Immunosuppressive Medications.
Immunosuppressive agents are commonly used in the nephrologist's practice in the treatment of autoimmune and immune-mediated diseases and transplantation, and they are investigational in the treatment of AKI and ESRD. Drug development has been rapid over the past decades as mechanisms of the immune response have been better defined both by serendipity (the discovery of agents with immunosuppressive activity that led to greater understanding of the immune response) and through mechanistic study (the study of immune deficiencies and autoimmune diseases and the critical pathways or mutations that contribute to disease). Toxicities of early immunosuppressive agents, such as corticosteroids, azathioprine, and cyclophosphamide, stimulated intense investigation for agents with more specificity and less harmful effects. Because the mechanisms of the immune response were better delineated over the past 30 years, this specialty is now bestowed with a multitude of therapeutic options that have reduced rejection rates and improved graft survival in kidney transplantation, provided alternatives to cytotoxic therapy in immune-mediated diseases, and opened new opportunities for intervention in diseases both common (AKI) and rare (atypical hemolytic syndrome). Rather than summarizing clinical indications and clinical trials for all currently available immunosuppressive medications, the purpose of this review is to place these agents into mechanistic context together with a brief discussion of unique features of development and use that are of interest to the nephrologist.


A highly specific PCR assay for detecting the fish ectoparasite Amyloodinium ocellatum.
Amyloodiniosis, caused by the dinoflagellate ectoparasite Amyloodinium ocellatum, is one of the most serious diseases affecting marine fish in warm and temperate waters. Current diagnostic methods rely entirely on the microscopic identification of parasites on the skin or gills of infested fish. However, subclinical infestations usually go undetected, while no method of detecting the free-swimming, infective (dinospore) stage has been devised. Targeting the parasite's ribosomal DNA region, we have developed a sensitive and specific PCR assay that can detect as little as a single cell from any of the 3 stages of the parasite's life cycle (trophont, tomont, dinospore). This assay performs equally well in a simple artificial seawater medium and in natural seawater containing a plankton community assemblage. The assay is also not inhibited by gill tissue. Sequence analysis of the internal transcribed spacer region of 5 A. ocellatum isolates, obtained from fish in the Red Sea (Israel), eastern Mediterranean Sea (Israel), Adriatic Sea (Italy), Gulf of Mexico (Florida), and from an unknown origin, revealed insignificant variation, indicating that all isolates were the same species. However, 3 of these isolates propagated in cell culture varied in behavior and morphology, and these differences were consistent during at least 2 yr in culture. Thus, our findings do not eliminate the possibility that different strains are in fact 'subspecies' or lower taxa, which may also differ in pathogenic and immunogenic characteristics, environmental tolerance, and other features.


In vivo antiplasmodial potentials of the combinations of four nigerian antimalarial plants.
Various combinations of Nauclea latifolia root, Artocarpus altilis stem bark, Murraya koenigii leaf and Enantia chlorantha stem bark used in African ethnomedicine as decoctions for malaria and fevers, and combinations with standard drugs, were investigated for antiplasmodial activities using Plasmodium berghei berghei-infected mice. The respective prophylactic and curative ED50 values of 189.4 and 174.5 mg/kg for N. latifolia and chemosuppressive ED50 value of 227.2 mg/kg for A. altilis showed that they were the best antimalarial herbal drugs. A 1.6-fold increase of the survival time given by the negative control was elicited by M. koenigii, thereby confirming its curative activity. Pyrimethamine with an ED50 of 0.5 ± 0.1 mg/kg for the prophylactic, and chloroquine with ED50 = 2.2 ± 0.1 and 2.2 ± 0.0 mg/kg for the chemosuppressive and curative tests, respectively, were significantly (p < 0.05) more active. Co-administrations of N. latifolia with the standard drugs significantly reduced their prophylactic, chemosuppressive and curative actions, possibly increasing the parasites' resistance. Binary combinations of N. latifolia or M. koenigii with any of the other plants significantly increased the prophylactic and suppressive activities of their individual plants, respectively. Also, E. chlorantha with A. altilis or N. latifolia enhanced their respective prophylactic or curative activities, making these combinations most beneficial against malaria infections. Combinations of three and four extracts gave varied activities. Hence, the results justified the combinations of ethnomedicinal plants in antimalarial herbal remedies and showed the importance of the three in vivo models in establishing antimalarial activity.


Self-coalescing flows in microfluidics for pulse-shaped delivery of reagents.
Microfluidic systems can deliver portable point-of-care diagnostics without the need for external equipment or specialist operators, by integrating all reagents and manipulations required for a particular assay in one device1. A key approach is to deposit picogram quantities of dried reagents in microchannels with micrometre precision using specialized inkjet plotters2-5. This means that reagents can be stored for long periods of time and reconstituted spontaneously when adding a liquid sample. But it is challenging to carry out complex operations using multiple reagents, because shear flow enhances their dispersion and they tend to accumulate at moving liquid fronts, resulting in poor spatiotemporal control over the concentration profile of the reconstituted reagents6. One solution is to limit the rate of release of reagents into the liquid7-10. However, this requires the fine-tuning of different reagents, conditions and targeted operations, and cannot readily produce the complex, time-dependent multireagent concentration pulses required for sophisticated on-chip assays. Here we report and characterize a capillary flow phenomenon that we term self-coalescence, which is seen when a confined liquid with a stretched air-liquid interface is forced to 'zip' back onto itself in a microfluidic channel, thereby allowing reagent reconstitution with minimal dispersion. We provide a comprehensive framework that captures the physical underpinning of this effect. We also fabricate scalable, compact and passive microfluidic structures-'self-coalescence modules', or SCMs-that exploit and control this phenomenon in order to dissolve dried reagent deposits in aqueous solutions with precise spatiotemporal control. We show that SCMs can reconstitute multiple reagents so that they either undergo local reactions or are sequentially delivered in a flow of liquid. SCMs are easily fabricated in different materials, readily configured to enable different reagent manipulations, and readily combined with other microfluidic technologies, so should prove useful for assays, diagnostics, high-throughput screening and other technologies requiring efficient preparation and manipulation of small volumes of complex solutions.


Over-expression of specific HvCslF cellulose synthase-like genes in transgenic barley increases the levels of cell wall (1,3;1,4)-β-d-glucans and alters their fine structure.
Cell walls in commercially important cereals and grasses are characterized by the presence of (1,3;1,4)-β-d-glucans. These polysaccharides are beneficial constituents of human diets, where they can reduce the risk of hypercholesterolemia, type II diabetes, obesity and colorectal cancer. The biosynthesis of cell wall (1,3;1,4)-β-d-glucans in the Poaceae is mediated, in part at least, by the cellulose synthase-like CslF family of genes. Over-expression of the barley CslF6 gene under the control of an endosperm-specific oat globulin promoter results in increases of more than 80% in (1,3;1,4)-β-d-glucan content in grain of transgenic barley. Analyses of (1,3;1,4)-β-d-glucan fine structure indicate that individual CslF enzymes might direct the synthesis of (1,3;1,4)-β-d-glucans with different structures. When expression of the CslF6 transgene is driven by the Pro35S promoter, the transgenic lines have up to sixfold higher levels of (1,3;1,4)-β-d-glucan in leaves, but similar levels as controls in the grain. Some transgenic lines of Pro35S:CslF4 also show increased levels of (1,3;1,4)-β-d-glucans in grain, but not in leaves. Thus, the effects of CslF genes on (1,3;1,4)-β-d-glucan levels are dependent not only on the promoter used, but also on the specific member of the CslF gene family that is inserted into the transgenic barley lines. Altering (1,3;1,4)-β-d-glucan levels in grain and vegetative tissues will have potential applications in human health, where (1,3;1,4)-β-d-glucans contribute to dietary fibre, and in tailoring the composition of biomass cell walls for the production of bioethanol from cereal crop residues and grasses.


A randomized controlled trial of pretransplant antiviral therapy to prevent recurrence of hepatitis C after liver transplantation.
Hepatitis C virus (HCV) infection recurs in liver recipients who are viremic at transplantation. We conducted a randomized, controlled trial to test the efficacy and safety of pretransplant pegylated interferon alpha-2b plus ribavirin (Peg-IFN-α2b/RBV) for prevention of post-transplant HCV recurrence. Enrollees had HCV and were listed for liver transplantation, with either potential living donors or Model for End-Stage Liver Disease upgrade for hepatocellular carcinoma. Patients with HCV genotypes (G) 1/4/6 (n = 44/2/1) were randomized 2:1 to treatment (n = 31) or untreated control (n = 16); HCV G2/3 (n=32) were assigned to treatment. Overall, 59 were treated and 20 were not. Peg-IFN-α2b, starting at 0.75 μg/kg/week, and RBV, starting at 600 mg/day, were escalated as tolerated. Patients assigned to treatment versus control had similar baseline characteristics. Combined virologic response (CVR) included pretransplant sustained virologic response and post-transplant virologic response (pTVR), defined as undetectable HCV RNA 12 weeks after end of treatment or transplant, respectively. In intent-to-treat analyses, 12 (19%) assigned to treatment and 1 (6%) assigned to control achieved CVR (P = 0.29); per-protocol values were 13 (22%) and 0 (0%) (P = 0.03). Among treated G1/4/6 patients, 23 of 30 received transplant, of whom 22% had pTVR; among treated G2/3 patients 21 of 29 received transplant, of whom 29% had pTVR. pTVR was 0%, 18%, and 50% in patients treated for <8, 8-16, and >16 weeks, respectively (P = 0.01). Serious adverse events (SAEs) occurred with similar frequency in treated versus untreated patients (68% versus 55%; P = 0.30), but the number of SAEs per patient was higher in the treated group (2.7 versus 1.3; P = 0.003). Pretransplant treatment with Peg-IFN-α2b/RBV prevents post-transplant recurrence of HCV in selected patients. Efficacy is higher with >16 weeks of treatment, but treatment is associated with increased risk of potentially serious complications.


Internal manual rotation in intrapartal arrest of fetal head engagement.
The success of internal manual or digital rotation of the head in mechanical dystocia due to malpresentation, malposition or malrotation is presented in this paper on our own clinical material with reference of today's research and clinical recommendations. Through a retrospective bicentric clinical study, we investigated the success of internal head rotation in two University Clinics for gynecology and obstetrics from year 2017 to 2023. In 152 singleton term (37-42 weeks) in cases of persistens intrapartum arrest of the fetal head. After palpatory and ultrasonographically verified arrest of fetal head engagement, a therapeutic manual (Liepmann) or digital rotation was performed. In 152 cases, manual rotation was performed in 108 (71.05 %) and digital rotation in 44 (28.94 %) cases in 73 (48.02 %) primiparous and 79 (51.97 %) multiparous. Intrapartum identification by digital palpation was done in all cases, and the following are: persistent occipital posterior position in 68 (44.73%), persistent deep transverse head presentation in 12 (7.89%), persistent high (longitudinal) occipital presentation in 64 (42.10 %) and persistent anterior asynclitism in 8 (5.26 %) cases. Episiotomy was used in 36 (23.68%) cases. Vacuum extraction was completed in 14 (9.21 %) deliveries, and cesarean section due to unsuccessful internal rotation in 15 (9.8 %) cases (%) without other indication. We did not record any intrapartum complications or cardiotocographic abnormalities. Cervical lacerations were treated with sutures in 4 cases (2.63 %). Successful correction of internal rotation procedure with spontaneous vaginal delivery was found in 80.92 % of cases. If we exclude delivery assisted by vacuum extraction whose indications were fetal hypoxia or dystocia after successful internal head rotation procedure, then the success rate of this method was 90.13 %. Internal head rotation is a simple, safe and successful obstetric manual intervention that directly increases the rate of vaginal deliveries after correction of the birth mechanism anomaly and directly reduces the percentage of cesarean section. Manual or digital head rotation is an established midwifery/obstetric skill in several centers which, based on numerous clinical researches and experience, should become protocolized and included in the guidelines of professional associations.


Low-level laser therapy promotes vascular endothelial growth factor receptor-1 expression in endothelial and nonendothelial cells of mice gastrocnemius exposed to snake venom.
Crotalinae snake venoms cause severe local myonecrosis and microvasculature failure at the bite site. We evaluated whether low-level laser therapy (LLLT) could accelerate angiogenesis and myoregeneration in male Swiss mice injected with Bothrops moojeni venom through immunohistochemistry of the vascular endothelial growth factor receptor-1 (VEGFR-1). Envenomed gastrocnemius was either unirradiated (V) or irradiated with HeNe (VHN, 632.8 nm) or GaAs (VGA, 904 nm, 10000 Hz). Animals sacrificed at 3 and 12 h were irradiated once (4 J cm(-2)), at 24 h (twice) and at 3, 7, 21 days (4, 8, 22 times, respectively). At 3 days, LLLT increased angiogenesis (80%:HeNe vs 40%:GaAs), decreased neutrophils and increased proliferation of regenerating cells. However, after 21 days, myoregeneration observed in the VHN group appeared delayed compared with the V group. As LLLT improved revascularization, the suggestive delay in myoregeneration could be a dose-response inhibitory effect caused by multiple irradiations in myogenesis. The immunodetection of VEGFR-1 in neutrophils, macrophages, satellite cells, fibroblasts, Schwann cells and skeletal and smooth muscle fibers (not seen in saline-controls) at only the acute stages of envenoming suggests a mediator role for VEGFR-1 in local alterations. This is the first time that VEGFR-1 expression, and its modulation by photostimulation, has been demonstrated in endothelial and nonendothelial cells of snake envenomed skeletal muscle.


Genomic inbreeding estimation in small populations: evaluation of runs of homozygosity in three local dairy cattle breeds.
In the local breeds with small population size, one of the most important problems is the increase of inbreeding coefficient (F). High levels of inbreeding lead to reduced genetic diversity and inbreeding depression. The availability of high-density single nucleotide polymorphism (SNP) arrays has facilitated the quantification of F by genomic markers in farm animals. Runs of homozygosity (ROH) are contiguous lengths of homozygous genotypes and represent an estimate of the degree of autozygosity at genome-wide level. The current study aims to quantify the genomic F derived from ROH (F ROH) in three local dairy cattle breeds. F ROH values were compared with F estimated from the genomic relationship matrix (F GRM), based on the difference between observed v. expected number of homozygous genotypes (F HOM) and the genomic homozygosity of individual i (F MOL i ). The molecular coancestry coefficient (f MOL ij ) between individuals i and j was also estimated. Individuals of Cinisara (71), Modicana (72) and Reggiana (168) were genotyped with the 50K v2 Illumina BeadChip. Genotypes from 96 animals of Italian Holstein cattle breed were also included in the analysis. We used a definition of ROH as tracts of homozygous genotypes that were >4 Mb. Among breeds, 3661 ROH were identified. Modicana showed the highest mean number of ROH per individual and the highest value of F ROH, whereas Reggiana showed the lowest ones. Differences among breeds existed for the ROH lengths. The individuals of Italian Holstein showed high number of short ROH segments, related to ancient consanguinity. Similar results showed the Reggiana with some extreme animals with segments covering 400 Mb and more of genome. Modicana and Cinisara showed similar results between them with the total length of ROH characterized by the presence of large segments. High correlation was found between F HOM and F ROH ranged from 0.83 in Reggiana to 0.95 in Cinisara and Modicana. The correlations among F ROH and other estimated F coefficients were generally lower ranged from 0.45 (F MOL i -F ROH) in Cinisara to 0.17 (F GRM-F ROH) in Modicana. On the basis of our results, recent inbreeding was observed in local breeds, considering that 16 Mb segments are expected to present inbreeding up to three generations ago. Our results showed the necessity of implementing conservation programs to control the rise of inbreeding and coancestry in the three Italian local dairy cattle breeds.


Increasing incidence rates, distribution and histological characteristics of primary gastrointestinal non-Hodgkin lymphoma in a North American population.
The incidence of primary extranodal non-Hodgkin lymphoma (NHL) of the gastrointestinal (GI) tract has been on the rise. To determine the incidence of primary GI NHL and distribution according to site and histological type in a large North American adult population over a 10-year period. All diagnoses of GI NHL made between January 1999 and January 2009 were reviewed using a regional pathology database. Patients ≥18 years of age living within health region boundaries were included. Age- and sex-adjusted incidence rates of GI NHL according to GI site and histological type over a 10-year period were calculated and compared. A total of 149 cases of primary GI NHL were identified during the study period. Age- and sex-adjusted yearly incidence rates ranged from 0.13 per 100,000 in 1999, to 2.39 per 100,000 in 2007. Histological distribution (47% diffuse large B cell lymphoma, 24% extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue type, 8% follicular and 5% mantle cell) and site distribution (47% stomach, 26% small bowel, 17% colon) were obtained with increasing annualized incidence rates for each of these sites over time. Remaining cases included multiple GI sites of involvement (9%) and esophagus (0.7%). Population-based GI NHL incidence rates in the present study were higher than those described elsewhere in North America and Europe. Nearly one-half showed high-grade (diffuse large B cell lymphoma) histology at diagnosis. Incidence rates for the colon exceed those described in other studies worldwide. Because the majority of GI NHL are diagnosed on endoscopic biopsy, clinicians and pathologists must be vigilant of this entity.


Recombinant β-Carotene Production by Yarrowia lipolytica - Assessing the Potential of Micro-Scale Fermentation Analysis in Cell Factory Design and Bioreaction Optimization.
The production of β-carotene has become increasingly interesting within the biotechnological industry due to a rising demand for safer and more natural colorants, nutritional supplements, and antioxidants. A recent study has described the potential of Yarrowia lipolytica as a β-carotene-producing cell factory, reporting the highest titer of recombinant β-carotene produced to date. Finding the best conditions to maximize production and scaling up the process to full scale, a costly and time-consuming process, it is often a bottleneck in biotechnology. In this work, we explored the benefits of using micro-fermentation equipment to significantly reduce the time spent on design and optimization of bioreaction conditions, especially in the early stages of process development. In this proof-of-concept study, a β-carotene producing Y. lipolytica strain was tested in micro-fermentations partly to assess the robustness of the cell factory design and partly to perform media optimization. The medium optimization led us to an improvement of up to 50% in the yield of β-carotene production in the best of the conditions. Overall, the micro-fermentation system had a high degree of reliability in all tests.


The use of Mycophenolate Mofetil (Cellcept) in renal transplantation.
Mycophenolate Mofetil (MMF), the morpholinoethyl ester of the active compound mycophenolic acid, is a new immunosuppressive agent which blocks the de novo synthesis of guanine nucleotides. As this pathway is essential for purine synthesis of both T and B lymphocytes, the blocking effect on cell division is lymphocyte-selective with little effect on the division of other cell types. In three recent randomized multicenter trials, MMF in conjunction with ciclosporin and steroids has been shown to reduce the incidence of early acute rejection after renal transplantation by almost 50%. The toxicity profile is comparable to that of azathioprine but with a significantly higher efficacy. Major side effects are gastrointestinal with a slight increase of the incidence of mainly viral (CMV) infections. In contrast to ciclosporin and FK-506, MMF is not nephrotoxic. Future studies have to show whether the use of MMF in combination with other immunosuppressants may allow to reduce the doses of the most toxic ones. In addition, long-term studies are needed to prove its potential protective effect on the occurrence of chronic rejection as suggested by several experimental studies and by its effect on the incidence of acute rejection.


Measures of growth, development, and sexual maturation in mongolian gerbils (Meriones unguiculatus): effects of photic period during ontogeny.
We examined, in Mongolian gerbils, the adequacy of two commonly used indices of rate of development, age at eye-opening and age at vaginal introitus, as predictors of biologically important events: initiation of feeding on solid food and age at first parturition. We then enquired as to the interrelationships among body weight, eye-opening, and vaginal introitus in Mongolian gerbils whose rates of development were manipulated by altering the extent of their exposure to illumination during ontogeny. We found that (1) age at vaginal introitus was significantly, but weakly, correlated with age at first parturition (Experiment I); (2) age at eye-opening strongly predicted age of initiation of gnawing on solid food (Experiment II); (3) eye-opening occurred at markedly different ages, but at the same body weight in gerbils reared under 0/24-, 6/18-, 12/12-, and 24/0-hr day-night cycles (Experiment III); and (4) vaginal introitus occurred at both different ages and different body weights in females reared under different day-night cycles (Experiment III).


Nanofluidic and monolithic environmental cells for cryogenic microscopy.
We present a device capable of combining nanofluidics and cryogenic transmission electron microscopy (cryo-TEM) to allow inspection of water-soluble samples under near-native conditions. The devices can be produced in a multitude of designs, but as a general rule, they consist of channels or chambers enclosed between two electron-transparent silicon nitride windows. With the appropriate design, those devices can allow screening of multiple samples in parallel and remove the interaction between the sample and the environment (no air-water interface). We demonstrate channel sizes from 80 to 500 nm in height and widths from 100 to 2000 μm. The presented fabrication flow allows producing hollow devices on a single wafer eliminating the need of aligning or bonding two half-cavities from separate wafers, which provides additional resistance to thermal stress. Taking advantage of a single-step through-membrane exposure with a 100 keV electron beam, we introduced arrays of thin (10-15 nm) electron-transparent silicon nitride membrane windows aligned between top and bottom (200-250 nm) carrier membranes. Importantly, the final devices are compatible with standard TEM holders. Furthermore, they are compatible with rapid freezing of samples, which is crucial for the formation of vitreous water, hence avoiding the formation of crystalline ice, that is detrimental for TEM imaging. To demonstrate the potential of this technology, we tested those devices in imaging experiments verifying their applicability for cryo-TEM applications and proved that vitreous water could be prepared through conventional plunge freezing of the chips.


Memory books: helping children cope with loss.
Children who have a parent or significant family member who is ill or dying or who has died handle their grief in different ways. Memory books have helped keep children mentally healthier and kept them at home rather than being hospitalized for the behaviors by which they express their grief.


The newly proposed Metabolic Score for Visceral Fat is a reliable tool for identifying non-alcoholic fatty liver disease, requiring attention to age-specific effects in both sexes.
The newly proposed Metabolic Visceral Fat Score (METS-VF) is considered a more effective measure for visceral adipose tissue (VAT) than other obesity indicators. This study aimed to reveal the association between METS-VF and non-alcoholic fatty liver disease (NAFLD), and its variations across age groups within both sexes. Data from 14,251 medical examiners in the NAGALA project were employed in this study. 3D fitted surface plots were constructed based on multivariate logistic regression models to visualize the isolated and combined effects of aging and METS-VF on NAFLD. Receiver operating characteristic curve (ROC) analysis was conducted to compare the diagnostic performance of METS-VF with other VAT surrogate markers in predicting NAFLD. The results of multivariate logistic regression analysis showed that each unit increase in METS-VF was independently associated with a 333% and 312% increase in the odds of NAFLD in males and females, respectively. Additionally, the 3D fitted surface plot showed that age significantly influenced the association between METS-VF and the odds of NAFLD in both sexes, as follows: (i) In males, when METS-VF was less than 6.2, the METS-VF-related odds of NAFLD increased gradually with age in the 20-45 age group, reached a plateau in the 45-65 age group, and then decreased in the group above 65 years old; however, when male METS-VF exceeded 6.2, aging and METS-VF combined to further increase the odds of NAFLD in all age groups, particularly in the 45-65 age group. (ii) In females, aging seemed to reduce METS-VF-related odds of NAFLD in the 18-40 age group, but significantly increased it in the 40-60 age group, particularly for those with higher METS-VF levels. Further ROC analysis revealed that compared to other VAT surrogate markers, METS-VF showed the highest diagnostic accuracy for NAFLD in females, especially in those under 45 years of age [area under the curve (AUC) = 0.9256]. This study firstly revealed a significant positive correlation between METS-VF and the odds of NAFLD, with METS-VF surpassing other VAT surrogate markers in NAFLD diagnosis. Moreover, age significantly influenced the METS-VF-related odds of NAFLD and METS-VF's diagnostic efficacy for NAFLD in both sexes.


Minimally invasive video-assisted thyroidectomy and parathyroidectomy with intraoperative recurrent laryngeal nerve monitoring.
Objective. Our goal is to study the feasibility of using intraoperative neuromonitoring (IONM) in minimally invasive video-assisted thyroidectomy and parathyroidectomy (MIVAT/P) with emphasis given to the identification of recurrent laryngeal nerve (RLN). Methods. Consecutive series of forty-seven patients with seventy-seven recurrent laryngeal nerves at risk undergoing both MIVAT/P and IONM were enrolled in this retrospective, nonrandomized analysis study. All operations were performed by the same surgeon within an academic institution setting. All patients underwent vocal cord evaluation postoperatively. Demographics and intraoperative and postoperative complications following surgery were collected. Results. Out of seventy-seven RLNs, there was one permanent unilateral RLN injury (1.29%) in a patient with advanced papillary thyroid cancer, managed by cord injection. There was another transient RLN paresis that resolved spontaneously (1.29%). There were no instances of equipment malfunction or interference. Conclusions. To our knowledge, this is the first reported MIVAT/P series from the United States of America with a standardized IONM technique. The technical feasibility of IONM seems acceptable and may serve as a meaningful adjunct to the visual identification of nerves. Neuromonitoring during MIVAT/P is effective in providing identification of laryngeal nerves and enables surgeons to feel more comfortable with MIVAT/P. Comparative series are needed for further evaluation.


Advances in mediation analysis can facilitate nursing research.
Two recent advances in the statistical methods for testing hypotheses about mediation effects are important for nursing science. First, bootstrap sampling distributions provide more accurate tests of hypotheses about mediated effects. Second, methods for testing statistical hypotheses about subgroup differences in mediation models (moderated mediation) are now well developed. The aims of this study were to demonstrate the use and relatively simple computation of bootstrap sampling distributions in tests of mediation effects and to demonstrate a recently refined method for testing hypotheses about moderated mediation. Using hypothetical data, a step-by-step demonstration was provided of the construction of a bootstrap sampling distribution for a correlation coefficient. Then, tests of mediation and moderated mediation were demonstrated using data from a clinical trial of an intervention for caregivers of patients with Parkinson's disease or Alzheimer's disease. In a model hypothesizing that mutuality between caregiver and care recipient mediates the effect of objective on subjective levels of caregiver burden, the bootstrap sampling distribution was calculated of the mediation effect and, from that, two types of 95% confidence intervals for it. Then the hypothesis was tested that the mediating effect of mutuality was stronger for caregivers of patients with Parkinson's disease than for caregivers of patients with Alzheimer's disease. Statistical hypothesis testing should never dictate all conclusions. However, the statistical advances in mediation analysis described here will facilitate nursing research as both nurse scientists and methodologists understand their assumptions and logic.


Effect of Embryo Vitrification on Rabbit Foetal Placenta Proteome during Pregnancy.
Very limited information on the post-implantatory effects of vitrification has been published till now. We observed in a previous study that the vitrification procedure for the cryopreservation of embryos introduced transcriptomic and proteomic modifications in the rabbit foetal placenta at the middle of gestation. Now, we have conducted a proteomic study to determine whether protein alterations in the foetal placenta induced by the vitrification procedure remain during pregnancy. In this study, we used 2D-DIGE and mass spectrometry (MALDI-TOF-TOF and LC-MS/MS analysis) to identify the protein changes during middle and late stages of gestation (Day 14 and Day 24, respectively) in rabbit foetal placenta. We identified 11 differentially expressed proteins at Day 14 and 13 proteins at Day 24. Data are available via ProteomeXchange with identifiers PXD001840 and PXD001836. In addition, we demonstrate the presence of three proteins, serum albumin, isocitrate dehydrogenase 1 [NADP+], and phosphoglycerate mutase 1, which were altered during pregnancy. We demonstrate the existence of changes in foetal placental protein during pregnancy induced by the vitrification procedure, which brings into question whether vitrification effects observed during foetal development could lead to physiological and metabolic disorders in adulthood. This effect, taken together with other effects reported in the literature, suggests that embryo cryopreservation is not neutral.


Modeling cascading diffusion of new energy technologies: case study of residential solid oxide fuel cells in the US and internationally.
Subsidy programs for new energy technologies are motivated by the experience curve: increased adoption of a technology leads to learning and economies of scale that lower costs. Geographic differences in fuel prices and climate lead to large variability in the economic performance of energy technologies. The notion of cascading diffusion is that regions with favorable economic conditions serve as the basis to build scale and reduce costs so that the technology becomes attractive in new regions. We develop a model of cascading diffusion and implement via a case study of residential solid oxide fuel cells (SOFCs) for combined heating and power. We consider diffusion paths within the U.S. and internationally. We construct market willingness-to-pay curves and estimate future manufacturing costs via an experience curve. Combining market and cost results, we find that for rapid cost reductions (learning rate = 25%), a modest public subsidy can make SOFC investment profitable for 20-160 million households. If cost reductions are slow however (learning rate = 15%), residential SOFCs may not become economically competitive. Due to higher energy prices in some countries, international diffusion is more favorable than domestic, mitigating much of the uncertainty in the learning rate.


Carfilzomib, cyclophosphamide, and dexamethasone in patients with newly diagnosed multiple myeloma: a multicenter, phase 2 study.
This multicenter, open-label phase 2 trial determined the safety and efficacy of carfilzomib, a novel and irreversible proteasome inhibitor, in combination with cyclophosphamide and dexamethasone (CCyd) in patients with newly diagnosed multiple myeloma (NDMM) ≥65 years of age or who were ineligible for autologous stem cell transplantation. Patients (N = 58) received CCyd for up to 9 28-day cycles, followed by maintenance with carfilzomib until progression or intolerance. After a median of 9 CCyd induction cycles (range 1-9), 95% of patients achieved at least a partial response, 71% achieved at least a very good partial response, 49% achieved at least a near complete response, and 20% achieved stringent complete response. After a median follow-up of 18 months, the 2-year progression-free survival and overall survival rates were 76% and 87%, respectively. The most frequent grade 3 to 5 toxicities were neutropenia (20%), anemia (11%), and cardiopulmonary adverse events (7%). Peripheral neuropathy was limited to grades 1 and 2 (9%). Fourteen percent of patients discontinued treatment because of adverse events, and 21% of patients required carfilzomib dose reductions. In summary, results showed high complete response rates and a good safety profile. This trial was registered at clinicaltrials.gov as #NCT01346787.


The development of clinical reasoning and interprofessional behaviors: service-learning at a student-run free clinic.
This article examines the benefits of a student run free clinic (SRFC) as a service learning experience for students in medicine, pharmacy, occupational therapy, physical therapy and physician assistant programs. We hypothesized that students who participate in an interprofessional service learning course and volunteer at a SRFC would demonstrate significant increases in perceptions and attitudes for working in interprofessional health care teams and clinical reasoning skills compared to students who did not participate. Three assessments were administered to an experimental and control group of pre-clinical students from medical, occupational therapy, physical therapy, pharmacy and physician assistant programs before and after participation in an interprofessional service-learning course and volunteering at the SRFC. The tools were the Interdisciplinary Education Perception Scale (IEPS), Readiness for Interprofessional Learning Scale (RIPLS) and the Self-Assessment of Clinical Reflection and Reasoning (SACRR). Students who completed the course had improvements in interprofessional perceptions and attitudes (p = 0.03) and perceptions of clinical reasoning skills when compared to the control group (p = 0.002). This study is novel as it examined students' perceptions of interprofessional attitudes and clinical reasoning following participation in an interprofessional service-learning course and participation in a SRFC.


Modeling complex genetic and environmental influences on comorbid bipolar disorder with tobacco use disorder.
Comorbidity of psychiatric and substance use disorders represents a significant complication in the clinical course of both disorders. Bipolar Disorder (BD) is a psychiatric disorder characterized by severe mood swings, ranging from mania to depression, and up to a 70% rate of comorbid Tobacco Use Disorder (TUD). We found epidemiological evidence consistent with a common underlying etiology for BD and TUD, as well as evidence of both genetic and environmental influences on BD and TUD. Therefore, we hypothesized a common underlying genetic etiology, interacting with nicotine exposure, influencing susceptibility to both BD and TUD. Using meta-analysis, we compared TUD rates for BD patients and the general population. We identified candidate genes showing statistically significant, replicated, evidence of association with both BD and TUD. We assessed commonality among these candidate genes and hypothesized broader, multi-gene network influences on the comorbidity. Using Fisher Exact tests we tested our hypothesized genetic networks for association with the comorbidity, then compared the inferences drawn with those derived from the commonality assessment. Finally, we prioritized candidate SNPs for validation. We estimate risk for TUD among BD patients at 2.4 times that of the general population. We found three candidate genes associated with both BD and TUD (COMT, SLC6A3, and SLC6A4) and commonality analysis suggests that these genes interact in predisposing psychiatric and substance use disorders. We identified a 69 gene network that influences neurotransmitter signaling and shows significant over-representation of genes associated with BD and TUD, as well as genes differentially expressed with exposure to tobacco smoke. Twenty four of these genes are known drug targets. This work highlights novel bioinformatics resources and demonstrates the effectiveness of using an integrated bioinformatics approach to improve our understanding of complex disease etiology. We illustrate the development and testing of hypotheses for a comorbidity predisposed by both genetic and environmental influences. Consistent with our hypothesis, the selected network models multiple interacting genetic influences on comorbid BD with TUD, as well as the environmental influence of nicotine. This network nominates candidate genes for validation and drug testing, and we offer a panel of SNPs prioritized for follow-up.


[Clinical distribution and antimicrobial resistance analysis of 754 pathogenic bacteria in diabetic foot infection].
To explore the microbiological profiles and antibiotic susceptibility patterns of organisms isolated from diabetic foot ulcers so as to provide selection rationales of antibiotics. A retrospective study was conducted on the microbiological profiles and antibiotic susceptibilities in 754 strains of pathogens isolated from 519 patients with diabetic foot ulcers at our hospital from January 2010 to August 2013. The inter-group data were compared by Chi-square test. There were 322 (62.0%) males and 197 (38.0%) females. Their mean age was (67.7 ± 12.3) (30-93) years, duration of diabetes 10 (0-40) years, duration of lower-limb lesion 1.0 (0.0-72.0) months and HbA1c (9.09% ± 2.28%). Among 444 (85.5%) cases, a total of 754 strains of pathogens were isolated. Gram-positive aerobes were the most frequently isolated (47.3%, 357 strains) and followed by gram-negative aerobes and fungus (40.3% vs 12.3%, 304 vs 93 strains respectively). With rising Wagner's grades, bacterial floras transformed from Gram-positive cocci to Gram-negative rods while fungus and composite infections increased. And 122 strains were of multi drug resistant organisms (MDRO). Among 357 strains of Gram-positive bacteria, Staphylococcus aureus, Staphylococcus epidermidis and Enterococcus faecalis were dominating floras. Staphylococcus was highly resistant to penicillin G, erythromycin, and oxacillin while vancomycin and linezolid were the most effective agents against gram-positive bacteria. Among 304 strains of gram-negative bacteria, enterobacteria were the most prevalent, including 48 strains of Escherichia coli, 34 strains of Proteus mirabilis and 31 strains of Proteus vulgaris. And there were 29 strains of Pseudomonas aeruginosa. Enterobacteria were highly resistant to ampicillin, followed by bactrim and furadantin while meropenem, imipenem, piperacillin/sulbactam, sulperazone and cefepime were the most effective agents. The predominant fungus was Blastomyces albicans. In patients with severe diabetic foot ulcers, Gram-negative rods predominate while the prevalence of fungus and composite infections increases. Vancomycin and imipenem maintain highly antibacterial activity. It is essential to pay attention to pathogen survey and use antibiotics more rationally.


Blast-induced traumatic brain injury: the experience from a level I trauma center in southern Thailand.
OBJECTIVEIn the ongoing conflict in southern Thailand, the improvised explosive device (IED) has been a common cause of blast-induced traumatic brain injury (bTBI). The authors investigated the particular characteristics of bTBI and the factors associated with its clinical outcome.METHODSA retrospective cohort study was conducted on all patients who had sustained bTBI between 2009 and 2017. Collected data included clinical characteristics, intracranial injuries, and outcomes. Factors analysis was conducted using a forest plot.RESULTSDuring the study period, 70 patients met the inclusion criteria. Fifty individuals (71.4%) were military personnel. One-third of the patients (32.9%) suffered moderate to severe bTBI, and the rate of intracerebral injuries on brain CT was 65.7%. Coup contusion was the most common finding, and primary blast injury was the most common mechanism of blast injury. Seventeen individuals had an unfavorable outcome (Glasgow Outcome Scale score 1-3), and the overall mortality rate for bTBI was 11.4%. In the univariate analysis, factors associated with an unfavorable outcome were preoperative coagulopathy, midline shift of the brain ≥ 5 mm, basal cistern effacement, moderate to severe TBI, hypotension, fixed and dilated pupils, surgical site infection, hematocrit < 30% on admission, coup contusion, and subdural hematoma. In the multivariable analysis, midline shift ≥ 5 mm (OR 29.1, 95% CI 2.5-328.1) and coagulopathy (OR 28.7, 95% CI 4.5-180.3) were the only factors predicting a poor outcome of bTBI.CONCLUSIONSbTBIs range from mild to severe. Midline shift and coagulopathy are treatable factors associated with an unfavorable outcome. Hence, in cases of bTBI, reversing an abnormal coagulogram is required as soon as possible to improve clinical outcomes. The management of brain shift needs further study.


mTOR (Mechanistic Target of Rapamycin) Inhibition Decreases Mechanosignaling, Collagen Accumulation, and Stiffening of the Thoracic Aorta in Elastin-Deficient Mice.
Elastin deficiency because of heterozygous loss of an ELN allele in Williams syndrome causes obstructive aortopathy characterized by medial thickening and fibrosis and consequent aortic stiffening. Previous work in Eln-null mice with a severe arterial phenotype showed that inhibition of mTOR (mechanistic target of rapamycin), a key regulator of cell growth, lessened the aortic obstruction but did not prevent early postnatal death. We investigated the effects of mTOR inhibition in Eln-null mice partially rescued by human ELN that manifest a less severe arterial phenotype and survive long term. Thoracic aortas of neonatal and juvenile mice with graded elastin deficiency exhibited increased signaling through both mTOR complex 1 and 2. Despite lower predicted wall stress, there was increased phosphorylation of focal adhesion kinase, suggestive of greater integrin activation, and increased transforming growth factor-β-signaling mediators, associated with increased collagen expression. Pharmacological blockade of mTOR by rapalogs did not improve luminal stenosis but reduced mechanosignaling (in delayed fashion after mTOR complex 1 inhibition), medial collagen accumulation, and stiffening of the aorta. Rapalog administration also retarded somatic growth, however, and precipitated neonatal deaths. Complementary, less-toxic strategies to inhibit mTOR via altered growth factor and nutrient responses were not effective. In addition to previously demonstrated therapeutic benefits of rapalogs decreasing smooth muscle cell proliferation in the absence of elastin, we find that rapalogs also prevent aortic fibrosis and stiffening attributable to partial elastin deficiency. Our findings suggest that mTOR-sensitive perturbation of smooth muscle cell mechanosensing contributes to elastin aortopathy.


[Kawasaki disease. Report of two cases].
We present two reports of Kawasaki's disease. The first a five months old male, which was diagnosed through autopsy developed cardiovascular complications (myocarditis, coronary aneurysms, myocardial infarction) which led to death. The second one a two years and ten months old female, which was diagnosed at the fifth day of illness, was treated with aspirin at rate of 100 mg/kg/day and healed without consequences. It is interesting to emphasize the different development as a function of age, sex and precocity of diagnosis.


The Investigation and Prediction of Voriconazole-Associated Hepatotoxicity under Therapeutic Drug Monitoring.
Voriconazole is commonly used as the first-line agent to treat invasive fungal infections (IFIs), but the induction of hepatotoxicity limits its use. To improve the treatment outcomes and minimize toxicity, doctors often administer Therapeutic Drug Monitoring (TDM) to patients receiving voriconazole treatment. Here, we conducted a real-world clinical investigation of voriconazole-treated patients and found significant differences between the TDM (n=318) and non-TDM cohort (n=6,379), and such incidence of hepatotoxicity showed 10.6% in the non-TDM cohort, compared with 21.5% in the TDM cohort. Based on our previous investigation, we presented and compared several machine learning models (including AdaBoost, decision tree, GBDT, logistic regression, neural networks, and random forest) for the early warning of voriconazole-associated hepatoxicity. Through the five-fold cross validation, the logistic model outperformed other models with a mean AUC of 0.7933±0.0934. Our findings offer important insights into the safe and effective application of voriconazole.


Multiple mechanisms explain loss of anthocyanins from betalain-pigmented Caryophyllales, including repeated wholesale loss of a key anthocyanidin synthesis enzyme.
In this study, we investigate the genetic mechanisms responsible for the loss of anthocyanins in betalain-pigmented Caryophyllales, considering our hypothesis of multiple transitions to betalain pigmentation. Utilizing transcriptomic and genomic datasets across 357 species and 31 families, we scrutinize 18 flavonoid pathway genes and six regulatory genes spanning four transitions to betalain pigmentation. We examined evidence for hypotheses of wholesale gene loss, modified gene function, altered gene expression, and degeneration of the MBW (MYB-bHLH-WD40) trasnscription factor complex, within betalain-pigmented lineages. Our analyses reveal that most flavonoid synthesis genes remain conserved in betalain-pigmented lineages, with the notable exception of TT19 orthologs, essential for the final step in anthocyanidin synthesis, which appear to have been repeatedly and entirely lost. Additional late-stage flavonoid pathway genes upstream of TT19 also manifest strikingly reduced expression in betalain-pigmented species. Additionally, we find repeated loss and alteration in the MBW transcription complex essential for canonical anthocyanin synthesis. Consequently, the loss and exclusion of anthocyanins in betalain-pigmented species appear to be orchestrated through several mechanisms: loss of a key enzyme, downregulation of synthesis genes, and degeneration of regulatory complexes. These changes have occurred iteratively in Caryophyllales, often coinciding with evolutionary transitions to betalain pigmentation.


Immunoglobulin VH gene expression in human aging.
Immune responses change in aging humans, but it is not known whether there is an age-associated change in the expressed B cell repertoire. We compared Ig VH cDNA libraries from circulating B cells of five elderly and three young human adults. As in young persons, nearly two-thirds of the cDNA clones from older subjects had zero to three V(H) mutations, although there was more individual variation among the elderly. V(H)4 family expression increased in older subjects, both in unmutated and in mutated cDNA clones, whereas V(H)3 family expression predominated in young adults. To test for bias toward activated cells in the cDNA libraries, we studied two older persons by both cDNA library analysis and single-cell RT-PCR. In one subject, more than 85% of VH segments were unmutated by either analysis. In the second, mutated Ig segments were much more frequent in cDNA clones than in consecutive single cells; however, V(H) family usage and high representation of particular genes were similar in both analyses. While aging humans continue to produce naive B cells with unmutated Ig genes, a shift to greater use of the V(H)4 family members and expression of particular genes may reflect changes in selection of developing B cells before affinity maturation toward reactivity with foreign antigen.


Development and testing of a scale to assess interprofessional education (IPE) facilitation skills.
Interprofessional education (IPE) is interactive and constructivist in nature and requires specific facilitation skills to engage participants in a unique body of content, interpersonal interaction, and learning from each other. This article describes the development and testing of a scale, the Interprofessional Facilitation Scale (IPFS), to assess educators' skills in facilitating IPE. Following participation in an Interprofessional Facilitator Development Program, facilitators provided interprofessional workshops for health professionals caring for patients with cancer. Workshop participants (311 community health professionals) assessed IPE facilitation skills with the use of the IPFS. Psychometric testing of the scale demonstrated high reliability and strong construct and content validity. Factor analysis produced a 2-factor solution that explained 62.1% of the scale variance. The factors "Encouraging IP interaction" and "Contextualizing IPE" were psychometrically rigorous and supported by the literature as being critical to facilitating successful IPE. The IPFS can be used in facilitator development as a concise guide to IPE facilitation skills and for assessment and further enhancement of IP facilitation competencies. Further study is required to assess the scale in diverse settings, with preservice learners, and over the longer term.


Modulation of cell cycle progression in the spermatocyte cell line [GC-2spd(ts) Cell-Line] by cigarette smoke condensate (CSC) via arylhydrocarbon receptor-nuclear factor erythroid 2-related factor 2 (Ahr-Nrf2) pathway.
Prior studies in our laboratory have demonstrated that cigarette smoke condensate (CSC) activates arylhydrocarbon receptor (Ahr) leading to upregulation of several antioxidant enzymes in murine spermatocytes. In this study, we show that exposure of the spermatocyte cell line GC-2spd(ts) to CSC induces an increase in Cyp1a1, demonstrating AHR activation, and simultaneous expression and nuclear translocation of nuclear factor erythroid 2-related factor 2 (NRF2), where it is believed to modulate Ahr expression by a feedback mechanism. Pharmacological inhibition by the AHR-antagonist CH223191 and interference by Ahr- and Nrf2-small interfering RNA followed by quantitative real-time PCR implicate the Ahr-Nrf2 pathway in the modulation of DNA damage and growth suppression genes such as Gadd45a and P21 and oxidative stress-related genes Cyp1a1, Nrf2, and Ahrr. Flow cytometry accompanied with cell proliferation assay indicate the CSC induces accumulation of spermatocytes at the S-G2/M phase of the cell cycle. Thus, the data obtained suggest that CSC contains several AHR-agonists that are capable of altering the growth pattern of spermatocytes in vitro through the Ahr-Nrf2 signaling mechanism.


Exposure to sexually explicit media in early adolescence is related to risky sexual behavior in emerging adulthood.
Sexually explicit media exposure during early adolescence has been found to be associated with risky sexual behavior. However, previous study suffered from methodological issue, such as selection bias. Furthermore, little is known about the effect of multi-modality sexually explicit media exposure on risky sexual behavior, and how this relationship can be applied to non-western societies. This study aimed to improve upon previous studies by using instrumental variable estimation. In addition, this study also included multi-modality of sexually explicit media and three risky sexual behavior measure from a sample of Taiwanese adolescents. Participants were recruited from a prospective longitudinal study (Taiwan Youth Project). All were in 7th grade (mean age = 13.3) when the study was initiated in 2000. Sexually explicit media exposure, including ever-exposure and number of modalities exposed to, was measured in wave 2 (8th grade). Risky sexual behavior was measured in waves 8 (mean age = 20.3) and 10 (mean age = 24.3). A two-stage least squares regression was employed, with pubertal timing as the instrumental variable. About 50% of participants had been exposed to sexual media content by 8th grade, from an average of one modality. Sexually explicit media exposure predicted early sexual debut, unsafe sex, and multiple sexual partners (all: p < .05). Furthermore, exposure to more media modalities increased the likelihood of risky sexual behaviors. However, only the effect on early sexual debut was gender invariant. Exposure to sexually explicit media in early adolescence had a substantive relationship with risky sexual behavior in the emerging adulthood. Knowledge of this causal like effect provides a basis for building better preventive programs in early adolescence. One prominent way is early education on media literacy, and physicians themselves may need to be familiar with such content to initiate it.


[Sentinel ganglion in breast cancer: selective biopsy compared with axillary lymphadectomy].
Selective biopsy of the sentinel ganglion (SBSG) has replaced axillary lymphadectomy (AL) as the procedure of choice in staging breast cancer in its initial stages and in clinically negative axilla. The aim of this study is to compare global event-free survival of those patients subjected to SBSG followed by AL, during the period of validation of the technique, with respect to those subjected to SBSG and AL if the sentinel ganglion (SG) showed metastasis. One hundred and forty-eight patients were included, 81 belonging to the period of validation and 67 to the clinical application group. Radiocoloid was administered intraperitumorally, obtaining images up until the visualisation of the SG; its identification and extirpation were carried out subsequently in the surgical intervention. The efficacy of the technique in the validation group was 92.5%, sensitivity was 95.6% and the rate of false negatives was 4%. Of the 81 patients, 75 are free of disease (92.6%). Of the 67 patients belonging to the clinical application group, 63 (94%) are free of disease. No patient has presented axillary ganglion recurrence. In the validation of the technique we obtained values that fall within the demands of generally accepted quality. With an average follow up of 6 years we did not observe axillary ganglion recurrence in any of the two groups. There is no statistically significant difference in global and event free survival between the two groups.


[Effects of three kinds of inorganic dusts on lipid peroxidation of erythrocytes].
This paper reports on the effects of erythrocytes when incubated with three kinds of inorganic dusts, i.e. quartz (free SiO2 content 97%), alpha-Al2O3 and TiO2. Malonaldehyde as a product of lipid peroxidation was determined as it reacted with thiobarbituric acid, and haemolytic activity was analysed spectrophotometrically. The results showed that quartz presented a stronger stimulative effect on lipid peroxidation of erythrocyte at doses of 1-8 mg than the control (P less than 0.01). No change was observed with alpha-Al2O3 and TiO2 at doses of 1-9 mg. The results suggest that the effects of dusts on lipid peroxidation of erythrocyte are similar to hemolytic reaction, directly reflecting the degree of their damage to the cell membrane. Therefore lipid peroxidation on cell membrane caused by dusts was one of the parameters of cytotoxic effects, and could be considered as one of the measures used in the assessment of cytotoxicity of dusts.


Prevention of measles spread on a paediatric ward.
Since measles is a highly contagious respiratory infection with significant airborne transmission risk in hospitals, effective prevention measures are crucial. After a mother accompanying her child on a paediatric ward lacking a negative pressure room was diagnosed with measles, exposed persons without evidence of immunity (documentary evidence of receiving two doses of measles-mumps-rubella vaccine) were treated with vaccination or intravenous immunoglobulin (IVIG). The interruption of transmission with these treatments was evaluated. There were 44 children and 101 adults exposed to the index patient. Twenty-five children and 88 adults were considered immune, providing evidence of immunity. Nineteen children and 13 adults were either given vaccination or IVIG for post-exposure prophylaxis (PEP). There were no additional cases of measles after 3 weeks follow-up. We conclude that measles is highly preventable by adequate PEP with vaccination or IVIG in a healthcare setting that lacks the benefit of a negative pressure room.


Free energy of conformational transition paths in biomolecules: the string method and its application to myosin VI.
A set of techniques developed under the umbrella of the string method is used in combination with all-atom molecular dynamics simulations to analyze the conformation change between the prepowerstroke (PPS) and rigor (R) structures of the converter domain of myosin VI. The challenges specific to the application of these techniques to such a large and complex biomolecule are addressed in detail. These challenges include (i) identifying a proper set of collective variables to apply the string method, (ii) finding a suitable initial string, (iii) obtaining converged profiles of the free energy along the transition path, (iv) validating and interpreting the free energy profiles, and (v) computing the mean first passage time of the transition. A detailed description of the PPS↔R transition in the converter domain of myosin VI is obtained, including the transition path, the free energy along the path, and the rates of interconversion. The methodology developed here is expected to be useful more generally in studies of conformational transitions in complex biomolecules.


[The action of diuretics in renal ischemia].
A protective action of lasix, dichlothiazide, and triampur (dichlothiazide + triamterene) was studied in experiments on rats. Ischemia was simulated by obstruction of kidney vessels and ureter for 90 min. Lasix and dichlothiazide produced a protective effect in renal ischemia and at the same time resulted decrease of lifetime of experimental rats. Triampur increased the lifetime and decreased the losses of potassium in kidney tissue.


The effect of structured health promotion education given to adolescents on health literacy and health-promoting behaviors.
This study was conducted to determine the effect of structured health promotion education given to adolescents on health literacy and health promotion level. The research was designed according to the pretest-posttest control group model and conducted using the quasi-experimental method. The population of the study consisted of students studying in the 6th, 7th, and 8th grades of a secondary school. A total of 191 students were assigned to the intervention group and the control group. The data were collected using the Sociodemographic Characteristics Form, the Adolescent Health Promotion Scale, and the School Age Health Literacy Scale. Structured Health Promotion Education consisting of six modules was given to the intervention group. The control group did not receive any intervention. Of the students, 33.9% were in the 8th grade, 76.6% were born in Turkey, 31.6% had a father and mother who were secondary school graduates, 72.5% were from moderate-income families, and 83% had no chronic health problems. The groups' health literacy and adolescent health-promoting behaviors pretest mean scores before the intervention were homogeneous (p > 0.05). There was a significant difference between the groups' health literacy and adolescent health-promoting behaviors posttest mean scores after the intervention (p < 0.05). While structured health promotion education was found to increase health literacy and health-promoting behaviors, nutrition, stress management, life satisfaction, and health responsibility behaviors in adolescents, it did not affect social support and exercise behaviors. Nurses should provide training to increase students' health literacy and health-promoting behaviors.


[Gene expression profiling in human thyroid tumors].
Our work aims at defining by microarray gene expression profiles in different thyroid tumors: hyperfunctioning autonomous adenomas, and sporadic and post-Chernobyl papillary cancers. Gene expression analysis in hyperfunctioning autonomous adenomas allowed us to identify genes involved in various physiological processes of the thyroid. Concerning papillary cancers, gene expression profiling in post-Chernobyl and sporadic papillary carcinomas could not identify a specific gene expression signature allowing to distinguish both tumors although such a signature exists for autonomous adenomas and carcinomas. This suggests that both cancers represent the same disease, and that there is unlikely to be a molecular signature for radiation-induced thyroid cancer.


Subclinical ketosis leads to lipid metabolism disorder by down-regulating the expression of Acetyl-CoA acetyltransferase 2 (ACAT2) in dairy cows.
Ketosis is a metabolic disease that often occurs in dairy cows postpartum and results from disordered lipid metabolism. Acetyl-coenzyme A (CoA) acetyltransferase 2 (ACAT2) is important for balancing cholesterol and triglyceride (TG) metabolism; however, its role in subclinical ketotic dairy cows is unclear. This study aimed to explore the potential correlation between ACAT2 and lipid metabolism disorders in subclinical ketotic cows through in vitro and in vivo experiments. In the in vivo experiment, liver tissue and blood samples were collected from healthy cows (CON, n = 6, β-hydroxybutyric acid [BHBA] concentration < 1.0 mM) and subclinical ketotic cows (subclinical ketosis [SCK], n = 6, BHBA concentration = 1.2-3.0 mM) to explore the effect of ACAT2 on lipid metabolism disorders in SCK cows. For the in vitro experiment, bovine hepatocytes (BHEC) were used as the model. The effects of BHBA on ACAT2 and lipid metabolism were investigated via BHBA concentration gradient experiments. Subsequently, the relation between ACAT2 and lipid metabolism disorder was explored by transfection with siRNA of ACAT2. Transcriptomics showed an upregulation of differentially expression genes (DEGs) during lipid metabolism and significantly lower ACAT2 mRNA levels in the SCK group. Compared with the CON group in vivo, the SCK group showed significantly higher expression levels of peroxisome proliferator-activated receptor γ (PPARγ) and sterol regulator element binding protein 1c (SREBP1c) and significantly lower expression levels of peroxisome proliferator-activated receptor α (PPARα), carnitine palmitoyl-transferase 1A (CPT1A), sterol regulatory element binding transcription factor 2 (SREBP2), and 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR). Moreover, the SCK group had a significantly higher liver TG content and significantly lower plasma total cholesterol (TC) and free cholesterol (FC) content. These results were indicative of TG and cholesterol metabolism disorders in the liver of dairy cows with SCK. Additionally, the SCK group showed an increased expression of Perilipin-2 (PLIN2), decreased expression of apolipoprotein B (APOB), and decreased plasma concentration of very low-density lipoproteins (VLDL) and low-density lipoproteins-cholesterol (LDL-C) by downregulating ACAT2, which indicated an accumulation of TG in liver. In vitro experiments showed that BHBA induced an increase in the TG content of BHEC, decreased content TC, increased expression of PPARγ and SREBP1c, and decreased expression of PPARα, CPT1A, SREBP2, and HMGCR. Additionally, BHBA increased the expression of PLIN2 in BHEC, decreased the expression and fluorescence intensity of ACAT2, and decreased the VLDL and LDL-C contents. Furthermore, silencing ACAT2 expression increased the TG content; decreased the TC, VLDL, and LDL-C contents; decreased the expression of HMGCR and SREBP2; and increased the expression of SREBP1c; but had no effect on the expression of PLIN2. These results suggest that ACAT2 downregulation in BHEC promotes TG accumulation and inhibits cholesterol synthesis, leading to TG and cholesterol metabolic disorders. In conclusion, ACAT2 downregulation in the SCK group inhibited cholesterol synthesis, increased TG synthesis, and reduced the contents of VLDL and LDL-C, eventually leading to disordered TG and cholesterol metabolism.


Dynamics and retention of misfolded proteins in native ER membranes.
When co-translationally inserted into endoplasmic reticulum (ER) membranes, newly synthesized proteins encounter the lumenal environment of the ER, which contains chaperone proteins that facilitate the folding reactions necessary for protein oligomerization, maturation and export from the ER. Here we show, using a temperature-sensitive variant of vesicular stomatitis virus G protein tagged with green fluorescent protein (VSVG-GFP), and fluorescence recovery after photobleaching (FRAP), the dynamics of association of folded and misfolded VSVG complexes with ER chaperones. We also investigate the potential mechanisms underlying protein retention in the ER. Misfolded VSVG-GFP complexes at 40 degrees C are highly mobile in ER membranes and do not reside in post-ER compartments, indicating that they are not retained in the ER by immobilization or retrieval mechanisms. These complexes are immobilized in ATP-depleted or tunicamycin-treated cells, in which VSVG-chaperone interactions are no longer dynamic. These results provide insight into the mechanisms of protein retention in the ER and the dynamics of protein-folding complexes in native ER membranes.


Basal metabolic rate of Brazilians living in the Southwestern United States.
Estimation of energy requirements relies on adequate values of basal metabolic rate (BMR). Prediction equations recommended for international use have been shown to overestimate BMR in populations living in the tropics. We have previously shown the inadequacy of these equations in samples of Brazilians living in tropical and temperate regions of the country. We sought to investigate whether BMR could adequately be estimated by prediction equations in a sample of Brazilians living in a different setting: the Sonoran desert of the Southwestern USA. BMR was measured under standard conditions in 33 subjects (14 men). Mean bias (estimated-measured) varied from 404.4 to 708.6 kJ day(-1) in women and 566.8 to 1122.8 kJ day(-1) in men, representing 8.5-15 and 8.9-17.6% overestimation, respectively, using the Schofield equations. Bland and Altman analyses showed large, relevant limits of agreement. The results using the recommended equations for the American population (IOM, 2005) were only 2% different from the Schofield equations. The Harris and Benedict equations yielded higher overestimations (15.0 and 16.8% for women and men, respectively) and the Henry and Rees equations also overestimated BMR (8.5 and 8.9%) even though they were developed for populations from the tropics, although to a lesser degree. It is concluded that the equations currently recommended for international use are not appropriate for Brazilians living in the Southwestern USA.


Canine pancreatic islet cell tumours secreting insulin-like growth factor type 2: a rare entity.
Insulin-like growth factor type II (IGF-II) is the main cause of non-islet cell tumour hypoglycaemia (NICTH) and insulin is thought to be the only factor causing hypoglycaemia in insulinomas. However, two case reports of pancreatic neuroendocrine tumours (PNETs) producing IGF-II have been previously published: a human and a canine patient. In this study, we investigated clinical, histopathological, immunohistochemical and ultrastructural features, and biological behaviour of canine pancreatic IGF-II-omas, a subgroup of PNETs that has not been previously characterized. Case records of 58 dogs with confirmed PNETs and hypoglycaemia were reviewed: six patients were affected by IGF-II-omas. Surgery was performed in all cases and two dogs had metastases. Four patients remained alive and in remission at 370, 440, 560 and 890 days post-diagnosis; two died of non-tumour-related causes. IGF-II-omas can be differentiated from insulinomas through hypoinsulinaemia, IGF-II positive and insulin negative immunostaining. The prevalence of this neoplasia is low, accounting for just 6% of PNETs.


Simple form of multimode laser diode rate equations incorporating the band filling effect.
To derive a simple form of the multimode laser diode rate equations incorporating the band filling effect, the laser diode gain in the direct bandgap model is introduced into the conventional multimode laser diode rate equations. By numerically examining each modal gain under the gain-switching condition, it is found that both the differential gain coefficient and the carrier density at transparency show an approximately linear dependency on the oscillation frequency. As a result, it is possible to derive a simple form of the multimode laser diode rate equations with linearized gain, which can be used to simulate the behaviors of a gain-switched laser diode characterized by the band filling effect, in both the multimode and single-mode oscillation cases.


The effectiveness of suggestive techniques in reducing postoperative side effects: a meta-analysis of randomized controlled trials.
Suggestive interventions such as hypnosis and therapeutic suggestions are frequently used to alleviate surgical side effects; however, the effectiveness of therapeutic suggestion intervention has not yet been systematically evaluated. In the present study, we tested the hypotheses that (1) suggestive interventions are useful for reducing postoperative side effects; (2) therapeutic suggestions are comparable in effectiveness to hypnosis; (3) live presentation is more effective than recordings; and (4) suggestive interventions would be equally effective used in minor and major surgeries. We performed random effect meta-analysis with meta-regression and sensitivity analysis by moderating factors on a pool of 26 studies meeting the inclusion criteria (N = 1890). Outcome variables were postoperative anxiety, pain intensity, pain medication requirement, and nausea. Suggestive interventions reduced postoperative anxiety (g = 0.40; 99% confidence interval [CI] = 0.13-0.66; P < 0.001) and pain intensity (g = 0.25; 99% CI = 0.00-0.50; P = 0.010), but did not significantly affect postoperative analgesic drug consumption (g = 0.16; 99% CI = -0.16 to 0.47; P = 0.202) and nausea (g = 0.38; 99% CI = -0.06 to 0.81; P = 0.026). No significant differences were found for intervention type, presentation method, and severity of surgery; however, sensitivity analysis only supported the effectiveness of hypnosis (g = 0.62; 99% CI = 0.31-0.92; P < 0.001) and live presentation (g = 0.55; 99% CI = 0.23-0.88; P < 0.001) for decreasing postoperative anxiety, and that of live presentation for alleviating postoperative pain (g = 0.44; 99% CI = 0.07-0.82; P = 0.002). Sensitivity analyses also suggested that suggestive interventions are only effective for decreasing pain intensity during minor surgical procedures (g = 0.39; 99% CI = 0.00-0.78; P = 0.009). Suggestive techniques might be useful tools to alleviate postoperative anxiety and pain; however, strength of the evidence is weak because of possible bias in the reviewed articles. The lack of access to within-subjects data and the overlap between moderator conditions also limit the scope of the analysis. More methodologically correct studies are required with sensitivity to moderating factors and to within-subjects changes. For clinical purposes, we advise the use of hypnosis with live presentation to reduce postoperative anxiety and pain, until convincing evidence is uncovered for the effectiveness of therapeutic suggestions and recorded presentation. Pain management with adjunct suggestive interventions is mostly encouraged in minor rather than major surgeries.


Carcinoma Showing Thymus-Like Elements: A Rare Malignant Tumor of the Thyroid Gland.
Carcinoma showing thymus-like elements (CASTLE) is a rare tumor arising in the thyroid gland. Fewer than 100 patients with this tumor, those predominantly from Eastern Asia, have been reported before. We present the first CASTLE case from Turkey. A 51-year-old male was admitted with a complaint of a neck mass and hoarseness. A laryngoscopic examination revealed left vocal cord paralysis. Neck ultrasonography showed a tumor which was compressing the esophagus and had invaded the left recurrent laryngeal nerve (RLN). The patient underwent a total thyroidectomy, a unilateral central-compartment neck dissection, and following adjuvant radiotherapy. Intraoperative nerve monitoring was performed during the operation to preserve the contralateral nerve. He completed a 3-year follow-up period after the completion radiotherapy, and no recurrence was observed. The treatment is controversial, although surgery with or without adjuvant radiotherapy appears to be the best choice. In cases of RLN destruction due to tumor invasion, we recommend using intraoperative nerve monitoring to preserve the contralateral RLN to avoid devastating complications, such as a tracheostomy.


Cytoarchitecture of sensorimotor areas in the cat cerebral cortex.
The organization of multiple motor areas in the cerebral cortex has been investigated frequently in primates but rarely in nonprimate species. To compare sensorimotor areas in cats and primates, the cytoarchitecture of frontal and parietal areas of the cat cerebral cortex was described and mapped from coronal sections stained with cresyl violet or thionine. Multiple subdivisions of areas 4 and 6 were recognized; of these, the cytoarchitecture of area 4 gamma is similar to that of area 4 described in other carnivores and in primates and is characterized by giant pyramidal cells in multiple rows or clusters in lamina V. In other subdivisions of area 4 (4 delta, 4sfu, and 4fu), giant pyramidal cells are few or absent in lamina V, and these subdivisions resemble area 6 of primates. Area 6 of the cat cortex is heterogeneous, and differences in laminar appearance and size of pyramidal cells in lamina V distinguish its four subdivisions (6a alpha, 6a beta, 6a gamma, and 6iffu). The adjoining prefrontal areas are distinguishable from area 6 by the presence of a thin internal granular lamina (lamina IV) and the reduced size of pyramidal cells in lamina V. Laminae are poorly differentiated in the cingulate areas, where a rostral and caudal subdivision can be distinguished on the basis of the absence or presence of lamina IV. Area 3a is characterized by a thin lamina IV and is located between frontal agranular and parietal granular (well-defined lamina IV) fields (3b, 1, 2, 2pri, 5, and 7). Insular cortex can be subdivided into granular and agranular fields.


Myocardial infarction does not affect circulating haematopoietic stem and progenitor cell self-renewal ability in a rat model.
What is the central question of this study? Although peripheral blood haematopoietic stem and progenitor cells are potentially important in regeneration after acute myocardial infarction, their self-renewal ability in the post-acute phase has not yet been addressed. What is the main finding and its importance? In rat peripheral blood, we show that myocardial infarction does not negatively affect circulating haematopoietic stem and progenitor cell self-renewal ability 2 weeks after acute infarction, which suggests a constant regenerative potential in the myocardial infarction post-acute phase. Given the importance of peripheral blood haematopoietic stem and progenitor cells (HPCs) in post-acute regeneration after acute myocardial infarction (MI), the aim of the present study was to investigate the number and secondary replating capacity/self-renewal ability of HPCs in peripheral blood before and 2 weeks after MI. In female Lewis inbred rats (n = 9), MI was induced by ligation of the left coronary artery, and another nine underwent sham surgery, without ligation, for control purposes. Myocardial infarction was confirmed by troponin I concentrations 24 h after surgery. Peripheral blood was withdrawn and fractional shortening and ejection fraction of the left ventricle were assessed before (day 0) and 14 days after MI or sham surgery (day 14). After mononuclear cell isolation, primary and secondary functional colony-forming unit granulocyte-macrophage (CFU-GM) assays were performed in order to detect the kinetics of functional HPC colony counts and cell self-renewal ability in vitro. The CFU-GM counts and cell self-renewal ability remained unchanged (P > 0.05) in both groups at day 14, without interaction between groups. In the intervention group, higher day 0 CFU-GM counts showed a relationship to lower fractional shortening on day 14 (ρ = -0.82; P < 0.01). Myocardial infarction did not negatively affect circulating HPC self-renewal ability, which suggests a constant regenerative potential in the post-acute phase. A relationship of cardiac contractile function 14 days after MI with circulating CFU-GM counts on day 0 might imply functional colony count as a predictive factor for outcome after infarction.


Treatment of keratoconjunctivitis sicca in dogs with cyclosporine ophthalmic ointment: a European clinical field trial.
The results are reported of a six-week clinical trial of the efficacy of 0.2 per cent cyclosporine ophthalmic ointment for the treatment of chronic idiopathic keratoconjunctivitis sicca in dogs in the United Kingdom, Germany and France. The 87 dogs were referral cases with a history of chronic unresponsive keratoconjunctivitis sicca of which the aetiology was unknown, and they had to meet stringent criteria before being included in the trial. The clinical response to the therapy was monitored after seven, 21 and 42 days and the results for the right and left eyes were analysed separately. There was a statistically significant increase in lacrimal secretion throughout the study, with most of the increase occurring during the first week of treatment. The percentage of eyes with improved lacrimal secretion was higher in the dogs with initial Schirmer tear test values > or = 2 mm/min than in those with initial values of 0 or 1 mm/min. The observed steady improvement in conjunctival health was not always correlated with an improvement in lacrimal secretion. The incidence of blepharospasm, other signs of discomfort and corneal oedema decreased significantly during the study. No improvement in corneal vascularisation or pigmentation was observed during the six-week trial. Overall, 76 per cent of the left eyes and 87 per cent of the right eyes were considered to have improved at the end of the treatment period. No serious adverse reactions were observed and only mild irritation was noticed by the owners immediately after the application of the ointment. This irritation resulted in the recording of an adverse reaction at the scheduled observations in only three cases.


Frequency, clinical characteristics and outcome of diabetic ketoacidosis in children with type-1 diabetes at a tertiary care hospital.
To observe the frequency, demographic data and outcome of diabetic ketoacidosis (DKA) in children with established type 1 diabetes and newly diagnosed diabetes at a tertiary care hospital. The case record review was done of children admitted with the diagnosis of DKA at The National Institute of Child Health, Karachi from 1st June 2008 till 31st May 2009. All records with the diagnosis of DKA were reviewed and those children with only hyperglycaemia, or who did not fulfill the criteria of DKA were excluded. The demographic data and laboratory investigations which included blood sugar monitoring, arterial blood gases, urine analysis especially for ketones, serum electrolytes, complete blood count and blood culture 'were reviewed. The previous numbers of admissions in children with established DKA were also noted with reasons. The duration of symptoms and fluids required, time of recovery, complications, and outcome were noted and compared between those with established diabetes and children with newly diagnosed diabetes. Data was entered and analyzed on SPSS version 15. Out of 124 case records, 117 were included which fulfilled the criteria of DKA. A large number, 65 (55.5%) children were in the > 10 years age group with a female predominance. Out of 117 children 50 (42.7%) had established Type 1 diabetes and 67 (57.2 %) children had newly diagnosed diabetes. The commonest presenting complaints in both groups were respiratory distress (87.1%) and vomiting (77.7%). The symptoms of polyuria, polydipsia and nocturia were more among the newly diagnosed children as compared to those with established diabetes with a significant p value <0.001. The comparison of clinical features and laboratory investigations of the two groups showed no difference except that those children with established diabetes improved earlier, required lesser duration of intravenous fluids and their insulin was changed to subcutaneous in less time compared with newly diagnosed children. (p < 0.001). The commonest complication in both groups was hypoglycaemia followed by hyponatraemia, more in newly diagnosed diabetic children. These soaring numbers are just from one center, highlighting the issue of this much neglected disease in our country. More studies on a larger scale are needed to assess the prevalence/incidence in our children and also more emphasis with educational programmes on prevention of recurrent attacks of DKA.


Beta-catenin accelerates human papilloma virus type-16 mediated cervical carcinogenesis in transgenic mice.
Human papilloma virus (HPV) is the principal etiological agent of cervical cancer in women, and its DNA is present in virtually all of these tumors. However, exposure to the high-risk HPV types alone is insufficient for tumor development. Identifying specific collaborating factors that will lead to cervical cancer remains an unanswered question, especially because millions of women are exposed to HPV. Our earlier work using an in vitro model indicated that activation of the canonical Wnt pathway in HPV-positive epithelial cells was sufficient to induce anchorage independent growth. We therefore hypothesized that constitutive activation of this pathway might function as the "second hit." To address this possibility, we developed two double-transgenic (DT) mouse models, K14-E7/ΔN87βcat and K14-HPV16/ΔN87βcat that express either the proteins encoded by the E7 oncogene or the HPV16 early region along with constitutively active β-catenin, which was expressed by linking it to the keratin-14 (K14) promoter. We initiated tumor formation by treating all groups with estrogen for six months. Invasive cervical cancer was observed in 11% of the K14-ΔN87βcat mice, expressing activated β-catenin and in 50% of the animals expressing the HPV16 E7 oncogene. In double-transgenic mice, coexpression of β-catenin and HPV16 E7 induced invasive cervical cancer at about 7 months in 94% of the cases. We did not observe cervical cancer in any group unless the mice were treated with estrogen. In the second model, K14-HPV16 mice suffered cervical dysplasias, but this phenotype was not augmented in HPV16/ΔN87βcat mice. In summary, the phenotypes of the K14-E7/ΔN87βcat mice support the hypothesis that activation of the Wnt/β-catenin pathway in HPV-associated premalignant lesions plays a functional role in accelerating cervical carcinogenesis.


Anomalous anatomy of identified neurons in the larval prawn: spontaneous and induced by microlesions.
The abdominal ganglia of the prawn Macrobrachium rosenbergii undergo developmental changes of fundamental interest between the time of hatching and metamorphosis. These changes include an increase in cell numbers and changes in the connectivity between identified neurons. The giant motoneurons involved in the escape response, which form a syncytium in the adult, are observed as separate neurons with crossed axons in early larvae. Anomalous growth and connections of identified neurons were studied in order to gain some understanding of the rules and mechanisms governing normal development. Spontaneous anomalies included: supernumerary axons and abnormal axonal trajectories. The plasticity and specificity of identified neurons were studied by following the anatomical effects of deletions of giant neurons. Microlesions were inflicted reproducibly by means of a focused beam of visible and ultraviolet light. Within a day, irradiated cell bodies are eliminated; complete disappearance of the axon takes about 10 days, indicating that the remarkable ability of some invertebrate neurons to survive without a soma is not present in the larval prawn. As a result of the removal of an axon, the most common effect found in central connections was the absence of the collaterals or axons deprived of their targets. No collateral sprouting was detected in the central nervous system. In about a third of the ganglia where a giant motoneuron was killed and structure was analyzed 2 or more weeks after irradiation, anomalous connections were found. They usually involved contacts between an interneuron deprived of its normal target and the contralateral motoneuron which remained intact. The restricted types of anomalies observed support the notion of a hierarchical order in the rules governing formation of central synapses, in which neuron type ranks higher than laterality.


A Novel Topical Wound Therapy Delivery System.
Wound care dressings have evolved over time, from bandaging to the development of occlusive dressings to negative pressure wound therapy. A novel therapeutic delivery system dressing has been cleared by the United States Food and Drug Administration. This semi occlusive wound dressing has been developed to provide local, continuous delivery of aqueous topical agents, such as therapeutics (anesthetics, antiseptics, antibiotics, steroids, topical beta-blockers, immune modulatory agents, growth factors, and fibrinolytic agents, among others), at a rate of about ¾ mL per day, thus maintaining a hydrated environment and providing topical treatment. This type of system may be beneficial in situations where systemic therapies cannot be used, wounds are small and few, wounds may need frequent application of medication or moisture, or low and steady delivery of medications is needed. The authors assessed a delivery system dressing with different types of liquid medications for the management of hard-to-heal, chronic lower extremity wounds. Patients aged ≥ 18 and ≤ 90 years with stalled chronic wounds > 30 days' duration were selected for the use of a topical delivery system, which consists of a semi occlusive wound dressing and fluid delivery unit that can provide local application of small therapeutic quantities of medication directly to the wound. Several successful cases with the use of this device are presented in which pain relief, enhancement of epithelial migration, inflammation reduction, bacterial control, and wound size reduction were achieved. This delivery system dressing is an effective and safe treatment option for wounds. Advantages include reduced potential of systemic side effects, flexibility in what can be delivered, constant rate of medication delivery, and convenience.


Efficient biomass saccharification using a novel cellobiohydrolase from Clostridium clariflavum for utilization in biofuel industry.
The present study describes the cloning of the cellobiohydrolase gene from a thermophilic bacterium Clostridium clariflavum and its expression in Escherichia coli BL21(DE3) utilizing the expression vector pET-21a(+). The optimization of various parameters (pH, temperature, isopropyl β-d-1-thiogalactopyranoside (IPTG) concentration, time of induction) was carried out to obtain the maximum enzyme activity (2.78 ± 0.145 U ml-1) of recombinant enzyme. The maximum expression of recombinant cellobiohydrolase was obtained at pH 6.0 and 70 °C respectively. Enzyme purification was performed by heat treatment and immobilized metal anionic chromatography. The specific activity of the purified enzyme was 57.4 U mg-1 with 35.17% recovery and 3.90 purification fold. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) showed that the molecular weight of cellobiohydrolase was 78 kDa. Among metal ions, Ca2+ showed a positive impact on the cellobiohydrolase enzyme with increased activity by 115%. Recombinant purified cellobiohydrolase enzyme remained stable and exhibited 77% and 63% residual activity in comparison to control in the presence of n-butanol and after incubation at 80 °C for 1 h, respectively. Our results indicate that our purified recombinant cellobiohydrolase can be used in the biofuel industry.


Antigen recognition and immunomodulation by gamma delta T cells in bovine tuberculosis.
This report describes the in vitro proliferative responses of peripheral blood gammadelta T cells to defined mycobacterial protein Ags and the immunomodulatory effect of gammadelta T cells in cattle infected with Mycobacterium bovis. gammadelta T cell responses were specific to M. bovis infection because they were detected in cattle either experimentally or naturally infected with M. bovis, but were not present in uninfected controls. Proliferating gammadelta T cell cultures produced enhanced levels of IFN-gamma and TGF-beta, but not IL-2 in response to the more immunodominant mycobacterial AGS: Depletion of gammadelta T cells from PBMC resulted in an increased Ag-specific proliferation in half the animals tested, indicating a suppressive effect of gammadelta T cells upon other (alphabeta) T cell responses. Because gammadelta T cells constitute a major T cell population in the peripheral blood of cattle, the activities of gammadelta T cells described in this report could make a significant contribution to the immune response in bovine tuberculosis.


Protein disulphide-isomerase reduces ricin to its A and B chains in the endoplasmic reticulum.
Cells expressing ricin B chain within the secretory pathway are significantly more resistant to intoxication by ricin holotoxin but not to other cytotoxins that exploit similar endocytic routes to the cytosol. Furthermore, cells expressing the related B chain of abrin are protected against both incoming abrin and ricin. These phenotypes can be correlated with the abilities of the respective B chains to form disulphide-linked A-B holotoxins, since abrin B chain forms heterodimers with either abrin or ricin A chains, whereas ricin B chain forms heterodimers with ricin A chain only. In the ricin B-expressing cells, this newly made lectin disappears with biphasic kinetics comprising a retention phase followed by slow turnover and disposal after disengagement from calnexin cycle components. Interference with ricin cytotoxicity occurs during the early retention phase when ricin B chain is associated with PDI (protein disulphide-isomerase). The data show that retrotranslocation of incoming toxin is impeded by PDI-catalysed formation of heterodimers between endogenous B and A chains derived from reduced holotoxin, thus proving that reduction of ricin occurs in the endoplasmic reticulum. In contrast with other toxins, ricin does not appear to require either proteolytic cleavage or unfolding for PDI-catalysed reduction.


Normalisation of surfactant protein -A and -B expression in the lungs of low birth weight lambs by 21 days old.
Intrauterine growth restriction (IUGR) induced by placental restriction (PR) in the sheep negatively impacts lung and pulmonary surfactant development during fetal life. Using a sheep model of low birth weight (LBW), we found that there was an increase in mRNA expression of surfactant protein (SP)-A, -B and -C in the lung of LBW lambs but no difference in the protein expression of SP-A or -B. LBW also resulted in increased lysosome-associated membrane glycoprotein (LAMP)-3 mRNA expression, which may indicate an increase in either the density of type II Alveolar epithelial cells (AEC) or maturity of type II AECs. Although there was an increase in glucocorticoid receptor (GR) and 11β-hydroxysteroid dehydrogenase (11βHSD)-1 mRNA expression in the lung of LBW lambs, we found no change in the protein expression of these factors, suggesting that the increase in SP mRNA expression is not mediated by increased GC signalling in the lung. The increase in SP mRNA expression may, in part, be mediated by persistent alterations in hypoxia signalling as there was an increase in lung HIF-2α mRNA expression in the LBW lamb. The changes in the hypoxia signalling pathway that persist within the lung after birth may be involved in maintaining SP production in the LBW lamb.


On-line monitoring of gas-phase bioreactors for biogas treatment: hydrogen sulfide and sulfide analysis by automated flow systems.
Biogas is produced by biological processes under anaerobic conditions and may contain up to 20,000 ppm(v) hydrogen sulfide (H(2)S), a corrosive substance that attacks power engines and can affect the health of the industrial staff. H(2)S must be removed from the biogas, especially in co-generation facilities where the biogas is burnt for energy production. Nowadays, biofiltration is being studied and considered as an interesting alternative for removing H(2)S from the biogas besides classical chemical processes. The novelty of this work is the design and construction of an automated H(2)S on-line analyser to assess the composition of the liquid and gas phases of gas-phase bioreactors. The analyser is made of two parallel flow configurations which share the same detection device. The first configuration is a single-channel flow injection analyser (FIA) to detect S(2-) in the liquid phase. The second configuration is a continuous flow analyser (CFA) with a gaseous diffusion step (GD-CFA) for detecting H(2)S in the gas phase. The diffusion step enables separation of the H(2)S((g)) from the sample and its conversion into a detectable chemical species (S(2-)). S(2-) detection was performed with an Ag(2)S ion-selective electrode (ISE) selective to S(2-)(aq). The main response parameters of the FIA system are a linear range between 3 x 10(-5) and 1 x 10(-1) mol L(-1) S(2-) (0.61-3,200 mg L(-1)), with a sensitivity of 27.9 mV decade(-1) and a detection limit of 1.93 x 10(-5) mol L(-1) S(2-). The GD-CFA configuration presents a linear range between 400 and 10,000 ppm(v) H(2)S((g)) with a sensitivity of 26.1 mV decade(-1) and a detection limit of 245 ppm(v) H(2)S. The proposed analyser was used by analysing real gas and liquid samples with optimal results at a full-scale biotrickling filter for biogas treatment at a municipal wastewater treatment plant.