LH release is facilitated by agents that alter cyclic AMP-generating system.
The issue of whether the adenosine 3',5'-monophosphate (cAMP)-generating system contributes to luteinizing hormone (LH) release was addressed by using several complementary probes in vitro. Pertussis toxin is considered to modify covalently an inhibitory adenylate cyclase regulatory protein. Treatment of gonadotrophs with this toxin increased both basal LH release and the efficacy of gonadotropin-releasing hormone (GnRH)-stimulated LH release with no apparent effect on GnRH potency. Cholera toxin, which probably activates adenylate cyclase by covalently altering another regulatory protein, forskolin, which directly stimulates the catalytic subunit of adenylate cyclase, and the cAMP analogue 8-Br-cAMP amplified both basal LH release (in a dose-dependent manner) and GnRH-stimulated LH release after a lag of 1 (cholera toxin and 8-Br-cAmP) and 4 (forskolin) h. It is noteworthy that these belated effects occurred in spite of the fact that cellular cAMP accumulation was markedly increased within 30 min after cholera toxin and at 1 min after forskolin addition. There was no change in total radioimmunoassayable LH (cellular + released) in either the basal or GnRH-treated cells after cholera toxin and forskolin for up to 24 h. Finally, the forskolin-amplified LH release was reversible and calcium dependent because D-600, EDTA, and calcium-free medium inhibited this effect. These results, generated with three complementary probes that affect integral proteins of the adenylate cyclase complex, suggest a function for cAMP in modulating LH release.


Detection of Retinal Pigment Epithelium-Specific Antibody in iPSC-Derived Retinal Pigment Epithelium Transplantation Models.
Antibody-mediated rejection is characterized by donor-specific antibody produced by B cells. However, to our knowledge, B cell invasion and antibody in the inflamed retina after transplantation of retinal pigment epithelial (RPE) cells has not been reported. To determine if RPE transplantation could be performed using allografts, we established in vivo immune rejection models with induced pluripotent stem cell (iPSC)-RPE allografts and determined whether RPE-specific antibody could be detected in these models. We detected alloantibodies in the serum from recipient monkeys that had immune attacks in the retina in an immunofluorescent assay using the transplanted iPSC-RPE cells as the antigen. In addition to T cell and antigen-presenting cell immunity, peripheral blood cells and lymph nodes in animal models with allogeneic iPSC-RPE cells also had activated B cells, which were probably secreting alloantibodies. Using serum and transplanted cells, alloreactive antibody can be detected for the diagnosis of immune rejection after transplantation.


A Review of Success Factors for Piglet Fostering in Lactation.
Piglet movement from one sow to another, or fostering, is required in modern pig farming but there is little available literature on the most effective strategy. In this review, we focus on the behavioural and physiological mechanisms responsible for piglet survival and growth, and have identified six key principles. (1) Colostrum provides piglets with warmth, energy and immunity. It is most accessible during the first 12 h from the birth sow, therefore no piglet should be moved before this; (2) To ensure even intake of birth sow colostrum, techniques such as split suckling prior to piglet movement should be implemented; (3) Udder assessment for functional teats should occur at farrowing, with number of fostered piglets not exceeding teat number; (4) Primiparous sows should receive as many piglets as the udder allows to maximise mammary stimulation, although older parities should be assessed for rearing ability; (5) Piglet fostering should occur between 12 and 24 h and movement kept to a minimum to prevent transfer of disease; Litter outliers should be moved and relocated to a litter of similar size; (6) Piglet movement after 24 h should be minimised. When required, strategies such as nurse usage should be employed. These principles will result in improved farrowing house performance by increasing the litter weight weaned per sow.


DNA methylation and expression of HLA-DR alpha.
B-cell lines established from two individuals with T-cell acute lymphocytic leukemia (T-ALL) express HLA-DR antigens, whereas the isogenic T-cells do not. The lack of expression correlates with a lack of detectable HLA-DR mRNA. All of the DR alpha DNA sequences detected by a cloned DR alpha cDNA probe are contained in a BglII fragment which varies slightly in size (4.0 to 4.8 kilobases) from one individual to another. In DNA from the T-cells not expressing DR alpha mRNA, all of the potential HpaII sites within the BglII fragment appeared to be methylated. In contrast, at least some of these sites were not methylated in DNA from the B-cells expressing high levels of DR alpha mRNA. Treatment of these T-cells with 5-azacytidine resulted in the induction of DR surface antigen expression, the appearance of DR alpha mRNA, and the partial demethylation of the DR alpha DNA sequences. T-cell lines established from human T-cell leukemia-lymphoma virus associated T-cell neoplasias, in contrast to the T-cell acute lymphocytic leukemia cell lines, expressed both DR antigens and DR alpha mRNA; the HpaII sites within the BglII fragment of DR alpha DNA of these human T-cell leukemia-lymphoma virus-positive T-cell lines were in all cases at least partially unmethylated. Uncultured peripheral blood T-cells from human T-cell leukemia-lymphoma virus-infected individuals expressed DR antigens at a low level, and the DR alpha locus was partially unmethylated. After 48 h in culture, DR antigen expression was substantially increased, but no significant changes were observed in methylation of the DR alpha locus or in the amount of DR mRNA which was present. This suggests that expression of DR antigens also can be modulated post-transcriptionally.


Bacterial community composition and diversity of two different forms of an organic residue of bioenergy crop.
The use of residue of sugarcane ethanol industry named vinasse in fertirrigation is an established and widespread practice in Brazil. Both non-concentrated vinasse (NCV) and concentrated vinasse (CV) are used in fertirrigation, particularly to replace the potassium fertilizer. Although studies on the chemical and organic composition of vinasse and their impact on nitrous oxide emissions when applied in soil have been carried out, no studies have evaluated the microbial community composition and diversity in different forms of vinasse. We assessed the bacterial community composition of NCV and CV by non-culturable and culturable approaches. The non-culturable bacterial community was assessed by next generation sequencing of the 16S rRNA gene and culturable community by isolation of bacterial strains and molecular and biochemical characterization. Additionally, we assessed in the bacterial strains the presence of genes of nitrogen cycle nitrification and denitrification pathways. The microbial community based on 16S rRNA sequences of NCV was overrepresented by Bacilli and Negativicutes while CV was mainly represented by Bacilli class. The isolated strains from the two types of vinasse belong to class Bacilli, similar to Lysinibacillus, encode for nirK gene related to denitrification pathway. This study highlights the bacterial microbial composition particularly in CV what residue is currently recycled and recommended as a sustainable practice in sugarcane cultivation in the tropics.


Endothelial Progenitor Cells as Mediators of the Crosstalk between Vascular Repair and Immunity: Lessons from Systemic Autoimmune Diseases.
From the discovery of Endothelial Progenitor Cells (EPC), these bone marrowderived precursors have been placed as crucial mediators of the endothelial repair. Accordingly, altered levels and function of EPC have been found in different scenarios of CV risk. Despite the fact that EPC exhibit important characteristics which support a link of this cell subset with a number of inflammatory and immune networks, little is known on the actual mediators involved and the clinical relevance of these features. Systemic diseases are generally hallmarked by a vascular repair failure and increased cardiovascular disease occurrence, EPC impairment having a pivotal role. Because of their immunemediated etiology, this group of conditions represents an invaluable scenario to unravel the connections between immune dysregulation and EPC dysfunction. In the present review, we summarize the current knowledge regarding the cutting-edge area of the modulation of EPC levels and function by inflammatory cytokines in systemic diseases. We also address the possibility of the available immunomodulatory drugs to counteract this situation. Finally, due to the emerging role of the vitamin D as a common mediator in the immune system and the cardiovascular axis, we cover the topic of the role of vitamin D as a potential player in the inflammatory-mediated EPC dysfunction in systemic diseases.


A Pilot Trial of the Combination of Transgenic NY-ESO-1-reactive Adoptive Cellular Therapy with Dendritic Cell Vaccination with or without Ipilimumab.
Transgenic adoptive cell therapy (ACT) targeting the tumor antigen NY-ESO-1 can be effective for the treatment of sarcoma and melanoma. Preclinical models have shown that this therapy can be improved with the addition of dendritic cell (DC) vaccination and immune checkpoint blockade. We studied the safety, feasibility, and antitumor efficacy of transgenic ACT with DC vaccination, with and without CTLA-4 blockade with ipilimumab. Freshly prepared autologous NY-ESO-1-specific T-cell receptor (TCR) transgenic lymphocytes were adoptively transferred together with NY-ESO-1 peptide-pulsed DC vaccination in HLA-A2.1-positive subjects alone (ESO, NCT02070406) or with ipilimumab (INY, NCT01697527) in patients with advanced sarcoma or melanoma. Six patients were enrolled in the ESO cohort, and four were enrolled in the INY cohort. Four out of six patients treated per ESO (66%), and two out of four patients treated per INY (50%) displayed evidence of tumor regression. Peripheral blood reconstitution with NY-ESO-1-specific T cells peaked within 2 weeks of ACT, indicating rapid in vivo expansion. Tracking of transgenic T cells to the tumor sites was demonstrated in on-treatment biopsies via TCR sequencing. Multiparametric mass cytometry of transgenic cells demonstrated shifting of transgenic cells from memory phenotypes to more terminally differentiated effector phenotypes over time. ACT of fresh NY-ESO-1 transgenic T cells prepared via a short ex vivo protocol and given with DC vaccination, with or without ipilimumab, is feasible and results in transient antitumor activity, with no apparent clinical benefit of the addition of ipilimumab. Improvements are needed to maintain tumor responses.


Enhanced Viability and Anti-rotavirus Effect of Bifidobacterium longum and Lactobacillus plantarum in Combination With Chlorella sorokiniana in a Dairy Product.
Microalgae and probiotics such as Bifidobacterium and Lactobacillus genera are associated with human beneficial effects. The aim of this study was to evaluate the activity of Chlorella sorokiniana on Bifidobacterium longum and Lactobacillus plantarum viability in a dairy product (flan) and its microbial effect against rotavirus, which is one of the major diarrhea-causing pathogens worldwide. Microalge were isolated from a Mexican river and characterized by molecular tools. Their prebiotic activity was evaluated by determining Bifidobacterium longum and Lactobacillus plantarum shelf-life after incorporation in the food matrix. In addition, HT-29 cells were infected with rotavirus Wa and treated with 1 × 109 CFU/mL L. plantarum and B. longum metabolites alone or in combination with 1 × 109 cells/mL Chlorella sorokiniana; viral titers in probiotics- and/or microalgae-treated cells were evaluated for antiviral activity. Results indicated that C. sorokiniana not only significantly (p < 0.05) improved L. plantarum and B. longum viability in flan, but also increased their antiviral activity; potent anti-rotavirus effect of C. sorokiniana alone was observed. Although more studies are needed, results suggest that incorporation of this microalga into a dairy product confers enhanced viability and antiviral effects, which indicates that C. sorokiniana might be used as an ingredient to design products with additional health benefits.


[Therapeutic observance in haematology: the particular case of chronic myeloid leukemia].
Therapeutic adhesion is essential because it modulates the efficiency of treatment, the occurrence of side effects and the socioeconomic costs possibly associated with it. In haematology, in the particular case of Chronic Myeloid Leukemia, the "ADAGIO" study performed by Noens and co-workers demonstrated that therapeutic observance is lower than the practitioners, the patients and the family members believe it is. This lack of adhesion can engender a decrease of the efficiency of treatment, in particular by imatinib. Factors such as the chronicity of the disease and the length of treatment modify therapeutic adhesion over time. Simple methods, addressing both the patient and the doctor should be developed to improve this therapeutic adhesion.


[Problems of aortocoronary bypass. Indications for coronary angiography and ventriculography; results of direct bypassing coronary surgery].
The introduction of selective coronary arteriography and aorto-to-coronary saphenous vein bypass surgery (a.-c bypass) has fundamentally altered our understanding of ischemic heart disease (IHD). The indications for the effective diagnostic procedure and the results of the new and increasingly important surgical technique are summarized. Selective coronary arteriography should be performed (a) in patients with known IHD in order to furnish the anatomical and functional information necessary to assess the indication for surgery, i.e. in patients below 60 years with intractable stable or unstable (impending infarction) angina. It is rarely indicated in patients with an old myocardial infarction who are free from symptoms. It is debatable in patients during the acute stage of infarction; (b) in patients with questionable IHD, with the aim of ruling out or confirming the condition, i.e. mainly in patients with atypical chest pain or with equivocal ecg findings. The risks of the procedure, if carried out by experienced personnel, are small. Selective arteriography will always be supplemented by a selective left ventricular angiography yielding important information concerning the functional behaviour of the myocardium. In judging the therapeutic value of a.-c. bypass surgery it should be noted that postoperatively 60 to 70 percent of the patients present without symptoms and 80 to 95 percent feel markedly better, and that physical performance is enhanced in about the same proportions. An improvement in left ventricular function under exercise conditions seems to be rare. Hospital mortality of a.-c. bypass operation is small and below 5 percent if patients with stable angina and without myocardial failure, previous infarctions or mitral regurgitation are considered. In the presence of an ischemic cardiomyopathy, on the other hand, the mere surgical risk soon reaches prohibitive limits. The incidence of early complicating myocardial infarctions ranges around 10 percent. Bypass occlusion occurs in some 5 to 15 percent during the early postoperative phase, while in the following months and years the patency rate diminishes but little. If the survival rates of operated and non-operated patients with IHD are compared it becomes evident that a prolongation of life is possible whenever surgery aims at a correction of two- and three-vessel disease (including the prognostically unfavourable isolated stenosis of the left anterior descending branch and stenosis of left main artery) whereas the natural course of isolated lesions of the right coronary artery and the left circumflex branch seems to balance the effect of corresponding surgical interventions. It should be borne in mind, however, that the follow-up periods on which these statements are based do not exceed 3-4 years.


Revealing the effect of 6-gingerol, 6-shogaol and curcumin on mPGES-1, GSK-3β and β-catenin pathway in A549 cell line.
In our study, anticancer effects of 6-gingerol, 6-shogaol from ginger and curcumin from turmeric were investigated and the results were compared with each other. We aimed to reveal their effects on microsomal prostaglandine E2 synthase 1 (mPGES-1) which is related with cancer progression and inflammation as well as β-catenin and glycogen synthase kinase 3β (GSK-3β) that are the main components of Wnt/GSK3 pathway. As it is known activation of GSK-3β and high levels of mPGES-1 pathway leads to cell proliferation and aggravates cancer progression. Therefore both of them are potential targets for cancer therapy. 6-shogaol and 6-gingerol' s effect on this pathway is not known very well up to now while curcumin that is known as an mPGES-1 inhibitor has anticancer properties via this pathway and many other pathways. Besides being in Zingiberaceae family, ginger's 6-gingerol and 6-shogaol have a molecular similarity with turmeric's curcumin. In our study we investigated their effects using a popular non small lung cancer cell line named A549 which expresses mPGES-1 and has active GSK3β pathway. IL-1β was used for inducing mPGES-1 and enabling the cancer characteristics such as cell proliferation. So compounds that inactivates or decreases the level of these components might be potential anticancer agents. A549 cells were incubated with interleukin 1β (IL-1β) for 24 h in order to maintain mPGES-1 enzyme induction. Experiments were performed both on IL-1β and non-IL-1β group. Real time cell analysis was performed to determine the cytotoxicity. Samples for western blotting and RT-PCR were collected after 24 h incubation with compounds to determine the amount of mPGES-1, GSK-3β, p-GSK-3β, β-catenin protein and mRNA. PGE2 which is the end product of mPGES-1 was measured by using ELISA kit. As a result of cell profile assay, cells exposed to IL-1β proliferate faster than non-IL-1β ones. This shows that induced mPGES-1 might play a role through GSK3β pathway and 24 h IC50 value of 6-shogaol is 62 μM. IL-1β increased protein and mRNA levels of mPGES-1, p-GSK-3β, β-catenin and GSK-3β in control group. Effects of curcumin and 6-shogaol on these parameters were against IL-1β's effect while 6-gingerol was not effective at all. Furthermore, 6-shogaol and curcumin might be effective on GSK3β pathway via lowering PGE2 levels. We saw that 6-shogaol is as effective as curcumin on this pathway and our study shows that 6-shogaol might show its anticancer properties via mPGES-1 and GSK3β pathway. May be these results might used for designing in vivo studies in future.


Disposition of a CYP2C9 phenotyping agent, losartan, is not influenced by the common 3435C > T variation of the drug transporter gene ABCB1 (MDR1).
Losartan is oxidized to E3174 by cytochrome P450 2C9 (CYP2C9); it has been suggested as a useful probe drug for CYP2C9 activity. It has also been shown to be a substrate for the drug-efflux transporter ATP-binding cassette sub-family B member 1 (ABCB1, MDR1). Both CYP2C9 and ABCB1 genes are polymorphic. The aim of the study was to determine if losartan disposition was influenced by the 3435C > T polymorphism of ABCB1 in healthy persons. These participants (n = 58) whose CYP2C9 genotypes and phenotypes were determined previously were genotyped for 3435C > T polymorphism in ABCB1. The concentrations of losartan and E3174 were compared across genotypes for ABCB1 3435C > T variation. For persons with the ABCB1 3435 CC, CT, TT genotypes, the concentrations (microM, means +/- S.D.) of neither losartan (1.76 +/- 0.87, 1.68 +/- 0.84 and 1.80 +/- 0.85, respectively, P = 0.70) nor E3174 (2.97 +/- 2.49, 2.53 +/- 2.09 and 3.18 +/- 2.75, respectively, P = 0.65) were significantly different. These results suggest that ABCB1 3435C > T polymorphism does not have any influence on losartan disposition. Therefore, ABCB1 3435C > T polymorphism is probably not a confounding factor in the prediction of CYP2C9 activity by using losartan as a probe agent.


What maintains signal honesty in animal colour displays used in mate choice?
Many of the colour displays of animals are proposed to have evolved in response to female mate choice for honest signals of quality, but such honest signalling requires mechanisms to prevent cheating. The most widely accepted and cited mechanisms for ensuring signal honesty are based on the costly signalling hypothesis, which posits that costs associated with ornamentation prevent low-quality males from being highly ornamented. Alternatively, by the index hypothesis, honesty can be achieved via cost-free mechanisms if ornament production is causally linked to core physiological pathways. In this essay, we review how a costly signalling framework has shaped empirical research in mate choice for colourful male ornaments and emphasize that alternative interpretations are plausible under an index signalling framework. We discuss the challenges in both empirically testing and distinguishing between the two hypotheses, noting that they need not be mutually exclusive. Finally, we advocate for a comprehensive approach to studies of colour signals that includes the explicit consideration of cost-free mechanisms for honesty.This article is part of the themed issue 'Animal coloration: production, perception, function and application'.


First-in-man phase I study of GC33, a novel recombinant humanized antibody against glypican-3, in patients with advanced hepatocellular carcinoma.
GC33 is a novel recombinant fully humanized monoclonal antibody that binds to human glypican-3 (GPC3). The antitumor activity of GC33 was shown in preclinical models of hepatocellular carcinoma (HCC). This first-in-man clinical trial was conducted to evaluate the safety, pharmacokinetic characteristics, and preliminary efficacy of GC33 in patients with advanced HCC. Patients with measurable, histologically proven, advanced HCC were enrolled to a dose-escalation study of GC33 (2.5-20 mg/kg) given intravenously weekly. The primary endpoint was to determine the maximum tolerated dose of GC33 for further development. Pharmacokinetic characteristics were measured in serum samples. Immunohistochemistry was conducted on tumor biopsies to evaluate GPC3 expression. Tumor response was assessed every 8 weeks using Response Evaluation Criteria in Solid Tumors criteria. Twenty patients were enrolled and treated with GC33. A maximum tolerated dose was not reached as there were no dose-limiting toxicities (DLT) up to the highest planned dose level. Common adverse events with all grades included fatigue (50%), constipation (35%), headache (35%), and hyponatremia (35%). The incidence of adverse events seemed not to be dose dependent. Trough serum concentrations at steady state were in excess of target concentration at doses of 5 mg/kg or greater. Median time to progression (TTP) was 26.0 weeks in the GPC3 high expression group and 7.1 weeks in the low expression group (P = 0.033). This study shows that GC33 was well tolerated in advanced HCC and provides preliminary evidence that GPC3 expression in HCC may be associated with the clinical benefit to GC33 that warrants prospective evaluation.


Detection by immunofluorescence assay of Bartonella henselae in lymph nodes from patients with cat scratch disease.
Laboratory diagnosis of Bartonella henselae infections can be accomplished by serology or PCR assay on biopsy samples. The purpose of our work was to assess immunofluorescence detection (IFD) in lymph node smears using a specific monoclonal antibody directed against B. henselae and a commercial serology assay (IFA) compared with PCR detection. Among 200 lymph nodes examined from immunocompetent patients, 54 were positive for B. henselae by PCR, of which 43 were also positive by IFD. Among the 146 PCR-negative lymph nodes, 11 were positive by IFD. Based on PCR results, the specificity of this new technique was 92.5%, the sensitivity was 79.6%, and the positive predictive value was 79.6%. At a cutoff titer of 64, the sensitivity of the IFA was 86.8% and the specificity was 74.1%. Diagnosis of cat scratch disease (CSD) may be improved, with a specificity of 100%, when the two tests (IFD and IFA) were negative; the sensitivity was 97.4% if one of the two tests was positive. Since PCR-based detection with biopsy samples is available only in reference laboratories, we suggest using IFD coupled with the commercial serology test for the diagnosis of CSD.


How can medical schools create the general practice workforce? An international perspective.
This article was migrated. The article was marked as recommended. Medical schools are responsible for producing the future medical workforce that is fit for purpose to meet the needs of patients. The burden of care largely falls upon primary care, a specialty which has been under-doctored, under-resourced, culturally not valued, and whose status has been subordinate to secondary care. In addition, the demographic changes of increasing longevity and chronic diseases, have increased the demand for healthcare at a time when governments' finances are stretched more than ever. The poor state of recruitment to the primary care workforce is a global phenomenon as evidenced by similar issues in North America, UK and the Middle East including Turkey. This contradicts the fact that the strength of primary care has a direct effect on the health of the nation. This paper presents an international perspective of why primary care is important, the reasons why new graduates shy away from a career in primary care, and what steps medical schools, residency programmes and governments should undertake to encourage medical students to make primary care a preferred career choice.


Comparison of pregnancy and neonatal outcomes in a retrospective full pregnancy history survey versus population-based prospective records: a validation study in rural Sarlahi District, Nepal.
Countries without complete civil registration and vital statistics systems rely on retrospective full pregnancy history surveys (FPH) to estimate incidence of pregnancy and mortality outcomes, including stillbirth and neonatal death. Yet surveys are subject to biases that impact demographic estimates, and few studies have quantified these effects. We compare data from an FPH vs. prospective records from a population-based cohort to estimate validity for maternal recall of live births, stillbirths, and neonatal deaths in a rural population in Sarlahi District, Nepal. We used prospective data, collected through frequent visits of women from early pregnancy through the neonatal period, from a population-based randomized trial spanning 2010-2017. We randomly selected 76 trial participants from three pregnancy outcome groups: live birth (n = 26), stillbirth (n = 25), or neonatal death (n = 25). Data collectors administered the Nepal 2016 Demographic and Health Surveys (DHS)-VII pregnancy history survey between October 22, 2021, and November 18, 2021. We compared total pregnancy outcomes and numbers of pregnancy and neonatal outcomes between the two data sources. We matched pregnancy outcomes dates in the two sources within ± 30 days and calculated measures of validity for adverse outcomes. Among 76 participants, we recorded 122 pregnancy outcomes in the prospective data and 104 outcomes in the FPH within ± 30 days of each woman's total observation period in the trial. Among 226 outcomes, we observed 65 live births that survived to 28 days, 25 stillbirths, and 32 live births followed by neonatal death in the prospective data and participants reported 63 live births that survived to 28 days, 15 stillbirths, and 26 live births followed by neonatal death in the pregnancy history survey. Sixty-two FPH outcomes were matched by date within ± 30 days to an outcome in prospective data. Stillbirth, neonatal death, higher parity, and delivery at a health facility were associated with likelihood of a non-matched pregnancy outcome. Stillbirth and neonatal deaths were underestimated overall by the FPH, potentially underestimating the burden of mortality in this population. There is a need to develop tools to reduce or adjust for biases and errors in retrospective surveys to improve reporting of pregnancy and mortality outcomes.


Liver Histopathology in Late Protocol Biopsies after Pediatric Liver Transplantation.
Liver transplantation has become a routine treatment for children with end stage liver failure. Recently, the long term survival of pediatric patients after liver transplantation has improved, with a life expectancy much longer than that of adult recipients, but also with longer exposition of the graft to various injuries, including immunological, inflammatory and others. Biochemical tests, although important, do not always reflect graft injury. The aim of our study was to analyze the histopathology of the graft in late protocol biopsies and correlate it with the clinical and biochemical status of these patients. We analyzed 61 protocol liver biopsies taken from 61 patients. Biopsies were taken 9.03-17.09 years (mean 12.68, median 11.74 years) after transplantation. Liver specimens were examined particularly for the presence and stage of liver fibrosis, inflammation, steatosis, and acute or chronic cellular and humoral rejection. We did not find any abnormalities in 26 (42.6%) liver specimens. None of the patients had signs of cellular or antibody mediated rejection or chronic rejection. In 23 liver biopsies (37.7%), we found non-specific lymphoid infiltrates. Another problem was fibrosis (equal to or more than three on the Ishak scale)-we found it in 17 patients, including seven liver specimens (11.5%) with severe fibrosis (Ishak 5-6). Conclusions: Various pathomorphological abnormalities were found in more than half of patients with a median 11.74 years post-transplant follow-up. Most of them presented normal laboratory liver tests at the same time, suggesting a slow subclinical process leading to pathomorphological abnormalities. No single factor for the development of these abnormalities was found, but our study supports the need for protocol liver biopsies even in patients with normal/almost normal biochemical liver tests.


Targeting MEK/MAPK signal transduction module potentiates ATO-induced apoptosis in multiple myeloma cells through multiple signaling pathways.
We demonstrate that blockade of the MEK/ERK signaling module, using the small-molecule inhibitors PD184352 or PD325901 (PD), strikingly enhances arsenic trioxide (ATO)-induced cytotoxicity in human myeloma cell lines (HMCLs) and in tumor cells from patients with multiple myeloma (MM) through a caspase-dependent mechanism. In HMCLs retaining a functional p53, PD treatment greatly enhances the ATO-induced p53 accumulation and p73, a p53 paralog, cooperates with p53 in caspase activation and apoptosis induction. In HMCLs carrying a nonfunctional p53, cotreatment with PD strikingly elevates the (DR4 + DR5)/(DcR1 + DcR2) tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) receptors ratio and caspase-8 activation of ATO-treated cells. In MM cells, irrespective of p53 status, the combined PD/ATO treatment increases the level of the proapoptotic protein Bim (PD-mediated) and decreases antiapoptotic protein Mcl-1 (ATO-mediated). Moreover, Bim physically interacts with both DR4 and DR5 TRAIL receptors in PD/ATO-treated cells, and loss of Bim interferes with the activation of both extrinsic and intrinsic apoptotic pathways in response to PD/ATO. Finally, PD/ATO treatment induces tumor regression, prolongs survival, and is well tolerated in vivo in a human plasmacytoma xenograft model. These preclinical studies provide the framework for testing PD325901 and ATO combination therapy in clinical trials aimed to improve patient outcome in MM.


Comparing Provider and Client Preferences for HIV Prevention Services in South Africa among Men Who Have Sex with Men.
Combination prevention efforts are now recommended toward reducing HIV incidence among men who have sex with men (MSM). Understanding the perceptions of both MSM and service providers is critical to informing the development of prevention packages and ultimately improving intervention effectiveness. This study assessed the preferences of MSM and health service providers in the administration of HIV-prevention efforts. Qualitative data were gathered from a series of separate MSM and health care provider focus groups in 2 South African cities. Participants discussed HIV-prevention services and MSM client experiences within South Africa and identified the 3 most important clinic characteristics and 3 most important HIV-prevention services for MSM clients. Priorities indicated by both MSM and health care providers were confidentiality of visit, friendly staff, and condoms, while discrepancies existed between MSM and providers regarding provider consistency and the provision of pre-exposure prophylaxis/post-exposure prophylaxis (PrEP/PEP) and lubricant as prevention methods. Effective interventions must address these discrepancies through the design of intervention and provider training to optimally accommodate MSM.


Managing emerging diseases borne by fruit bats (flying foxes), with particular reference to henipaviruses and Australian bat lyssavirus.
Since 1994, a number of novel viruses have been described from bats in Australia and Malaysia, particularly from fruit bats belonging to the genus Pteropus (flying foxes), and it is probable that related viruses will be found in other countries across the geographical range of other members of the genus. These viruses include Hendra and Nipah viruses, members of a new genus, Henipaviruses, within the family Paramyxoviridae; Menangle and Tioman viruses, new members of the Rubulavirus genus within the Paramyxoviridae; and Australian bat lyssavirus (ABLV), a member of the Lyssavirus genus in the family Rhabdoviridae. All but Tioman virus are known to be associated with human and/or livestock diseases. The isolation, disease associations and biological properties of the viruses are described, and are used as the basis for developing management strategies for disease prevention or control. These strategies are directed largely at disease minimization through good farm management practices, reducing the potential for exposure to flying foxes, and better disease recognition and diagnosis, and for ABLV specifically, the use of rabies vaccine for pre- and post-exposure prophylaxis. Finally, an intriguing and long-term strategy is that of wildlife immunization through plant-derived vaccination.


Gender- and age-dependencies of oxidative stress, as detected based on the steady state concentrations of different biomarkers in the MARK-AGE study.
Recently, Weber et al. published a thorough investigation of the age-dependency of oxidative stress (OS) determined by the steady state concentrations of different compounds - oxidation products and antioxidants - that are in common use as biomarkers of OS in 2207 healthy individuals of the cross-sectional MARK-AGE Project. The correlations among biomarkers were significant but weak. These findings may indicate different manifestations of OS and must further be evaluated. Here, we report a refined analysis of OS based on the above-mentioned original data. We show that malondialdehyde (MDA) appears to be sensitive to both gender and age. It is significantly lower and shows a greater age-dependence in women than in men. The age-dependency of MDA in women arises in a stepwise fashion. The age-dependent slope of the steady state concentration is maximal at the age between 50 and 55 years, indicating that it may be attributed to the change of metabolism in the post-menopause. Interestingly, total glutathione (GSH) decreased with age simultaneously with the increase in MDA. Different biomarkers yield different gender- and age-dependencies. Unlike the concentration of MDA, the concentrations of the other two oxidation products, i.e. protein carbonyls and 3-nitrotyrosine were similar in men and women and appeared to be independent of age in the healthy study population. The analyzed antioxidants exhibited different gender- and age-dependencies. In conclusion, it appears that all the biomarkers assessed here reflect different types of OS and that MDA and GSH reflect the same type of OS.


CD61 identifies a superior population of aged murine HSCs and is required to preserve quiescence and self-renewal.
Aging leads to a decline in function of hematopoietic stem cells (HSCs) and increases susceptibility to hematological disease. We found CD61 to be highly expressed in aged murine HSCs. Here we investigate the role of CD61 in identifying distinct subpopulations of aged HSCs and assess how expression of CD61 affects stem cell function. We show that HSCs with high expression of CD61 are functionality superior and retain self-renewal capacity in serial transplantations. In primary transplantations aged CD61High HSCs function similarly compared to young HSCs. CD61High HSCs are more quiescent compared to their CD61Low counterparts. We also show that in aged BM CD61High and CD61Low HSCs are transcriptomically distinct populations. Collectively, our research identifies CD61 as a key player in maintaining stem cell quiescence, ensuring the preservation of their functional integrity and potential during aging. Moreover, CD61 emerges as a marker to prospectively isolate a superior, highly dormant population of young and aged HSCs, making it a valuable tool both in fundamental and clinical research.


Endovascular treatment of fusiform aneurysms with stents and coils: technical feasibility in a swine model.
To assess the biomechanical feasibility of treating experimental fusiform aneurysms endovascularly with a combination of stents and coils. An experimental model was surgically constructed in the necks of nine swine to simulate intracranial fusiform aneurysms possessing important "perforators" or side branches. Balloon-expandable metal stents were positioned across the aneurysms in eight swine. In five of these, additional treatment was intraaneurysmal placement of detachable microcoils. Attempts were made to deposit these coils strategically away from the origin of the side branch. Stent placement was successful in seven swine but failed in one swine because of stent-aneurysm size mismatch. Two swine treated with only stents showed no significant alterations in blood filling of the aneurysm or side branch. Satisfactory coil placement (outside the stent, within the aneurysm sac, and away from the orifice of the side branch) was achieved in four of the five swine treated with stents and coils. Careful fluoroscopic monitoring and controlled coil delivery were necessary to avoid covering the sidebranch origin. These aneurysms could not be packed densely after detachment of the first coil because of the resultant radiographic overlap of multiple coil loops on the stent and its lumen in all projections. In one swine there was inadvertent untoward reentry of the coil tip into the expanded stent lumen during its delivery. Endovascular treatment of experimental fusiforms aneurysms using stents and coils is technically feasible. The stent maintains patency of the parent artery while allowing strategic coil placement in the aneurysm sac away from the origin of side branches. This technique may prove useful in the future treatment of intracranial fusiform aneurysms. However, potential sources of technical difficulties have been identified, and further longterm studies using an appropriate intracranial stent will be necessary before human application.


Phables: from fragmented assemblies to high-quality bacteriophage genomes.
Microbial communities have a profound impact on both human health and various environments. Viruses infecting bacteria, known as bacteriophages or phages, play a key role in modulating bacterial communities within environments. High-quality phage genome sequences are essential for advancing our understanding of phage biology, enabling comparative genomics studies and developing phage-based diagnostic tools. Most available viral identification tools consider individual sequences to determine whether they are of viral origin. As a result of challenges in viral assembly, fragmentation of genomes can occur, and existing tools may recover incomplete genome fragments. Therefore, the identification and characterization of novel phage genomes remain a challenge, leading to the need of improved approaches for phage genome recovery. We introduce Phables, a new computational method to resolve phage genomes from fragmented viral metagenome assemblies. Phables identifies phage-like components in the assembly graph, models each component as a flow network, and uses graph algorithms and flow decomposition techniques to identify genomic paths. Experimental results of viral metagenomic samples obtained from different environments show that Phables recovers on average over 49% more high-quality phage genomes compared to existing viral identification tools. Furthermore, Phables can resolve variant phage genomes with over 99% average nucleotide identity, a distinction that existing tools are unable to make. Phables is available on GitHub at https://github.com/Vini2/phables.


Differential Effects of Tango Versus Dance for PD in Parkinson Disease.
Over half of the general population does not achieve recommended daily levels of physical activity, and activity levels in people with Parkinson disease (PD) are lower than in healthy older adults. Dance can serve as an adjunct to traditional treatments to improve gait, balance, and quality of life in people with PD. This study directly compares a tango dance intervention and a dance intervention based on the Dance for PD model, which integrates multiple dance styles. Eleven people with PD participated in a community-based mixed styles dance intervention called Dance for Parkinson's (D4PD). Participants in the D4PD group were matched to participants in an ongoing community-based exercise study who participated in tango dance. The groups received 12 weeks of intervention, attending 1-h group classes twice a week. Participants were evaluated off anti-PD medication before and after intervention. Measures of balance, repeated sit-to-stand performance and endurance (mini-balance evaluation systems test, four square step test, five times sit to stand, 6-min walk time) improved from pre to post similarly in both groups. Motor sign severity (movement disorders society unified Parkinson disease rating scale motor subsection) and functional mobility (timed up and go) improved in the tango group and worsened in the D4PD group. Gait velocity was not affected by either intervention. Direct comparisons of different interventions are -critical for developing optimal exercise interventions designed to specifically target motor impairments in PD. Tango dance interventions may preferentially improve mobility and motor signs in people with PD, compared to D4PD.


Aplastic anemia in eosinophilic fasciitis: responses to immunosuppression and marrow transplantation.
Eosinophilic fasciitis (EF) is a rare connective tissue disorder which is frequently associated with hematologic disorders, especially aplastic anemia (AA) and variants (amegakaryocytic thrombocytopenia). The prognosis for AA with EF has generally been poor, but a few reports suggest a role for immunosuppressive therapy. We have seen four cases of AA complicating EF. All received corticosteroids and anti-thymocyte globulin without any benefit. One patient died of bleeding and infection. A second achieved unmaintained partial remission after two courses of cyclosporine A, although he had difficulty with side effects. Two patients received bone marrow transplants and both initially engrafted well. One had received marrow from a phenotypically HLA-matched parent and died of late graft failure. The second transplanted patient appears to be the only reported case of long term cure of both the AA and EF. Our four patients constitute the largest reported series of AA with EF and shed light on clinical aspects of the disease, and on the pathogenesis, particularly on responsiveness to different therapies; furthermore, there are implications to the treatment of AA in general.


Neighborhoods and obesity in later life.
We examined the influence of neighborhood environment on the weight status of adults 55 years and older. We conducted a 2-level logistic regression analysis of data from the 2002 wave of the Health and Retirement Study. We included 8 neighborhood scales: economic advantage, economic disadvantage, air pollution, crime and segregation, street connectivity, density, immigrant concentration, and residential stability. When we controlled for individual- and family-level confounders, living in a neighborhood with a high level of economic advantage was associated with a lower likelihood of being obese for both men (odds ratio [OR] = 0.86; 95% confidence interval [CI] = 0.80, 0.94) and women (OR = 0.83; 95% CI = 0.77, 0.89). Men living in areas with a high concentration of immigrants and women living in areas of high residential stability were more likely to be obese. Women living in areas of high street connectivity were less likely to be overweight or obese. The mechanisms by which neighborhood environment and weight status are linked in later life differ by gender, with economic and social environment aspects being important for men and built environment aspects being salient for women.


Acupuncture in reproductive medicine: the motivations of infertile women to participate in a randomised controlled trial.
Randomised controlled trials are the gold standard in medical research and are challenging to conduct successfully since high numbers of participants are needed to produce robust results. Therefore, it is important to understand what motivates patients to participate in one, particularly in Reproductive Medicine where the conduct of RCTs is rare. Just as it is important to evaluate medical interventions, it is equally important that adjuvant therapies are properly assessed. There has been an increased interest in adding acupuncture to in vitro fertilisation (IVF) in the hope of increasing the chance of pregnancy and a live birth. However, evidence that acupuncture assists IVF outcomes is conflicted and insight into the experiences and motivations of infertile women is important. This paper describes how an invitation to participate in an RCT of acupuncture as an adjuvant to IVF was received by infertile women and how they processed their decision to participate. In-depth interviews were conducted with 50 infertile women recruited from the RCT sample cohort. Recruitment aimed for maximum variation in social demographics. The data were saturated. Data pertaining to the theme of motivations to participate in an RCT were subjected to semantic thematic analysis. Two subthemes contained categories related to (a) the reasons women put forward for participation in an RCT, and (b) the rationale that underpinned and surrounded their decision. Women described themselves as active agents searching for a better outcome for their infertility or improved outcomes for women in the future. Their decision to participate in an RCT was motivated by factors such as opportunity, novelty and a value of science and was made after weighing various risks and benefits. The decision to participate in an RCT was an informed one. Infertile women in a stressful treatment situation participated in an RCT in the hope of finding a therapy to improve IVF outcomes for themselves and for other infertile women.


Detection of Circulating Tumor Cells (CTCs) in Malignant Pleural Mesothelioma (MPM) with the "Universal" CTC-Chip and An Anti-Podoplanin Antibody NZ-1.2.
Circulating tumor cell (CTC) is a potentially useful surrogate of micro-metastasis, but detection of rare tumor cells contaminated in a vast majority of normal hematologic cells remains technical challenges. To achieve effective detection of a variety of CTCs, we have developed a novel microfluidic system (CTC-chip) in which any antibody to capture CTCs is easily conjugated. In previous studies, we employed an antibody (clone E-1) against podoplanin that was strongly expressed on mesothelioma cells. The CTC-chip coated by the E-1 antibody (E1-chip) provided a modest sensitivity in detection of CTCs in malignant pleural mesothelioma (MPM). Here, to achieve a higher sensitivity, we employed a novel anti-podoplanin antibody (clone NZ-1.2). In an experimental model, MPM cells with high podoplanin expression were effectively captured with the CTC-chip coated by the NZ-1.2 antibody (NZ1.2-chip). Next, we evaluated CTCs in the peripheral blood sampled from 22 MPM patients using the NZ1.2-chip and the E1-chip. One or more CTCs were detected in 15 patients (68.2%) with the NZ1.2-chip, whereas only in 10 patients (45.5%) with the E1-chip. Of noted, in most (92.3%, 12/13) patients with epithelioid MPM subtype, CTCs were positive with the NZ1.2-chip. The CTC-count detected with the NZ1.2-chip was significantly higher than that with the E1-chip (p = 0.034). The clinical implications of CTCs detected with the NZ1.2-chip will be examined in a future study.


XDR tuberculosis and the Indian-subcontinent: effective prevention strategies needed.
The Indian-subcontinent is becoming, what could be, the next big battle ground for the control of Extensively Drug Resistant Tuberculosis (XDR-TB). The region caters to not only an ever-increasing incidence of new TB cases, but also a huge burden of already existing TB cases, thereby facing a dual threat for spread of XDR-TB. Most of the health-care facilities are devoid of essential laboratory-based infra-structure which is essential for diagnosing XDR-TB. Coupled to this is the lack of knowledge regarding TB diagnosis and treatment amongst the General Practitioners of India and Pakistan, who still treat the bulk of the TB cases. The element of AIDS is also slowly creeping into the equation. Questions have already been raised about the effectiveness of the governmental responses to curb the TB threat. What is needed right away is an effective strategy, incorporating a broad-tiered approach between the government and social sector. This can facilitate greatly in preventing further spread of these resistant strains.


A Comprehensive Histological Assessment of Osteoarthritis Lesions in Mice.
Accurate histological assessment of osteoarthritis (OA) is critical in studies evaluating the effects of interventions on disease severity. The purpose of the present study was to develop a histological grading scheme that comprehensively and quantitatively assesses changes in multiple tissues that are associated with OA of the stifle joint in mice. Two representative midcoronal sections from 158 stifle joints, including naturally occurring and surgically induced OA, were stained with H&E and Safranin-O stains. All slides were evaluated to characterize the changes present. A grading scheme that includes both measurements and semiquantitative scores was developed, and principal components analysis (PCA) was applied to the resulting data from the medial tibial plateaus. A subset of 30 tibial plateaus representing a wide range of severity was then evaluated by 4 observers. Reliability of the results was evaluated using intraclass correlation coefficients (ICCs) and area under the receiver operating characteristic (ROC) curve. Five factors were retained by PCA, accounting for 74% of the total variance. Interobserver and intraobserver reproducibilities for evaluations of articular cartilage and subchondral bone were acceptable. The articular cartilage integrity and chondrocyte viability factor scores were able to distinguish severe OA from normal, minimal, mild, and moderate disease. This newly developed grading scheme and resulting factors characterize a range of joint changes in mouse stifle joints that are associated with OA. Overall, the newly developed scheme is reliable and reproducible, characterizes changes in multiple tissues, and provides comprehensive information regarding a specific site in the stifle joint.


Predictive factors for a good prognosis following surgery for temporal lobe epilepsy: a cohort study in Spain.
To investigate the outcome of temporal lobe epilepsy surgery and identify the variables which predict a good prognosis with respect to seizures in postoperative follow-up after two and four years. This retrospective study included 115 selected patients who underwent surgery for temporal lobe epilepsy between 1996 and 2007. In the second year after surgery 86.1% of patients had a good prognosis for seizure control (73.9% Engel class I and 12.2% Engel class II) and 89.2% (76.3% Engel class I and 12.9% Engel class II) in the fourth year. Sixty-four of 93 (68.8%) patients were free of disabling seizures (Engel class I) during the entire period and 78 (83.8%) had good prognosis (Engel class I and II). For the second year, logistic regression analysis revealed the following variables to be independently predictive of good seizure control: absence of two or more seizure episodes in the first year after surgery, normal postoperative video-EEG, and age at surgery of less than 35 years. In the fourth year, mesial temporal sclerosis, female sex and normal postoperative video-EEG were the predictive factors. For the group with a good prognosis in both the second and the fourth year, the predictive variables were: absence of two or more seizure episodes in the first year after surgery (OR: 13.762, CI 95%: 2.566-73.808, p<0.002) and normal postoperative video-EEG (OR: 16.301, CI 95%: 3.704-71.740, p<0.001). This study illustrates the sustained benefit of temporal lobe epilepsy surgery. The multivariate logistic regression analysis failed to identify a good predictive model composed of preoperative variables alone, although it was possible to build such a model with either pre- and postoperative variables or only postoperative variables.


[Fast detection of DNA of hepatitis B virus by real time PCR].
90 blood plasma samples from patients with suspected chronic viral hepatitis B (CVHB) were analyzed by real-time polymerized chain reaction (PCR). The findings were compared with the results obtained by 2 PCR electrophoresis-based test-systems; the sensitivity limit for the quantification of DNA of hepatitis B virus (HBV) was determined; in the present case, the limit corresponded to 30 replications of HBV DNA to reaction (600 GE/ml). The positive result of real-time PCR was registered in 53.3% of cases. The quantity of HBV DNA replications in blood plasma samples varied from 30 to 3.9 x 10(6) per reaction (600-7.8 x 10(7) GE/ml). Serological profiles were analyzed in 18 patients with the verified diagnosis of CVHB. HBV DNA was detected in blood of 65% of HBsAg(+)-patients. The markers of HBeAg replication were noted in 35.5% of patients; it is noteworthy, that HBeAg(+)-samples were characterized by a higher level of viral loads (> or = 10(6) GE/ml) versus HBeAg(-)-samples (> or = 6 x 10(3) GE/ml). An analysis of blood-plasma samples dynamically obtained from one patient with chronic renal insufficiency and CVHB showed a decreased level of viral load from 1.7 x 10(7) GE/ml to a negative finding of real-time PCR registered after a therapy course by zeffix. Hence, the automated and standardized real-time PCR, when used at a multi-field patient-care facility, ensures a better diagnosis of viral hepatitis B.


Letter regarding "Chronic mechanical irritation and oral squamous cell carcinoma: A systematic review and meta-analysis".
Dear Editor: We have read with a great interest the article by Gupta et al. [1] who performed a meta-analysis exploring the association between chronic mechanical irritation and oral squamous cell carcinoma. The conclusion of the meta-analysis is that chronic oral mucosa irritation has a significant association with oral squamous cell carcinoma, and the nature of association could be that of a potential co-factor (dependent risk factor) rather than an independent risk factor. Read more in PDF.


Processing dewatered sewage sludge using electrokinetic technology.
High content of heavy metals and presence of pathogens in the dewatered sewage sludge have been the main obstacles for land application of sewage sludge-made fertilizer. The aim of this study was to examine the effects of the innovative electrokinetic (EK) technology on removal of heavy metals from sewage sludge, on the reduction of pathogens, and on sludge chemical characteristics. The results showed that the removal efficiencies for Zn, Cu, Ni, Cr, As and Pb were 94.9%, 95.4%, 89.7%, 67.8%, 31.2% and 18.7%, respectively. Acidification pretreatment of the dewatered sludge for 29 h decreased the content of heterotrophic bacteria from 1.5 x 10(8) c.f.u./g of wet sludge to 1.1 x 10(4) c.f.u./g of wet sludge. Although the initial content of total coliforms and fecal coliforms in sewage sludge were 5.8 x 10(5) c.f.u./g of wet sludge and 4.0 x 10(5) c.f.u./g of wet sludge, respectively, no viable cells were detected. Minor losses of K and N were detected, but the loss of P was found to be significant in EK treated sewage sludge. The treated sludge was technically considered as very stable based on the carbon dioxide evolution rate.


Fibrin degradation products and ovarian tumours.
Fibrin degradation products (F.D.P.) were determined in the serum of 163 women in whom ovarian tumours had been suspected on palpation at gynaecological examination and who were afterwards examined by laparoscopy or subjected to laparotomy. F.D.P. were found in the serum (0.5-30 mg/100 ml) of 23 (72%) out of 32 patients with malignant tumours. Of 131 patients with benign findings F.D.P. (traces to 2 mg/100 ml) were found in six (4.5%), and in most of these the occurrence of F.D.P. could be explained on other clinical grounds. The findings suggest that the examination of F.D.P. in suspected malignant ovarian tumour may be of diagnostic value.Determination of F.D.P. in malignant ascitic fluid showed very high values, ranging between 40 and 350 mg/ 100 ml. This argues for the occurrence of F.D.P. in the blood being due to an extravascular breakdown of fibrin caused by tumour cells, but they may also be due to thromboplastic and fibrinolytic agents from the tumour entering the blood stream.


Efficacy of transport media use versus direct inoculation of blood agar plates in the microbiologic evaluation of experimental Streptococcus pneumoniae keratitis.
To compare the microbiologic yield of cultures obtained by direct inoculation of blood agar plates (BAP) from corneal ulcer swabbings versus indirect inoculation via transport media in a rabbit model of Streptococcus pneumoniae bacterial keratitis. The corneas of 12 rabbits were inoculated with S. pneumoniae. Keratitis was confirmed 18 hours later. Sampling was performed at four 2.5-hour intervals. At each interval, corneal swabs were directly applied to BAP and placed into transport medium: thioglycollate and Amies medium without charcoal. Swabbings were then subcultured onto BAP at two time points: 2 and 24 hours after collection in transport medium. Plates were evaluated 48 hours later. Organism recovery rates were measured in terms of the number of positive culture plates observed and the bacterial colony counts on each plate. The rate of positive cultures overall was 69%. The recovery rates were similar for direct inoculation, inoculation via Amies held for 2 hours, and inoculation via Amies held for 24 hours. Direct inoculation yielded fewer colonies than indirect inoculation via Amies held for 24 hours (p = 0.008). Direct inoculation yielded a higher rate of positive cultures than did thioglycollate held for 2 hours (p = 0.004) or 24 hours (p < 0.001). The rate of nonpneumococcal contaminants ranged from 6% of BAP subcultured from thioglycollate held for 24 hours to 28% of directly inoculated BAP. Amies medium without charcoal may be used as a transport medium for up to 24 hours in the recovery of S. pneumoniae from corneal ulcers in this rabbit model. Thioglycollate appears to be less effective as a transport medium. Results of this study may justify studies of other transport media and/or other corneal pathogens. Altogether, such studies may provide justification for human clinical trials.


The effects of homocysteine and folic acid on angiogenesis and VEGF expression during chicken vascular development.
Homocysteine (Hcy) has been implicated in the development of cardiovascular developmental defects. Additionally, in experimental studies, vasculotoxic properties of Hcy have been described. Although Hcy has been identified as a vascular pathogen, little is known about the direct effects Hcy exerts during early embryonic vascular development. Angiogenesis is a critical process involved in embryo survival and development. There are limited studies on the effects of Hcy on early embryonic vasculogenesis and angiogenesis. Folic acid (FA) is a B vitamin essential in embryo development, and FA supplementation may lead to reduced Hcy levels. Therefore, the purpose of our study was to explore the effects of Hcy and FA on early embryonic vascular development. Embryonic day (E) 3.5 chicken embryos were treated with a sham, Hcy or FA solution. We developed a computational program for systematic analysis of microscopic images obtained from the extra embryonic vascular beds. These results were combined with real-time PCR data on the expression of VEGF-A and its receptor in these vascular beds. Our data show that Hcy exposure inhibits early vascular development, displayed by a significant reduction of vessel area and altered composition of the vascular beds. Vascular beds of Hcy embryos for the greater part consisted of vessels of the smallest diameters, compared to middle size vessels in control and FA embryos. Hcy also reduced expression of VEGF-A and VEGFR-2. No significant effects of FA were found. We conclude that Hcy exposure causes impaired early extra embryonic vascular development, shown by altered composition of the vascular beds as well as reduced expression of VEGF-A and VEGFR-2. These effects of Hcy, and the consecutive cascade of events, may be involved in the development of cardiovascular developmental defects.


Notes on valence bond structures for S2N2 and related systems.
For the D2h E2N2 and D4h E4(2+) (E=S, Se and Te) systems with one valence-shell π-electron atomic orbital per atomic centre, the ground-state resonance between two increased-valence structures with one-electron and fractional electron-pair π bonds is equivalent to resonance between the six Lewis-type valence-bond structures considered in this journal (B. Braïda, A. Lo, P. C. Hiberty, ChemPhysChem 2012, 13, 811-819). Approximate values are deduced for the one-electron bond polarity parameters. Three types of electron transfer processes are used to convert one increased-valence structure into the other. Theory is developed to show that one component of the associated electron transfer matrix element is not dependent on atomic orbital overlap. Brief consideration is also given to other types of VB descriptions for the S2N2 ground state.


Sadistic personality disorder and comorbid mental illness in adolescent psychiatric inpatients.
Sadistic personality disorder (SPD) is a controversial diagnosis proposed in the DSM-III-R, but not included in the DSM-IV. Few studies have focused on this disorder in adolescents. This article describes the results of a study that sought to determine the presence of sadistic personality characteristics in psychiatrically hospitalized adolescents and of comorbid Axis I or personality disorder patterns in those youth with SPD or SPD traits. Fifty-six adolescents were assessed for sadistic and other personality disorders with the Structured Interview for DSM-III-R Personality Disorders-Revised (SIDP-R). Axis I disorders were assessed using the Diagnostic Interview for Children and Adolescents, Adolescent Version (DICA-R-A) and portions of the Schedule for Affective Disorders and Schizophrenia for School Age Children, Epidemiologic (K-SADS-E). The youth were divided into those with SPD and SPD traits, the Sadistic Group (n = 18), and the Nonsadistic Group (n = 38). A significant proportion of the adolescents in this study met full DSM criteria for SPD (14%). The Sadistic Group (32%) had significantly more Axis I and personality pathology than did the Nonsadistic Group. However, all but one in the Sadistic Group met criteria for other personality disorders, confounding the interpretation of these findings and consistent with adult literature studies. Subjects with sadistic personality characteristics were identified in this adolescent inpatient sample, and they had more extensive Axis I and II psychopathology than the comparison group. The validity of this disorder in younger populations requires further study. Future studies should also explore the impact that the mandatory use of the pleasure/gratification criterion has on the validity of the SPD diagnosis and whether the requisite presence of this criterion decreases the overlap currently noted between SPD and other Axis II diagnoses.


Prediction of disability-free survival in healthy older people.
Prolonging survival in good health is a fundamental societal goal. However, the leading determinants of disability-free survival in healthy older people have not been well established. Data from ASPREE, a bi-national placebo-controlled trial of aspirin with 4.7 years median follow-up, was analysed. At enrolment, participants were healthy and without prior cardiovascular events, dementia or persistent physical disability. Disability-free survival outcome was defined as absence of dementia, persistent disability or death. Selection of potential predictors from amongst 25 biomedical, psychosocial and lifestyle variables including recognized geriatric risk factors, utilizing a machine-learning approach. Separate models were developed for men and women. The selected predictors were evaluated in a multivariable Cox proportional hazards model and validated internally by bootstrapping. We included 19,114 Australian and US participants aged ≥65 years (median 74 years, IQR 71.6-77.7). Common predictors of a worse prognosis in both sexes included higher age, lower Modified Mini-Mental State Examination score, lower gait speed, lower grip strength and abnormal (low or elevated) body mass index. Additional risk factors for men included current smoking, and abnormal eGFR. In women, diabetes and depression were additional predictors. The biased-corrected areas under the receiver operating characteristic curves for the final prognostic models at 5 years were 0.72 for men and 0.75 for women. Final models showed good calibration between the observed and predicted risks. We developed a prediction model in which age, cognitive function and gait speed were the strongest predictors of disability-free survival in healthy older people.Trial registration Clinicaltrials.gov (NCT01038583).


Administration of bifidobacterium and lactobacillus strains modulates experimental myasthenia gravis and experimental encephalomyelitis in Lewis rats.
Probiotics beneficial effects on the host are associated with regulation of the intestinal microbial homeostasis and with modulation of inflammatory immune responses in the gut and in periphery. In this study, we investigated the clinical efficacy of two lactobacillus and two bifidobacterium probiotic strains in experimental autoimmune myasthenia gravis (EAMG) and experimental autoimmune encephalomyelitis (EAE) models, induced in Lewis rats. Treatment with probiotics led to less severe disease manifestation in both models; ex vivo analyses showed preservation of neuromuscular junction in EAMG and myelin content in EAE spinal cord. Immunoregulatory transcripts were found differentially expressed in gut associated lymphoid tissue and in peripheral immunocompetent organs. Feeding EAMG animals with probiotics resulted in increased levels of Transforming Growth Factor-β (TGFβ) in serum, and increased percentages of regulatory T cells (Treg) in peripheral blood leukocyte. Exposure of immature dendritic cells to probiotics induced their maturation toward an immunomodulatory phenotype, and secretion of TGFβ. Our data showed that bifidobacteria and lactobacilli treatment effectively modulates disease symptoms in EAMG and EAE models, and support further investigations to evaluate their use in autoimmune diseases.


Muscle activity pattern regulates postnatal development of acetylcholinesterase molecular forms in normal mice and mice with motor endplate disease.
We have studied the relative contributions of muscle activity and nerve-supplied materials to the regulation of AChE molecular forms during postnatal development of muscles in normal mice and in mice with motor endplate disease (med mice). Onset of this hereditary disease causes a progressive failure of evoked release of ACh from the motor neuron, which prevents contraction in muscles such as biceps and soleus. In these innervated but inactive muscles, one can examine the consequences of inactivity on the distribution of AChE forms. In normal mouse biceps the distribution of AChE forms, as shown by sucrose-gradient analysis, change substantially after birth; the most dramatic alteration is an increase in G4 AChE from 15 to 45% of total AChE during the third postnatal week. AChE profiles in normal or med biceps are indistinguishable until 10-12 d after birth, but the changes in distribution of AChE forms does not occur in med biceps nor in normal biceps denervated 2 weeks after birth. In contrast, the distributions of AChE forms in a predominantly slow muscle, the soleus, are similar in med and normal mice both early (10 d) and late (20 d) in the course of the disease, and the distributions are affected little by denervation. The profiles of AChE forms seen in normal soleus at all times studied resembled those seen in newborn biceps or biceps inactivated by denervation or the med disease. We conclude that neither innervation, age-dependent changes intrinsic to muscle, nor muscle activity is sufficient to induce the changes we seen in AChE forms in biceps. These results support the hypothesis that neonatal, inactive, or tonically active muscles produce an intrinsic pattern of AChE molecular forms, and that a phasic pattern of activity induces a postnatal redistribution of the AChE molecular forms expressed by the muscle.


The Use of Social Networking Sites in Mental Health Interventions for Young People: Systematic Review.
The onset of mental health problems peaks between adolescence and young adulthood; however, young people face barriers to treatment and are often reluctant to seek professional help. Many are instead seeking support and information regarding their mental health via the Web, especially via social networking sites (SNSs), and hence, there is a promising opportunity to use SNSs to deliver or integrate with youth-focused online mental health interventions. Previous reviews have evaluated the effectiveness of SNSs for specific disorders in young people; however, none of the reviews have covered the breadth of SNS-based youth mental health interventions available across all mental health issues. This review aimed to systematically identify available evidence regarding the use of SNS-based interventions to support the mental health of young people aged up to 25 years, to evaluate their effectiveness, suitability, and safety, and identify gaps and opportunities for future research. The PubMed and PsycINFO databases were searched using Medical Subject Headings terms and exploded keywords and phrases. Retrieved abstracts (n=974) were double screened, yielding 235 articles for screening at the full-text level. Of these, 9 articles met the review inclusion criteria. Given the small number of studies, and the variety of outcome measures used, a quantitative meta-analysis was not possible. The 9 articles (quantitative studies, qualitative studies, and descriptions of the iterative design process) covered 5 separate interventions. Of the 5 interventions, 2 interventions used purpose-built platforms based on the moderated online social therapy (MOST) model, 2 used Facebook, and 1 evaluated a purpose-built mobile app. The 2 MOST interventions targeted specific mental health issues (depression and psychosis), whereas the others focused on improving mental health literacy, social support, and general well-being. Only 3 quantitative studies were identified, and all used a pre-post design (without a control group) to establish proof of concept. Of the outcome variables assessed, there were significant improvements in mental health knowledge and number of depressive symptoms but no improvement in anxiety or psychosis symptoms. Acceptability of and engagement with the SNS platforms were generally high, as were perceptions of usefulness and safety. Moderation by clinical experts was identified as a key component of the more successful interventions. When offered a choice, users showed a preference for mobile apps over Web-based interfaces. The evidence reviewed suggests young people find SNS-based interventions highly usable, engaging, and supportive. However, future studies need to address the current lack of high-quality evidence for their efficacy in reducing mental health symptoms. Given young people are already turning to SNSs to engage in knowledge seeking and peer-to-peer support, SNS-based youth mental health interventions provide an opportunity to address some of the barriers young people face in accessing qualified mental health support and information.


Preparative HPLC method for the purification of sulforaphane and sulforaphane nitrile from Brassica oleracea.
An extraction and preparative HPLC method has been devised to simultaneously purify sulforaphane and sulforaphane nitrile from the seed of Brassica oleracea var. italica cv. Brigadier. The seed was defatted with hexane, dried, and hydrolyzed in deionized water (1:9) for 8 h. The hydrolyzed seed meal was salted and extracted with methylene chloride. The dried residue was redissolved in a 5% acetonitrile solution and washed with excess hexane to remove nonpolar contaminants. The aqueous phase was filtered through a 0.22-microm cellulose filter and separated by HPLC using a Waters Prep Nova-Pak HR C-18 reverse-phase column. Refractive index was used to detect sulforaphane nitrile, and absorbance at 254 nm was used to detect sulforaphane. Peak identification was confirmed using gas chromatography and electron-impact mass spectrometry. Each kilogram of extracted seed yielded approximately 4.8 g of sulforaphane and 3.8 g of sulforaphane nitrile. Standard curves were developed using the purified compounds to allow quantification of sulforaphane and sulforaphane nitrile in broccoli tissue using a rapid GC method. The methodology was used to compare sulforaphane and sulforaphane nitrile content of autolyzed samples of several broccoli varieties.


[Vandetanib, in the management of patients with locally advanced or metastatic medullary thyroid carcinomas].
Medullary thyroid cancer (MTC) is a rare cancer. Vandetanib, a RET tyrosine-kinase inhibitor, significantly increased progression free survival and is the first treatment approved in France for unresectable, locally advanced or metastatic MTC that is symptomatic or progressive. Most frequents adverse events are diarrhea, folliculitis and asthenia. The prolongation of the QT interval on electrocardiogram is frequent but in the most of cases not clinically relevant. A good selection of the patients who could benefit from this treatment and management of side effect are important for the risk-benefit assessment.


Indistinguishable synaptic pharmacodynamics of the N-methyl-D-aspartate receptor channel blockers memantine and ketamine.
Memantine and ketamine, voltage- and activation-dependent channel blockers of N-methyl-d-aspartate (NMDA) receptors (NMDARs), have enjoyed a recent resurgence in clinical interest. Steady-state pharmacodynamic differences between these blockers have been reported, but it is unclear whether the compounds differentially affect dynamic physiologic signaling. In this study, we explored nonequilibrium conditions relevant to synaptic transmission in hippocampal networks in dissociated culture and hippocampal slices. Equimolar memantine and ketamine had indistinguishable effects on the following measures: steady-state NMDA currents, NMDAR excitatory postsynaptic current (EPSC) decay kinetics, progressive EPSC inhibition during repetitive stimulation, and extrasynaptic NMDAR inhibition. Therapeutic drug efficacy and tolerability of memantine have been attributed to fast kinetics and strong voltage dependence. However, pulse depolarization in drug presence revealed a surprisingly slow and similar time course of equilibration for the two compounds, although memantine produced a more prominent fast component (62% versus 48%) of re-equilibration. Simulations predicted that low gating efficacy underlies the slow voltage-dependent relief from block. This prediction was empirically supported by faster voltage-dependent blocker re-equilibration with several experimental manipulations of gating efficacy. Excitatory postsynaptic potential-like voltage commands produced drug differences only with large, prolonged depolarizations unlikely to be attained physiologically. In fact, we found no difference between drugs on measures of spontaneous network activity or acute effects on plasticity in hippocampal slices. Despite indistinguishable synaptic pharmacodynamics, ketamine provided significantly greater neuroprotection from damage induced by oxygen glucose deprivation, consistent with the idea that under extreme depolarizing conditions, the biophysical difference between drugs becomes detectable. We conclude that despite subtle differences in voltage dependence, during physiologic activity, blocker pharmacodynamics are largely indistinguishable and largely voltage independent.


Variational analysis of the mouse and rat eye optical parameters.
Rodent models are increasingly used to study refractive eye development and development of refractive errors; however, there is still some uncertainty regarding the accuracy of the optical models of the rat and mouse eye primarily due to high variability in reported ocular parameters. In this work, we have systematically evaluated the contribution of various ocular parameters, such as radii of curvature of ocular surfaces, thicknesses of ocular components, and refractive indices of ocular refractive media, using variational analysis and a computational model of the rodent eye. Variational analysis revealed that not all variation in ocular parameters has critical impact on the refractive status of the eye. Variation in the depth of the vitreous chamber, thickness of the lens, radius of the anterior surface of the cornea, radius of the anterior surface of the lens, as well as refractive indices for the lens and vitreous, appears to have the largest impact on the refractive error. The radii of the posterior surfaces of the cornea and lens have much smaller contributions to the refractive state. These data provide the framework for further refinement of the optical models of the rat and mouse eye and suggest that extra efforts should be directed towards increasing the linear resolution of the rodent eye biometry and obtaining more accurate data for the refractive indices of the lens and vitreous.


In-hospital mortality after acute myocardial infarction in patients with diabetes mellitus.
To examine in-hospital mortality after acute myocardial infarction in patients with diabetes mellitus. All patients in an 800-bed teaching hospital who had a discharge diagnosis of myocardial infarction, verified by creatine kinase levels at admission, between 1991 and 1993 made up the study population. All 118 such patients who died during this period made up the case group. Two control subjects (n = 236), survivors of the hospitalization, matched by sex, age, and length of hospitalization, were selected randomly for each case. Information on the presence of diabetes mellitus, medical history, and data related to myocardial infarction were obtained through retrospective chart review. The mean age of all subjects in the study was 76 years. Thirty-three percent of the patients in the case group and 31% of the control subjects had a history of diabetes mellitus (odds ratio = 1.04; 95% CI, 0.64-1.70), indicating that diabetes mellitus was not associated with an increased risk of in-hospital death. The adjusted odds ratio was 1.10 (95% CI, 0.48-2.51) in patients with non-insulin-treated diabetes mellitus and 0.80 (95% CI, 0.34-1.86) in insulin-treated patients. Multivariate analysis, with conditional logistic regression, confirmed that known prognostic factors for myocardial infarction, rather than diabetic status, are predictive of in-hospital mortality. Once the effects of age are accounted for, the risk of in-hospital mortality is not greater in patients with diabetes mellitus than in patients without diabetes; however, diabetes mellitus may be an important factor for long-term survival.


Phase I and pharmacokinetic study of KRN8602 alone and with filgrastim in patients with advanced cancer.
To determine the recommended dose, toxicity profile, and pharmacokinetics of KRN8602 (MX2-hydrochloride), a novel morpholino anthracycline with potent cytotoxicity against anthracycline-sensitive and resistant experimental tumors in vitro and in vivo. KRN8602 was administered alone in increasing doses to patients with advanced cancer or high-grade gliomas until dose-limiting toxicity (DLT) was observed in three or more of five patients treated in a dose level. Because neutropenia was dose limiting, further escalation was investigated with filgrastim support. Fifty-six assessable patients completed at least one cycle of chemotherapy. The recommended dose of KRN8602 alone was 40 mg/m2. Dose escalation was limited by neutropenia. The recommended dose of KRN8602 with filgrastim was 70 mg/m2, and limiting toxicities were neutropenia, diarrhea, and vomiting. The most commonly experienced nonhematologic toxicity was nausea and vomiting. Alopecia and mucositis were infrequent and mild. Pharmacokinetic parameters showed substantial variation, although the area under the plasma concentration-time curve (AUC) and maximum concentration both increased with dose. There was no relationship between pharmacokinetic parameters and toxicity. KRN8602 at doses of 40 mg/m2 when administered alone and 70 mg/m2 when administered with filgrastim appeared to be manageable. The major DLTs were neutropenia and, at higher doses, diarrhea and vomiting. The efficacy of this drug is currently being tested in phase II studies.


The Refugee Health Partnership: A Longitudinal Experiential Medical Student Curriculum in Refugee/Asylee Health.
In 2017, there were 25.4 million refugees worldwide, of whom 33,400 were resettled in the United States. In fiscal year 2016, 20,455 individuals were granted permanent asylum status in the United Sates. Both in the United States and overseas, refugees/asylees face significant disparities in accessing needed medical, mental health, and social support. The Refugee Health Partnership (RHP) was developed by Johns Hopkins University School of Medicine students and colleagues at a local refugee resettlement agency in 2011. The program pairs teams of preclinical medical students with recently resettled refugees/asylees who have special health care needs. After receiving training, students conducted monthly home visits and accompanied patients to appointments to assist them in navigating the health care system over one year. Students participated in monthly reflection exercises to process experiences and attended monthly seminars facilitated by expert faculty and guests. From 2012 to 2016, the RHP served 20 refugee families and engaged 60 students across four cohorts. Refugee participant retention was 20/22 (90.9%), and student retention was 57/60 (95.0%). In surveys completed at the end of their programs, students reported improvement in all measures, including understanding of different patient perspectives as well as comfort in communicating with patients across cultures and language barriers. The authors plan to integrate more objective measures of students' progress into the evaluations. They are scaling this model up both locally and beyond and plan to gather data from refugee/asylee participants to more accurately assess how they benefit from the program.


Assessing multiple factors affecting the gut microbiome structure of very preterm infants.
The composition and diversity of the gut microbiota are essential for the health and development of the immune system of infants. However, there is limited information on factors that influence the gut microbiota of very preterm infants. In this study, we analyzed factors that affect the gut microbiota of very preterm infants. The stool samples from 64 very preterm infants with a gestational age less than 32 weeks were collected for 16S rRNA gene sequencing. The infants were divided according to the delivery mode, antibiotic use during pregnancy, and feeding methods. The abundance of Proteobacteria was high in both cesarean (92.7%) and spontaneous (55.5%) delivery groups and then shifted to Firmicutes after the first week of birth. In addition, Proteobacteria was also the dominant phylum of infant gut microbiome for mothers with antibiotic use, with more than 50% after the first week of birth. In comparison, the dominant phylum for mothers without antibiotic use was Firmicutes. Proteobacteria level was also high in breastfeeding and mixed-feeding groups, consisting of more than 90% of the community. By contrast, Proteobacteria was the dominant phylum at the first week of birth but then shifted to Firmicutes for the formula-fed group. The alterations of gut microbiota in infants can affect their health condition during growth. This study confirmed that the different feeding types, delivery modes, and use of antibiotics during pregnancy can significantly affect the composition of the gut microbiota of very preterm infants.


Tuning the doping type and level of graphene with different gold configurations.
Au nanoparticles and films are deposited onto clean graphene surfaces to study the doping effect of different Au configurations. Micro-Raman spectra show that both the doping type and level of graphene can be tuned by fine control of the Au deposition. The morphological structures of Au on graphene are imaged by transmission electron microscopy, which indicate a size-dependent electrical characteristic: isolated Au nanoparticles produce n-type doping of graphene, while continuous Au films produce p-type doping. Accordingly, graphene field-effect transistors are fabricated, with the in situ measurements suggesting the tunable conductivity type and level by contacting with different Au configurations. For interpreting the experimental observations, the first-principles approach is used to simulate the interaction within graphene-Au systems. The results suggest that, different doping properties of Au-graphene systems are induced by the chemical interactions between graphene and the different Au configurations (isolated nanoparticle and continuous film).


Associations Between Sleep and Metabolic Outcomes in Preadolescent Children.
Growing evidence suggests an important role for sleep for the metabolic health of children. We aimed to determine how sleep is related to insulin sensitivity, insulin secretion, beta-cell function, and adiposity (BMI z-scores, body fat %, waist to height ratio) using objectively measured sleep and oral glucose tolerance test (OGTT)-derived measures. Sixty-two children aged 7-11 years, born at Kaiser Permanente Southern California, wore wrist accelerometers for 7 days to objectively measure sleep, completed an OGTT, and had anthropometric measures (height [cm], weight [kg], waist [cm], body fat [%]) collected. Using linear regression, associations between Matsuda insulin sensitivity index (ISI), insulinogenic index (IGI), disposition index (DI), BMI z-score, waist to height ratio, and body fat % with sleep parameters [total sleep time (TST; min), sleep efficiency (SE; %), time in bed (TIB; min), wake after sleep onset (WASO; min), and sleep latency (SL; min)] were assessed. Body fat % was tested as a mediator of the relationship between TST and ISI. Longer TST was associated with better insulin sensitivity (P = 0.02), but not after adjusting for body fat %. Sleep parameters were not associated with IGI or DI. Longer TST was associated with lower % body fat (P = 0.01) and lower waist-to-height-ratios (P = 0.05). Body fat % explained 62% (P = 0.01) of the relationship between TST and ISI. Longer TIB was associated with lower adiposity measures (P < 0.05). There were no associations between SE, WASO, or SL and metabolic outcomes. Objectively measured sleep duration was associated with lower adiposity, and the relationship between sleep duration and ISI appeared partly through adiposity levels in preadolescent children. Longer sleep duration may be important for metabolic health.


Genipin as a novel chemical activator of EBV lytic cycle.
Epstein-Barr virus (EBV) is a ubiquitous gammaherpesvirus that causes acute infection and establishes life-long latency. EBV causes several human cancers, including Burkitt's lymphoma, nasopharyngeal and gastric carcinoma. Antiviral agents can be categorized as virucides, antiviral chemotherapeutic agents, and immunomodulators. Most antiviral agents affect actively replicating viruses, but not their latent forms. Novel antiviral agents must be active on both the replicating and the latent forms of the virus. Gardenia jasminoides is an evergreen flowering plant belonging to the Rubiaceae family and is most commonly found growing wild in Vietnam, Southern China, Taiwan, Japan, Myanmar, and India. Genipin is an aglycone derived from an iridoid glycoside called geniposide, which is present in large quantities in the fruit of G. jasminoides. In this study, genipin was evaluated for its role as an antitumor and antiviral agent that produces inhibitory effects against EBV and EBV associated gastric carcinoma (EBVaGC). In SNU719 cells, one of EBVaGCs, genipin caused significant cytotoxicity (70 μM), induced methylation on EBV C promoter and tumor suppressor gene BCL7A, arrested cell-cycle progress (S phases), upregulated EBV latent/lytic genes in a dose-dependent manner, stimulated EBV progeny production, activated EBV F promoter for EBV lytic activation, and suppressed EBV infection. These results indicated that genipin could be a promising candidate for antiviral and antitumor agents against EBV and EBVaGC.


Lateralization of low-frequency tones and narrow bands of noise.
It is well known and universally accepted that people's ability to use ongoing interaural temporal disparities conveyed via pure tones is limited to frequencies below 1600 Hz. We wish to determine if this limitation is the result of the constant amplitude and periodic axis-crossings which characterize pure tones. To this end, an acoustic pointing task was employed in which listeners varied the interaural intensitive difference of a 500-Hz narrow-band noise (the pointer) so that the position of its intracranial image matched that of a second, experimenter-controlled stimulus (the target). Targets were either pure tones or narrow bands of noise (50 or 100 Hz wide). The narrow bands of noise were delayed interaurally in two distinct manners: Either the entire waveform or only the carrier was delayed. In the latter case, the envelopes and phase-functions of the bands of noise were identical interaurally. This resulted in noises which resemble the pure tone case in that the interaural delay is manifested as a constant phase-shift and resemble ordinary noises in that the envelope and phase are random functions of time. Surprisingly, it appears that all three targets were lateralized virtually identically regardless of frequency or bandwidth. Apparently, the dynamically changing envelopes and phases did not affect the listeners' use of interaural temporal disparities in any discernible fashion.


Femoral Neck Non-Union - A Biomechanical Solution with Valgus Sliding Subtrochanteric Osteotomy: A Case Report.
Avascular necrosis and non-union are two most dreaded complications of femoral neck fracture fixations. Hip replacement seems to be a simple solution for this complex problem. However, the long-term efficacy of prosthetic replacement in the young population with higher functional demand is still questionable. Femoral head preserving valgus subtrochanteric osteotomies in properly selected cases have strong support from literature. The conventional technique of valgus subtrochanteric osteotomy involves lateral based wedge resection. Alternatively, a simpler sliding oblique subtrochanteric osteotomy without any wedge removal can also be performed. We hereby describe a successful case of sliding subtrochanteric osteotomy with 135° dynamic hip screw (DHS) plate fixation in treating non-union neck of femur fracture in a young gentleman.


Hypertriglyceridemia independent propofol-induced pancreatitis.
Propofol can cause acute pancreatitis related to hypertriglyceridemia, however, other mechanisms may also exist. A 71-year-old male on propofol infusion in the intensive care setting, developed acute pancreatitis as confirmed biochemically and by imaging. He did not have any elevation of triglycerides and on propofol re-challenge, had recurrence of pancreatitis with the absence of triglyceride elevation. We report a case of hypertriglyceridemia independent propofol induced pancreatitis possibly related to an idiosyncratic reaction and propose that this drug to be reassigned as a class Ia drug as an etiologic factor for acute pancreatitis.


Impact of location of inpatient cancer care on patients with Ewing sarcoma and osteosarcoma-A population-based study.
Ewing sarcoma (EWS) and osteosarcoma (OS) require multidisciplinary treatment. Care at specialized cancer centers (SCC: Children's Oncology Group affiliated and/or National Cancer Institute-designated cancer center) has been found to improve outcomes in patients with leukemia, but studies have not considered location of care and outcomes in EWS and OS patients, an ideal group to evaluate given their specialized multidisciplinary treatment needs. Patients hospitalized with primary EWS and OS (2000-2014) were identified using the California Cancer Registry linked with hospitalization data. Patients were divided into age groups (0-18, 19-39, ≥40 years), and classified on whether they received all versus part/none of their inpatient treatment at a SCC within 1 year of diagnosis. Multivariable Cox proportional hazards regression identified factors associated with survival. There were 531 ES and 959 OS patients. Five-year overall survival was better for patients with EWS (all: 63% vs. part/none: 42%) and OS (all: 64% vs. part/none: 47%) who received all of their treatment at a SCC. After adjusting for sociodemographic and clinical factors, receiving all inpatient cancer treatment at a SCC was associated with superior overall survival (EWS HR: 0.49, CI 0.37-0.67; OS HR: 0.78, CI 0.63-0.97). Our results suggest that treatment for EWS and OS at a SCC is associated with significantly improved survival even after adjustment for known prognostic factors. The superior survival among those treated at SCCs may be due to having greater access to clinical trials and services at SCCs.


Multi-biomarker panel prediction model for diagnosis of pancreatic cancer.
The current study aimed to develop a prediction model using a multi-marker panel as a diagnostic screening tool for pancreatic ductal adenocarcinoma. Multi-center cohort of 1991 blood samples were collected from January 2011 to September 2019, of which 609 were normal, 145 were other cancer (colorectal, thyroid, and breast cancer), 314 were pancreatic benign disease, and 923 were pancreatic ductal adenocarcinoma. The automated multi-biomarker Enzyme-Linked Immunosorbent Assay kit was developed using three potential biomarkers: LRG1, TTR, and CA 19-9. Using a logistic regression model on a training data set, the predicted values for pancreatic ductal adenocarcinoma were obtained, and the result was classification into one of the three risk groups: low, intermediate, and high. The five covariates used to create the model were sex, age, and three biomarkers. Participants were categorized into four groups as normal (n = 609), other cancer (n = 145), pancreatic benign disease (n = 314), and pancreatic ductal adenocarcinoma (n = 923). The normal, other cancer, and pancreatic benign disease groups were clubbed into the non-pancreatic ductal adenocarcinoma group (n = 1068). The positive and negative predictive value, sensitivity, and specificity were 94.12, 90.40, 93.81, and 90.86, respectively. This study demonstrates a significant diagnostic performance of the multi-marker panel in distinguishing pancreatic ductal adenocarcinoma from normal and benign pancreatic disease states, as well as patients with other cancers.