Distinguishing Mast Cell Progenitors from Mature Mast Cells in Mice.
Mast cells originate from the bone marrow and develop into c-kit(+) FcɛRI(+) cells. Both mast cell progenitors (MCp) and mature mast cells express these cell surface markers, and ways validated to distinguish between the two maturation forms with flow cytometry have been lacking. Here, we show that primary peritoneal MCp from naïve mice expressed high levels of integrin β7 and had a low side scatter (SSC) light profile; whereas mature mast cells expressed lower levels of integrin β7 and had a high SSC light profile. The maturation statuses of the cells were confirmed using three main strategies: (1) MCp, but not mature mast cells, were shown to be depleted by sublethal whole-body γ-irradiation. (2) The MCp were small and immature in terms of granule formation, whereas the mature mast cells were larger and had fully developed metachromatic granules. (3) The MCp had fewer transcripts of mast cell-specific proteases and the enzyme responsible for sulfation of heparin than mature mast cells. Moreover, isolated peritoneal MCp gave rise to mast cells when cultured in vitro. To summarize, we have defined MCp and mature mast cells in naïve mice by flow cytometry. Using this strategy, mast cell maturation can be studied in vivo.


Method for computing the scene-illuminant chromaticity from specular highlights.
The perception of an unchanging surface color under different illuminations requires the computation of the scene-illuminant color either directly or indirectly. A possible source for the computation is the specular highlight of the surface reflection. Some issues related to color constancy are discussed, and a theory for computing the scene-illuminant chromaticity from specular highlight is described. An interesting result of the theory is that in an ideal situation, two surfaces of different colors will be sufficient for the computation.


Vasodepressor effects of arachidonic acid and prostacyclin (PGI2) in hypertensive rats.
1. Vasodepressor responses to prostacyclin and nitroprusside were compared in anaesthetized, spontaneously hypertensive rats of the Okamoto strain and Wistar--Kyoto controls, and also in one-kidney, one-clip hypertensive rats and unilaterally nephrectomized controls of the Sprague--Dawley strain. The responses, measured as a percentage of resting blood pressure, did not differ significantly between the hypertensive rats and the normotensive controls within each strain. 2. The effects of intravenous injections of arachidonic acid were also studied in each strain. 3. The vasodepressor effects of high doses of arachidonic acid (1 or 3 mg/kg) were much greater and more prolonged in both groups of hypertensive rats. These differences were abolished by indomethacin (2 mg/kg). 4. Comparisons between the strains showed that whereas Okamoto rats have significantly greater depressor responsiveness to nitroprusside and prostacyclin than Sprague--Dawley rats, the depressor effects of high doses of arachidonic acid (1 and 3 mg/kg) were smaller in the normotensive Wistar--Kyoto than in the Sprague--Dawley rats. 5. It is concluded that hypertensive rats have enhanced ability to transform exogenous arachidonic acid into vasodilator prostanoids. This occurs both in spontaneous hypertension and in experimental renal hypertension. However, rats of the Okamoto strain appear to have reduced ability to form prostacyclin when compared with Sprague--Dawley rats.


An efficient high-throughput screening method for MYST family acetyltransferases, a new class of epigenetic drug targets.
Epigenetic aberrations are increasingly regarded as key factors in cancer progression. Recently, deregulation of histone acetyltransferases (HATs) has been linked to several types of cancer. Monocytic leukemia zinc finger protein (MOZ) is a member of the MYST family of HATs, which regulate gene expression in cell proliferation and differentiation. Deregulation of these processes through constitutively active MOZ fusion proteins gives rise to the formation of leukemic stem cells, rendering MOZ an excellent target for treating myeloid leukemia. The authors implemented a hit discovery campaign to identify small-molecule inhibitors of MOZ-HAT activity. They developed a robust, homogeneous assay measuring the acetylation of synthetic histone peptides. In a primary screening campaign testing 243 000 lead-like compounds, they identified inhibitors from several chemical classes. Secondary assays were used to eliminate assay-interfering compounds and prioritize confirmed hits. This study establishes a new high-throughput assay for HAT activity and could provide the foundation for the development of a new class of drugs for the treatment of leukemias.


Computational studies of molecular hydrogen binding affinities: the role of dispersion forces, electrostatics, and orbital interactions.
Intermolecular interactions between H2 and ligands, metals, and metal-ligand complexes determine the binding affinities of potential hydrogen storage materials (HSM), and thus their extent of potential for practical use. A brief survey of current activity on HSM is given. The key issue of binding strengths is examined from a basic perspective by surveying the distinct classes of interactions (dispersion, electrostatics, orbital interactions) in first a general way, and then in the context of calculated binding affinities for a range of model systems.


Economics and industry do not mean ethical conduct in clinical trials.
Clinical trials present an ethical dilemma for pharmaceutical companies. While companies may want to undertake and report these trials in an ethical manner, negative results can significantly affect product sales. There is accumulating evidence that company-financed trials are biased in favor of the product that the company makes. Ethical conduct in this article is defined as whether the trials are conducted in the best interests of the participants and/or reported in the best interests of patients. Nine examples of how clinical trials are violating multiple articles in the Declaration of Helsinki are discussed using concrete case reports from the literature. The recognition of ethical problems in company run trials is not something new, but to date no meaningful action has been taken to resolve this issue. What is necessary is to separate the financing of clinical trials from their conduct.


Choline and betaine intake and colorectal cancer risk in Chinese population: a case-control study.
Few studies have examined the association of choline and betaine intake with colorectal cancer risk, although they might play an important role in colorectal cancer development because of their role as methyl donors. The aim of this study was to examine the relationship between consumption of choline and betaine and colorectal cancer risk in a Chinese population. A case-control study was conducted between July 2010 and December 2013 in Guangzhou, China. Eight hundred and ninety consecutively recruited colorectal cancer cases were frequency matched to 890 controls by age (5-year interval) and sex. Dietary information was assessed with a validated food frequency questionnaire by face-to-face interviews. The logistic regression model was used to estimate multivariate odds ratios (ORs) and 95% confidence intervals (CIs). Total choline intake was inversely associated with colorectal cancer risk after adjustment for various lifestyle and dietary factors. The multivariate-adjusted OR was 0.54 (95%CI = 0.37-0.80, Ptrend <0.01) comparing the highest with the lowest quartile. No significant associations were observed for betaine or total choline+betaine intakes. For choline-containing compounds, lower colorectal cancer risk was associated with higher intakes of choline from phosphatidylcholine, glycerophosphocholine and sphingomyelin but not for free choline and phosphocholine. The inverse association of total choline intake with colorectal cancer risk was observed in both men and women, colon and rectal cancer. These inverse associations were not modified by folate intake. These results indicate that high intake of total choline is associated with a lower risk of colorectal cancer.


Purification and characterisation of tubulin from the parasitic nematode, Ascaridia galli.
We have developed a method for the purification of tubulin from a parasitic nematode using DEAE-Sephadex column chromatography and temperature-dependent assembly. The resulting microtubules were morphologically similar to those obtained from mammalian brain. The nematode tubulin showed similar properties to mammalian tubulin on one and two dimensional polyacrylamide gels, although certain electrophoretic conditions revealed a slight difference in the alpha-tubulins from mammals and nematodes. This was confirmed by limited proteolytic peptide mapping. The beta subunit of nematode tubulin appeared almost identical to that of mammals. Peptide maps of these tubulins were also compared with those of eukaryotic micro-organisms and these results interpreted in terms of the evolution of the tubulin polypeptides and the sensitivity of helminths to antimicrotubular agents.


Phenotypic analysis of hMSH2 mutations in mouse cells carrying human chromosomes.
Conversion of diploidy to haploidy is a method that allows the generation of stable murine/human hybrid cell lines carrying selected human chromosomes in only a single copy. In this setting, it is possible to detect genetic mutations with greater sensitivity and reliability than in diploid cells. Using this method, we were able to identify mutations in the human mismatch repair (MMR) gene hMSH2 in hereditary nonpolyposis colon cancer families, which have escaped detection by the conventional methods. In this report, we show that such hybrid cell lines can also be a valuable tool in the study of the mutated MMR proteins, in particular the variants found in hereditary nonpolyposis colon cancer families that carry missense mutations and where it is unclear whether they predispose to colon cancer. This analysis is made possible by the fact that the human hMSH2 protein is able to complement the MMR defect in the host murine cell line.


Clinical Guidelines and Standardization of Practice to Improve Outcomes: ACOG Committee Opinion Summary, Number 792.
Protocols and checklists have been shown to reduce patient harm through improved standardization and communication. Implementation of protocols and guidelines often is delayed because of lack of health care provider awareness or difficult clinical algorithms in medical institutions. However, the use of checklists and protocols clearly has been demonstrated to improve outcomes and their use is strongly encouraged. Checklists and protocols should be incorporated into systems as a way to help practitioners provide the best evidence-based care to their patients.


Targeted gene mutation in Phytophthora spp.
The genus Phytophthora belongs to the oomycetes and is composed of plant pathogens. Currently, there are no strategies to mutate specific genes for members of this genus. Whole genome sequences are available or being prepared for Phytophthora sojae, P. ramorum, P. infestans, and P. capsici and the development of molecular biological techniques for functional genomics is encouraged. This article describes the adaptation of the reverse-genetic strategy of targeting induced local lesions in genomes (TILLING) to isolate gene-specific mutants in Phytophthora spp. A genomic library of 2,400 ethylnitrosourea (ENU) mutants of P. sojae was created and screened for induced point mutations in the genes encoding a necrosisinducing protein (PsojNIP) and a Phytophthora-specific phospholipase D (PsPXTM-PLD). Mutations were detected in single individuals and included silent, missense, and nonsense changes. Homozygous mutant isolates carrying a potentially deleterious missense mutation in PsojNIP and a premature stop codon in PsPXTM-PLD were identified. No phenotypic effect has yet been found for the homozygous mutant of PsojNIP. For those of PsPXTM-PLD, a reduction in growth rate and an appressed mycelial growth was observed. This demonstrates the feasibility of target-selected gene disruption for Phytophthora spp. and adds an important tool for functional genomic investigation.


Bereaved Siblings' Advice to Health Care Professionals Working With Children With Cancer and Their Families.
Siblings of children with cancer experience psychosocial distress during the illness and after bereavement, but often stand outside the spotlight of attention and care. This study explored bereaved siblings' advice to health care professionals (HCPs) working with children with cancer and their families. In a nationwide Swedish survey of bereaved siblings, 174/240 (73%) participated. Of these, 108 answered an open-ended question about what advice they would give to HCPs working with children with cancer and their families. In this study, responses to this single question were analyzed using content analysis. The most common advice, suggested by 56% of siblings, related to their own support. One third suggested giving better medical information to siblings. Some siblings wanted to be more practically involved in their brother's/sister's care and suggested that HCPs should give parents guidance on how to involve siblings. Other common advice related to psychosocial aspects, such as the siblings' wish for HCPs to mediate hope, yet also realism, and the importance of asking the ill child about what care they wanted. Information, communication, and involvement should be emphasized by HCPs to support siblings' psychosocial needs in both the health care setting and within the family.


Sarcolemmal nNOS anchoring reveals a qualitative difference between dystrophin and utrophin.
Duchenne muscular dystrophy (DMD) is a lethal muscle disease caused by dystrophin deficiency. In normal muscle, dystrophin helps maintain sarcolemmal stability. Dystrophin also recruits neuronal nitric oxide synthase (nNOS) to the sarcolemma. Failure to anchor nNOS to the membrane leads to functional ischemia and aggravates muscle disease in DMD. Over the past two decades, a great variety of therapeutic modalities have been explored to treat DMD. A particularly attractive approach is to increase utrophin expression. Utrophin shares considerable sequence, structural and functional similarity with dystrophin. Here, we test the hypothesis that utrophin also brings nNOS to the sarcolemma. Full-length utrophin cDNA was expressed in dystrophin-deficient mdx mice by gutted adenovirus or via transgenic overexpression. Subcellular nNOS localization was determined by immunofluorescence staining, in situ nNOS activity staining and microsomal preparation western blot. Despite supra-physiological utrophin expression, we did not detect nNOS at the sarcolemma. Furthermore, transgenic utrophin overexpression failed to protect mdx muscle from exercise-associated injury. Our results suggest that full-length utrophin cannot anchor nNOS to the sarcolemma. This finding might have important implications for the development of utrophin-based DMD therapies.


Maintenance of Voluntary Self-regulation Learned through Real-Time fMRI Neurofeedback.
Neurofeedback based on real-time functional magnetic resonance imaging (fMRI) is an emerging technique that allows for learning voluntary control over brain activity. Such brain training has been shown to cause specific behavioral or cognitive enhancements, and even therapeutic effects in neurological and psychiatric patient populations. However, for clinical applications it is important to know if learned self-regulation can be maintained over longer periods of time and whether it transfers to situations without neurofeedback. Here, we present preliminary results from five healthy participants who successfully learned to control their visual cortex activity and who we re-scanned 6 and 14 months after the initial neurofeedback training to perform learned self-regulation. We found that participants achieved levels of self-regulation that were similar to those achieved at the end of the successful initial training, and this without further neurofeedback information. Our results demonstrate that learned self-regulation can be maintained over longer periods of time and causes lasting transfer effects. They thus support the notion that neurofeedback is a promising therapeutic approach whose effects can last far beyond the actual training period.


A new methodology for assessment of the performance of heartbeat classification systems.
The literature presents many different algorithms for classifying heartbeats from ECG signals. The performance of the classifier is normally presented in terms of sensitivity, specificity or other metrics describing the proportion of correct versus incorrect beat classifications. From the clinician's point of view, such metrics are however insufficient to rate the performance of a classifier. We propose a new methodology for the presentation of classifier performance, based on Bayesian classification theory. Our proposition lets the investigators report their findings in terms of beat-by-beat comparisons, and defers the role of assessing the utility of the classifier to the statistician. Evaluation of the classifier's utility must be undertaken in conjunction with the set of relative costs applicable to the clinicians' application. Such evaluation produces a metric more tuned to the specific application, whilst preserving the information in the results. By way of demonstration, we propose a set of costs, based on clinical data from the literature, and examine the results of two published classifiers using our method. We make recommendations for reporting classifier performance, such that this method can be used for subsequent evaluation. The proportion of misclassified beats contains insufficient information to fully evaluate a classifier. Performance reports should include a table of beat-by-beat comparisons, showing not-only the number of misclassifications, but also the identity of the classes involved in each inaccurate classification.


'Trifecta' outcomes of robot-assisted partial nephrectomy: a large Japanese multicenter study.
The objective of this study was to evaluate the early surgical outcomes of robot-assisted partial nephrectomy (RAPN) for small renal masses in a large Japanese multicenter series. A total of 804 consecutive cases of RAPN were examined at 42 institutes between 2011 and 2016. Medical records for clinical, pathological characteristics and perioperative outcomes were retrospectively reviewed. Univariable and multivariable analyses were performed to determine factors predicting Trifecta achievement. The median tumor size was 2.6 cm. The median RENAL score was 7. The median warm ischemia time was 21 min. The median estimated blood loss was 30 mL. Eight patients (1.0%) were converted to radical nephrectomy. The overall and Clavien-Dindo grade ≥ 3 complication rates were 13.0% and 5.8%, respectively. Pathologically, 91.4% of tumors were malignant and the positive surgical margin (PSM) rate was 1.1%. During the median 27.1-month observation period, the recurrence rate was 1.6%. Postoperative preservation rates of eGFR at 1, 6, 12 and 24 months were 90.3, 89.8, 89.4 and 89.2%, respectively. Trifecta was achieved in 62.1%. Multivariable analysis demonstrated that tumor diameter, estimated blood loss and hilar location of the tumor were significant negative factors predicting Trifecta achievement. The rate of Trifecta achievement for T1b tumors and hilar tumors was significantly lower (48.4% and 50.0%, respectively). RAPN was safely performed with acceptable oncological and functional outcomes, but the rate of Trifecta accomplishment for T1b or hilar tumors was significantly lower than that for T1a or non-hilar tumors, respectively.


Nuclear protein phosphatase-1: an epigenetic regulator of fear memory and amygdala long-term potentiation.
Complex brain diseases and neurological disorders in human generally result from the disturbance of multiple genes and signaling pathways. These disturbances may derive from mutations, deletions, translocations or rearrangements of specific gene(s). However, over the past years, it has become clear that such disturbances may also derive from alterations in the epigenome affecting several genes simultaneously. Our work recently demonstrated that epigenetic mechanisms in the adult brain are in part regulated by protein phosphatase 1 (PP1), a protein Ser/Thr phosphatase that negatively regulates hippocampus-dependent long-term memory (LTM) and synaptic plasticity. PP1 is abundant in brain structures involved in emotional processing like the amygdala, it may therefore be involved in the regulation of fear memory, a form of memory related to post-traumatic stress disorder (PTSD) in human. Here, we demonstrate that PP1 is a molecular suppressor of fear memory and synaptic plasticity in the amygdala that can control chromatin remodeling in neurons. We show that the selective inhibition of the nuclear pool of PP1 in amygdala neurons significantly alters posttranslational modifications (PTMs) of histones and the expression of several memory-associated genes. These alterations correlate with enhanced fear memory, and with an increase in long-term potentiation (LTP) that is transcription-dependent. Our results underscore the importance of nuclear PP1 in the amygdala as an epigenetic regulator of emotional memory, and the relevance of protein phosphatases as potential targets for therapeutic treatment of brain disorders like PTSD.


Expression of binding of plasminogen, thrombospondin, vitronectin, and fibrinogen, and adhesive properties by Escherichia coli strains isolated from patients with colonic diseases.
Escherichia coli strains isolated from patients with colonic disorders (n = 27) and strains isolated from the rectal mucosa of healthy subjects (n = 24) were compared with respect to expression of cell surface hydrophobicity, carriage of intestinal virulence factors, adhesion to tissue culture cells, and expression of binding of extracellular matrix proteins and plasma proteins. Strains isolated from patients with colonic disease did not express a more hydrophobic cell surface than strains from healthy subjects. Few strains from both groups carried genes encoding for recognised virulence factors of E coli. Only one strain, carrying the eae gene induced actin polymerisation in tissue culture cells. Strains from patients with colonic diseases adhered to HT29 cells, which are of intestinal origin, to a higher extent than E coli from healthy subjects. Significantly more strains from patients with colonic disorders than E coli from healthy subjects expressed binding of fibronectin, collagens, laminin, vitronectin, plasminogen, throbospondin, and fibrinogen. Expression of binding of these proteins may influence the pathogenesis of colonic disease by mediating binding to ulcerated tissue, preventing complement induced lysis of bacteria and by exerting proteolytic activity. There was no correlation between serotype, expression of cell surface hydrophobicity, and binding of extracellular matrix and plasma proteins.


Avoiding admission and facilitating early discharge through OPAT.
Fifteen years ago, only a small minority of hospital inpatients were considered suitable for outpatient parenteral antimicrobial therapy (OPAT) services (Wiselka and Nicholson, 1997). Now, almost 70% of those treated with intravenous (IV) antibiotics as hospital inpatients are considered suitable (Hitchcock et al, 2009). Around 38-53% of those requiring OPAT are able to self-administer (Hills et al, 2012). The demand for IV antimicrobial therapy is increasing and the way it is being delivered is changing. The delivery of IV anti-microbial therapy in the community has the potential to make a huge difference to the way health care is delivered. It can enable people who would once have been admitted to hospital to be treated in the community. It can facilitate early hospital discharge (Nazarko, 2013a). Potentially, there are huge benefits in developing OPAT but there are also risks. People may be subjected to an invasive treatment when it is not necessary; therapy may be continued when it is no longer necessary (Nazarko, 2013b; Conant et al, 2014); and patients may be selected for OPAT who would be best treated in hospital. The key to providing safe and effective care is to work together to maximise benefits and minimise risk (Duncan et al, 2013). This article examines how acute and community services can work together to ensure appropriate selection, treatment and follow-up of patients.


How Barunga Aboriginal community implemented and sustained an anaemia program - A case study evaluation.
Anaemia persists as a public health issue in many Aboriginal communities despite having standard practice guidelines. This case study reveals how Barunga Aboriginal Community in the Northern Territory (NT), Australia, implemented an Anaemia Program (1998-2016) which contributed to low anaemia prevalence in children aged under 5 years. This retrospective qualitative case study used purposive sampling to describe the Anaemia Program and factors influencing its implementation. Themes were developed from convergence of three data sources: interviews, program observation and document review. Data were inductively analysed by an Aboriginal and non-Aboriginal researcher and themes were validated by Barunga community health practitioners and compared to practice guidelines and implementation literature. Health practitioners reported that the Anaemia Program contributed to a marked reduction in childhood anaemia prevalence over time. This was supported by available prevalence data. The locally adapted Anaemia Program was unique in the NT with a novel approach to community supplementation for anaemia prevention in addition to anaemia treatment. Supportive implementation influences included: Aboriginal leadership and the use of culturally supportive processes which reinforced the development of trust and strong relationships facilitating community acceptance of the Program. Routine, opportunistic and flexible health care practice, a holistic approach and a stable, skilled and experienced team sustained program implementation. The holistic and successful Barunga Anaemia Program is supported by evidence and guidelines for treating and preventing childhood anaemia. The contextualisation of these guidelines aligned with the literature on effective Aboriginal primary health care implementation. SO WHAT?: This Anaemia Program provides a model for implementation of evidence-informed guidelines in an Aboriginal primary health care setting.


Two simple leg net devices designed to protect lower-extremity skin grafts and donor sites and prevent decubitus ulcer.
Burn therapists routinely are tasked to position the lower extremities of burn patients for pressure ulcer prevention, skin graft protection, donor site ventilation, and edema reduction. We developed two durable and low-maintenance devices that allow effective positioning of the lower extremities. The high-profile and low-profile leg net devices were simple to fabricate and maintain. The frame was assembled using a three-quarter-inch diameter copper pipe and copper fittings (45 degrees, 90 degrees, and tees). A double layer of elasticized tubular netting was pulled over the frame and doubled back for leg support to complete the devices. The devices can be placed on any bed surface. The netting can be exchanged when soiled and the frame can be disinfected between patients using standard techniques. Both devices were used on approximately 250 patients for a total of 1200 treatment days. No incidence of pressure ulcer was observed, and graft take was not adversely affected. The devices have not required repairs or replacement. Medical providers reported they are easy to apply and effectively maintain proper positioning throughout application. Neither device interfered with the application of other positioning devices. Both devices were found to be an effective method of positioning lower extremities to prevent pressure ulcer, minimize graft loss and donor site morbidity, and reduce edema. The devices allowed for proper wound ventilation and protected grafted lower extremities on any bed surface. The devices are simple to fabricate and maintain. Both devices can be effectively used simultaneously with other positioning devices.


Monoclonal antibodies to Pgp-1/CD44 block lympho-hemopoiesis in long-term bone marrow cultures.
A new panel of mAbs was prepared to a stromal cell line known to support lymphocytes in Whitlock-Witte type long-term bone marrow cultures. These antibodies were then screened with a cell adhesion assay and four were selected that inhibited the binding of B lineage cells to stromal cell monolayers. Immunofluorescent and biochemical analyses revealed that these new antibodies detected epitopes of the previously described Pgp-1/CD44 antigen complex. Addition of Pgp-1/CD44 antibodies to Dexter-type long-term bone marrow cultures completely prevented emergence of myeloid cells and they also blocked lymphocyte growth in Whitlock-Witte type cultures. mAbs MEL-14, LFA-1, and CD45R did not inhibit under the same conditions and there was no apparent relationship to Ig isotype. Adherent layers in treated cultures were not unusual in terms of morphology and the antibodies did not affect factor-dependent replication of lymphoid or myeloid progenitor cells. Therefore, the mechanism of inhibition may not involve direct toxicity to precursors or microenvironmental elements. Previous studies in humans and mice have implicated Pgp-1/CD44-related glycoproteins in the migration of peripheral lymphoid cells, as well as interactions of cells with the extracellular matrix. These findings suggest that they may also be critical for formation of lymphoid and myeloid cells within bone marrow.


Green bright squeezed light from a cw periodically poled KTP second harmonic generator.
We present the experimental observation of bright amplitude squeezed light from a singly resonant second harmonic generator (SHG) based on a periodically poled potassium titanyl phosphate (KTP) crystal. Contrary to conventional SHG, the interacting waves in this device couple efficiently using quasi phase matching (QPM) and more importantly QPM allows access to higher valued elements of the nonlinear tensor than is possible under the constraint of birefringence phase matching. We observe a noise reduction of 13% below the shot noise limit in the generated second harmonic field. This noise reduction is greater than what could be expected using normal birefringence phase matched KTP with the same experimental parameters. Excellent agreement between experiment and theory is found.


Cardiac ankyrin repeat protein attenuates cardiac hypertrophy by inhibition of ERK1/2 and TGF-β signaling pathways.
It has been reported that cardiac ankyrin repeat protein is associated with heart development and diseases. This study is aimed to investigate the role of CARP in heart hypertrophy in vivo. We generated a cardiac-specific CARP-overexpressing transgenic mouse. Although such animals did not display any overt physiological abnormality, they developed less cardiac hypertrophy in response to pressure overload than did wildtype mice, as indicated by heart weight/body weight ratios, echocardiographic and histological analyses, and expression of hypertrophic markers. These mice also exhibited less cardiac hypertrophy after infusion of isoproterenol. To gain a molecular insight into how CARP attenuated heart hypertrophy, we examined expression of the mitogen-activated protein kinase cascade and found that the concentrations of phosphorylated ERK1/2 and MEK were markedly reduced in the hearts of transgenic mice subjected to pressure overload. In addition, the expressions of TGF-β and phosphorylated Smad3 were significantly downregulated in the hearts of CARP Tg mice in response to pressure overload. Furthermore, addition of human TGF-β1 could reverse the inhibitory effect of CARP on the hypertrophic response induced by phenylephrine in cardiomyocytes. It was also evidenced that the inhibitory effect of CARP on cardiac hypertrophy was not attributed to apoptosis. CARP attenuates cardiac hypertrophy, in which the ERK and TGF-β pathways may be involved. Our findings highlight the significance of CARP as an anti-hypertrophic factor in therapy of cardiac hypertrophy.


Comparing ambient, air-convection, and fluid-convection heating techniques in treating hypothermic burn patients, a clinical RCT.
Hypothermia in burns is common and increases morbidity and mortality. Several methods are available to reach and maintain normal core body temperature, but have not yet been evaluated in critical care for burned patients. Our unit's ordinary technique for controlling body temperature (Bair Hugger®+ radiator ceiling + bed warmer + Hotline®) has many drawbacks e.g.; slow and the working environment is hampered.The aim of this study was to compare our ordinary heating technique with newly-developed methods: the Allon™2001 Thermowrap (a temperature regulating water-mattress), and Warmcloud (a temperature regulating air-mattress). Ten consecutive burned patients (> 20% total burned surface area and a core temperature < 36.0°C) were included in this prospective, randomised, comparative study. Patients were randomly exposed to 3 heating methods. Each treatment/measuring-cycle lasted for 6 hours. Each heating method was assessed for 2 hours according to a randomised timetable. Core temperature was measured using an indwelling (bladder) thermistor. Paired t-tests were used to assess the significance of differences between the treatments within the patients. ANOVA was used to assess the differences in temperature from the first to the last measurement among all treatments. Three-way ANOVA with the Tukey HSD post hoc test and a repeated measures ANOVA was used in the same manner, but included information about patients and treatment/measuring-cycles to control for potential confounding. Data are presented as mean (SD) and (range). Probabilities of less than 0.05 were accepted as significant. The mean increase, 1.4 (SD 0.6°C; range 0.6-2.6°C) in core temperature/treatment/measuring-cycle highly significantly favoured the Allon™2001 Thermowrap in contrast to the conventional method 0.2 (0.6)°C (range -1.2 to 1.5°C) and the Warmcloud 0.3 (0.4)°C (range -0.4 to 0.9°C). The procedures for using the Allon™2001 Thermowrap were experienced to be more comfortable and straightforward than the conventional method or the Warmcloud. The Allon™2001 Thermowrap was more effective than the Warmcloud or the conventional method in controlling patients' temperatures.


Cooperativity basis for small-molecule stabilization of protein-protein interactions.
A cooperativity framework to describe and interpret small-molecule stabilization of protein-protein interactions (PPI) is presented. The stabilization of PPIs is a versatile and emerging therapeutic strategy to target specific combinations of protein partners within the protein interactome. Currently, the potency of PPI stabilizers is typically expressed by their apparent affinity or EC50. Here, we propose that the effect of a PPI stabilizer be best described involving the cooperativity factor, α, between the stabilizer and binding partners in addition to the intrinsic affinity, K D II, of the stabilizer for one of the apo-proteins. By way of illustration, we combine fluorescence polarization measurements with thermodynamic modeling to determine the α and K D II for the PPI stabilization of 14-3-3 and TASK3 by fusicoccin-A (FC-A) and validate our approach by studying other PPI-partners of 14-3-3 proteins. Finally, we characterize a library of different stabilizer compounds, and perform structure-activity relationship studies in which molecular changes could be attributed to either changes in cooperativity or intrinsic affinity. Such insights should aid in the development of more effective protein-protein stabilizer drugs.


Causes and consequences of reciprocal translocations on sex chromosomes.
Under XY sex determination, the Y chromosome is only inherited via males, whereas the X chromosome is predominantly found in females. Thus, it is favourable when alleles with high male fitness become associated with the Y chromosome and when alleles with high female fitness become associated with the X chromosome. These favourable associations can be strengthened through linkage. Rearrangements, such as inversions and sex chromosome-autosome fusions, can increase linkage and thereby become favoured (Charlesworth, 2017). In a From the Cover article in this issue of Molecular Ecology, Toups, Rodrigues, Perrin, and Kirkpatrick (2019) present the first genomic analysis of a sex chromosome reciprocal translocation, a particularly dramatic chromosomal rearrangement that modifies linkage with the sex chromosome. As a result of reciprocal translocation, one studied population of the common frog (Rana temporaria, Figure 1) displays a remarkable sex-determining system in which there are two physically unlinked sex chromosomes that are exclusively cotransmitted (Figure 2a).


Homologous recombination rescues mismatch-repair-dependent cytotoxicity of S(N)1-type methylating agents in S. cerevisiae.
Resistance of mammalian cells to S(N)1-type methylating agents such as N-methyl-N'-nitro-N-nitrosoguanidine (MNNG) generally arises through increased expression of methylguanine methyltransferase (MGMT), which reverts the cytotoxic O(6)-methylguanine ((Me)G) to guanine, or through inactivation of the mismatch repair (MMR) system, which triggers cell death through aberrant processing of (Me)G/T mispairs generated during DNA replication when MGMT capacity is exceeded. Given that MMR and (Me)G-detoxifying proteins are functionally conserved through evolution, and that MMR-deficient Escherichia coli dam(-) strains are also resistant to MNNG, the finding that MMR status did not affect the sensitivity of Saccharomyces cerevisiae to MNNG was unexpected. Because (Me)G residues in DNA trigger homologous recombination (HR), we wondered whether the efficient HR in S. cerevisiae might alleviate the cytotoxic effects of (Me)G processing. We now show that HR inactivation sensitizes S. cerevisiae to MNNG and that, as in human cells, defects in the MMR genes MLH1 and MSH2 rescue this sensitivity. Inactivation of the EXO1 gene, which encodes the only exonuclease implicated in MMR to date, failed to rescue the hypersensitivity, which implies that scExo1 is not involved in the processing of (Me)G residues by the S. cerevisiae MMR system.


Decreased teicoplanin susceptibility of methicillin-resistant strains of Staphylococcus aureus.
Between February 1992 and February 1993, 12 patients were seen who were infected or colonized with methicillin-resistant strains of Staphylococcus aureus. The strains had decreased susceptibility to teicoplanin (MICs, 8-16 micrograms/mL). Field inversion gel electrophoresis showed the strains belonged to three related clones (A1, A2, and A3). The patients were though to have acquired the strains nosocomially. Consistent with results of laboratory studies of teicoplanin-resistant S. aureus, all but 1 strain expressed a 35-kDa membrane protein, and 9 strains expressed increased levels of penicillin-binding protein 2 complex. Six strains were isolated from patients treated with glycopeptides. These data suggest that nosocomial transmission of S. aureus with decreased teicoplanin susceptibility may occur during glycopeptide use and that such strains develop resistance by a mechanism associated with the appearance of a 35-kDa membrane protein. Surveillance is necessary to monitor for the potential selection of resistant clones that may be capable of dissemination.


Effect of zidovudine and Pneumocystis carinii pneumonia prophylaxis on progression of HIV-1 infection to AIDS. The Multicenter AIDS Cohort Study.
Although used widely, the effectiveness of zidovudine therapy and primary prophylaxis for Pneumocystis carinii pneumonia (PCP) in HIV-1-infected individuals, has not been assessed in a large cohort. We have done an observational study between October, 1986, and October, 1990, of a cohort of 2145 HIV-1-seropositive men and 371 who seroconverted during the study. A Markov chain transitional analysis was used to examine the effect of zidovudine and PCP prophylaxis on the probability of progression of HIV-1 infection to AIDS (after 6, 12, 18, and 24 months) after follow-up visits categorised into one of six disease states. The six starting states were based on CD4+ lymphocyte counts and the presence of HIV-related symptoms. Use of pre-AIDS zidovudine and PCP prophylaxis was associated with significant reductions in rates of progression to AIDS at 6, 12, 18, and 24 months for participants starting with less than 350 CD4+ lymphocytes/microliter. For those starting with 350 or more CD4+ lymphocytes/microliter, non-significant protective trends were seen during 12, 18, and 24 month intervals. In multivariate log-linear models virtually all the treatment effect was due to zidovudine. However, after adjusting for the effects of zidovudine, PCP prophylaxis reduced significantly the probability of progression to a first episode of PCP during 6, 12, 18, and 24 month intervals. This study suggests that early primary PCP prophylaxis is effective in preventing first episodes of PCP, and that the efficacy of zidovudine demonstrated in clinical trials can be translated to the population level.


Highly sensitive detection of five typical fluoroquinolones in low-fat milk by field-enhanced sample injection-based CE in bubble cell capillary.
Fluoroquinolones are a group of synthetic antibiotics with a broad activity spectrum against mycoplasma, Gram-positive, and Gram-negative bacteria. Due to the extensive use of fluoroquinolones in farming and veterinary science, there is a constant need in the analytical methods able to efficiently monitor their residues in food products of animal origin, regulated by Commission Regulation (European Union) no. 37/2010. Herein, field-enhanced sample injection for sample stacking prior the CZE separation was developed inside a bubble cell capillary for highly sensitive detection of five typical fluoroquinolones in bovine milk. Ethylenediamine was proposed as the main component of BGE for the antibiotics separation. The effect of BGE composition, injection parameters, and water plug length on the field-enhanced sample injection-based CE with UV detection was investigated. Under the optimized conditions, described field-enhanced sample injection-based CE-UV analysis of fluoroquinolones provides LODs varying from 0.4 to 1.3 ng/mL. These LOD values are much lower (from 460 to 1500 times) than those obtained by a conventional CE in a standard capillary without bubble cell. The developed method was finally applied for the analysis of fluoroquinolones in low-fat milk from a Swiss supermarket. Sample recovery values from 93.6 to 106.0% for different fluoroquinolones, and LODs from 0.7 to 2.5 μg/kg, were achieved. Moreover, the proposed ethylenediamine-based BGE as volatile and compatible with MS system, enabled the coupling of the field-enhanced sample injection-based CE with a recently introduced electrostatic spray ionization MS via an iontophoretic fraction collection interface for qualitative fluoroquinolones identification.


The activation of the CGRP receptor.
The CGRP (calcitonin gene-related peptide) receptor is a family B GPCR (G-protein-coupled receptor). It consists of a GPCR, CLR (calcitonin receptor-like receptor) and an accessory protein, RAMP1 (receptor activity modifying protein 1). RAMP1 is needed for CGRP binding and also cell-surface expression of CLR. CLR is an example of a family B GPCR. Unlike family A GPCRs, little is known about how these receptors are activated by their endogenous ligands. This review considers what is known about the activation of family B GPCRs and then considers how this might be applied to CLR, particularly in light of new knowledge of the crystal structures of family A GPCRs.


Failure to rescue as a source of variation in hospital mortality after rectal surgery: The Italian experience.
Failure to rescue (FTR) patients from postoperative complications could contribute to the variability in surgical mortality seen among hospitals with different volumes. We sought to examine the impact of complications and FTR on mortality following rectal surgery. The National Italian Hospital Discharge Dataset allowed to identify 75,280 patients who underwent rectal surgery between 2002 and 2014. Hospital volume was stratified into tertiles. Rates of major complications, FTR from complications and mortality following rectal surgery were compared. During the study period, both the incidence of complications (2002, 23.7% versus 2014, 21.2%), and FTR decreased overtime (2002, 6.9% versus 2014, 3.8%) (both P < 0.001). The complication rate was 24.4% in low-, 21.6% in intermediate- and 20.4% in high-volume hospitals (P < 0.001). Complications were less common in minimally invasive surgery (MIS) versus open cases (18.2% versus 23.2%; P < 0.001). The most frequent complications included prolonged ileus or small bowel obstruction (5.3%), and anemia requiring blood transfusions (5.3%). The rate of FTR was 5.5%, 5.6% and 3.7% for low-, intermediate- and high-volume hospitals, respectively (P < 0.001). FTR after MIS was 2.6% vs. 5.5% after open surgery (P < 0.001). After accounting for patient and hospital characteristics, patients treated at low-volume hospitals were 23% more likely to die after a complication, compared to patients at high-volume hospitals (OR 1.23, 95%CI 1.13-1.33). Hospital volume is the strongest predictor of complication and FTR. The reduction in mortality in high-volume hospitals could be determined by the better ability to rescue patients. These findings support the centralization policy of rectal cancer treatment.


Knowledge, attitudes and beliefs of first-degree relatives of patients with chronic kidney disease toward kidney donation in Nigeria.
In most parts of Sub-Saharan Africa, kidney transplant programs are dependent on the willingness of relatives of patients with kidney failure to donate kidneys. This study assessed the attitudes of relatives of patients with chronic kidney disease (CKD) toward kidney donation. This was a cross-sectional survey of relatives of patients with CKD attending the nephrology service of our hospital. The respondents' socio-demographic characteristics and knowledge and beliefs about kidney transplantation, as well as their willingness to donate a kidney, were assessed using a self-administered questionnaire. There were 161 respondents who returned completed questionnaires; the mean age of the respondents was 34.8±12.6 years and 52.2% of them were female. About 85.1% of the respondents were aware that kidney transplantation was a treatment option for end-stage renal failure, while 70% of them believed that kidney transplantation resulted in an improvement in the quality of life of these patients. However, 25.5% of the respondents believed that kidney donors were at risk of developing kidney failure in the future. Overall, 77.6% of the respondents were willing to donate a kidney, especially if the affected individual was their offspring. The majority of the respondents were willing to donate a kidney to a relative with CKD.


Membrane delivery to the yeast autophagosome from the Golgi-endosomal system.
While many of the proteins required for autophagy have been identified, the source of the membrane of the autophagosome is still unresolved with the endoplasmic reticulum (ER), endosomes, and mitochondria all having been evoked. The integral membrane protein Atg9 is delivered to the autophagosome during starvation and in the related cytoplasm-to-vacuole (Cvt) pathway that occurs constitutively in yeast. We have examined the requirements for delivery of Atg9-containing membrane to the yeast autophagosome. Atg9 does not appear to originate from mitochondria, and Atg9 cannot reach the forming autophagosome directly from the ER or early Golgi. Components of traffic between Golgi and endosomes are known to be required for the Cvt pathway but do not appear required for autophagy in starved cells. However, we find that pairwise combinations of mutations in Golgi-endosomal traffic components apparently only required for the Cvt pathway can cause profound defects in Atg9 delivery and autophagy in starved cells. Thus it appears that membrane that contains Atg9 is delivered to the autophagosome from the Golgi-endosomal system rather than from the ER or mitochondria. This is underestimated by examination of single mutants, providing a possible explanation for discrepancies between yeast and mammalian studies on Atg9 localization and autophagosome formation.


Single-Nucleotide-Polymorphism-Panel Population-Genetics Approach Based on the 1000 Genomes Database and Elite Soccer Players.
Purpose: Soccer is one of the most popular sports worldwide, a physical activity of great physiological demand and complexity. Currently, numerous trials involving physiological responses such as hypertrophy, energy expenditure, vasodilation, cardiac output, VO2max, and recovery have supported the possibility of genomic predictors' affecting performance. In a complementary way to association studies with single nucleotide polymorphisms (SNPs), the objective was to evaluate if the use of population genetics data from human-genomics databases can provide information for a better understanding of the relationship between heritability and sport performance. Methods: The study included 25 healthy male professional soccer players (25.5 [4.3] y, 177.4 [6.4] cm, 76.4 [6.4] kg, body fat 10.5% [4.3%]) from the Brazilian first-division soccer club. Anthropometric measurements and field and isokinetic tests were performed to evaluate performance and physiologic parameters of subjects. Moreover, 10 genetic polymorphisms previously related to performance were genotyped. The genotypes of the same polymorphisms were obtained for 2504 individuals from the populations deposited in the 1000 Genomes database. A principal-component analysis and matrix genetic-distances approach (Fst) were evaluated. Results: As expected, the admixture Brazilian population has numerous genetic similarities with the European and American populations from genomic databases. Although the African component is absolutely recognized in genomes from the Brazilian population, using the specific performance-related SNPs, surprisingly the African population was one of the most genetically distant of the players (P < .00001). Conclusions: The early results suggest a selective pressure on genes of elite soccer players, possibly related simultaneously to physical-performance, environmental, cognitive, and sociocultural aspects.


Lumbar disc herniation in adolescence.
Lumbar disc herniation very rarely occurs in adolescence. The aim of this study was to assess the radiological, clinical and surgical features and case outcomes for adolescents with lumbar disc herniation, and to compare with adult cases. The cases of 17 adolescents (7 girls and 10 boys, age range 13-17 years) who were surgically treated for lumbar disc herniation in our clinic between 1998 and 2003 were retrospectively reviewed. The mean follow-up time for these cases was 60 months. The collected histories revealed that 14 (82%) of the 17 cases involved trauma or intense sports activity. Low back pain was the most common complaint (15 cases, 88%). None of the 17 patients had major symptoms during follow-up, and most were engaged in intense sports or heavy work-related activities during this period. The main features of lumbar disc herniation in adolescents are different from those seen in adults. Careful assessment is vital to avoid misdiagnosis and prevent undesirable results from inappropriate surgery. When surgery is indicated and patients are carefully selected, the results of lumbar discectomy in adolescence can be satisfactory.


Deficiency of terminal ADP-ribose protein glycohydrolase TARG1/C6orf130 in neurodegenerative disease.
Adenosine diphosphate (ADP)-ribosylation is a post-translational protein modification implicated in the regulation of a range of cellular processes. A family of proteins that catalyse ADP-ribosylation reactions are the poly(ADP-ribose) (PAR) polymerases (PARPs). PARPs covalently attach an ADP-ribose nucleotide to target proteins and some PARP family members can subsequently add additional ADP-ribose units to generate a PAR chain. The hydrolysis of PAR chains is catalysed by PAR glycohydrolase (PARG). PARG is unable to cleave the mono(ADP-ribose) unit directly linked to the protein and although the enzymatic activity that catalyses this reaction has been detected in mammalian cell extracts, the protein(s) responsible remain unknown. Here, we report the homozygous mutation of the c6orf130 gene in patients with severe neurodegeneration, and identify C6orf130 as a PARP-interacting protein that removes mono(ADP-ribosyl)ation on glutamate amino acid residues in PARP-modified proteins. X-ray structures and biochemical analysis of C6orf130 suggest a mechanism of catalytic reversal involving a transient C6orf130 lysyl-(ADP-ribose) intermediate. Furthermore, depletion of C6orf130 protein in cells leads to proliferation and DNA repair defects. Collectively, our data suggest that C6orf130 enzymatic activity has a role in the turnover and recycling of protein ADP-ribosylation, and we have implicated the importance of this protein in supporting normal cellular function in humans.


HER2-Overexpressing Breast Cancers Amplify FGFR Signaling upon Acquisition of Resistance to Dual Therapeutic Blockade of HER2.
Purpose: Dual blockade of HER2 with trastuzumab and lapatinib or pertuzumab has been shown to be superior to single-agent trastuzumab. However, a significant fraction of HER2-overexpressing (HER2+) breast cancers escape from these drug combinations. In this study, we sought to discover the mechanisms of acquired resistance to the combination of lapatinib + trastuzumab.Experimental Design: HER2+ BT474 xenografts were treated with lapatinib + trastuzumab long-term until resistance developed. Potential mechanisms of acquired resistance were evaluated in lapatinib + trastuzumab-resistant (LTR) tumors by targeted capture next-generation sequencing. In vitro experiments were performed to corroborate these findings, and a novel drug combination was tested against LTR xenografts. Gene expression and copy-number analyses were performed to corroborate our findings in clinical samples.Results: LTR tumors exhibited an increase in FGF3/4/19 copy number, together with an increase in FGFR phosphorylation, marked stromal changes in the tumor microenvironment, and reduced tumor uptake of lapatinib. Stimulation of BT474 cells with FGF4 promoted resistance to lapatinib + trastuzumab in vitro Treatment with FGFR tyrosine kinase inhibitors reversed these changes and overcame resistance to lapatinib + trastuzumab. High expression of FGFR1 correlated with a statistically shorter progression-free survival in patients with HER2+ early breast cancer treated with adjuvant trastuzumab. Finally, FGFR1 and/or FGF3 gene amplification correlated with a lower pathologic complete response in patients with HER2+ early breast cancer treated with neoadjuvant anti-HER2 therapy.Conclusions: Amplification of FGFR signaling promotes resistance to HER2 inhibition, which can be diminished by the combination of HER2 and FGFR inhibitors. Clin Cancer Res; 23(15); 4323-34. ©2017 AACR.


Anti-atherogenic properties of date vs. pomegranate polyphenols: the benefits of the combination.
Hydrolysable tannin polyphenols in pomegranate and phenolic acids in date fruit and seeds are potent antioxidants and anti-atherogenic agents, and thus, in the present study we investigated the possible benefits of combining them in vivo in atherosclerotic apolipoprotein E KO (E(0)) mice, compared with the individual fruit. In vitro studies revealed that the date seed extract contains more polyphenols than Amari or Hallawi date extracts, and possesses a most impressive free radical scavenging capacity. Similarly, pomegranate juice (PJ), punicalagin, punicalain, gallic acid, and urolithins A and B are very potent antioxidants. E(0) mice consumed 0.5 μmol gallic acid equivalents (GAE) per mouse per day of PJ, Hallawi extract, date seed extract, or a combination for 3 weeks. Consumption of the combination was the most potent treatment, as it decreased serum cholesterol and triglyceride levels, and increased serum paraoxonase 1 (PON1) activity. Consumption of the combination also significantly reduced mouse peritoneal macrophage (MPM) oxidative stress, MPM cholesterol content, and MPM LDL uptake. Finally, the lipid peroxide content in the aortas of the mice significantly decreased, and the PON lactonase activity of the aortas increased after treatment with the combination. We thus conclude that consumption of pomegranate, together with date fruit and date seeds, has the most beneficial anti-atherogenic effects on E(0) mice serum, macrophages, and aortas, probably due to their unique and varied structures.


Tissue microarray construction from gross specimens: development of a novel simple technique.
The tissue microarray (TMA) technique is becoming a useful tool for research and quality control methods in surgical pathology. However, the widespread use of this technique in routine surgical pathology practice, especially in developing nations, is hampered by the high cost of commercially available instruments. The authors describe a novel, inexpensive technique of construction of TMA from gross specimen. The authors used a 16-gauge bone marrow trephine biopsy needle to obtain cores from 150 specimens of breast carcinoma. These cores were processed in the tissue processor as per the routine processing techniques of the department. A recipient block was prepared, followed by drilling of a suitable number of holes. The processed tissue cores were arrayed into the recipient block. These blocks were used for immunohistochemistry and compared with conventional whole tissue sections. Of the 150 cases, only two showed suboptimal preservation in the TMA block, and this was also reflected in the conventional tissue sections. Immunohistochemical staining for oestrogen receptor, progesterone receptor, Her2.neu and Ki-67 was found to be satisfactory in 148 cases with the staining intensity comparable with that performed on the conventional sections. This study describes a satisfactory technique for construction of TMA from a gross tumour specimen. Such a technique is well suited to centres which handle a high number of cases of a particular type, such as breast carcinoma. Further studies on other tumour types would help in validating these results.


Distortion product otoacoustic emissions in an industrial setting.
Distortion product otoacoustic emissions (DPOAEs) is an objective sensitive test of cochlear function. The aim of this study was the evaluation of noise-induced hearing loss in a group of industrial workers, using this method in conjunction with standard puretone audiometry (PTA). One hundred and five subjects (210 ears) were included in the study. PTA, tympanometry, and DPOAEs were performed. Results were analyzed using a mixed analysis of variance model, and compared with the data of 34 normal persons of similar age and sex. We found statistically significant lower DPOAE levels in the noise-exposed group than in the control group. Additionally, the effect of frequency was significant, indicating that amplitude varied across frequency, with lower responses observed at 4 and 6 kHz, and maximum response found at 2 kHz. PTA showed a statistically significant effect of Group, owed to elevated puretone thresholds in the noise-exposed subjects, but a Frequency main effect was not found, although the interaction between Frequency and Group was statistically significant, as well as the interaction between Frequency and Ear. A main effect for Ear was found only in puretone thresholds, due to better thresholds in the left ears of the subjects, and not in DPOAE measurements. DPOAE levels were selectively affected at the higher frequencies, whereas puretone thresholds were affected at all frequencies. Direct comparison of the number of significantly affected ears between the two methods at 1, 2, and 4 kHz showed statistically significant differences at all comparisons, with more ears affected in PTA in comparison with DPOAEs at 4 kHz, whereas more ears were affected in DPOAEs at the lower frequencies (1 and 2 kHz). Therefore, it may be concluded that DPOAEs and PTA are both sensitive methods in detecting noise-induced hearing loss, with DPOAEs tending to be more sensitive at lower frequencies.


Elimination of cross talk in waveguide intersections.
We present general criteria for crossing perpendicular waveguides with nearly 100% throughput and 0% cross talk. Our design applies even when the waveguide width is of the order of the wavelength. The theoretical basis for this phenomenon is explained in terms of symmetry considerations and resonant tunneling and is then illustrated with numerical simulations for both a two-dimensional photonic crystal and a conventional high-index-contrast waveguide crossing. Cross-talk reduction by up to 8 orders of magnitude is achieved relative to unmodified crossings.


Start-Up and Ongoing Practice Expenses of Behavioral Health and Primary Care Integration Interventions in the Advancing Care Together (ACT) Program.
Provide credible estimates of the start-up and ongoing effort and incremental practice expenses for the Advancing Care Together (ACT) behavioral health and primary care integration interventions. Expenditure data were collected from 10 practice intervention sites using an instrument with a standardized general format that could accommodate the unique elements of each intervention. Average start-up effort expenses were $44,076 and monthly ongoing effort expenses per patient were $40.39. Incremental expenses averaged $20,788 for start-up and $4.58 per patient for monthly ongoing activities. Variations in expenditures across practices reflect the differences in intervention specifics and organizational settings. Differences in effort to incremental expenditures reflect the extensive use of existing resources in implementing the interventions. ACT program incremental expenses suggest that widespread adoption would likely have a relatively modest effect on overall health systems expenditures. Practice effort expenses are not trivial and may pose barriers to adoption. Payers and purchasers interested in attaining widespread adoption of integrated care must consider external support to practices that accounts for both incremental and effort expense levels. Existing knowledge transfer mechanisms should be employed to minimize developmental start-up expenses and payment reform focused toward value-based, Triple Aim-oriented reimbursement and purchasing mechanisms are likely needed.


Nerve injury induced by vibration: prevention of the effect of a conditioning lesion by D600, a Ca2+ channel blocker.
Exposing a hind leg of a rat to vibration induces an injury to the sciatic nerve--a so called conditioning lesion. After such injury induced by vibration the regenerative capacity of the nerve is improved and can be detected as an increased axonal outgrowth from a test crush lesion to the same nerve. The purpose was to study whether the effect of a conditioning lesion induced by vibration can be prevented by local treatment with a Ca2+ channel blocker D600. D600 (methoxyverapamil) or Ringer's solution was locally applied to the sciatic nerve on one side through a silicone tube connected to a miniosmotic pump, which was implanted subcutaneously. During the same period the hind leg was exposed to vibration (80 Hz; 32 m/s2 root mean squared) for five hours daily for five consecutive days. The other hind leg was not vibrated. After the end of exposure to vibration the sciatic nerves were crushed bilaterally (test crush lesions) and three or six days later the regeneration distances of sensory axons were measured by the pinch reflex test. Nerves in the control animals (without implanted miniosmotic pumps and nerves on to which Ringer's solution was locally applied) that were exposed to vibration showed a significantly increased outgrowth length of sensory axons from the test crush lesion compared with the non-vibrated side. Such an effect of a conditioning lesion from the exposure to vibration was suppressed by local application of D600. Local administration of a Ca2+ channel blocker D600 can prevent the effect of a conditioning lesion-that is, the nerve injury induced by vibration can be inhibited by D600. This may have implications for the treatment of patients with neuropathy of the hand induced by vibration.


Epidemic of chronic kidney disease in India -what can be done?
The exact prevalence of chronic kidney disease in India is not clear in the absence of regular national registry data and provided only by small observational series or rely on reports from personal experience, but the quality of data is quiet uneven. There are only three population based studies in India commenting on the magnitude of chronic kidney disease. In a prevention program started at community level in Chennai, the reported prevalence is 0.86% in the project population and 1.39% in the control region. The second study is based on Delhi involving 4972 urban patients. The prevalence of chronic renal failure (defined as serum creatinine more than 1.8 mg/dL) to be 0.79 % or 7852 per million/population. The third study perhaps the only longitudinal study to identify the incidence of end stage renal disease is based on 572,029 subjects residing in city of Bhopal suggests that the average crude and age adjusted incidence rates of end stage renal disease were 151 and 232 per million population respectively. The resources and skill for taking care of this large case load, both in terms of personal and health care infrastructure do not exist currently and would need to be created. To tackle the problem of limited access to renal replacement therapy, an important method would be to try and reduce the incidence of end stage renal disease and the need of renal replacement therapy by preventive measures. It is clear that treatment of chronic kidney disease and its advanced stage end stage renal disease is expensive and beyond the reach of average Indian. Thus it is crucial that prevention of chronic kidney disease has to be the goal of medical fraternity, government of India and the general public. This article suggests a series of primary, secondary and tertiary preventive measures for prevention of chronic kidney disease. Clearly there are already many effective and attractive interventions for the treatment and prevention of chronic kidney disease exist and many more surely be developed.


Exploring the migration decisions of health workers and trainees from Africa: a meta-ethnographic synthesis.
The migration of healthcare workers from Africa depletes countries already suffering from substantial staffing shortages and considerable disease burdens. The recruitment of such individuals by high income countries has been condemned by the World Health Organisation. However, understanding the reasons why healthcare workers migrate is essential, in order to attempt to alter migration decisions. We aimed to systematically analyse factors influencing healthcare workers' decisions to migrate from Africa. We systematically searched CINAHL (1980-Nov 2010), Embase (1980-Nov 2010), Global Health (1973-Nov 2010) and Medline (1950-Nov 2010) for qualitative studies of healthcare workers from Africa which specifically explored views about migration. Two reviewers identified articles, extracted data and assessed quality of included studies. Meta-ethnography was used to synthesise new lines of understanding and meaning from the data. The search identified 1203 articles from which we included six studies of healthcare workers trained in seven African countries, namely doctors or medical students (two studies), nurses (three), and pharmacy students (one study). Using meta-ethnographic synthesis we produced six lines of argument relating to the migration decisions of healthcare workers: 1) Struggle to realise unmet material expectations of self, family and society, 2) Strain and emotion, interpersonal discord, and insecurity in workplace, 3) Fear from threats to personal or family safety, in and out of workplace, 4) Absence of adequate professional support and development, 5) Desire for professional prestige and respect, 6) Conviction that hopes and goals for the future will be fulfilled overseas. We conclude that a complex interaction of factors contribute to the migration decisions of healthcare workers from Africa. Some of the factors identified are more amenable to change than others, and addressing these may significantly affect migration decisions of African healthcare workers in the future.


Structure of active dimeric human telomerase.
Telomerase contains a large RNA subunit, TER, and a protein catalytic subunit, TERT. Whether telomerase functions as a monomer or dimer has been a matter of debate. Here we report biochemical and labeling data that show that in vivo-assembled human telomerase contains two TERT subunits and binds two telomeric DNA substrates. Notably, catalytic activity requires both TERT active sites to be functional, which demonstrates that human telomerase functions as a dimer. We also present the three-dimensional structure of the active full-length human telomerase dimer, determined by single-particle EM in negative stain. Telomerase has a bilobal architecture with the two monomers linked by a flexible interface. The monomer reconstruction at 23-Å resolution and fitting of the atomic structure of the TERT subunit from beetle Tribolium castaneum into the EM density reveals the spatial relationship between RNA and protein subunits, providing insights into telomerase architecture.


Weighted trade network in a model of preferential bipartite transactions.
Using a model of wealth distribution where traders are characterized by quenched random saving propensities and trade among themselves by bipartite transactions, we mimic the enhanced rates of trading of the rich by introducing the preferential selection rule using a pair of continuously tunable parameters. The bipartite trading defines a growing trade network of traders linked by their mutual trade relationships. With the preferential selection rule this network appears to be highly heterogeneous characterized by the scale-free nodal degree and the link weight distributions and presents signatures of nontrivial strength-degree correlations. With detailed numerical simulations and using finite-size scaling analysis we present evidence that the associated critical exponents are continuous functions of the tuning parameters. However the wealth distribution has been observed to follow the well-known Pareto law robustly for all positive values of the tuning parameters.


Muscle glucose metabolism in exercising rats: comparison with insulin stimulation.
We describe here a technique using bolus 2-[3H]deoxyglucose (2-[3H]DG) administration for estimating the glucose metabolic rate (R'g) in individual tissues of the rat during exercise. After 50 min of treadmill running at a moderate work load various tissues were analyzed for accumulation of phosphorylated 2-[3H]DG and/or glycogen mass. There was considerable heterogeneity in R'g in response to exercise among different muscles, despite similar basal values. R'g increased 32-fold above basal in soleus (to 90.4 +/- 5.7 mumol X 100 g-1 X min-1), 42-fold in red gastrocnemius (to 71.2 +/- 1.6), 5-fold in white gastrocnemius (to 7.0 +/- 0.8), and 2-fold in extensor digitorum longus (to 4.1 +/- 1.0) during exercise. There was a close relationship between muscle glycogen depletion and R'g within different muscles. In view of the magnitude of the increase in R'g during exercise, this method provides a very sensitive index of muscle fiber recruitment during exercise. The pattern of exercise-stimulated R'g in muscles of different fiber composition was similar to that seen with insulin stimulation. However, moderate exercise alone produced increments in R'g (soleus and red gastrocnemius) that were significantly higher (P less than 0.005) than that observed during maximal insulin stimulation even though whole body glucose utilization was slightly higher during maximal insulin stimulation. These data emphasize the heterogeneity of the response in different muscles during exercise and suggest that intracellular events beyond glucose transport may be rate limiting under the influence of one or both of these stimuli.


Claudin-1, A Double-Edged Sword in Cancer.
Claudins, a group of membrane proteins involved in the formation of tight junctions, are mainly found in endothelial or epithelial cells. These proteins have attracted much attention in recent years and have been implicated and studied in a multitude of diseases. Claudins not only regulate paracellular transepithelial/transendothelial transport but are also critical for cell growth and differentiation. Not only tissue-specific but the differential expression in malignant tumors is also the focus of claudin-related research. In addition to up- or down-regulation, claudin proteins also undergo delocalization, which plays a vital role in tumor invasion and aggressiveness. Claudin (CLDN)-1 is the most-studied claudin in cancers and to date, its role as either a tumor promoter or suppressor (or both) is not established. In some cancers, lower expression of CLDN-1 is shown to be associated with cancer progression and invasion, while in others, loss of CLDN-1 improves the patient survival. Another topic of discussion regarding the significance of CLDN-1 is its localization (nuclear or cytoplasmic vs perijunctional) in diseased states. This article reviews the evidence regarding CLDN-1 in cancers either as a tumor promoter or suppressor from the literature and we also review the literature regarding the pattern of CLDN-1 distribution in different cancers, focusing on whether this localization is associated with tumor aggressiveness. Furthermore, we utilized expression data from The Cancer Genome Atlas (TCGA) to investigate the association between CLDN-1 expression and overall survival (OS) in different cancer types. We also used TCGA data to compare CLDN-1 expression in normal and tumor tissues. Additionally, a pathway interaction analysis was performed to investigate the interaction of CLDN-1 with other proteins and as a future therapeutic target.


Visfatin and resistin serum levels in normal-weight and obese women with polycystic ovary syndrome.
Polycystic ovary syndrome (PCOS) is the most common endocrinopathy among women of reproductive age that is linked to insulin resistance and obesity. While studies have shown that plasma levels of resistin and visfatin increase with obesity, the association between PCOS and these markers has not been described well. This case-control study aimed to compare the serum levels of visfatin and resistin in women with PCOS in comparison with the healthy controls matched for age and body mass index (BMI). A total of 80 women consisted of 40 women with PCOS and 40 matched eumenorrheic women without hyperandrogenism enrolled in the study. They were subcategorized into obese and normal-weight women according to their BMI. Serum visfatin and resistin levels were assessed using sandwich enzyme-linked Immunosorbent assay (ELISA). Serum levels of resistin were higher among both obese and normal-weight women with PCOS in comparison with the controls (2.36 and 1.58 ng/mL in normal-weight women with PCOS and controls, respectively; and 2.10 and 1.91 ng/mL in obese women with PCOS and controls, respectively). Serum visfatin levels was higher in both obese women with PCOS and controls (3.46 and 3.49 ng/mL PCOS and control groups, respectively) in comparison with normal-weight women in both groups (3.16 and 3.15 in PCOS and control groups, respectively); however; there were no statistically significant differences in serum resistin and visfatin levels between PCOS and control groups (P > 0.05). While the expression of visfatin and resistin may be upregulated in women with PCOS, it is not translated at serum level.


Effects of acute nutritional ketosis during exercise in adults with glycogen storage disease type IIIa are phenotype-specific: An investigator-initiated, randomized, crossover study.
Glycogen storage disease type IIIa (GSDIIIa) is an inborn error of carbohydrate metabolism caused by a debranching enzyme deficiency. A subgroup of GSDIIIa patients develops severe myopathy. The purpose of this study was to investigate whether acute nutritional ketosis (ANK) in response to ketone-ester (KE) ingestion is effective to deliver oxidative substrate to exercising muscle in GSDIIIa patients. This was an investigator-initiated, researcher-blinded, randomized, crossover study in six adult GSDIIIa patients. Prior to exercise subjects ingested a carbohydrate drink (~66 g, CHO) or a ketone-ester (395 mg/kg, KE) + carbohydrate drink (30 g, KE + CHO). Subjects performed 15-minute cycling exercise on an upright ergometer followed by 10-minute supine cycling in a magnetic resonance (MR) scanner at two submaximal workloads (30% and 60% of individual maximum, respectively). Blood metabolites, indirect calorimetry data, and in vivo 31 P-MR spectra from quadriceps muscle were collected during exercise. KE + CHO induced ANK in all six subjects with median peak βHB concentration of 2.6 mmol/L (range: 1.6-3.1). Subjects remained normoglycemic in both study arms, but delta glucose concentration was 2-fold lower in the KE + CHO arm. The respiratory exchange ratio did not increase in the KE + CHO arm when workload was doubled in subjects with overt myopathy. In vivo 31 P MR spectra showed a favorable change in quadriceps energetic state during exercise in the KE + CHO arm compared to CHO in subjects with overt myopathy. Effects of ANK during exercise are phenotype-specific in adult GSDIIIa patients. ANK presents a promising therapy in GSDIIIa patients with a severe myopathic phenotype. TRIAL REGISTRATION NUMBER: ClinicalTrials.gov identifier: NCT03011203.


Photophysical characterization of natural cis-carotenoids.
By means of steady-state fluorescence spectroscopy we explore the photophysics of two lowest lying singlet excited states in two natural 15-cis-carotenoids, namely phytoene and phytofluene, possessing three and five conjugated double bonds (N), respectively. The results are interpreted in relation to the photophysics of all-transcarotenoids with varying N. The fluorescence of phytofluene is more Stokes-shifted relative to that of phytoene, and is ascribed to the forbidden S1-->S0 transition, with its first excited singlet state (S1) lying 3340 cm-1 below the dipole allowed second excited singlet state (S2), at 77 K. For phytoene the S2 and S1 potential surfaces are closer in energy, probably giving rise to the mixed S2 and S1 fluorescence characteristics. The origin of phytoene fluorescence is discussed and is suggested to be due to the S1-->S0 transition; with the S1 state located 1100 cm-1 below S2 at 77 K. The dependence of the fluorescence quantum yield on temperature and viscosity shows that large amplitude molecular motions are involved in the radiationless relaxation process of phytoene. The transition dipole moment of absorption and emission are parallel in phytoene and nonparallel in phytofluene.


HIV p24-specific helper T cell clones from immunized primates recognize highly conserved regions of HIV-1.
We have investigated Th cell recognition of the HIV core protein p24 by using CD4+ T cell clones derived from cynomolgus macaques immunized with hybrid HIV p24: Ty virus-like particles (VLP). T cell lines from two immunized animals responded to p24: Ty-VLP, control Ty-VLP, purified p24, and whole inactivated HIV, indicating the presence of T cells specific for p24 as well as the Ty carrier protein. The HIV determinants recognized by the T cell lines were identified by using a series of overlapping peptides synthesized according to the sequence of p24. Both T cell lines recognized peptide 11 (amino acids 235-249) and peptide 14 (amino acids 265-279). In addition, one T cell line also responded to peptide 9 (amino acids 215-229). Definitive identification of two T cell epitopes on p24 was confirmed at the clonal level: from a total of four T cell clones generated from one of the T cell lines, two respond specifically to peptide 11 and two to peptide 14. The T cell clones were CD4+ and MHC class II-restricted and secreted IL-2 in response to stimulation with purified p24, inactivated HIV or a single synthetic peptide. The specificity of the Th clones for variant peptides demonstrated cross-reactivity with two simian immunodeficiency virus isolates, but only limited responses to HIV-2 sequences. However, the Th cell epitopes identified on p24 are highly conserved between 12 HIV-1 isolates and were recognized by both of the immunized primates. These sequences may therefore be useful for priming a broadly reactive immune response to HIV-1.


Contribution of effluent organic matter (EfOM) to ultrafiltration (UF) membrane fouling: isolation, characterization, and fouling effect of EfOM fractions.
EfOM has been regarded as a major organic foulant resulting in UF membrane fouling in wastewater reclamation. To investigate fouling potential of different EfOM fractions, the present study isolated EfOM into hydrophobic neutrals (HPO-N), colloids, hydrophobic acids (HPO-A), transphilic neutrals and acids (TPI), and hydrophilics (HPI), and tested their fouling effect in both salt solution and pure water during ultrafiltration (UF). Major functional groups and chemical structure of the isolates were identified using Fourier transform infrared spectroscopy (FT-IR) and solid-state carbon nuclear magnetic resonance ((13)C NMR) analysis. The influence of the isolation process on the properties of EfOM fractions was minor because the raw and reconstituted secondary effluents were found similar with respect to UV absorbance, molecular size distribution, and fluorescence character. In membrane filtration tests, unified membrane fouling index (UMFI) and hydraulic resistance were used to quantify irreversible fouling potential of different water samples. Results show that under similar DOC level in feed water, colloids present much more irreversible fouling than other fractions. The fouling effect of the isolates is related to their size, chemical properties, and solution chemistry. Further investigations have identified that the interaction between colloids and other fractions also influences the performance of colloids in fouling phenomena.


A new HDV mouse model identifies mitochondrial antiviral signaling protein (MAVS) as a key player in IFN-β induction.
Studying hepatitis delta virus (HDV) and developing new treatments is hampered by the limited availability of small animal models. Herein, a description of a robust mouse model of HDV infection that mimics several important characteristics of the human disease is presented. HDV and hepatitis B virus (HBV) replication competent genomes were delivered to the mouse liver using adeno-associated viruses (AAV; AAV-HDV and AAV-HBV). Viral load, antigen expression and genomes were quantified at different time points after AAV injection. Furthermore, liver pathology, genome editing, and the activation of the innate immune response were evaluated. AAV-HDV infection initiated HDV replication in mouse hepatocytes. Genome editing was confirmed by the presence of small and large HDV antigens and sequencing. Viral replication was detected for 45days, even after the AAV-HDV vector had almost disappeared. In the presence of HBV, HDV infectious particles were detected in serum. Furthermore, as observed in patients, co-infection was associated with the reduction of HBV antigen expression and the onset of liver damage that included the alteration of genes involved in the development of liver pathologies. HDV replication induced a sustained type I interferon response, which was significantly reduced in immunodeficient mice and almost absent in mitochondrial antiviral signaling protein (MAVS)-deficient mice. The animal model described here reproduces important characteristics of human HDV infection and provides a valuable tool for characterizing the viral infection and for developing new treatments. Furthermore, MAVS was identified as a main player in HDV detection and adaptive immunity was found to be involved in the amplification of the innate immune response. Lay summary: Co-infection with hepatitis B and D virus (HBV and HDV, respectively) often causes a more severe disease condition than HBV alone. Gaining more insight into HDV and developing new treatments is hampered by limited availability of adequate immune competent small animal models and new ones are needed. Here, a mouse model of HDV infection is described, which mimics several important characteristics of the human disease, such as the initiation and maintenance of replication in murine hepatocytes, genome editing and, in the presence of HBV, generation of infectious particles. Lastly, the involvement of an adaptive immunity and the intracellular signaling molecule MAVS in mounting a strong and lasting innate response was shown. Thus, our model serves as a useful tool for the investigation of HDV biology and new treatments.


Androgen-responsive gene database: integrated knowledge on androgen-responsive genes.
Androgen signaling plays an important role in many biological processes. Androgen Responsive Gene Database (ARGDB) is devoted to providing integrated knowledge on androgen-controlled genes. Gene records were collected on the basis of PubMed literature collections. More than 6000 abstracts and 950 original publications were manually screened, leading to 1785 human genes, 993 mouse genes, and 583 rat genes finally included in the database. All the collected genes were experimentally proved to be regulated by androgen at the expression level or to contain androgen-responsive regions. For each gene important details of the androgen regulation experiments were collected from references, such as expression change, androgen-responsive sequence, response time, tissue/cell type, experimental method, ligand identity, and androgen amount, which will facilitate further evaluation by researchers. Furthermore, the database was integrated with multiple annotation resources, including National Center for Biotechnology Information, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes pathway, to reveal the biological characteristics and significance of androgen-regulated genes. The ARGDB web site is mainly composed of the Browse, Search, Element Scan, and Submission modules. It is user friendly and freely accessible at http://argdb.fudan.edu.cn. Preliminary analysis of the collected data was performed. Many disease pathways, such as prostate carcinogenesis, were found to be enriched in androgen-regulated genes. The discovered androgen-response motifs were similar to those in previous reports. The analysis results are displayed in the web site. In conclusion, ARGDB provides a unified gateway to storage, retrieval, and update of information on androgen-regulated genes.


Learning Parameter-Advising Sets for Multiple Sequence Alignment.
While the multiple sequence alignment output by an aligner strongly depends on the parameter values used for the alignment scoring function (such as the choice of gap penalties and substitution scores), most users rely on the single default parameter setting provided by the aligner. A different parameter setting, however, might yield a much higher-quality alignment for the specific set of input sequences. The problem of picking a good choice of parameter values for specific input sequences is called parameter advising. A parameter advisor has two ingredients: (i) a set of parameter choices to select from, and (ii) an estimator that provides an estimate of the accuracy of the alignment computed by the aligner using a parameter choice. The parameter advisor picks the parameter choice from the set whose resulting alignment has highest estimated accuracy. In this paper, we consider for the first time the problem of learning the optimal set of parameter choices for a parameter advisor that uses a given accuracy estimator. The optimal set is one that maximizes the expected true accuracy of the resulting parameter advisor, averaged over a collection of training data. While we prove that learning an optimal set for an advisor is NP-complete, we show there is a natural approximation algorithm for this problem, and prove a tight bound on its approximation ratio. Experiments with an implementation of this approximation algorithm on biological benchmarks, using various accuracy estimators from the literature, show it finds sets for advisors that are surprisingly close to optimal. Furthermore, the resulting parameter advisors are significantly more accurate in practice than simply aligning with a single default parameter choice.


Prevalence of diabetes, impaired fasting glucose and impaired glucose tolerance in patients with thalassemia major in Iran: A meta-analysis study.
This study aimed to investigate the prevalence of diabetes, impaired fasting glucose (IFG) and impaired glucose tolerance (IGT) in Iranian patients with thalassemia major. The current study has been conducted based on PRISMA guideline. To obtain the documents, Persian and English scientific databases such as Magiran, Iranmedex, SID, Medlib, IranDoc, Scopus, PubMed, ScienceDirect, Cochrane, Web of Science, Springer, Wiley Online Library as well as Google Scholar were searched until December 2015. All steps of the study were conducted by two authors independently. To the high heterogeneity of the studies, the random effect model was used to combine studies. Data were analyzed using STATA Version 11.1 software. Thirty-two studies involving 3959 major thalassemia patients with mean age of 16.83 years were included in the meta-analysis. The prevalence of diabetes in Iranian patients with thalassemia major was estimated as 9% (95% CI: 6.8-10.5) and estimated rate was 12.6% (95% CI: 6.1-19.1) for males and 10.8% (95% CI: 8.2-14.5) for females. The prevalence of IFG and IGT were 12.9% (95% CI: 7-18.8) and 9.6% (95% CI: 6.6-12.5) respectively. No relationship between serum ferritin and development of diabetes was noted. The prevalence of diabetes, IFG, and IGT in patients with thalassemia major in Iran is high and accordingly requires new management strategies and policies to minimize endocrine disorders in Iranian patients with thalassemia major. Screening of patients for the early diagnosis of endocrine disorders particularly diabetes, IFG, and IGT is recommended.


Bioaerosols chemometric characterization by laser-induced fluorescence: air sample analysis.
A laser-induced fluorescence (LIF) system was optimized using a solution of Micrococcus luteus in ethanol/water 50% (v/v) to obtain spectra in the gas phase of 46 bioaerosols. Experimental designs such as Plackett-Burman and factorial design were applied. The fluorescence spectra were treated chemometrically by principal component analysis, linear discriminant analysis and hierarchical cluster analysis to classify the microorganisms according to family, morphology and gram. The best results were obtained using LDA. The method was applied to air samples and the LIF results allowed to characterize bioaerosols reliability. The robustness of the technique was demonstrated by the identification of many bacteria.


Transient astrocyte-like NG2 glia subpopulation emerges solely following permanent brain ischemia.
NG2 glia display wide proliferation and differentiation potential under physiological and pathological conditions. Here, we examined these two features following different types of brain disorders such as focal cerebral ischemia (FCI), cortical stab wound (SW), and demyelination (DEMY) in 3-month-old mice, in which NG2 glia are labeled by tdTomato under the Cspg4 promoter. To compare NG2 glia expression profiles following different CNS injuries, we employed single-cell RT-qPCR and self-organizing Kohonen map analysis of tdTomato-positive cells isolated from the uninjured cortex/corpus callosum and those after specific injury. Such approach enabled us to distinguish two main cell populations (NG2 glia, oligodendrocytes), each of them comprising four distinct subpopulations. The gene expression profiling revealed that a subpopulation of NG2 glia expressing GFAP, a marker of reactive astrocytes, is only present transiently after FCI. However, following less severe injuries, namely the SW and DEMY, subpopulations mirroring different stages of oligodendrocyte maturation markedly prevail. Such injury-dependent incidence of distinct subpopulations was also confirmed by immunohistochemistry. To characterize this unique subpopulation of transient astrocyte-like NG2 glia, we used single-cell RNA-sequencing analysis and to disclose their basic membrane properties, the patch-clamp technique was employed. Overall, we have proved that astrocyte-like NG2 glia are a specific subpopulation of NG2 glia emerging transiently only following FCI. These cells, located in the postischemic glial scar, are active in the cell cycle and display a current pattern similar to that identified in cortical astrocytes. Astrocyte-like NG2 glia may represent important players in glial scar formation and repair processes, following ischemia.


Contribution of hydrogen peroxide to the cytotoxicity of green tea and red wines.
Green tea and red wine are claimed to have health benefits because of their high content of polyphenolic compounds, but they have also been reported as mutagenic in some test systems. In this paper, we show that a commonly used cell culture medium, Dulbecco's modified Eagle's medium (DMEM), catalyses oxidation of green tea and red wines to generate H(2)O(2). The level of H(2)O(2) produced from green tea accounted for all of the cytotoxic effects of this beverage on PCl2 cells. By contrast, H(2)O(2) was only responsible for part of the cytotoxicity of the red wines examined. Our data illustrate the danger of extrapolating from cell culture studies to predict the effects of complex beverages in vivo.


Efficacy of a 2-month dietary supplementation with polyunsaturated fatty acids in dry eye induced by scopolamine in a rat model.
This study was conducted to evaluate the efficacy of dietary n-6 and n-3 polyunsaturated fatty acids (PUFAs) in dry eye in a rat model. Female Lewis rats were fed with diets containing (1) gamma-linolenic acid (GLA), (2) eicosapentaenoic acid (EPA) + docosahexaenoic acid (DHA), or (3) GLA + EPA + DHA, for 2 months before the induction of dry eye using a continuous delivery of scopolamine and during scopolamine treatment. Two, 10 and 28 days after dry-eye induction, clinical signs of corneal dryness were evaluated in vivo using fluorescein staining. MHC II expression and mucin rMuc5AC production in the conjunctival epithelium were evaluated by immunostaining. Lipids and prostaglandins (PGs) E(1) and E(2) were analysed from the exorbital lacrimal gland (LG). Dietary PUFAs minimised the occurrence of corneal keratitis 28 days after induction of dry eye. The decrease in mucin production observed on the conjunctival epithelium was partially prevented by EPA + DHA supplementation after 2 days of scopolamine treatment, as well as by GLA and GLA + EPA + DHA diets after 10 days of treatment. The overexpression of MHC II in the conjunctival epithelium caused by dry eye induction was significantly reduced only with the GLA + EPA + DHA diet after 28 days of treatment. Dietary PUFAs were incorporated into phospholipids of the exorbital LG. Induction of dry eye was associated with a significant increase in PGE(1) and PGE(2) levels in the exorbital LG, which was inhibited by dietary EPA + DHA at 10 days (for PGE(2)) and 28 days (for PGE(1)). Dietary GLA, EPA and DHA significantly interfered with lipid homeostasis in the exorbital LG and partially prevented the course of dry eye. In particular, our results demonstrate the efficacy of the combination of n-6 and n-3 PUFAs.


Clinical Effectiveness of High-Flow Nasal Cannula in Hypoxaemic Patients during Bronchoscopic Procedures.
Bronchoscopy is a useful diagnostic and therapeutic tool. However, the clinical use of high-flow nasal cannula (HFNC) in adults with acute respiratory failure for diagnostic and invasive procedures has not been well evaluated. We present our experiences of well-tolerated diagnostic bronchoscopy as well as cases of improved saturation in hypoxaemic patients after a therapeutic bronchoscopic procedure. We retrospectively reviewed data of hypoxaemic patients who had undergone bronchoscopy for diagnostic or therapeutic purposes from October 2015 to February 2017. Ten patients (44-75 years of age) were enrolled. The clinical purposes of bronchoscopy were for diagnosis in seven patients and for intervention in three patients. For the diagnoses, we performed bronchoalveolar lavage in six patients. One patient underwent endobronchial ultrasonography with transbronchial needle aspiration of a lymph node to investigate tumour involvement. Patients who underwent bronchoscopy for therapeutic interventions had endobronchial mass or blood clot removal with cryotherapy for bleeding control. The mean saturation (SpO₂) of pre-bronchoscopy in room air was 84.1%. The lowest and highest mean saturation with HFNC during the procedure was 95% and 99.4, respectively. The mean saturation in room air post-bronchoscopy was 87.4%, which was 3.3% higher than the mean room air SpO₂ pre-bronchoscopy. Seven patients with diagnostic bronchoscopy had no hypoxic event. Three patients with interventional bronchoscopy showed improvement in saturation after the procedure. Bronchoscopy was well tolerated in all 10 cases. This study suggests that the use of HFNC in hypoxaemic patients during diagnostic and therapeutic bronchoscopy procedures has clinical effectiveness.