Subcutaneous granuloma caused by Phialophora richardsiae: case report and review of the literature.
A phaeomycotic cyst in a 47-year-old man, caused by Phialophora richardsiae, was treated successfully by excision. A critical review of the literature indicates that the pathological course of P. richardsiae infections usually follows a similar pattern to that of the present case, viz. generation of a well-defined and limited nodule following traumatic subcutaneous introduction of the fungus. The nodule typically accumulates a viscous yellow fluid. The infection remains localized, often being encapsulated by a fibrous layer; adenopathy is not observed. Concomitant bacterial infections are also common; in the present case Pseudomonas stutzeri was identified. Neither bacterial nor fungal infection recurred after surgery.


RNA-Guided RNA Modifications: Biogenesis, Functions, and Applications.
ConspectusPost-transcriptional modifications are ubiquitous in both protein-coding and noncoding RNAs (ncRNAs), playing crucial functional roles in diverse biological processes across all kingdoms of life. These RNA modifications can be achieved through two distinct mechanisms: RNA-independent and RNA-guided (also known as RNA-dependent). In the RNA-independent mechanism, modifications are directly introduced onto RNA molecules by enzymes without the involvement of other RNA molecules, while the cellular RNA-guided RNA modification system exists in the form of RNA-protein complexes, wherein one guide RNA collaborates with a set of proteins, including the modifying enzyme. The primary function of guide RNAs lies in their ability to bind to complementary regions within the target RNAs, orchestrating the installation of specific modifications. Both mechanisms offer unique advantages and are critical to the diverse and dynamic landscape of RNA modifications. RNA-independent modifications provide rapid and direct modification of RNA molecules, while RNA-guided mechanisms offer precise and programmable means to introduce modifications at specific RNA sites. Recently, emerging evidence has shed light on RNA-guided RNA modifications as a captivating area of research, providing precise and programmable control over RNA sequences and functions.In this Account, we focus on RNA modifications synthesized in an RNA-guided manner, including 2'-O-methylated nucleotides (Nm), pseudouridine (Ψ), N4-acetylcytidine (ac4C), and inosine (I). This Account sheds light on the intricate processes of biogenesis and elucidates the regulatory roles of these modifications in RNA metabolism. These roles include pivotal functions such as RNA stability, translation, and splicing, where each modification contributes to the diverse and finely tuned regulatory landscape of RNA biology. In addition to elucidating the biogenesis and functions of these modifications, we also provide an overview of high-throughput methods and their underlying biochemical principles used for the transcriptome-wide investigation of these modifications and their fundamental interactions in RNA-guided systems. This includes exploring RNA-protein interactions and RNA-RNA interactions, which play crucial roles in the dynamic regulatory networks of RNA-guided modifications. The ever-advancing methodologies have greatly enhanced our understanding of the dynamic and widespread nature of RNA-guided RNA modifications and their regulatory functions. Furthermore, the applications of RNA-guided RNA modifications are discussed, illuminating their potential in diverse fields. From basic research to gene therapy, the programmable nature of RNA-guided modifications presents exciting opportunities for manipulating gene expression and developing innovative therapeutic strategies.


Distinct Longitudinal Changes in Immunoglobulin G N-Glycosylation Associate with Therapy Response in Chronic Inflammatory Diseases.
Immunosuppressants and biologicals are widely used therapeutics for various chronic inflammatory diseases (CID). To gain more detailed insight into their downstream effects, we examined their impact on serum immunoglobulin G (IgG) glycosylation. We analyzed IgG subclass-specific fragment crystallizable (Fc) N-glycosylation in patients suffering from various CID using the LC-MS approach. Firstly, we compared IgG Fc N-glycosylation between 128 CID patients and 204 healthy controls. Our results replicated previously observed CID-related decrease in IgG Fc galactosylation (adjusted p-value range 1.70 × 10-2-5.95 × 10-22) and sialylation (adjusted p-value range 1.85 × 10-2-1.71 × 10-18). Secondly, to assess changes in IgG Fc N-glycosylation associated with therapy and remission status, we compared 139 CID patients receiving either azathioprine, infliximab, or vedolizumab therapy. We observed an increase in IgG Fc galactosylation (adjusted p-value range 1.98 × 10-2-1.30 × 10-15) and sialylation (adjusted p-value range 3.28 × 10-6-4.34 × 10-18) during the treatment. Furthermore, patients who reached remission displayed increased Fc galactosylation levels (p-value range 2.25 × 10-2-5.44 × 10-3) in comparison to patients with active disease. In conclusion, the alterations in IgG Fc glycosylation and the fact these changes are even more pronounced in patients who achieved remission, suggest modulation of IgG inflammatory potential associated with CID therapy.


More operations, more deaths? Relationship between operative intervention rates and risk-adjusted mortality at trauma centers.
The Trauma Quality Improvement Project has demonstrated significant variations in risk-adjusted mortality rates across the designated trauma centers. It is not known whether the outcome differences are related to provider-level clinical decision making. We hypothesized that centers with good outcomes undertake critical operative interventions aggressively, thereby avoiding complications and deaths. The previously validated Trauma Quality Improvement Project risk-adjustment algorithm was used to measure observed-to-expected mortality rates (O/E with 90% confidence intervals [CI]) for 152 Level I and II trauma centers participating in the National Trauma Data Bank (version 7.0). Adult patients (>or=16 years) with at least one severe injury (Abbreviated Injury Scale score >or=3) were included (N = 135,654). Operative intervention rates for solid organ injuries (spleen, liver, and kidney) were compared between the centers classified as high mortality (O/E with CI > 1, n = 35 centers) versus low mortality (O/E with CI < 1, n = 37 centers) using nonparametric tests. Low- and high-mortality trauma centers were similar in designation level, hospital and intensive care unit beds, teaching status, and number of trauma, orthopedic, and neurosurgeons. Despite a similar incidence and severity of solid organ injuries, low-mortality centers were less likely to undertake operative interventions. Trauma centers with higher risk-adjusted mortality rates are more likely to undertake operative interventions for solid organ injuries. Hence, there is a need to focus quality improvement efforts on medical decision-making and perioperative processes of care.


Twinned or not twinned, that is the question: crystallization and preliminary crystallographic analysis of the 2F1(3)F1 module pair of human fibronectin.
Human fibronectin (Fn) is a large multidomain protein found in the extracellular matrix and plasma. It is involved in many cellular processes, including cell adhesion and migration during embryogenesis and wound healing. The ability to bind Fn is a characteristic that has been demonstrated for a number of pathogens. For Staphylococcus aureus and Streptococcus pyogenes in particular, Fn-binding bacterial proteins (FnBPs) have been shown to mediate not only bacterial adhesion to host cells but also the uptake of bacteria by the cells. FnBPs interact with the amino-terminal region of Fn, where five type I ((1-5)F1) Fn modules are located. Although the structures of two F1 module pairs have been determined by NMR, no X-ray structures have been reported. To explore the conformational interactions between modules and the binding properties of FnBPs, the (2)F1(3)F1 module pair was crystallized using the vapour-diffusion method at 298 K. 12 X-ray diffraction data sets have been collected: six on an in-house rotating anode (three native, one Pt derivative and two peptide-bound) and six at synchrotron-radiation sources (two native and four derivative). Following analysis of these data, some of which have very high multiplicity (up to 50), probable space-group assignments were made (P42(1)2, P4(1)2(1)2 or P4(3)2(1)2) and the possibly twinned nature of the crystals was investigated using six different tests. The results presented here suggest that the crystals are not twinned.


Opioid Agonist Therapy During Hospitalization Within the Veterans Health Administration: a Pragmatic Retrospective Cohort Analysis.
Hospitalization of patients with opioid use disorder (OUD) is increasing, yet little is known about opioid agonist therapy (OAT: methadone and buprenorphine) administration during admission. Describe and examine patient- and hospital-level characteristics associated with OAT receipt during hospitalization in the Veterans Health Administration (VHA). A total of 12,407 unique patients, ≥ 18 years old, with an OUD-related ICD-10 diagnosis within 12 months prior to or during index hospitalization in fiscal year 2017 from 109 VHA hospitals in the continental U.S. OAT received during hospitalization. Few admissions received OAT (n = 1914; 15%) and when provided it was most often for withdrawal management (n = 834; 7%). Among patients not on OAT prior to admission who survived hospitalization (n = 10,969), 2.0% (n = 203) were newly initiated on OAT with linkage to care after hospital discharge. Hospitals varied in the frequency of OAT delivery (range, 0 to 43% of qualified admissions). Patients with pre-admission OAT (adjusted odds ratio [AOR] = 15.30; 95% CI [13.2, 17.7]), acute OUD diagnosis (AOR = 2.3; 95% CI [1.99, 2.66]), and male gender (AOR 1.52; 95% CI [1.16, 2.01]) had increased odds of OAT receipt. Patients who received non-OAT opioids (AOR 0.53; 95% CI [0.46, 0.61]) or surgical procedures (AOR 0.75; 95% CI [0.57, 0.99]) had decreased odds of OAT receipt. Large-sized (AOR = 2.0; 95% CI [1.39, 3.00]) and medium-sized (AOR = 1.9; 95% CI [1.33, 2.70]) hospitals were more likely to provide OAT. In a sample of VHA inpatient medical admissions, OAT delivery was infrequent, varied across the health system, and was associated with specific patient and hospital characteristics. Policy and educational interventions should promote hospital-based OAT delivery.


Immunoarchitecture of the regenerating rat spleen: effects of partial splenectomy and heterotopic autotransplantation.
To investigate the microstructure of in situ (eutopic) and autotransplanted (ectopic) splenic remnants, adult Sprague-Dawley rats were studied 60 days after subtotal (approximately 80%) splenectomy, total splenectomy followed by single or multiple remnant intraperitoneal autotransplantation, or sham operation. Total nucleated cell counts were determined in excised splenic remnants, and immunohistochemical staining using monoclonal antibodies to rat B- and T-cell antigens was performed in serial tissue sections. Immunoarchitecture of eutopic remnants was indistinguishable from that of intact spleens and total nucleated cell counts remained proportional to weight. In contrast, ectopic remnants showed sparsity and abnormal mixing of B and T lymphocyte subpopulations with widespread loss of follicles and periarteriolar lymphoid sheaths in addition to lower density and marked reduction of total nucleated cells. These findings provide immunohistologic evidence that preservation of intact vasculature is critical to splenic architecture, which may account in part for the demonstrable functional inferiority of ectopic remnants.


YY-1224, a terpene trilactone-strengthened Ginkgo biloba, attenuates neurodegenerative changes induced by β-amyloid (1-42) or double transgenic overexpression of APP and PS1 via inhibition of cyclooxygenase-2.
Ginkgo biloba has been reported to possess free radical-scavenging antioxidant activity and anti-inflammatory properties. In our pilot study, YY-1224, a terpene trilactone-strengthened extract of G. biloba, showed anti-inflammatory, neurotrophic, and antioxidant effects. We investigated the pharmacological potential of YY-1224 in β-amyloid (Aβ) (1-42)-induced memory impairment using cyclooxygenase-2 (COX-2) knockout (-/-) and APPswe/PS1dE9 transgenic (APP/PS1 Tg) mice. Repeated treatment with YY-1224 significantly attenuated Aβ (1-42)-induced memory impairment in COX-2 (+/+) mice, but not in COX-2 (-/-) mice. YY-1224 significantly attenuated Aβ (1-42)-induced upregulation of platelet-activating factor (PAF) receptor gene expression, reactive oxygen species, and pro-inflammatory factors. In addition, YY-1224 significantly inhibited Aβ (1-42)-induced downregulation of PAF-acetylhydrolase-1 (PAF-AH-1) and peroxisome proliferator-activated receptor γ (PPARγ) gene expression. These changes were more pronounced in COX-2 (+/+) mice than in COX-2 (-/-) mice. YY-1224 significantly attenuated learning impairment, Aβ deposition, and pro-inflammatory microglial activation in APP/PS1 Tg mice, whereas it significantly enhanced PAF-AH and PPARγ expression. A preferential COX-2 inhibitor, meloxicam, did not affect the pharmacological activity by YY-1224, suggesting that the COX-2 gene is a critical mediator of the neuroprotective effects of YY-1224. The protective activity of YY-1224 appeared to be more efficacious than a standard G. biloba extract (Gb) against Aβ insult. Our results suggest that the protective effects of YY-1224 against Aβ toxicity may be associated with its PAF antagonistic- and PPARγ agonistic-potential as well as inhibition of the Aβ-mediated pro-inflammatory switch of microglia phenotypes through suppression of COX-2 expression.


Circumcision of HIV-infected men and transmission of human papillomavirus to female partners: analyses of data from a randomised trial in Rakai, Uganda.
Male circumcision reduces the transmission of high-risk human papillomavirus (HPV) in HIV-uninfected men and their female partners. We assessed whether circumcision of HIV-infected men would reduce the transmission of high-risk HPV to their female partners. Female partners of HIV-infected men (aged 15-49 years) in Rakai, Uganda, with CD4 counts of greater than 350 cells per mL who were randomly assigned to undergo circumcision immediately (intervention group) and after 24 months (control group) were assessed for infection with high-risk HPV. Randomisation was done in blocks of 20, stratified by community, with computer-generated random numbers. Laboratory technicians and female fieldworkers were masked to the circumcision status of male participants. The main outcome assessed in this study was the effects of circumcision of HIV-infected men on transmission of HPV to their female partners. Analysis was by intention to treat. The trial is registered with ClinicalTrials.gov, number NCT00124878. 474 men were assigned to the intervention group, 448 to the control group. 211 women were in consensual relationships with 193 men in the intervention group, and 171 women were in consensual unions with 155 men in the control group. High-risk HPV at the 2-year follow-up was prevalent in 88 female partners (55%) of 159 men in the intervention group and 68 (52%) of 131 female partners of men in the control group (prevalence risk ratio 1·07, 95% CI 0·86-1·32, p=0·64). Incidence of high-risk HPV over 2 years was 32·0 per 100 person-years in the female partners of men in the intervention group and 30·6 per 100 person-years in the female partners of men in the control group (incidence rate ratio 1·05, 0·77-1·43, p=0·78). No difference was noted in the clearance of genotype-specific high-risk HPV between the intervention group (196 [46%] of 424) and control group (167 [48%] of 347; rate ratio 0·96, 0·83-1·12; p=0·61). Because circumcision of HIV-infected men did not affect transmission of high-risk HPV to their female partners, promotion of consistent safe sexual practices for HIV-infected men remains important. Bill & Melinda Gates Foundation and National Institutes of Health.


"Crisis management in the hospitality sector SMEs in Pakistan during COVID-19″.
The COVID-19 pandemic has delivered one of the worst economic shocks in modern history and the hospitality sector has been severely affected. Since small businesses from the hospitality sector are known to be relatively more susceptible to the economic complications arising from a crisis, we explore the underlying factors and management practices that influence their continuity of operations as they continue to struggle with the on-going COVID-19 crisis in Pakistan. Using a phenomenological approach, in-depth interviews were conducted to comprehend the experiences of owners-managers. The findings show that government support, cordial relationships with stakeholders, self-determination of entrepreneurs and formal planning are the most crucial factors that shaped the immediate adjustments of operational activities in response to COVID-19. These resilient practices are hygiene concerns, increased promotion through social media, innovative marketing practices (e.g., revised offerings), operational cost-cutting and employee training to comply with changing standard operating procedures from the government and industry. The practical and theoretical implications are also discussed.


Nap sleep preserves associative but not item memory performance.
Many studies have shown that sleep improves memory performance, and that even short naps during the day are beneficial. Certain physiological components of sleep such as spindles and slow-wave-sleep are thought to be particularly important for memory consolidation. The aim of this experiment was to reveal the role of naps for hippocampus-dependent associative memory (AM) and hippocampus-independent item memory (IM) alongside their corresponding ERP old/new effects. Participants learnt single words and word-pairs before performing an IM- and an AM-test (baseline). One group was subsequently allowed to nap (∼90min) while the other watched DVDs (control group). Afterwards, both groups performed a final IM- and AM-test for the learned stimuli (posttest). IM performance decreased for both groups, while AM performance decreased for the control group but remained constant for the nap group, consistent with predictions concerning the selective impact of napping on hippocampus-dependent recognition. Putative ERP correlates of familiarity and recollection were observed in the IM posttest, whereas only the later recollection-related effect was present in the AM test. Notably, none of these effects varied with group. Positive correlations were observed between spindle density during slow-wave-sleep and AM posttest performance as well as between spindle density during non-REM sleep and AM baseline performance, showing that successful learning and retrieval both before and after sleep relates to spindle density during nap sleep. Together, these results speak for a selective beneficial impact of naps on hippocampus-dependent memories.


[Cytochemical studies of T- and B-lymphocyte subpopulations in normal conditions and in malignant lymphoproliferative diseases].
Data from literature and results of the authors' own investigations concerning cytochemical peculiarities of T- and B-lymphocyte subpopulations at the antigen-independent and antigen-dependent differentiation stages are reviewed. Enzyme-cytochemical characteristics for human lymphoid cells at early ontogenesis are presented, peculiarities of cells are given in the thymus-dependent and thymus-independent sites of lymphopoiesis organs and certain types of patch-forming cells are discussed. Data on the enzyme topography in the functionally different subpopulations of lymphocytes identified by a set of the monoclonal antibodies are elucidated. It is shown that the combined application of immunological and cytochemical methods permits establishing more precisely the cell origin and maturity level for different forms and variants of tumour diseases of the lymphoid tissue.


Prepartum milking effects on parlour behaviour, endocrine and immune responses in Holstein heifers.
Transition of primiparous heifers to the milking herd is a period with multiple stressors. The objective of these studies was to determine effects of parlour experience and prepartum milking (pre-milking) on behavioural and physiological indicators of stress after calving. Two experiments were conducted, one was in a free-stall housing confinement system and the second was in a modified grazing system. Forty-eight first-calf heifers were assigned to three treatments: control; experienced heifers taken through the parlour without milking; or pre-milk heifers milked for 3 weeks prior to estimated parturition. Blood was collected within 24 h of parturition and on days 3, 5, 7, 10 and 14 following parturition for cortisol and acute phase protein determination. In the grazing system, 20 heifers were assigned to a prepartum milked or control group as in the confinement system and behaviour observations included days -21, -14, -7, -5, -3 and -1 relative to calving and days 1, 3, 7, 9, 14, and 16 post-calving. Milk production was greatest for prepartum milked heifers in both housing systems. However, somatic cell score was reduced by prepartum milking only in the confinement system. Balking occurred least in parlour-experienced heifers. In confinement housing, shifting while in the parlour was the only behaviour that was greater at first milking in control heifers. Kicking was most frequent for parlour experienced heifers on day 2. Grazing system pre-milked heifers shifted more at their first milking (day -21) than did the controls at their first milking (day 1). Shifting within cow was greatest on day -21 compared with day -5 (P<0.05). Pre-milked heifers shifted more on day 1 post-calving than did the control heifers (P<0.05). These results showed that shifting was the most indicative behaviour of restlessness, was transient, and decreased by day 5 prior to calving. Cortisol and alpha1-acid glycoprotein concentrations were not different; however, haptoglobin increased for all treatments up to and including day 3 and haptoglobin concentrations of pre-milked heifers began to decrease by day 5 post-calving. Pre-milked heifers had lower haptoglobin concentrations than the control heifers and tended to have lower concentrations than experienced heifers on day 10 post partum. By day 14 post partum, all haptoglobin concentrations were <200 microg/ml, but the haptoglobin concentration of control heifers was greater than that of pre-milked and experienced heifers. These results showed that prepartum milking and parlour experience shorten some acute phase protein responses, but minimally affect early parlour behaviours.


Tumor-specific DNA in plasma of breast cancer patients.
The presence of DNA fragments circulating in cancer patients was described a number of years ago. The mere presence of DNA in the circulation is not indicative of cancer. However, there are reports that apoptosis and necrosis of the cancer cells can increase the levels of circulating DNA. The study of plasma DNA with the detection of genetic abnormalities associated with specific cancers has produced some promising results. Primary cancer often harbors ras or p53 mutations and the detection of these mutations in free circulating DNA could indicate the presence of cancer. Other approaches have included detection of specific losses of heterozygosity (LOH), microsatellite instability (MI) and promoter hyper-methylation. For breast cancer, studies published to date have focused on detecting LOH, MI and methylation of the p16INK4A promoter. Good concordance between alterations in the primary tumor and detection of the same alterations in the circulation has been observed. Also, it is encouraging to note that DNA alterations have been detected in patients with small or even in situ lesions, indicating that circulating tumor DNA is shed early in the disease process. If 'universal' breast-specific DNA alterations can be identified, this approach may hold significant promise for early detection of breast cancer.


Predicting Ca2+ -binding sites using refined carbon clusters.
Identifying Ca(2+) -binding sites in proteins is the first step toward understanding the molecular basis of diseases related to Ca(2+) -binding proteins. Currently, these sites are identified in structures either through X-ray crystallography or NMR analysis. However, Ca(2+) -binding sites are not always visible in X-ray structures due to flexibility in the binding region or low occupancy in a Ca(2+) -binding site. Similarly, both Ca(2+) and its ligand oxygens are not directly observed in NMR structures. To improve our ability to predict Ca(2+) -binding sites in both X-ray and NMR structures, we report a new graph theory algorithm (MUG(C) ) to predict Ca(2+) -binding sites. Using carbon atoms covalently bonded to the chelating oxygen atoms, and without explicit reference to side-chain oxygen ligand co-ordinates, MUG(C) is able to achieve 94% sensitivity with 76% selectivity on a dataset of X-ray structures composed of 43 Ca(2+) -binding proteins. Additionally, prediction of Ca(2+) -binding sites in NMR structures was obtained by MUG(C) using a different set of parameters, which were determined by the analysis of both Ca(2+) -constrained and unconstrained Ca(2+) -loaded structures derived from NMR data. MUG(C) identified 20 of 21 Ca(2+) -binding sites in NMR structures inferred without the use of Ca(2+) constraints. MUG(C) predictions are also highly selective for Ca(2+) -binding sites as analyses of binding sites for Mg(2+) , Zn(2+) , and Pb(2+) were not identified as Ca(2+) -binding sites. These results indicate that the geometric arrangement of the second-shell carbon cluster is sufficient not only for accurate identification of Ca(2+) -binding sites in NMR and X-ray structures but also for selective differentiation between Ca(2+) and other relevant divalent cations.


3D fiber tractography with susceptibility tensor imaging.
Gradient-echo MRI has revealed anisotropic magnetic susceptibility in the brain white matter. This magnetic susceptibility anisotropy can be measured and characterized with susceptibility tensor imaging (STI). In this study, a method of fiber tractography based on STI is proposed and demonstrated in the mouse brain. STI experiments of perfusion-fixed mouse brains were conducted at 7.0T. The magnetic susceptibility tensor was calculated for each voxel with regularization and decomposed into its eigensystem. The major eigenvector is found to be aligned with the underlying fiber orientation. Following the orientation of the major eigenvector, we are able to map distinctive fiber pathways in 3D. As a comparison, diffusion tensor imaging (DTI) and DTI fiber tractography were also conducted on the same specimens. The relationship between STI and DTI fiber tracts was explored with similarities and differences identified. It is anticipated that the proposed method of STI tractography may provide a new way to study white matter fiber architecture. As STI tractography is based on physical principles that are fundamentally different from DTI, it may also be valuable for the ongoing validation of DTI tractography.


Origin of the Difference in Reactivity between Ir Catalysts for the Borylation of C-H Bonds.
A mechanistic study on the origin of the difference in reactivity between Ir catalysts for C-H borylation reactions is reported. Catalytic reactions of B2pin2 with a series of substrates that require high temperatures and long reaction times were conducted. These reactions catalyzed by the combination of [Ir(COD)(OMe)]2 and 3,4,7,8-tetramethylphenanthroline (tmphen) occur in yields that are substantially higher than those of reactions catalyzed by [Ir(COD)(OMe)]2 and 4,4'-di-tert-butylbipyridine (dtbpy). The electronic properties of Ir catalysts ligated by dtbpy or tmphen and their stoichiometric reactivity were investigated. It was found that a longer lifetime rather than higher reactivity of the catalyst leads to higher yields of reactions catalyzed by Ir-tmphen. The catalyst ligated by dtbpy decomposes principally by dissociation of the ligand and rapid borylation at the positions alpha to nitrogen. Thus, the greater stability of the catalyst containing tmphen results from its greater binding constant.


The hepatotoxicity of Polygonum multiflorum: The emerging role of the immune-mediated liver injury.
Herbal and dietary supplements (HDS)-induced liver injury has been a great concern all over the world. Polygonum multiflorum Thunb., a well-known Chinese herbal medicine, is recently drawn increasing attention because of its hepatotoxicity. According to the clinical and experimental studies, P. multiflorum-induced liver injury (PM-DILI) is considered to be immune-mediated idiosyncratic liver injury, but the role of immune response and the underlying mechanisms are not completely elucidated. Previous studies focused on the direct toxicity of PM-DILI by using animal models with intrinsic drug-induced liver injury (DILI). However, most epidemiological and clinical evidence demonstrate that PM-DILI is immune-mediated idiosyncratic liver injury. The aim of this review is to assess current epidemiological, clinical and experimental evidence about the possible role of innate and adaptive immunity in the idiosyncratic hepatotoxicity of P. multiflorum. The potential effects of factors associated with immune tolerance, including immune checkpoint molecules and regulatory immune cells on the individual's susceptibility to PM-DILI are also discussed. We conclude by giving our hypothesis of possible immune mechanisms of PM-DILI and providing suggestions for future studies on valuable biomarkers identification and proper immune models establishment.


Experimental Evaluation of Faecal Escherichia coli and Hepatitis E Virus as Biological Indicators of Contacts Between Domestic Pigs and Eurasian Wild Boar.
Domestic pigs and Eurasian wild boar (Sus scrofa) share several important viral and bacterial pathogens. Therefore, direct and indirect contacts between domestic pigs and wild boar present a risk of pathogen spillover and can lead to long-term perpetuation of infection. Biological indicators could be a powerful tool to understand and characterize contacts between wild boar and domestic pigs. Here, faecal Escherichia coli and Hepatitis E virus (HEV) were explored as potential biological indicators under experimental conditions. The data gained in our pilot study suggest that faecal E. coli can be used as biological indicator of contact between wild boar and domestic pig. For HEV, faecal transmission was also confirmed. However, molecular studies on full-genome basis did not reveal markers that would allow tracing of transmission direction. Based on these promising results, future field studies will especially target the practicability of E. coli microbiome molecular typing as surrogate of contacts at the wildlife-livestock interface.


Trichinella pseudospiralis overcomes innate resistance of the Chinese hamster to Trichinella spiralis.
Infectivity of Trichinella pseudospiralis in Chinese hamsters was greater than that of Trichinella spiralis. Concurrent infection with these two species of parasite was accompanied by an increase in infectivity of T. spiralis. The infectivity of T. pseudospiralis, but not that of T. spiralis, was significantly influenced by host age. Regardless of host age, T. pseudospiralis was more infective for the Chinese hamster than was T. spiralis. Early during infection, enteritis (myeloperoxidase activity) in hamsters infected with T. pseudospiralis alone was significantly less than that seen in animals infected with T. spiralis alone or with both species concurrently. Myositis in hamsters infected with T. pseudospiralis alone was significantly less on days 14, 17 and 25 post-infection (p.i.) than that observed in concurrently infected or T. spiralis-infected animals. On days 11 and 14 p.i. inflammation in diaphragm muscle from concurrently infected hamsters was significantly less than that seen in animals infected with T. spiralis alone. Muscle histology supported these chemical data on myositis. Suppression of the host inflammatory response against early muscle-stage larvae of T. pseudospiralis underlies the greater infectivity of this parasite for the Chinese hamster.


Long-term results with the Wagner metal-on-metal hip resurfacing prosthesis.
The purpose of this study was to report the long-term results of the metal-on-metal Wagner resurfacing prosthesis. Between January 1991 and August 2004, 54 Wagner resurfacing prostheses were implanted in 52 patients. There were 45 female patients (1 bilateral case) and 7 male patients (1 bilateral case). All the operative procedures were performed using the anterior approach. Of those cases available, the average time to follow-up was nine years. The average Merle dAubign score improved from 8.3 preoperatively to 17.3 postoperatively (p-value < 0.001). Seventeen prostheses required revision. Radiographic evidence of femoral neck narrowing was seen in three prostheses that remained in situ. Combined with a stricter diagnosis and an exact operating technique, the Wagner metal-on-metal surface replacement of the hip joint may offer an alternative surface replacement option.


A partial knowledge of friends of friends speeds social search.
Milgram empirically showed that people knowing only connections to their friends could locate any person in the U.S. in a few steps. Later research showed that social network topology enables a node aware of its full routing to find an arbitrary target in even fewer steps. Yet, the success of people in forwarding efficiently knowing only personal connections is still not fully explained. To study this problem, we emulate it on a real location-based social network, Gowalla. It provides explicit information about friends and temporal locations of each user useful for studies of human mobility. Here, we use it to conduct a massive computational experiment to establish new necessary and sufficient conditions for achieving social search efficiency. The results demonstrate that only the distribution of friendship edges and the partial knowledge of friends of friends are essential and sufficient for the efficiency of social search. Surprisingly, the efficiency of the search using the original distribution of friendship edges is not dependent on how the nodes are distributed into space. Moreover, the effect of using a limited knowledge that each node possesses about friends of its friends is strongly nonlinear. We show that gains of such use grow statistically significantly only when this knowledge is limited to a small fraction of friends of friends.


Modulation of band gap by an applied electric field in silicene-based hetero-bilayers.
Electronic properties of the hetero-structures consisting of silicene, graphene and BN monolayers under the influence of an electric field were investigated using density functional theory. With no electric field, both silicene/graphene and silicene/BN were shown to have a finite gap of about ∼50 meV, though silicene is a zero-gap two-dimensional material. Application of the field perpendicular to the bilayer system was found to facilitate modulation of the band gap, exhibiting an approximately linear relationship with the gap energy, in contrast to what was seen for the constituent monolayers. Also, the degree of the modulation was mainly determined by the Si-pz electronic states at the interface of the silicene/graphene and silicene/BN bilayers.


Theoretical and empirical dimensions of the Aberdeen Glaucoma Questionnaire: a cross sectional survey and principal component analysis.
To develop patient-reported outcome instruments, statistical techniques (e.g., principal components analysis; PCA) are used to examine correlations among items and identify interrelated item subsets (empirical factors). However, interpretation and labelling of empirical factors is a subjective process, lacking precision or conceptual basis. We report a novel and reproducible method for mapping between theoretical and empirical factor structures. We illustrate the method using the pilot Aberdeen Glaucoma Questionnaire (AGQ), a new measure of glaucoma-related disability developed using the International Classification of Functioning and Disability (ICF) as a theoretical framework and tested in a sample representing the spectrum of glaucoma severity. We used the ICF to code AGQ item content before mailing the AGQ to a UK sample (N = 1349) selected to represent people with a risk factor for glaucoma and people with glaucoma across a range of severity. Reflecting uncertainty in the theoretical framework (items with multiple ICF codes), an exploratory PCA was conducted. The theoretical structure informed our interpretation of the empirical structure and guided the selection of theoretically-derived factor labels. We also explored the discrimination of the AGQ across glaucoma severity groups. 656 (49%) completed questionnaires were returned. The data yielded a 7-factor solution with a simple structure (using cut-off point of a loading of 0.5) that together accounted for 63% of variance in the scores. The mapping process resulted in allocation of the following theoretically-derived factor labels: 1) Seeing Functions: Participation; 2) Moving Around & Communication; 3) Emotional Functions; 4) Walking Around Obstacles; 5) Light; 6) Seeing Functions: Domestic & Social Life; 7) Mobility. Using the seven factor scores as independent variables in a discriminant function analysis, the AGQ scores resulted in correct glaucoma severity grading of 32.5% of participants (p < 0.001). This paper addresses a methodological gap in the application of classical test theory (CTT) techniques, such as PCA, in instrument development. Labels for empirically-derived factors are often selected intuitively whereas they can inform existing bodies of knowledge if selected on the basis of theoretical construct labels, which are more explicitly defined and which relate to each other in ways that are evidence based.


Health care professionals' attitudes towards evidence-based medicine in the workers' compensation setting: a cohort study.
Problems may arise during the approval process of treatment after a compensable work injury, which include excess paperwork, delays in approving services, disputes, and allegations of over-servicing. This is perceived as undesirable for injured people, health care professionals and claims managers, and costly to the health care system, compensation system, workplaces and society. Introducing an Evidence Based Medicine (EBM) decision tool in the workers' compensation system could provide a partial solution, by reducing uncertainty about effective treatment. The aim of this study was to investigate attitudes of health care professionals (HCP) to the potential implementation of an EBM tool in the workers' compensation setting. The study has a mixed methods design. The quantitative study consisted of an online questionnaire asking about self-reported knowledge, attitudes and behaviour to EBM in general. The qualitative study consisted of interviews about an EBM tool being applied in the workers' compensation process. Participants were health care practitioners from different clinical specialties. They were recruited through the investigators' clinical networks and the workers' compensation government regulator's website. Participants completing the questionnaire (n = 231) indicated they were knowledgeable about the evidence-base in their field, but perceived some difficulties when applying EBM. General practitioners reported having the greatest obstacles to applying EBM. Participants who were interviewed (n = 15) perceived that an EBM tool in the workers' compensation setting could potentially have some advantages, such as reducing inappropriate treatment, or over-servicing, and providing guidance for clinicians. However, participants expressed substantial concerns that the EBM tool would not adequately reflect the impact of psychosocial factors on recovery. They also highlighted a lack of timeliness in decision making and proper assessment, particularly in pain management. Overall, HCP are supportive of EBM, but have strong concerns about implementation of EBM based decision making in the workers' compensation setting. The participants felt that an EBM tool should not be applied rigidly and should take into account clinical judgement and patient variability and preferences. In general, the treatment approval process in the workers' compensation insurance system is a sensitive area, in which the interaction between HCP and claims managers can be improved.


Evaluation of the Predictive Validity of Thermography in Identifying Extravasation With Intravenous Chemotherapy Infusions.
Early detection of extravasation is important, but conventional methods of detection lack objectivity and reliability. This study evaluated the predictive validity of thermography for identifying extravasation during intravenous antineoplastic therapy. Of 257 patients who received chemotherapy through peripheral veins, extravasation was identified in 26. Thermography was performed every 15 to 30 minutes during the infusions. Sensitivity, specificity, positive predictive value, and negative predictive value using thermography were 84.6%, 94.8%, 64.7%, and 98.2%, respectively. This study showed that thermography offers an accurate prediction of extravasation.


Travel time to maternity care and its effect on utilization in rural Ghana: a multilevel analysis.
Rates of neonatal and maternal mortality are high in Ghana. In-facility delivery and other maternal services could reduce this burden, yet utilization rates of key maternal services are relatively low, especially in rural areas. We tested a theoretical implication that travel time negatively affects the use of in-facility delivery and other maternal services. Empirically, we used geospatial techniques to estimate travel times between populations and health facilities. To account for uncertainty in Ghana Demographic and Health Survey cluster locations, we adopted a novel approach of treating the location selection as an imputation problem. We estimated a multilevel random-intercept logistic regression model. For rural households, we found that travel time had a significant effect on the likelihood of in-facility delivery and antenatal care visits, holding constant education, wealth, maternal age, facility capacity, female autonomy, and the season of birth. In contrast, a facility's capacity to provide sophisticated maternity care had no detectable effect on utilization. As the Ghanaian health network expands, our results suggest that increasing the availability of basic obstetric services and improving transport infrastructure may be important interventions.


Molecular characterization of sarcomatous change in a granulosa cell tumor.
Sarcomatous transformation of a granulosa cell tumor (GCT) is a rare event. We describe the development of a rapidly progressive sarcomatous change in a woman who had initially presented with a classical GCT. A first recurrence occurred 23 months after the initial diagnosis when she was treated with external beam radiotherapy to her pelvis. A second recurrence 76 months following her initial surgery was consistent with a GCT. At 92 months, she presented with a further recurrence, outside of the radiotherapy field. This last recurrence had a different histologic appearance with features of sarcomatous change. Molecular analysis, using both reverse transcription-polymerase chain reaction and complementary DNA microarrays, has been used to analyze tissue obtained before and after the observed change in the tumor. The data show that GCT-specific genes, such as inhibin alpha, estrogen receptor, and follicle-stimulating hormone receptor, have been downregulated in the sarcomatous change. Significant upregulation of genes associated with an inflammatory response was also noted in the sarcoma, and this was consistent with the presence of a marked inflammatory infiltrate seen on histopathology. This study represents the novel application of microarray technology and demonstrates the unexpected finding of expression of the fibroblast activation protein gene in normal ovary. Although tumors such as this may be targets for the novel fibroblast activating protein-directed chemotherapeutic monoclonal antibody sibrotuzumab, the finding of expression in the normal ovary suggests the need for caution.


Biogenic Carbon Quantum Dots: Synthesis and Applications.
The new class of nanomaterials termed carbon dots: a quasi-spherical nanoparticle having a size less than 10 nm, possesses some unique characteristics like good aqueous solubility, colloidal stability, resistance to photobleaching, and fluorescence tunability, resulting in the unfolding of their various properties and their usage in different applications. Materials that are naturally derived or produced by living organisms are termed 'biogenic'. Over the past few years, there has been a gradual increase in the use of naturally derived materials in synthesizing carbon dots. Green precursors or biogenic materials are of low cost, readily available, renewable, and environmentally benign. Most importantly, they provide essential benefits not found in synthesized carbon dots. This review focuses on the use of biogenic materials for the synthesis of biogenic carbon dots developed in the past five years. It also briefly explains different synthetic protocols used, along with some significant findings. Thereafter, an overview of the use of biogenic carbon dots (BCDs) in different applications like chemo and biosensors, drug delivery, bioimaging, catalysis and energy applications, etc., is discussed. Thus biogenic carbon dots are future sustainable materials that are now fast replacing conventional carbon quantum prepared from other sources.


An Assessment of Sleep Disturbance in Patients before and after Carpal Tunnel Release.
Background: Night time numbness is a key characteristic of CTS and relief of night time symptoms is one of the outcomes most important to patients. This study tested the null hypothesis that there is no difference between sleep quality and night symptoms before and after carpal tunnel release (CTR). Methods: Forty-four, English-speaking adult patients requesting open CTR for electrodiagnostically confirmed carpal tunnel syndrome completed questionnaires before and after surgery. Average age was 59, 24 patients were men and 20 were women. Patient with a primary or secondary sleep disorder were excluded. Before surgery, patients completed the Pittsburg Sleep Quality index (PSQI). At an average of 3 months after surgery, participants completed PSQI questionnaires. Onset of sleep quality improvement was specifically addressed. Differences between preoperative and postoperative sleep quality were evaluated using the paired t-test. Spearman correlations were used to assess the relationship between continuous variables. Results: Of the 44 patients, 32 (72%) were classified as poor sleepers (PSQI > 5.5) prior to surgery. At 3 months follow up, there was a significant improvement PSQI global scores (7.8 ± 5.1 vs 4 ± 3.5, p < 0.001) as well as subdivisions. Daytime dysfunction (0.2 ± 0.4, p < 0.001) and medication use (1.0 ± 1.2 vs 0.9 ± 1.2, p < 0.045) secondary to sleep disturbance and was improved as well. In all patients, onset of improvement was within 24 hours of surgery. Conclusions: CTR is associated with improvement in sleep quality at 3 months follow-up. CTR improves daytime dysfunction related to the sleep disturbance. The onset of sleep improvement is 24 hours after surgery in most cases.


The role of antioxidant versus pro-oxidant effects of green tea polyphenols in cancer prevention.
Consumption of green tea (Camellia sinensis) may provide protection against chronic diseases, including cancer. Green tea polyphenols are believed to be responsible for this cancer preventive effect, and the antioxidant activity of the green tea polyphenols has been implicated as a potential mechanism. This hypothesis has been difficult to study in vivo due to metabolism of these compounds and poor understanding of the redox environment in vivo. Green tea polyphenols can be direct antioxidants by scavenging reactive oxygen species or chelating transition metals as has been demonstrated in vitro. Alternatively, they may act indirectly by upregulating phase II antioxidant enzymes. Evidence of this latter effect has been observed in vivo, yet more work is required to determine under which conditions these mechanisms occur. Green tea polyphenols can also be potent pro-oxidants, both in vitro and in vivo, leading to the formation of hydrogen peroxide, the hydroxyl radical, and superoxide anion. The potential role of these pro-oxidant effects in the cancer preventive activity of green tea is not well understood. The evidence for not only the antioxidant, but also pro-oxidant, properties of green tea is discussed in the present review.


Purification of bile acid-binding proteins from rat hepatic cytosol. Use of a photoaffinity label to detect novel Y' binders.
A 125I-labelled photolabile derivative of cholic acid has been used to investigate the organic anion-binding Y' fraction from rat liver, prepared by the method of Sugiyama, Y., Yamada, T. and Kaplowitz, N. (1982) Biochimica Biophysica Acta, 709, 342-352. The use of this photoaffinity probe led to the discovery of previously undescribed bile acid-binding proteins. A comprehensive purification scheme for the Y' proteins which allows the isolation of these novel binding species is described. Electrophoretic analysis shows that the Y' binders can be divided into two groups. The proteins in group 1 are dimeric and the 5B, 6E and 7F binding species consist of subunits with approximate molecular masses of 19.6, 15.6 and 14.9 kDa, respectively. The group 2 binding proteins, 5C, 5D and 8C, are monomeric and have molecular masses of approximately 36.2, 36.2 and 33 kDa, respectively. Calculation of the incorporation of 125I by these proteins showed that the group 1 proteins displayed a significantly greater specific incorporation of radioactivity than group 2. The specificity of 125I-labelled 3 beta-azidocholylhistamine is further demonstrated by analysis of tryptic digests of photoaffinity labelled Y' binders and glutathione S-transferases AA, A, D and F by reverse-phase high-performance liquid chromatography (HPLC) and thin-layer chromatography (TLC). The majority of the radioactivity was shown to be incorporated into a single component, which was not coincident with the free photoaffinity label.


[Pattern of wight gain during normal pregnancy].
The maternal weight gain during pregnancy has been established as a common acceptance criterion about 11 Kg., and its evaluation is of interest in Gynecology and Obstetrics; the objective of this study was to evaluate the Gestational Weight Gain Pattern (GWGP) during the normal pregnancy and its association with the maternal height, pregestional weight, Body Mass Index (BMI) schooling, the intergensic interval and the newborn weight. It was found through a longitudinal and prospective study in women of the urban area, under prenatal control in the Instituto Mexicano del Seguro Social (IMSS), in Morelia, Mich. México, that the average overall GWGP was of 9.3 Kg, and a normality band within 6.6 to 12 Kg which is smaller than the established by obstetrics criteria. It was not found a correlation between the GWGP and the newborn weight, now that the 94.3% of them had normal weight. The mother's age, weight, body mass index and schooling didn't show any correlation with the maternal weight gain during the pregnancy. It was shown that the mother's height and parity influence notably the GWGP. It's convenient the adoption during the prenatal control to determine that the GWGP normality is in agreement with the bio-psycho-social characteristics of the Mexican woman.


Evaluation of unmeshed and 1:1 meshed AlloDerm bolsters for stapled rectal anastomoses in a porcine model.
The major morbidity of colorectal anastomoses is leaks. The concept of staple-line reinforcement is a growing area of interest. In this study, we evaluated the feasibility and effect of utilizing AlloDerm to bolster end-to-end stapled rectal anastomoses in a porcine model. A total of 30 female 45-kg domestic pigs were studied, and each served as its own control by creating a bolstered and unbolstered anastomosis in each animal. All anastomoses were created with a 29-mm end-to-end stapling device. Bolstered anastomoses were randomized to proximal and distal positions along the rectum, and each rectorectal anastomosis was separated by an average of 10 cm. In 20 pigs, an unmeshed bolster of a 0.5-0.7-mm thickness was used. The remaining 10 pigs had a 1:1 meshed bolster that was 0.34-0.51 mm thick. The animals were survived for 14 days. Barium enemas were then performed and the two anastomotic sites harvested, and each anastomosis underwent burst testing. The internal diameter of each anastomosis was measured and a biochemical analysis was performed for matrix metalloproteinase (MMP), elastin and collagen content. The unmeshed bolstered anastomoses burst fewer times than the unbolstered anastomoses (P=0.004) and had higher burst pressures (P=0.023), though their anastomotic circumferences were smaller (P=0.007). Meshed bolsters offered no strength advantage to anastomoses and were significantly (P=0.009) smaller than unbolstered anastomoses in the same animal. No difference in elastin, collagen, or MMP content was observed between bolstered and unbolstered groups. No animals had clinical or radiographic leaks. The routine use of unmeshed and 1:1 meshed AlloDerm bolsters is safe and does not appear to inhibit healing in elective colorectal surgery on healthy subjects. AlloDerm may have a role as a tissue bolster in select patients who are more prone to develop anastomotic leaks.


Stroke and translational research - review of experimental models with a focus on awake ischaemic induction and anaesthesia.
Animal models are an indispensable tool in the study of ischaemic stroke with hundreds of drugs emerging from the preclinical pipeline. However, all of these drugs have failed to translate into successful treatments in the clinic. This has brought into focus the need to enhance preclinical studies to improve translation. The confounding effects of anaesthesia on preclinical stroke modelling has been raised as an important consideration. Various volatile and injectable anaesthetics are used in preclinical models during stroke induction and for outcome measurements such as imaging or electrophysiology. However, anaesthetics modulate several pathways essential in the pathophysiology of stroke in a dose and drug dependent manner. Most notably, anaesthesia has significant modulatory effects on cerebral blood flow, metabolism, spreading depolarizations, and neurovascular coupling. To minimise anaesthetic complications and improve translational relevance, awake stroke induction has been attempted in limited models. This review outlines anaesthetic strategies employed in preclinical ischaemic rodent models and their reported cerebral effects. Stroke related complications are also addressed with a focus on infarct volume, neurological deficits, and thrombolysis efficacy. We also summarise routinely used focal ischaemic stroke rodent models and discuss the attempts to induce some of these models in awake rodents.


Academic health centers.
Academic Health Centers (AHCs) are comprised of academic, hospital, and clinical practice components that play a key role in healthcare delivery by their special ability to identify and implement improvements in outcomes, safety, cost-benefit, and satisfaction. They do this by utilizing a wide range of academic and clinical health professionals and disciplines to provide cutting-edge, highly specialized patient care as well as disproportionate uncompensated care in communities nationwide; to identify the effectiveness of different diagnostic and therapeutic approaches through clinical research; to foster new discoveries in biomedical science and technology and their clinical application; and to educate future generations of health professionals who apply these improvements. As the traditional homes of innovation in health and healing through research, and as the major sites of implementing change through education, AHCs have been at the forefront of improving healthcare. To successfully improve the effectiveness and efficiency of healthcare delivery, it is critical that AHCs continue to serve as uniquely integrated models for improving quality and value through novel approaches in education, research, and service.


Genetics and beyond--the transcriptome of human monocytes and disease susceptibility.
Variability of gene expression in human may link gene sequence variability and phenotypes; however, non-genetic variations, alone or in combination with genetics, may also influence expression traits and have a critical role in physiological and disease processes. To get better insight into the overall variability of gene expression, we assessed the transcriptome of circulating monocytes, a key cell involved in immunity-related diseases and atherosclerosis, in 1,490 unrelated individuals and investigated its association with >675,000 SNPs and 10 common cardiovascular risk factors. Out of 12,808 expressed genes, 2,745 expression quantitative trait loci were detected (P<5.78x10(-12)), most of them (90%) being cis-modulated. Extensive analyses showed that associations identified by genome-wide association studies of lipids, body mass index or blood pressure were rarely compatible with a mediation by monocyte expression level at the locus. At a study-wide level (P<3.9x10(-7)), 1,662 expression traits (13.0%) were significantly associated with at least one risk factor. Genome-wide interaction analyses suggested that genetic variability and risk factors mostly acted additively on gene expression. Because of the structure of correlation among expression traits, the variability of risk factors could be characterized by a limited set of independent gene expressions which may have biological and clinical relevance. For example expression traits associated with cigarette smoking were more strongly associated with carotid atherosclerosis than smoking itself. This study demonstrates that the monocyte transcriptome is a potent integrator of genetic and non-genetic influences of relevance for disease pathophysiology and risk assessment.


Early detection of communication delays with the PEDS tools in at-risk South African infants.
Prevalence of communication delays or disorders is increasing, possibly because of various environmental risk factors. Selection and implementation of effective screening tools are important to detect at-risk infants as early as possible. This study aimed to evaluate the accuracy of the Parents' Evaluation of Developmental Status (PEDS), PEDS-Developmental Milestones and PEDS tools to detect communication delays in infants (6-12 months) in a South African primary healthcare context. A comparative study design evaluated the accuracy of the PEDS tools to detect communication delays, using an internationally accepted diagnostic assessment tool, the Rossetti Infant-Toddler Language Scale (RITLS). A convenience sample of 201 infants was selected at primary healthcare clinics. Expressive and receptive language sensitivity scores were low across all three screens (ranging between 14% and 44%). The PEDS tools had high sensitivity (71%) and specificity (73%) ratings for the receptive and expressive language and socio-emotional domain in combination. In the sample population, the PEDS tools did not accurately detect receptive and expressive language delays; however, communication delays in general were identified. Future research determining accuracy of the PEDS, PEDS-Developmental Milestones and PEDS tools for children aged 2-5 years in detecting communication delays should be prioritised.


Control of rectal gland secretion in the dogfish (Squalus acanthias): steps in the sequence of activation.
We measured the venous and arterial pressure, as well as the rate of secretion and content of cyclic AMP and high energy phosphate compounds, of the rectal gland of the anaesthetized dogfish, Squalus acanthias (L.). Intravenous infusion of isotonic solutions produced a very large increase in the rate of secretion by the rectal gland. The increase in secretion was preceded by an increase in venous blood pressure, but arterial blood pressure was not modified. Injections of small doses of veratridine stimulated gland secretion when given in the vicinity of the heart but not when given in the dorsal aorta. During volume expansion the creatine phosphate and ATP content of the gland were markedly reduced, while ADP and AMP as well as cyclic AMP content were increased. We conclude that: volume expansion leads to the release of a message that activates adenyl cyclase in the gland; the increased venous pressure may be the initial signal in the sequence that leads to the release of the activating messenger; there is a receptor mechanism in the atrial and cardiac region that triggers the sequence that activates glandular secretion; the reduction in the content of high energy phosphate compounds during volume expansion is caused by an increase in energy expenditure, probably due to gland secretion.


Evidence-based medicine in military orthopaedics: are we doing our part?
The clinical application of evidence-based medicine is influenced by the quality of the published literature. Journals assign objective levels of evidence to articles to describe the quality and reliability of individual articles. Studies assigned levels I to III are considered higher quality studies. This study reviewed 60 continuous years of the American edition of the Journal of Bone and Joint Surgery to analyze trends in the quality of research published by U.S. military orthopaedic surgeons. Three hundred and six of 371 identified articles were assigned a level of evidence from I to V. These were then analyzed to determine if military orthopaedic surgeons were producing higher quality studies over time. Over time, the total number of articles published decreased. However, the number of higher quality studies and average level of evidence per decade improved over time. This trend toward publishing higher quality studies is consistent with the general orthopaedic community and highlights the need for continued work by the military orthopaedic community to conduct higher quality studies.


Carriage of antibiotic resistant bacteria in endangered and declining Australian pinniped pups.
The rapid emergence of antimicrobial resistance (AMR) is a major concern for wildlife and ecosystem health globally. Genetic determinants of AMR have become indicators of anthropogenic pollution due to their greater association with humans and rarer presence in environments less affected by humans. The objective of this study was to determine the distribution and frequency of the class 1 integron, a genetic determinant of AMR, in both the faecal microbiome and in Escherichia coli isolated from neonates of three pinniped species. Australian sea lion (Neophoca cinerea), Australian fur seal (Arctocephalus pusillus doriferus) and long-nosed fur seal (Arctocephalus forsteri) pups from eight breeding colonies along the Southern Australian coast were sampled between 2016-2019. DNA from faecal samples (n = 309) and from E. coli (n = 795) isolated from 884 faecal samples were analysed for class 1 integrons using PCRs targeting the conserved integrase gene (intI) and the gene cassette array. Class 1 integrons were detected in A. p. doriferus and N. cinerea pups sampled at seven of the eight breeding colonies investigated in 4.85% of faecal samples (n = 15) and 4.52% of E. coli isolates (n = 36). Integrons were not detected in any A. forsteri samples. DNA sequencing of the class 1 integron gene cassette array identified diverse genes conferring resistance to four antibiotic classes. The relationship between class 1 integron carriage and the concentration of five trace elements and heavy metals was also investigated, finding no significant association. The results of this study add to the growing evidence of the extent to which antimicrobial resistant bacteria are polluting the marine environment. As AMR determinants are frequently associated with bacterial pathogens, their occurrence suggests that these pinniped species are vulnerable to potential health risks. The implications for individual and population health as a consequence of AMR carriage is a critical component of ongoing health investigations.


Impact of caffeic acid phenethyl ester treatment on vancomycin-induced pancreatic damage in rats.
This study investigates the preventive effect of caffeic acid phenethyl ester (CAPE) on pancreatic damage induced by vancomycin (VCM) in rats. Rats were equally divided into three groups: group I (control), group II (only VCM-treated group) and group III (VCM + CAPE-treated groups). VCM was intraperitoneally administered at a dose of 200 mg kg(-1)twice daily for 7 days. CAPE was administered orally at 10 µM mL(-1) kg(-1) dose once daily for 7 days. The first dose of CAPE administration was performed 24 h prior to VCM injection. Blood and pancreas tissue samples were removed and collected after the study. Serum alkaline phosphatase (ALP), amylase, γ-glutamyl transferase (GGT) and lipase activities were determined. Pancreas tissue samples were evaluated with the light microscope. Group II significantly increased serum ALP, amylase, GGT and lipase activities when compared with the control group. Group III significantly decreased serum ALP, amylase, GGT and lipase activities when compared with group II. In histopathological examination, it has been observed that there was a significant pancreatic damage in group II. CAPE exerted prominent structural protection against VCM-induced pancreatic damage and this effect was statistically significant. CAPE caused a marked reduction in the extent of pancreatic damage. We have concluded that it may play an important role in the VCM-induced pancreatic damage and reduce the pancreatic damage both at the biochemical and histopathological aspects.


Salivary immunoglobulin A responses in professional top-level futsal players.
The purpose of this study was to investigate the responses of salivary immunoglobulin A (SIgA) in 10 professional top-level Brazilian futsal players after 2 highly competitive games separated by 7 days. Unstimulated saliva was collected over a 5-minute period at PRE- and POST-match. The SIgA was measured by an enzyme-linked immunosorbent assay and expressed as the absolute concentration (SIgAabs) and secretion rate of IgA (SIgArate). Rate of perceived exertion and heart rate were used to monitor the exercise intensity. A 2-way analysis of variance with repeated measures showed nonsignificant differences between matches to SIgAabs, SIgArate, and saliva flow rate (p > 0.05). However, significant time differences were observed for all these parameters. In summary, we showed that a competitive training match induced a decrease in SIgA levels in top-level futsal players, which suggests an increment of the vulnerability to infections meditated by the training stimulus. This decrease suggests that the athletes were at an increased risk of developing an upper respiratory tract infection, and therefore, it could be necessary to take protective actions to minimize contact with cold viruses or even reduce the training load for athletes.


Genome-scale CRISPRa screening identifies MTX1 as a contributor for sorafenib resistance in hepatocellular carcinoma by augmenting autophagy.
Sorafenib is the standard first-line drug for the treatment of advanced hepatocellular carcinoma (HCC), however, its therapeutic efficacy is not satisfactory due to primary or secondary resistance of HCC cells. In the present study, we identified Metaxin 1 (MTX1) as a new regulator of sorafenib resistance in HCC through genome-scale CRISPR activation (CRISPRa) screening. We found that MTX1 was frequently upregulated in HCC tissues and overexpression of MTX1 promoted HCC cell proliferation in vitro and in vivo. As well, MTX1 overexpression increased cell growth rate and decreased cell apoptosis upon sorafenib treatment. Consistently, the resistance induced by MTX1 was also observed in subcutaneous xenograft tumor model. Clinically, high expression of MTX1 was closely related with poor outcomes in HCC patients who received sorafenib treatment. Mechanistically, overexpression of MTX1 could promote HCC cell autophagy via interacting with and inhibiting CDGSH iron sulfur domain 1 (CISD1), an autophagy negative regulator. Taken together, our findings suggest that MTX1 is upregulated in HCC and contributes to sorafenib resistance via a possible mechanism involving CISD1 mediated autophagy.


Comparative assessment of prognostic value for revised-Mayo risk model and Child-Pugh score in patients with primary sclerosing cholangitis.
The aim of this study was to compare the utility of the revised Mayo risk model (rMRM) and Child-Pugh scores (CPSs) for predicting the prognosis of disease in patients with primary sclerosing cholangitis (PSC). Patients were divided into 2 groups: Group I (37 patients; alive and not requiring liver transplantation) and Group II (8 patients; deceased or requiring liver transplantation). rMRM suggests the possible survival percentage over a 4-year period. Thus, rMRM scores and CPSs on the first visit were calculated from the data at the time of diagnosis for patients diagnosed with PSC <4 years ago. rMRM scores and CPSs of patients with >4 years of follow-up were calculated using data from the visit 4 years prior to their last follow-up. Bivariate analyses showed that need for liver transplantation/mortality was correlated with either first visit CPS (r=0.481, p=0.001) or rMRM (r=0.452, p=0.002). Analysis of the area under the curve showed that both models performed similarly in terms of predicting the need for liver transplantation/mortality (rMRM: 0.780; CPS: 0.762; p=0.8). There was a significant difference in Kaplan-Meier survival rates between Group I and Group II for both risk models (rMRM: p<0.001; CPS: p<0.001) when the decisive event was death or need for liver transplantation. Both rMRM and CPSs are useful in risk assessment of patients with PSC. The ability to predict prognosis is similar for both risk models.


Scanning electron microscopy of the wall of the third ventricle of the brain of Rana temporia. II. Electron microscopy of the ventricular surface of the pars ventralis of the tuber cinereum.
Scanning electrom microscopy of the ventricular surface of the pars ventralis of the tuber cinereum of the frog Rana temporaria shows that it can be divided in (1) a dorsolateral area, characterized by the presence of very numerous large, solid, bulbous protrusions (2) a medial area, where the large bulbous structures are completely absent and which is diffusely covered with very numerous cilia, (3) a transiotional area located between the two other regions. The large bulbs, shown by scanning electron microscopy, correspond with the end-bulbs of thick ventricular dendrites of nerve cells revealed by transmission electron microscopy. At least many of these intra-ventricular bulbs are dendritic endings of peptidergic neurosecretory neurons, which have been tentatively identified as adenohypophysiotropic neuro-hormone producing neurons (Derickx et al., 1972, 1973a, 1973b). The structural features of the dendritic endings plead in favour of a possible receptive role.


Alginate microcapsule as a 3D platform for propagation and differentiation of human embryonic stem cells (hESC) to different lineages.
Human embryonic stem cells (hESC) are emerging as an attractive alternative source for cell replacement therapy since they can be expanded in culture indefinitely and differentiated to any cell types in the body. Various types of biomaterials have also been used in stem cell cultures to provide a microenvironment mimicking the stem cell niche(1-3). The latter is important for promoting cell-to-cell interaction, cell proliferation, and differentiation into specific lineages as well as tissue organization by providing a three-dimensional (3D) environment(4) such as encapsulation. The principle of cell encapsulation involves entrapment of living cells within the confines of semi-permeable membranes in 3D cultures(2). These membranes allow for the exchange of nutrients, oxygen and stimuli across the membranes, whereas antibodies and immune cells from the host that are larger than the capsule pore size are excluded(5). Here, we present an approach to culture and differentiate hESC DA neurons in a 3D microenvironment using alginate microcapsules. We have modified the culture conditions(2) to enhance the viability of encapsulated hESC. We have previously shown that the addition of p160-Rho-associated coiled-coil kinase (ROCK) inhibitor, Y-27632 and human fetal fibroblast-conditioned serum replacement medium (hFF-CM) to the 3D platform significantly enhanced the viability of encapsulated hESC in which the cells expressed definitive endoderm marker genes(1). We have now used this 3D platform for the propagation of hESC and efficient differentiation to DA neurons. Protein and gene expression analyses after the final stage of DA neuronal differentiation showed an increased expression of tyrosine hydroxylase (TH), a marker for DA neurons, >100 folds after 2 weeks. We hypothesized that our 3D platform using alginate microcapsules may be useful to study the proliferation and directed differentiation of hESC to various lineages. This 3D system also allows the separation of feeder cells from hESC during the process of differentiation and also has potential for immune-isolation during transplantation in the future.


A radiologic sign of lateral subluxation of the distal tibial epiphyses.
Linear calcification of the interosseus membrane at the ankle in a juvenile who has sustained an eversion/abduction injury with otherwise normal radiographs indicates that a Type I injury has occurred to the distal tibial and fibular epiphyses, with displacement at the time of injury. This is a significant injury with the potential to cause growth arrest of the tibial epiphyseal plate.


Promoting the development of nurses' presentation and publication skills.
When nurses share their clinical knowledge with others through presentations and publications, new ideas emerge that could result in improved patient care. Nurses also gain opportunities for professional recognition and advancement through such pursuits. Managers are in a strategic position to promote the development of presentation and publication skills. This article describes a three-phase model that can help managers utilize appropriate strategies and resources to facilitate skills development. Each phase of development is designed to increase skill level, confidence, and experience. With managerial support, this model can help nurses learn to communicate effectively with colleagues through presentations and publications.


[Radicular low back pain: is there a relationship between the treatment regimen and outcome?].
To identify a possible relationship between the non-surgical treatment regimen and outcome. In a sample of 170 patients with sciatica due to a herniated disk the intensity of a conservative multimodal inpatient treatment in a neurological department was extracted. The outcome was examined using two prospective cohorts (183 patients). These results were compared with published data from orthopaedic inpatient rehabilitation in Germany. The neurological inpatient treatment regimen was more intense than the orthopaedic inpatient rehabilitation, especially with regard to physiotherapy. In contrast, physical therapy was applied more often in orthopaedic rehabilitation. A better short-term outcome with regard to pain intensity was found after multimodal conservative treatment. Further studies are needed to compare different treatment regimens in patients with sciatica.


Membrane insertion at the leading edge of motile fibroblasts.
We are concerned with the mechanisms involved in the directed migration of eukaryotic cells. Previously we found that, inside cells at the edge of an experimental wound, the Golgi apparatus and the microtubule-organizing center were rapidly repositioned forward of the nucleus in the direction of subsequent cell migration into the wound. This repositioning was proposed to serve the purpose of introducing new membrane mass at the leading edge of the cell, by directing Golgi apparatus-derived vesicles bound for the plasma membrane to that edge. We now provide evidence to support this proposal. Cultured fibroblastic cells at the edge of a wound were infected with a temperature-sensitive mutant (0-45) of vesicular stomatitis virus. It is known that the G-protein, an integral membrane protein of the virus, is synthesized and remains in the rough endoplasmic reticulum at the nonpermissive temperature, but when the infected cells are shifted to the permissive temperature, the G-protein moves through the Golgi apparatus to the plasma membrane. By immunofluorescence microscopy, we here show that the first appearance of the G-protein at the cell surface corresponds to the leading edge of the motile cell. These observations are incorporated into a coherent scheme for the mechanisms involved in cell migration.


Hybrid fuzzy cluster ensemble framework for tumor clustering from biomolecular data.
Cancer class discovery using biomolecular data is one of the most important tasks for cancer diagnosis and treatment. Tumor clustering from gene expression data provides a new way to perform cancer class discovery. Most of the existing research works adopt single-clustering algorithms to perform tumor clustering is from biomolecular data that lack robustness, stability, and accuracy. To further improve the performance of tumor clustering from biomolecular data, we introduce the fuzzy theory into the cluster ensemble framework for tumor clustering from biomolecular data, and propose four kinds of hybrid fuzzy cluster ensemble frameworks (HFCEF), named as HFCEF-I, HFCEF-II, HFCEF-III, and HFCEF-IV, respectively, to identify samples that belong to different types of cancers. The difference between HFCEF-I and HFCEF-II is that they adopt different ensemble generator approaches to generate a set of fuzzy matrices in the ensemble. Specifically, HFCEF-I applies the affinity propagation algorithm (AP) to perform clustering on the sample dimension and generates a set of fuzzy matrices in the ensemble based on the fuzzy membership function and base samples selected by AP. HFCEF-II adopts AP to perform clustering on the attribute dimension, generates a set of subspaces, and obtains a set of fuzzy matrices in the ensemble by performing fuzzy c-means on subspaces. Compared with HFCEF-I and HFCEF-II, HFCEF-III and HFCEF-IV consider the characteristics of HFCEF-I and HFCEF-II. HFCEF-III combines HFCEF-I and HFCEF-II in a serial way, while HFCEF-IV integrates HFCEF-I and HFCEF-II in a concurrent way. HFCEFs adopt suitable consensus functions, such as the fuzzy c-means algorithm or the normalized cut algorithm (Ncut), to summarize generated fuzzy matrices, and obtain the final results. The experiments on real data sets from UCI machine learning repository and cancer gene expression profiles illustrate that 1) the proposed hybrid fuzzy cluster ensemble frameworks work well on real data sets, especially biomolecular data, and 2) the proposed approaches are able to provide more robust, stable, and accurate results when compared with the state-of-the-art single clustering algorithms and traditional cluster ensemble approaches.


Accumulation of 8-oxo-2'-deoxyguanosine and increased expression of hMTH1 protein in brain tumors.
Oxidative DNA damage generated by an attack of reactive oxygen species causes mutation or cell death that may lead to various diseases and may be related to initiation or progression of carcinogenesis. 8-Oxo-2'-deoxyguanosine (8-oxo-dG) is a major oxidative DNA damage product that can result in mutation, and hMTH1, human MutT homolog protein 1, has been identified as an enzyme that hydrolyzes 8-oxo-dGTP to the monophosphate, thus preventing accumulation of 8-oxo-dG in DNA. With immunohistochemical approaches, we investigated accumulation of 8-oxo-dG and expression of hMTH1 in brain tumor tissues obtained from surgical and autopsy cases, including 42 neuroepithelial tumors, 5 meningiomas, 2 metastatic brain tumors, and 1 schwannoma. 8-Oxo-dG accumulation and hMTH1 expression were increased in various brain tumors. Nuclei of brain tumor cells were immunoreactive for 8-oxo-dG in all cases. In most cases, both nuclei and cytoplasm of the tumor cells were immunoreactive for hMTH1. Both 8-oxo-dG accumulation and hMTH1 expression were most evident in high-grade gliomas, indicating that oxidative stress was high in these gliomas. Thus, the defense mechanism against such oxidative stress may be enhanced as well. These results suggest that oxidative stress may play a role in tumor progression.


Role of Nucleophosmin Gene Mutation in Leukemogenesis of Acute Myeloid Leukemia.
Acute myeloid leukemia (AML) is a hematopoietic stem cell disorder that carries very poor prognosis. Understanding molecular basis of AML leukemogenesis could lead to the emergence of effective targeted therapies for AML. AML bearing nucleophosmin (NPM) gene mutation has distinct features. This study was conducted to investigate the role of mutated (m) NPM in pathogenesis of de novo AML through studying its contribution in proliferation of AML cell line cells. Two types of human leukemia cell lines were used. One of them was a model for AMLs with mNPM and the other for AMLs with wild type (wt) NPM. Assessment of the proliferative role of mNPM in AML was carried out using cell culture and viability studies. The obtained results were reaffirmed by immunocytochemical and immunoblotting techniques. Analysis of results was done with the appropriate computer software. It showed higher proliferative potential of cells with mNPM compared to those bearing wtNPM only. Furthermore, the immunocytochemical studies demonstrated subcellular localization of NPM isoforms during various phases of mitosis. Mitosis was associated with cytoplasmic translocation of wtNPM in certain phases, while localization of mNPM remained unchanged throughout the cell cycle. Results of immunoblotting showed little or no change in protein expression of either NPM moieties during mitosis. The current study demonstrated important contribution of NPM gene mutation in enhancing proliferation of AML cell lines. These results confirmed the role of mNPM in AML leukemogenesis, and highlighted the importance of targeting mNPM in new evolving AML therapies.


Symposium review: The Mozzarella/pasta filata years: A tribute to David M. Barbano.
In the decades that followed the end of World War II, Mozzarella cheesemaking in the United States grew steadily as the pizza restaurant established an ever-expanding footprint across America and beyond. By the 1980s, Mozzarella cheesemaking had attained unprecedented scales of production, yet even as new cheese plants were coming on line across the country and production capacities were reaching extraordinary levels, the scientific and technological knowledge base needed to standardize production schedules, control product quality, and maximize cheese yields and efficiency lagged far behind industry needs. It was within this historical context that David Barbano turned his systematic and meticulous research program toward the needs of the Mozzarella cheese industry during the 1980s. By the early 1990s, Barbano was leading a team of graduate students, post-docs, technical staff, and collaborators in a systematic evaluation of every step in the Mozzarella cheesemaking process. The end product of these studies was nothing less than the transformation of what had been (to a large degree) a poorly understood "black box" process into a precisely controlled make procedure that lent itself to precision tailoring of cheese functionality, tight control over manufacturing schedules and efficiency, and maximization of cheese yields. Barbano's international collaborators included research scientists from Italy, where Mozzarella originated. Working with them, Barbano led a systematic evaluation of the scientific and technological aspects of Ragusano cheese, a traditional Protected Designation of Origin pasta filata cheese from Sicily. In the process, Barbano's team demonstrated a new approach to traditional artisanal practices that merged both the art and science of cheesemaking, in effect combining the best of both worlds, toward the goal of sustaining traditional cheesemakers and the working landscapes that they support. Throughout all of these studies, Barbano's research led to innovations in cheesemaking technology (such as improved salting methods, preacidification treatments, and strategies to improve low-fat Mozzarella functionality) that have revolutionized the Mozzarella industry worldwide.


Catecholamines modulate human NK cell circulation and function via spleen-independent beta 2-adrenergic mechanisms.
Increases in catecholamines have been shown to induce changes in migration of lymphocytes, in particular NK cells. To analyze the mechanisms of catecholamine-induced NK cell trafficking, normal healthy male human subjects and splenectomized individuals were infused with either adrenaline (0.10 microgram/kg/min), noradrenaline (0.15 microgram/kg/min), or NaCl i.v. for 20 min. Lymphocyte subsets (CD3+, CD4+, CD8+) transiently increased after administration of both catecholamines, with most pronounced increases (up to 600%) in NK cell numbers (CD16+ or CD56+) after infusion of adrenaline. These changes in NK cell numbers and function were accompanied neither by alterations in expression of adhesion molecules (CD11a), CD11b, CD31, CD43, CD44, CD62L) on NK cells nor by changes in plasma concentrations of soluble (s) adhesion molecules (sVCAM-1, sICAM-1, sE-selectin). Comparable increases in lymphocyte subsets were observed in splenectomized subjects, suggesting lymphocyte recruitment from other sources than the spleen. Furthermore, catecholamine-induced increases in lymphocyte subsets could be inhibited by pretreatment with the nonselective beta-adrenoceptor antagonist propranolol, but not by the beta1-selective antagonist bisoprolol. These data demonstrate that adrenaline and noradrenaline modulate the migratory capacity of human NK cells via spleen-independent beta 2-adrenoceptor mechanism.


Solitary fibrous tumor (SFT) of the pelvis.
Solitary fibrous tumors (SFTs) are well recognized in the pleura, but their occurrence at other sites has only become appreciated in recent years, as a consequence of which extrapleural examples often go unrecognized and misdiagnosed. Because of their rarity, overall experience concerning this tumor has not been significant and reports detailing radiological findings are few. We herein report an unusual case of a large retroperitoneal pelvic SFT with features of high vascularity negating successful surgical resection.


Carbon Conductive Paste Smart Textile Strain Sensor for Sports and Rehabilitation.
Due to the growth observed in the wearable market, stretchable strain sensors have been the focus of several studies. However, combining high sensitivity and linearity with low hysteresis presents a difficult challenge.Here, we propose a stretchable strain sensor obtained with off-the-shelf materials by printing a carbon conductive paste into a piece of fabric to be integrated into a smart garment. This process is cheap and easily scalable, allowing its mass production. The sensor developed has a large sensitivity (GF=11.27), high linearity (R2>0.99), very low hysteresis (γH =4.23%) and brings an added value, for example, in sports or rehabilitation monitoring.


[Blood lipid levels in various forms of ischemic heart disease].
Enzyme assays of major blood lipid parameters detected no differences between peripheral venous blood specimens of normal subjects and functional cardiologic patients, nor between peripheral and central flow specimens of patients with neurocirculatory dystonia. Alpha cholesterol was typically decreased in all types of coronary disease, the decrease being particularly marked in central cardiac flow. Chronic coronary disease is associated with a rise of total cholesterol, its atherogenic fractions, the atherogenic coefficient and blood triglycerides, as compared to the respective parameters of functional cardiac patients. Acute and subacute coronary disease is associated with decreased blood lipid values, as compared to chronic coronary disease. Blood lipid phenotyping and secondary preventive procedures should rather be done in the absence of aggravations.


RNA secondary structure prediction using conditional random fields model.
Non-coding RNAs (ncRNAs) have important biological functions in living cells dependent on their conserved secondary structures. Here, we focus on computational RNA secondary structure prediction by exploring primary sequences and complementary base pair interactions using the Conditional Random Fields (CRFs) model, which treats RNA prediction as a sequence labelling problem. Proposing suitable feature extraction from known RNA secondary structures, we developed a feature extraction based on natural RNA's loop and stem characteristics. Our CRFs models can predict the secondary structures of the test RNAs with optimal F-score prediction between 56.61 and 98.20% for different RNA families.


Supplementation of L-arginine prevents glucocorticoid-induced reduction of bone growth and bone turnover abnormalities in a growing rat model.
The present study was designed to evaluate the effects of glucocorticoid (GC) treatment on bone turnover and bone mineral density in the growing rat. Because of the recent evidence that nitric oxide (NO) can counteract prednisolone-induced bone loss in mature rats, we examined the effect on bone of the NO donor L: -arginine in young male rats, in which bone mass is increased by the same biological mechanism as in children and adolescents. Thirty-six 10-week-old Sprague-Dawley male rats were assigned to six groups of six animals each, and treated for 4 weeks with either vehicle (once a week subcutaneous injection of 100 microl of sesame oil); prednisolone sodium succinate, 5 mg/kg, 5 days per week by intramuscular injection (i.m.); L-arginine, 10 mg/kg intraperitoneally (i.p.) once a day; N(G)-nitro-L-arginine methylester (L-NAME), 50 mg/kg subcutaneously once a day; prednisolone sodium succinate 5 mg/kg, 5 days per week i.m. +L-arginine 10 mg/kg i.p. once a day; or prednisolone sodium succinate, 5 mg/kg, 5 days per week i.m. +L-NAME 50 mg/kg subcutaneously once a day. Serum calcium, alkaline phosphatase (ALP), osteocalcin, and the C-terminal telopeptides of type I collagen (RatLaps) were measured at baseline conditions and after 2 and 4 weeks. Prior to treatment, and after 2 and 4 weeks, the whole body, vertebral, pelvic, and femoral bone mineral density (BMD) was measured by dual-energy X-ray absorptiometry (DXA) scanning. Prednisolone and prednisolone+L-NAME treated rats had significantly lower ALP and osteocalcin levels than controls at 2 and 4 weeks, and significantly higher levels of Rat-Laps than controls at 4 weeks. Prednisolone, L-NAME, and prednisolone+L-NAME produced a significant inhibition of bone accumulation and bone growth at all sites measured. Supplementation with L-arginine appeared to prevent the inhibition of bone growth and increase in bone resorption induced by prednisolone. These data would suggest, for the first time, that supplementation with an NO donor could be considered as a treatment for steroid-induced osteoporosis in the developing skeleton.


Passaging protocols for mammalian neural stem cells in suspension bioreactors.
Mammalian neural stem cells (NSC) offer great promise as therapeutic agents for the treatment of central nervous system disorders. As a consequence of the large numbers of cells that will be needed for drug testing and transplantation studies, it is necessary to develop protocols for the large-scale expansion of mammalian NSC. Neural stem cells and early progenitor cells can be expanded in vitro as aggregates in controlled bioreactors using carefully designed media. The first objective of this study was to determine if it is possible to maintain a population of murine neural stem and progenitor cells as aggregates in suspension culture bioreactors over extended periods of time. We discovered that serial passaging of a mixture of aggregates sizes resulted in high viabilities, high viable cell densities, and good control of aggregate diameter. When the NSC aggregates were serially subcultured three times without mechanical dissociation, a total multiplication ratio of 2.9 x 10(3) was achieved over a period of 12 days, whereas the aggregate size was controlled (mean diameter less than 150 microm) below levels at which necrosis would occur. Moreover, cell densities of 1.0 x 10(6) cells/mL were repeatedly achieved in batch culture with viabilities exceeding 80%. The second objective was to examine the proliferative potential of single cells shed from the surface of these aggregates. We found that the single cells, when subcultured, retained the capacity to generate new aggregates, gave rise to cultures with high viable cell densities and were able to differentiate into all of the primary cell phenotypes in the central nervous system.


Neural correlates of pair-bonding in a monogamous primate.
The neurobiology of social bonding, despite its relevance to human mental health, has been studied primarily in rodents. In this study we used position emission tomography (PET), registered with structural magnetic resonance imaging (MRI) to investigate central glucose uptake in 17 males of a monogamous primate species, the titi monkey (Callicebus cupreus). Twelve pair-bonded males (including six with a lesion of the prefrontal cortex) and five lone males were scanned. The five lone males were re-scanned 48 h after pairing with a female. Significant differences in glucose uptake were found between males in long-term pair-bonds and lone males in areas including the nucleus accumbens, ventral pallidum, medial preoptic area, medial amygdala, and the supraoptic nucleus of the hypothalamus. In paired before and after comparisons, males showed significant changes following pairing in the right nucleus accumbens and ventral pallidum but not in other areas. Lesioned males showed significantly higher uptake in the posterior cingulate cortex than all other males. These results indicate some basic similarities between rodents and primates in the formation and maintenance of selective social bonds, but emphasize the importance of studying long-term maintenance in addition to short-term formation of social bonds.


Cytohistopathological correlation of a case of squamous cell carcinoma of gallbladder with lymph node metastasis.
Primary carcinomas of the gallbladder are rare malignancies and adenocarcinoma is the more common subtype. Primary squamous cell carcinoma of the gallbladder has rarely been diagnosed by aspiration cytology. Here, we present a case of a 62-year old female patient suffering from abdominal complaints who underwent ultrasound-guided fine-needle aspiration cytology and was diagnosed as keratinizing squamous cell carcinoma of gallbladder. The diagnosis was confirmed on subsequent histopathological examination that also revealed metastasis in the cystic lymph node. Histogenesis and biological behavior of squamous cell carcinoma of gallbladder remains a matter of debate but the role of aspiration cytology in diagnosing these lesions cannot be undermined.


Long-range promoter-enhancer contacts are conserved during evolution and contribute to gene expression robustness.
Gene expression is regulated through complex molecular interactions, involving cis-acting elements that can be situated far away from their target genes. Data on long-range contacts between promoters and regulatory elements are rapidly accumulating. However, it remains unclear how these regulatory relationships evolve and how they contribute to the establishment of robust gene expression profiles. Here, we address these questions by comparing genome-wide maps of promoter-centered chromatin contacts in mouse and human. We show that there is significant evolutionary conservation of cis-regulatory landscapes, indicating that selective pressures act to preserve not only regulatory element sequences but also their chromatin contacts with target genes. The extent of evolutionary conservation is remarkable for long-range promoter-enhancer contacts, illustrating how the structure of regulatory landscapes constrains large-scale genome evolution. We show that the evolution of cis-regulatory landscapes, measured in terms of distal element sequences, synteny, or contacts with target genes, is significantly associated with gene expression evolution.


Bariatric Surgery and Liver Disease: General Considerations and Role of the Gut-Liver Axis.
Weight loss is a therapeutic solution for many metabolic disorders, such as obesity and its complications. Bariatric surgery aims to achieve lasting weight loss in all patients who have failed after multiple dietary attempts. Among its many benefits, it has been associated with the regression of non-alcoholic fatty liver disease (NAFLD), which is often associated with obesity, with evidence of substantial improvement in tissue inflammation and fibrosis. These benefits are mediated not only by weight loss, but also by favorable changes in systemic inflammation and in the composition of the gut microbiota. Changes in microbial metabolites such as short-chain fatty acids (SCFAs), capable of acting as endocrine mediators, and bile acids (BAs) as well as modifications of the gut-brain axis, are among the involved mechanisms. However, not all bariatric surgeries show beneficial effects on the liver; those leading to malabsorption can cause liver failure or a marked worsening of fibrosis and the development of cirrhosis. Nevertheless, there are still many unclear aspects, including the extent of the benefits and the magnitude of the risks of bariatric surgery in cirrhotic patients. In addition, the usefulness and the safety of these procedures in patients who are candidates to or who have undergone liver transplant need solid supporting evidence. This paper aims to review literature data on the use of bariatric surgery in the setting of chronic liver disease.