively. Epineurial capping by microsuture has been shown to be somewhat effective in the reduction of the size of stump neuromas and in their confinement away from a weightbearing area. The present study shows that epineurial capping by coagulation of the epineurium with a radiofrequency wave Surgitron forms an effective seal around the proximal stumps of severed axons in rats. This seal effectively reduced axon proliferation but did not completely inhibit neuroma formation. Removal of the distal stump did not significantly affect neuroma formation in the proximal stump.

A case report is presented involving a nutcracker fracture of the cuboid and concomitant fractures of the navicular and calcaneus. A literature review of the mechanism and treatments of the nutcracker fracture is discussed. Also presented are the methods used to radiographically evaluate and surgically correct the compression fracture of the cuboid and its associated fractures.

The authors present a concise review of age-related changes that occur in the skin and its derivatives, as they pertain to the podiatric practitioner. A brief discussion of wound healing and several common skin disorders that affect the elderly is also included.

Neuroma excision is a common surgical practice that typically leads to axon regeneration and/or Schwann cell overproliferation from the proximal nerve stump. The effectiveness of the epineurium as a barrier to this reaction to nerve severing was studied in 31 rats. The right sciatic nerve was severed in each case. Control groups that had no further treatment displayed typical neuromas with axon regeneration 3, 6, and 8 months postoperatively. Experimental groups that had a "cap" of epineurium microsutured tightly over the proximal stump displayed significantly reduced neuromas up to 8 months postoperatively.

A comparative study on the catalytic and allosteric properties of particulate and soluble forms of ox liver glutamate dehydrogenase has been carried out. The response of the bound enzyme to release by various effectors was investigated. The particulate enzyme was found to have catalytic activities similar to the free enzyme in contrast to its behaviour when bound to pure anionic phospholipids. Possible reasons for such outstanding differences are discussed.

To study changes in lipid-protein-interaction and fluidity in mitochondrial and microsomal membranes of brain during development and aging, the Arrhenius plots of marker enzymes for mitochondrial inner and outer membranes as well as those of microsomal membranes were compared at different ages. The enzymes were, beta-hydroxybutyrate dehydrogenase (BDH) for the inner mitochondrial membrane and rotenone-insensitive-NADH-cytochrome c reductase (Mit-NADH-CytR) for the outer membrane; also antimycin-insensitive-NADH-cytochrome c reductase (Micr-NADH-CytR) and NADPH-cytochrome c reductase for the microsomal membranes. The ages studied were, 1, 5, 30, 60 days postnatal and 2 years. In the microsomes, the plots of NADH-cytochrome c reductase were found to be biphasic at all ages except in the newborn where no break temperature was observed. The activation energy of this enzyme in the physiological range of temperature was found to be high in the newborn, declining with brain maturation. The plot of this enzyme in the old rat showed no difference when compared with the mature. The specific activity of the enzyme, however, was markedly reduced in the old brain microsomal fraction. In contrast, comparison of the plots of microsomal NADPH-cytochrome c reductase at the various ages revealed no break temperature and very similar energies of activation. For the inner mitochondrial membrane beta-hydroxybutyrate dehydrogenase, the Arrhenius plots were generally biphasic at all ages studied with a break temperature of about 20 degrees C. However, the newborn plot was only barely biphasic showing a high energy of activation in the physiological range of temperature. In contrast, for the outer membrane NADH-cytochrome c reductase, the newborn plots were definitely biphasic, exhibiting low activation energy above the breaks. There was also a significant decline in the break temperature with brain development. No significant differences in the plots of this enzyme were found between the old and mature brain cortex. It is concluded that the enzymes of inner and outer mitochondrial membranes as well as the oxidative enzymes of microsomal membranes may show differential patterns of change in lipid-protein-interaction during development and aging, the changes being more marked in development than in aging.