Power sources among district health managers in Ghana: a qualitative study.
In Ghana district directors of health services and district hospital medical superintendents provide leadership and management within district health systems. A healthy relationship among these managers is dependent on the clarity of formal and informal rules governing their routine duties. These rules translate into the power structures within which district health managers operate. However, detailed nuanced studies of power sources among district health managers are scarce. This paper explores how, why and from where district health directors and medical superintendents derive power in their routine functions. A multiple case study was conducted in three districts; Bongo, Kintampo North and Juaboso. In each case study site, a cross-sectional design was used to explore the research question. Purposive sampling technique was used to select study sites and 61 participants for interview and focus group discussion. A total of 11 interviews (3 in each district and 2 with deputy regional directors), and 9 focus group discussions (3 in each district) were conducted. Transcriptions of the voice-recordings were done verbatim, cleaned and imported into the Nvivo version 11 software for analysis using the inductive content analysis approach. The findings revealed that legitimacy provides formal power source for district health managers since they are formally appointed by the Director General of the Ghana Health Service after going through the appointment processes. These appointments serve as the primary power source for district health managers based on the existing legal and policy framework of the Ghana Health Service. Additionally, resource control especially finances and medical dominance are major informal sources of power that district health managers often employ for the management and administration of their functional areas in the health districts. The study concludes that district health managers derive powers primarily from their positions within the hierarchical structure (legitimacy) of the district health system. Secondary sources of power stems from resource control (medical dominance and financial dominance), and these power sources inform the way district health managers relate to each other. This paper recommends that district health managers are oriented to understand the power dynamics in the district health system.


Eukaryotic mismatch repair: an update.
The discovery that mutations in mismatch repair genes segregate with hereditary nonpolyposis colon cancer has awakened a great deal of interest in the study of the process of postreplicative mismatch repair. The characterisation of the principal players involved in this important metabolic pathway has been greatly facilitated by the amino acid sequence conservation among functional homologues of bacteria, yeast and mammals. The phenotypes of mismatch repair deficient mutants are also similar in many ways. In humans, mismatch repair malfunction demonstrates itself in the form of a mutator phenotype of the affected cells, an instability of microsatellite sequences and increased levels of somatic recombination. Moreover, mismatch repair deficient cells display also varying levels of tolerance to DNA damaging agents and are thought to be involved in the cell killing mediated by these agents. This article discusses some recent developments in this fast-moving field.


Regional specificity in degenerative changes in finger joints: an anatomical study using cadavers of the elderly.
Rehabilitation therapists and orthopedists regard degenerative changes (DCs) of the articular cartilage or osteoarthritis in the finger joint as the major factors that interfere with normal grasp and pinch. However, previous research has concentrated on DCs in the carpometacarpal joint (CM) of the thumb. The present study revealed some general tendencies concerning the occurrence of DCs: (1) distal joint dominance between joints in a thumb or finger; (2) distal surface dominance in a joint; (3) radioulnar dominance in a joint; and (4) fifth-finger specificity (i.e., high incidences of both DCs and osteophytes). Moreover, according to our evaluation of multiple and solitary occurrences (i.e., how many segments carried DCs) we consider that DCs in the radioulnar segments advance not only to the complementary surface (i.e., mirror-image lesion formation) but also to other adjacent segments in the early stage before progression to cartilage defects occurs in the primarily affected segment. In addition, osteophytes were not associated with the cartilage defect in the adjacent segment, but, rather, occurred at opposite or distant segments in the joint. These results are discussed in relation to lateral pinch and precision grasping, with special reference to the hypothetical stress on these movements in the finger joint.


Rational design and fabrication of MoSx nanoclusters decorated Mn0.3Cd0.7S nanorods with promoted interfacial charge transfer toward robust photocatalytic H2 generation.
In this work, we report a novel MoSx/Mn0.3Cd0.7S composite catalyst that has been designed and fabricated by in situ coupling MoSx nanoclusters with 1D Mn0.3Cd0.7S nanorods for photocatalytic H2 production. The catalyst features a 1D nanostructure with MoSx nanoclusters uniformly dispersed along the Mn0.3Cd0.7S nanorod. It was found that an intimate interface is built between MoSx nanoclusters and Mn0.3Cd0.7S nanorods thanks to the facile in situ photoreduction route, which contributes to a high-efficiency interfacial charge separation. The resulting MoSx/Mn0.3Cd0.7S photocatalyst shows a dramatically enhanced visible-light-driven photocatalytic H2 production activity compared with the control samples, owing to more efficient spatial charge separation as well as enriched active sites. This work is expected to provide an optimized structure model for rational design and constructing novel, inexpensive, efficient and stable cocatalyst/metal sulfide photocatalyst systems for H2 production.


Two nucleotide substitutions in the A-site of yeast 18S rRNA affect translation and differentiate the interaction of ribosomes with aminoglycoside antibiotics.
Two mutations in the A-site of 18S rRNA of Saccharomyces cerevisiae were investigated. The first, A1491G (rdn15), creates in yeast the same C1409-G1491 base pair as in Escherichia coli and has behaved as an antisuppressor in genetic studies. Ribosomes from rdn15 are error-restrictive but their peptidyltransferase activity remains unchanged. The second mutation, U1495C (rdnhyg1), was initially isolated as a hygromycin-resistant mutation in Tetrahymena thermophila. We show that rdnhyg1 ribosomes are slightly error prone. Mutation rdnhyg1 does not affect catalytic activity, but it affects translocation, confirming the importance of nucleotide 1495 in the ratchet-like movement of the two subunits during translation. Paromomycin, an aminoglycoside antibiotic that binds to the ribosomal A-site, induces translational misreading and causes sensitivity to yeast cells. Mutation rdn15 is shown to be highly sensitive to both effects of paromomycin, while mutation rdnhyg1 is relatively resistant. Tobramycin, another aminoglycoside, does not affect the growth of yeast cells. Like paromomycin, however, it increases the error rate in rdn15 ribosomes relative to wild-type and decreases it in rdnhyg1 ribosomes. These mutations help define the role of two crucial sites in ribosome function and distinguish the modes of action of two aminoglycosides, a useful fact in the search for new strategies in drug design.


A computer-assisted management information system for nutrition services.
A computer-assisted management information system was created to facilitate effective management of clinical nutrition services and labor resources in a 330-bed teaching and research hospital. Standards were developed for the quality and quantity of nutrition care, time required to provide nutrition care, and utilization of dietitians' time. Computer software was developed to report the volume of services provided, the need for services, and the utilization of labor hours. Data were evaluated to determine whether services were consistent with standards and to calculate a recommended number of clinical dietitian full-time equivalents for the hospital. Furthermore, the management information system was instrumental in developing a fee-for-service structure, for evenly distributing work loads among dietitians, and for monitoring adherence to standards of care.


Analysing differences in clinical outcomes between hospitals.
To examine the variation between hospitals in rates of severe intraventricular haemorrhage (IVH) in preterm babies adjusting for case mix and sampling variability. Cross sectional study of pooled data from 1995 to 1997. 24 neonatal intensive care units (NICUs) in the Australian and New Zealand Neonatal Network. 5413 infants of gestational age 24-30 weeks. Crude rates of severe (grades 3 and 4) IVH and rates adjusted for case mix using logistic regression, and for sampling variability using shrinkage estimators. The overall rate of severe IVH was 6.8%, but crude rates for individual units ranged from 2.9 to 21.4%, with interquartile range (IQR) 5.7-8.1%. Adjusting for the five significant predictor variables--gestational age at birth, 1 minute Apgar score, antenatal corticosteroids, transfer after birth, and sex--actually increased the variability in rates (IQR 5.9-9.7%). Shrinkage estimators, which adjust for differences in unit sizes and outcome rates, reduced the variation in rates (IQR 6.3-7.5%). Adjusting for case mix and using shrinkage estimators showed that one unit had a significantly higher adjusted rate than expected, while another was significantly lower. If all units could achieve an average rate equal to the 20th centile (5.74%), then 60 cases of severe IVH could be prevented in a 3 year period. The use of shrinkage estimators may have a greater impact on the variation in outcomes between hospitals than adjusting for case mix. Greater reductions in morbidity may be achieved by concentrating on the best rather than the worst performing hospitals.


Regional inhibition of monoamine oxidase activity by administration of clorgyline in rat brain.
The inhibiting effect of monoamine oxidase (MAO) activities towards 5-hydroxytryptamine, dopamine and beta-phenylethylamine by an acute and chronic administration of clorgyline was investigated in five locations of the rat brain. Inhibiting the percentage distributed unevenly in each hypothalamic nucleus and in the circumventricular areas, except the median eminence, the percentages were significantly low as compared with the hypothalamic lateral nucleus. A significant percentage of inhibition for type B MAO towards beta-phenylethylamine was noticed in the chronically administered group. In conclusion, the MAO-inhibiting effect of clorgyline administered intraperitoneally was unevenly distributed in the discrete brain nuclei and the regional difference depended upon the dose and the duration of the clorgyline administration.


Highly water soluble multi-layer graphene nanoribbons and related honey-comb carbon nanostructures.
Multi-layer graphene nanoribbons have been made highly water soluble (4.7 mg ml(-1)) and stable for the first time by repetitious derivatization with p-carboxyphenyldiazonium salt; similarly, single-walled carbon nanotubes (4.8 mg ml(-1)) and ultra-short carbon nanotubes (50 mg ml(-1)) can also be made highly soluble by the methodology.


Barriers and Facilitators to Engagement With a Weight Management Intervention in Asian Patients With Overweight or Obesity: A Systematic Review.
The obesity epidemic is a global health concern with Asian countries facing one of the most rapid rises in obesity rates. However, given the underwhelming long-term efficacy of weight loss strategies, especially in Asia, this review aimed to explore barriers and facilitators to weight management of patients with overweight and obesity in Asia. Medline, CINAHL, PsycINFO, and Web of Science were searched for articles discussing barriers and facilitators of treatment to obesity from the perspectives of both health care professionals (HCPs) and patients. Qualitative and mixed method studies from Asia were included. Key quotes were extracted, coded, and thematically analyzed according to the methodology of Thomas and Harden. A total of 26 articles were included in this review. From patient perspectives, 3 main themes were identified: factors influencing poor eating behavior, inhibiting lifestyle modifications, and facilitating lifestyle modifications. Patients highlighted several barriers including the lack of social support, physiologic limitations to exercise, and low health literacy. Rigid sociocultural norms and lack of accessible health care services, exercise facilities, and healthy food exacerbated the barriers. Facilitators to lifestyle modifications consisted of strong support systems and high health literacy. HCPs agreed that low health literacy, lack of social support, and patient motivation impeded patients' weight loss attempts but were unaware of the other barriers they faced. There are discrepancies between ideas of barriers and facilitators of HCPs and patients. A mixture of population level, primary care, and personal interventions are required to address this disparity, and enhanced health literacy can improve weight loss outcomes.


Medical image computing in diagnosis and intervention of spinal diseases.
Spinal image analysis and computer assisted intervention have emerged as new and independent research areas, due to the importance of treatment of spinal diseases, increasing availability of spinal imaging, and advances in analytics and navigation tools. Among others, multiple modality spinal image analysis and spinal navigation tools have emerged as two keys in this new area. We believe that further focused research in these two areas will lead to a much more efficient and accelerated research path, avoiding detours that exist in other applications, such as in brain and heart.


Administration of neuropsychological tests using interactive voice response technology in the elderly: validation and limitations.
Interactive voice response (IVR) systems are computer programs, which interact with people to provide a number of services from business to health care. We examined the ability of an IVR system to administer and score a verbal fluency task (fruits) and the digit span forward and backward in 158 community dwelling people aged between 65 and 92 years of age (full scale IQ of 68-134). Only six participants could not complete all tasks mostly due to early technical problems in the study. Participants were also administered the Wechsler Intelligence Scale fourth edition (WAIS-IV) and Wechsler Memory Scale fourth edition subtests. The IVR system correctly recognized 90% of the fruits in the verbal fluency task and 93-95% of the number sequences in the digit span. The IVR system typically underestimated the performance of participants because of voice recognition errors. In the digit span, these errors led to the erroneous discontinuation of the test: however the correlation between IVR scoring and clinical scoring was still high (93-95%). The correlation between the IVR verbal fluency and the WAIS-IV Similarities subtest was 0.31. The correlation between the IVR digit span forward and backward and the in-person administration was 0.46. We discuss how valid and useful IVR systems are for neuropsychological testing in the elderly.


Recent trends in methane to bioproduct conversion by methanotrophs.
Methane is an abundant and low-cost gas with high global warming potential and its use as a feedstock can help mitigate climate change. Variety of valuable products can be produced from methane by methanotrophs in gas fermentation processes. By using methane as a sole carbon source, methanotrophic bacteria can produce bioplastics, biofuels, feed additives, ectoine and variety of other high-value chemical compounds. A lot of studies have been conducted through the years for natural methanotrophs and engineered strains as well as methanotrophic consortia. These have focused on increasing yields of native products as well as proof of concept for the synthesis of new range of chemicals by metabolic engineering. This review shows trends in the research on key methanotrophic bioproducts since 2015. Despite certain limitations of the known production strategies that makes commercialization of methane-based products challenging, there is currently much attention placed on the promising further development.


Diets of large mammals in the woodlands around Lake Kariba, Rhodesia.
Large mammals that formerly occupied the Kariba basin of the Middle Zambezi Valley have, since it was flooded, been forced to live in a restricted range of deciduous woodland habitats. Feeding records in a study area on the shore of Lake Kariba showed that most herbivore species were browsers, and that only the tree components of their diets varied significantly between vegetation types. The common species varied a proportion of their diet seasonally. However, each depended upon a small range of food staples which differed from those of other species and acted as food refuges for part of the year. Diets overlapped during the wet season because of diversification, and in the late dry season because of common use of a restricted range of plants remaining green. There was a good correlation between the ability of a species to avoid dietary overlap and its biomass in the study area. The chance of interspecific competition occurring increased in the late dry season when most of the species would formerly have migrated from the study area to the flood plain. Despite the enforced occupation of only part of their former annual range the more common herbivores maintained a considerable degree of ecological separation through utilisation of different foods.


Neodymium:YAG laser phacofracture: an aid to phacoemulsification.
A series of approximately 1,000 cases of nucleus pretreatment prior to phacoemulsification is presented. Current results indicate that the technique eases phacoemulsification, especially of hard nuclei, and has little risk when done properly.


The effect of a multicomponent professional development training on the beliefs and behaviors of community health educators concerning food irradiation.
Beliefs have a significant effect on the health behaviors of individuals and educators; however, they can be difficult to change. The purpose of this study was to determine if exposing community health educators, specifically family and consumer sciences county extension educators, to a multicomponent professional development training on food irradiation could change their beliefs and behaviors. This study compared the food irradiation beliefs and educational programming of educators who participated in a professional development training with those who did not. Results indicated that the training significantly improved the food irradiation beliefs of participants. In addition, the number of participants who provided food irradiation education significantly improved compared with educators who had not attended the training. These results suggest that this type of professional development training format can significantly affect beliefs and could increase the amount of food irradiation information available to consumers through community health educators.


[Adult pulmonary Langerhans' cell histiocytosis: approach to the reality of the Spanish population].
Pulmonary Langerhans cell histiocytosis (PLCH) is a rare respiratory disease closely associated with smoking. The aim of the study is to know the reality of PLCH in the Catalonian population. We included all patients diagnosed of PLCH in 8 different Catalonian hospitals from April 2011 to December 2012. Epidemiologic, clinical, radiological, functional, diagnostic and therapeutic variables were collected, both at the beginning and at the last visit of the process. Thirty two patients were included. Overall, patients were middle-aged (38 years), with no gender predominance. Most of them were smokers (71.9%). The main pulmonary function values (forced vital capacity 75.9%, forced expiratory volume in one second 68.9%, diffusing capacity of the lung for carbon monoxide 59.6%) and radiological findings (cysts 89.7%, nodules 60.3%) showed no significant changes during evolution. However, the disease progressed with a worsening lung function in 34.4% of cases. The analysis of the functional outcome (forced vital capacity, forced expiratory volume in one second, and diffusing capacity of the lung for carbon monoxide) did not reveal significant differences according to the persistence or not of smoking (P<0.05). Our patients with PLCH had similar clinical and radiological characteristics to those described in other populations. The results observed regarding the relationship with smoking suggest that there may be other factors beyond smoking, which influence on the evolution of the disease. It therefore seems advisable to design prospective studies to investigate this aspect.


Protective autophagy promotes the resistance of HER2-positive breast cancer cells to lapatinib.
Lapatinib, a tyrosine kinase inhibitor of HER2/EGFR, can inhibit the proliferation of HER2-positive breast cancer cells. Additionally, the combination of lapatinib and chemotherapy can markedly prolong patient survival time. However, the clinical therapeutic effect of lapatinib is severely limited by drug resistance. We previously found that brief treatment with lapatinib induced both apoptosis and autophagy in HER2-positive breast cancer cells. Additionally, the apoptosis induced by lapatinib was dependent on autophagy. In our current study, however, we used extended treatment of HER2-positive breast cancer cells with lapatinib to confirm the presence of protective autophagy in the previously established lapatinib-resistant cells. Specifically, we found that inhibition of autophagy could reduce the proliferation, DNA synthesis, and colony-forming capacity of resistant cells. Thus, autophagy is a potential novel therapeutic target for reversing lapatinib resistance of HER2-positive breast cancer cells. Our data provide clear, novel evidence of both anti-apoptotic and pro-apoptotic functions of autophagy in breast cancer during lapatinib treatment.


Responses to questions by medical students about family practice.
Medical students frequently have questions about the specialty of family practice. Responses to 30 questions commonly asked about family practice are presented with a review of recent literature. These responses may assist medical students and their advisors in considering the choice of family practice as a career.


Construction of a novel Staphylokinase (SAK) mutant with low immunogenicity and its evaluation in rhesus monkey.
The heterologous nature of SAK, a thrombolytic drug, elicits high titers of neutralizing antibodies, which limits its clinical use. Here, we aim to establish a SAK mutant with equivalent activity to the wild type but reduced antigenicity, which may allow for multiple injections. Biosun software was used to predict SAK antigenic epitopes, and several main epitopes were modified by gene deletion and mutation. Ten SAK mutants were constructed, and their thrombolytic activity and immunogenicity were analyzed in vitro. SAK6, with a high expression level (45%), similar thrombolytic activity, and lower antibody reaction, was chosen for in vivo analysis in rhesus monkey. In the nearly 8-month experimental period, the antibody level of the SAK6 group was significantly lower than that of the SAK group. Moreover, only 5% of SAK activity was retained, whereas 75.6% of SAK6 activity was retained after incubating with respective antiserum. Overall, these results demonstrated that SAK6, established through comprehensive site-directed mutagenesis program, had identical thrombolytic activity to SAK, low immunogenicity, and less side effects, demonstrating its efficient clinical potential for thrombus disease.


Monitoring for Extra-Intestinal Cancers in IBD.
Multiple studies have demonstrated an increased risk for extra-intestinal cancers in inflammatory bowel disease (IBD) patients, mainly from treatment modalities. Prominent cancers that are related to IBD treatment include the following: lymphoproliferative disorders associated with thiopurine use, hepatosplenic T cell lymphoma primarily in younger male patients on thiopurines and anti-tumor necrosis factor (TNF) agents, non-melanoma skin cancers in patients treated with thiopurines and anti-TNF agents, and melanomas in patients who are on monotherapy with anti-TNF agents. In addition, women with IBD may have higher rates of cervical dysplasia and cervical cancer. The focus of this review is to provide a comprehensive overview on extra-intestinal cancers in IBD patients and how to monitor for these malignancies.


Adenosine deaminase 2 produced by infiltrative monocytes promotes liver fibrosis in nonalcoholic fatty liver disease.
Elevated circulating activity of adenosine deaminase 2 (ADA2) is associated with liver fibrosis in nonalcoholic fatty liver disease (NAFLD). In the liver of NAFLD patients, ADA2-positive portal macrophages are significantly associated with the degree of liver fibrosis. These liver macrophages are CD14- and CD16-positive and co-express chemokine receptors CCR2, CCR5, and CXCR3, indicating infiltrative monocyte origin. Human circulatory monocytes release ADA2 upon macrophage differentiation in vitro. When stimulated by recombinant human ADA2 (rhADA2), human monocyte-derived macrophages demonstrate upregulation of pro-inflammatory and pro-fibrotic genes, including PDGF-B, a key pro-fibrotic cytokine. This PDGF-B upregulation is reproduced by inosine, the enzymatic product of ADA2, but not adenosine, and is abolished by E359N, a loss-of-function mutation in ADA2. Finally, rhADA2 also stimulates PDGF-B production from Kupffer cells in primary human liver spheroids. Together, these data suggest that infiltrative monocytes promote fibrogenesis in NAFLD via ADA2-mediated autocrine/paracrine signaling culminating in enhanced PDGF-B production.


Elimination of paradoxical sleep by lesions of the pontine gigantocellular tegmental field in the cat.
Bilateral lesions of the pontine gigantocellular tegmental field in the cat resulted in the complete elimination of paradoxical sleep during 3 weeks postoperative recording. The tonic muscular atonia, normally characteristic of this state, was absent. The phasic components, rapid eye movements and ponto-geniculooccipital (PGO) spikes, did not occur in association with an activated EEG, as they normally do in paradoxical sleep. In fact, PGO spikes were virtually absent immediately after the lesion and were only secondarily apparent as isolated phenomena during slow wave sleep to represent in total daily number 5% of normal the first week and 15% of normal the third week after the lesion. These results indicate that neurons whose perikarya and/or processes are located within the pontine gigantocellular tegmental field and which are not part of the noradrenaline locus coeruleus complex, are critical for paradoxical sleep.


PTP1B up-regulates EGFR expression by dephosphorylating MYH9 at Y1408 to promote cell migration and invasion in esophageal squamous cell carcinoma.
Esophageal squamous cell carcinoma (ESCC) is one of the most common cancers worldwide. Protein tyrosine phosphatase 1B (PTP1B) is a member of protein tyrosine phosphatases (PTPs) family. In our previous work, PTP1B was found to be overexpressed in ESCC tissues and made contributions to the the cell migration and invasion as well as lung metastasis of ESCC. In this study, we explored the underlying molecular mechanisms. PTP1B enhanced cell migration and invasion by promoting epidermal growth factor receptor (EGFR) expression in ESCC, which was relied on phosphatase activity of PTP1B. Using GST-pulldown combined with LC/MS/MS, we found that nonmuscle myosin IIA (MYH9) was a novel substrate of PTP1B in ESCC cells. PTP1B dephosphorylated MYH9 at Y1408, by which PTP1B up-regulated EGFR expression and enhanced cell migration and invasion in ESCC. In conclusion, our study first reported that PTP1B was the positive regulator of EGFR by dephosphorylating MYH9 at Y1408 to promote cell migration and invasion, which revealed the regulatory mechanism of PTP1B-MYH9-EGFR axis in ESCC.


FBXO32 activates NF-κB through IκBα degradation in inflammatory and genotoxic stress.
In response to diverse stresses, the canonical NF-κB pathway gets activated primarily to protect the cells and maintain their genomic integrity. It activates the cell cycle checkpoints allowing the cells with limited damage to restore a normal life cycle. One of the key events in activation of the canonical NF-κB pathway is the selective proteasomal degradation of IκBα. It has been previously shown that F-box protein βTRCP1 has limited role in directing the proteasomal degradation of IκBα during stress conditions. Here, we report another member of F-box family proteins, FBXO32, as a potential activator of NF-κB signaling during genotoxic stress and inflammatory response. Following genotoxic or inflammatory stress, FBXO32 is stabilized, which leads to polyubiquitination and proteasome mediated degradation of IκBα. We also found that FBXO32 is required for physiological regulation of IκBα levels in unstressed cells. Thus, we decipher the new role of FBXO32 in regulation of NF-κB signaling pathway.


Quantitative morphometry of renal biopsies prior to cyclosporine in nephrotic syndrome.
Use of cyclosporine (CsA) in the management of children with steroid-resistant (SRNS) and steroid-dependent (SDNS) nephrotic syndrome has become increasingly popular in recent years. Although most children receive a renal biopsy prior to initiation of CsA, the relationship between initial renal histology and the subsequent clinical response to CsA is not known. We analyzed the correlation between pre-CsA segmental and global glomerular scarring and interstitial fibrosis and the subsequent response to CsA in 23 children (5.6+/-1.0 years, Mean+/-SEM) with SDNS (n=8) and SRNS (n=15) treated with CsA for 24.2+/-3.8 months and followed for 28.0+/-4.1 months. Complete remission was obtained in 78% of patients within 67.6+/-16 days, while 18% had a partial response and 4% no response. Quantitative histological analysis revealed a trend toward partial rather than complete response with increasing segmental glomerular (P=0.13), global glomerular (P=0.05), and interstitial (P=0.08) scarring, and among patients with minimal change nephrotic syndrome versus IgM nephropathy versus focal segmental glomerulosclerosis. Among complete responders, linear regression analyses revealed no correlation between time to response and pre-CsA glomerular or interstitial scarring. We conclude that increased glomerular or interstitial scarring on a pre-CsA renal biopsy tends to correlate with a partial, rather than complete, response to CsA in childhood nephrotic syndrome.


Determinants of reproductive success in female Columbian ground squirrels.
We studied the reproductive success of female Columbian ground squirrels (Spermophilus columbianus) in southwestern Alberta for nine years. We defined reproductive success as the number of offspring surviving their first hibernation, classified as yearlings. The number of weaned juveniles explained one third of the variance in number of yearlings at emergence from their first hibernation the following spring, and much of the variance in individual reproductive success originated after weaning. Weight of adult females at emergence from hibernation was correlated with annual reproductive success. The mother's survival beyond weaning and the subsequent winter's snow accumulation had positive effects on annual reproductive success, whereas population density and summer temperature had negative effects. We found no effects on annual reproductive success of date of litter emergence, weight at emergence as a yearling, presence or absence of adult kin, distance from the natal site, location within the study area, winter temperature or summer precipitation. Age of first breeding did not affect lifetime reproductive success, which ranged from 0 to 19 yearlings produced over a lifetime. The greatest source of variation in lifetime reproductive success for females surviving to breeding age was offspring survival, followed by reproductive lifespan.


Anti-inflammatory and antithrombotic effects of nicotine exposure in oral contraceptive-induced insulin resistance are glucocorticoid-independent.
Reports showed that estrogen-progestin oral contraceptive (COC) or tobacco smoking causes increased risk of cardiovascular diseases (CVD) in premenopausal women. Studies also suggest that nicotine, a major tobacco alkaloid, may worsen or improve atherothrombotic CVD. Altered hemorheology, prothrombotic and pro-inflammatory biomarkers, have been implicated in the development of atherothrombotic CVD events. However, the effect of non-smoking nicotine exposure on these biomarkers during COC treatment is not yet established. We therefore sought to determine the effects of nicotine exposure during COC treatment on these biomarkers, and also tested the hypothesis that the nicotine effects would be glucocorticoid-dependent. Female Sprague-Dawley rats aged 10 weeks were given (po) vehicle, low-dose nicotine (0.1mg/kg) or high-dose nicotine (1.0mg/kg) with or without COC steroids (5.0μg/kg ethinylestradiol and 25.0μg/kg levonorgestrel) daily for 6 weeks. COC treatment or nicotine exposure led to increased insulin resistance (IR), hemorheological (blood viscosity, hematocrit and plasma viscosity), prothrombotic (plasminogen activator inhibitor-1), pro-inflammatory (uric acid, C-reactive protein, neutrophil/lymphocyte and platelet/lymphocyte ratios) biomarkers and corticosterone. However, these effects except that on corticosterone were abrogated by nicotine exposure during COC treatment. Our study indicates that nicotine- or COC-induced IR may be mediated via inflammatory/thrombotic pathway. The results imply that nicotine exposure could impact negatively on atherothrombotic biomarkers in COC non-users, whereas the impact in COC users could be positive. The results also suggest that the anti-inflammatory, antithrombotic and blood viscosity-lowering effects of nicotine exposure during COC use is circulating glucocorticoid-independent.


Disposable electrochemical genosensor for the simultaneous analysis of different bacterial food contaminants.
This paper deals with the use of an electrochemical genosensor array for the rapid and simultaneous detection of different food-contaminating pathogenic bacteria. The method includes PCR amplification followed by analysis of the amplicons by hybridisation with toxin-specific oligonucleotide probes. A screen-printed array of four gold electrodes, modified using thiol-tethered oligonucleotide probes, was used. Unmodified PCR products were captured at the sensor interface via sandwich hybridisation with surface-tethered probes and biotinylated signaling probes. The resulting biotinylated hybrids were coupled with a streptavidin-alkaline phosphatase conjugate and then exposed to an alpha-naphthyl phosphate solution. Differential pulse voltammetry was finally used to detect the alpha-naphthol oxidation signal. Mixtures of DNA samples from different bacteria were detected at the nanomolar level without any cross-interference. The selectivity of the assay was also confirmed by the analysis of PCR products unrelated to the immobilised probes.


Increased uncoupling protein-2 and -3 gene expressions in skeletal muscle of STZ-induced diabetic rats.
Streptozotocin (STZ)-induced diabetic animals are vulnerable to cold stress. Uncoupling proteins (UCPs) play an important role in regulating thermogenesis. We investigated the gene expressions of UCPs in brown adipose tissue (BAT), white adipose tissue (WAT), liver and gastrocnemius muscle of STZ-diabetic rats using Northern blot. UCP-1, -2 and -3 mRNA expressions in BAT were all remarkably lower in STZ-diabetic rats than those in control rats. Both UCP-2 and -3 gene expressions in gastrocnemius muscle were substantially elevated in STZ-diabetic rats and insulin treatment restored UCP gene expressions to normal levels. These results suggest that in STZ-diabetic rats, the overexpression of UCP-2 and UCP-3 in skeletal muscle provides a defense against hypothermogenesis caused by decreased UCPs in BAT.


Immunological detection of penicillin-binding protein 2' in clinical isolates of methicillin-resistant Staphylococcus aureus and Staphylococcus epidermidis.
The additional penicillin-binding protein (PBP 2') that is important in determining intrinsic resistance in methicillin-resistant strains of Staphylococcus aureus (MRSA) has been detected immunologically in strains from a variety of world-wide locations. This additional protein has also been definitively identified both immunologically and as a PBP in methicillin-resistant strains of S. epidermidis (MRSE). The assay described is rapid, specific and sensitive and has been used to detect PBP 2' in S. haemolyticus but not in beta-lactam resistant Streptococci.


Cataract extraction in a major ophthalmic hospital: day-case or overnight stay?
Cataract surgery is increasing in Australia and represents a significant burden on limited health resources. This study examines the frequency and outcomes of cataract surgery for patients who were hospitalized overnight compared with those treated as day surgery cases. Medical records of 671 consecutive admissions for cataract surgery at the Royal Victorian Eye and Ear Hospital were reviewed. Data analysed included demographic features, insurance status, length of hospitalization, ophthalmic conditions, medical conditions, social problems and planned surgical technique. Ophthalmic and anaesthetic complications, active ophthalmic and medical interventions were also studied. Of the 671 patient admissions for cataract during the study period, 226 (33.4%) were hospitalized overnight. Factors significantly associated with overnight hospitalization in univariate analyses include older age, female sex, country residence, Veterans' Affairs insurance, monocular vision status, pre-existing ischaemic heart disease, pre-existing asthma/chronic obstructive lung disease, absence of carer, transportation problems, planned extra-capsular cataract extraction technique, ophthalmic complications and active ophthalmic and/or medical interventions. After adjusting for possible confounding factors using backwards stepwise multivariate logistic regression models all except pre-existing ischaemic heart disease and ophthalmic complications were significantly associated with overnight admission for cataract surgery. In total, 14 cases (2.1%) needed active ophthalmic and/or medical interventions, 13 overnight cases and one day case. These data suggest that many patients who are hospitalized overnight for cataract surgery could be safely treated as day cases. Such a shift in the pattern of care for cataract surgery could provide a significant potential for health care savings.


In vitro activity of cepharanthine hydrochloride against clinical wild-type and lamivudine-resistant hepatitis B virus isolates.
Hepatitis B virus (HBV) infection causes major public health problems worldwide. The clinical limitation of current antiviral drugs for HBV, such as lamivudine, is the emergence of drug-resistant viral strains during prolonged antiviral therapy. Cepharanthine hydrochloride (CH), a natural alkaloid-derived compound, has been reported to possess potent activity against various viruses. The present study was performed to evaluate the in vitro activity of CH against clinical wild-type and lamivudine-resistant HBV isolates in transiently transfected cells. HBV DNA was extracted from serum samples collected both before lamivudine therapy and at the time of viral breakthrough and was amplified by polymerase chain reaction (PCR). The amplicons were cloned into a novel expression vector, pHY106, which can initiate the intracellular HBV replication cycle after cell transfection. Following transfection of the cloned amplicon into HepG2 cells, a drug susceptibility assay was performed. The level of viral antigen, HBeAg, was determined by enzyme-linked immunosorbent assay (ELISA). Quantitative real-time PCR (Q-PCR) was used for determining the amount of intracellular HBV DNA. Heat stress cognate 70 (Hsc70), a host protein required for HBV replication, was also analyzed by reverse transcription PCR (RT-PCR) to explore the possible antiviral mechanism of CH. The results showed that CH inhibited replication and HBeAg production by either wild-type or lamivudine-resistant HBV clinical isolates in a dose-dependent manner. The Hsc70 mRNA was also downregulated significantly. In conclusion, CH is active against both wild-type and lamivudine-resistant HBV clinical isolates, and its activity may be associated with its inhibition of host Hsc70.


Mycobacterium tuberculosis inhibits human innate immune responses via the production of TLR2 antagonist glycolipids.
Mycobacterium tuberculosis is a major human pathogen that is able to survive inside host cells and resist immune clearance. Most particularly, it inhibits several arms of the innate immune response, including phagosome maturation or cytokine production. To better understand the molecular mechanisms by which M. tuberculosis circumvents host immune defenses, we used a transposon mutant library generated in a virulent clinical isolate of M. tuberculosis of the W/Beijing family to infect human macrophages, utilizing a cell line derivative of THP-1 cells expressing a reporter system for activation of the transcription factor NF-κB, a key regulator of innate immunity. We identified several M. tuberculosis mutants inducing a NF-κB activation stronger than that of the wild-type strain. One of these mutants was found to be deficient for the synthesis of cell envelope glycolipids, namely sulfoglycolipids, suggesting that the latter can interfere with innate immune responses. Using natural and synthetic molecular variants, we determined that sulfoglycolipids inhibit NF-κB activation and subsequent cytokine production or costimulatory molecule expression by acting as competitive antagonists of Toll-like receptor 2, thereby inhibiting the recognition of M. tuberculosis by this receptor. Our study reveals that producing glycolipid antagonists of pattern recognition receptors is a strategy used by M. tuberculosis to undermine innate immune defense. Sulfoglycolipids are major and specific lipids of M. tuberculosis, considered for decades as virulence factors of the bacilli. Our study uncovers a mechanism by which they may contribute to M. tuberculosis virulence.


Heritability of daytime cortisol levels and cortisol reactivity in children.
Individuals differ widely in cortisol output over the day and cortisol reactivity to challenge, both of which are relevant to disease risk. There is limited evidence concerning the heritability of these differences, so we evaluated the heritability of cortisol levels in the afternoon and cortisol reactivity using a twin design. The study involved 80 monozygotic (MZ) and 70 dizygotic (DZ) same-sex twin pairs aged 11.2 years on average. Salivary cortisol was measured in the afternoon at home before and after playing a computer game. Ratings of excitement and upset were also obtained, and objective task performance was assessed. Salivary cortisol levels averaged 4.08 (S.D. 2.3) nmol/l at pretask baseline, and declined on average over the session to 3.45 (1.9) nmol/l immediately after the tasks and 2.87 (1.6) nmol/l 10min later. There were, however, marked individual differences, with cortisol reactivity (difference between pretask baseline and post-task 1) ranging from +4.53 to -6.23nmol/l. Intra-class correlations for all the cortisol parameters were substantially greater for MZ (range 0.41-0.57) than for DZ (0.11-0.29) twin pairs. Quantitative genetic modelling confirmed significant heritability for pretask baseline cortisol (58%), the two post-task values (60 and 56%), and cortisol reactivity (44%). The study lacked power for assessing sex differences. Subjective reports of excitement were also somewhat heritable, but there was little covariation of cortisol and subjective responses, so genetic influences on covariation could not be tested. These findings indicate that individual differences in children's cortisol levels recorded before tasks and cortisol reactivity to behavioural challenges are influenced by genetic factors.


Crystal structure of bis-(aceto-nitrile-κN)bis-(4-benzoyl-pyridine-κN)bis-(thio-cyanato-κN)cobalt(II).
The crystal structure of the title compound, [Co(NCS)2(C2H3N)2(C12H9NO)2], consists of cobalt(II) cations that are octa-hedrally coordinated by the N atoms of two terminal thio-cyanate anions, two aceto-nitrile mol-ecules and two 4-benzoyl-pyridine ligands. The discrete complexes are located on centres of inversion. They are connected by weak inter-molecular C-H⋯O and C-H⋯S hydrogen-bonding inter-actions between one of the pyridine H atoms and the carbonyl O atom, and between one of the methyl H atoms of the aceto-nitrile mol-ecule and the thio-cyanate S atoms into layers parallel to (101). No pronounced inter-molecular inter-actions are observed between these layers.


The Prediction of Fissure Integrity by Quantitative Computed Tomography Analysis.
Incomplete interlobar fissure may increase the difficulty of thoracoscopic lobectomy. Herein, we compared the accuracy of visual versus quantitative analysis to predict fissure integrity in lung cancer patients undergoing thoracoscopic lobectomy and evaluated the effects of fissure integrity on surgical outcome. This was a single-center retrospective study including consecutive patients undergoing VATS (video-assisted thoracoscopic surgery) lobectomy for lung cancer. The target interlobar fissures were classified as complete or incomplete by visual and quantitative analysis. Using the intraoperative finding as the reference method, the diagnostic accuracy of the two methods to define fissure completeness (dependent variable) was calculated and statistically compared. Yet, we evaluated differences in postoperative outcomes between patients with complete and incomplete fissure integrity. A total of 93 patients were included in the study; 33/93 (36%) presented complete fissure. Visual and quantitative analyses correctly identified complete fissure in 19/33 (57%) and 29/33 (88%) patients, respectively, and incomplete fissure in 56/60 (93%) and 58/60 (96%) patients, respectively. Quantitative analysis had better diagnostic accuracy than visual analysis (81 vs. 93%; p = 0.01). Patients with incomplete fissure compared with those with complete fissure had a higher conversion rate (6 vs. 13%; p = 0.43), higher persistent air leak rate (0/33 vs. 14/60; p = 0.03), and longer hospitalization (12.6 ± 3.8 vs. 7.1 ± 2.4 days; p = 0.01). Quantitative analysis accurately predicted the fissures' integrity; it may be useful for selecting suitable cases for thoracoscopic lobectomy especially for surgeons with limited minimally invasive experience.


The treatment of migraine with variable frequency photo-stimulation.
Variable Frequency Photo-stimulation (VFP) goggles are a form of portable stroboscope, using red Light Emitting Diodes to alternately illuminate the right and left eyes, with the eyelids closed, at a rate of 0.5 to 50 Hz. VFP goggles were used in the treatment of 7 patients with migraine headache. Of 50 migraine headaches recorded, 49 were rated by the patient as being helped and 36 as being stopped by using the VFP goggles. Treatment reduced the median duration of headache in all patients. The interval between migraine headaches appeared to be increased in the two cases with a follow up of more than 18 months. We conclude in this preliminary study that VFP appears to be effective in the treatment of migraine.


Tissue-specific transcriptome analysis within the maturing sugarcane stalk reveals spatial regulation in the expression of cellulose synthase and sucrose transporter gene families.
Sugarcane (Saccharum spp. hybrids) accumulates high concentrations of sucrose in its mature stalk and a considerable portion of carbohydrate metabolism is also devoted to cell wall synthesis and fibre production. We examined tissue-specific expression patterns to explore the spatial deployment of pathways responsible for sucrose accumulation and fibre synthesis within the stalk. We performed expression profiling of storage parenchyma, vascular bundles and rind dissected from a maturing stalk internode of sugarcane, identifying ten cellulose synthase subunit genes and examining significant differences in the expression of their corresponding transcripts and those of several sugar transporters. These were correlated with differential expression patterns for transcripts of genes encoding COBRA-like proteins and other cell wall metabolism-related proteins. The sugar transporters genes ShPST2a, ShPST2b and ShSUT4 were significantly up-regulated in storage parenchyma while ShSUT1 was up-regulated in vascular bundles. Two co-ordinately expressed groups of cell wall related transcripts were also identified. One group, associated with primary cell wall synthesis (ShCesA1, ShCesA7, ShCesA9 and Shbk2l3), was up-regulated in parenchyma. The other group, associated with secondary cell wall synthesis (ShCesA10, ShCesA11, ShCesA12 and Shbk-2), was up-regulated in rind. In transformed sugarcane plants, the ShCesA7 promoter conferred stable expression of green fluorescent protein preferentially in the storage parenchyma of the maturing stalk internode. Our results indicate that there is spatial separation for elevated expression of these important targets in both sucrose accumulation and cell wall synthesis, allowing for increased clarity in our understanding of sucrose transport and fibre synthesis in sugarcane.


Dezocine prevents sufentanil-induced cough during general anesthesia induction: a meta-analysis of randomised controlled trials.
Sufentanil is one of the opioids currently used to induce general anesthesia, and cough is one of the most common complications. Many drugs have been used to prevent sufentanil-induced cough (SIC), and dezocine is one of them. Dezocine is an analgesic, acting as partial antagonist of κ-receptors and agonist of μ-receptors. The purpose of our meta-analysis is to evaluate the efficacy of dezocine on SIC. We searched multiple databases including PubMed, Embase, ScienceDirect, the Cochrane Library, and China National Knowledge Infrastructure databases (CNKI) to identify studies that met the inclusion criteria. This meta-analysis focused on the incidence and severity of SIC after dezocine intervention, as well as adverse effects. This meta-analysis was registered on PROSPERO with reference number ID: CRD 42020144943. Five randomised controlled trials (RCTs) were identified, including 890 patients. Each study was a comparison of dezocine with an equal volume of 0.9% saline. When the injection dose of dezocine was 0.1 mg/kg, the incidence (pooled risk ratio (RR) = 0.03, [95% CI: 0.02 to 0.07], P < 0.00001, I2 = 0%) and severity (mild: RR = 0.07, [95% CI: 0.03 to 0.18], P < 0.00001, I2 = 0%; moderate: RR = 0.05, [95% CI: 0.02 to 0.16], P < 0.00001, I2 = 0%; severe: RR = 0.04, [95% CI: 0.01 to 0.16], P < 0.00001, I2 = 0%) of SIC were significantly decreased. There were no statistically significant differences in vital signs between the two groups based on the results of the pooled analysis. This meta-analysis showed that dezocine significantly reduced the incidence and severity of SIC in the induction of general anesthesia, but had no significant effect on vital signs. More high-quality RCTs are needed to complement existing conclusions.


Kinetics of precursor cleavage at the dibasic sites. Involvement of peptide dynamics.
The presence in the P'1 position relative to the LysArg doublet of either Phe, Tyr or Trp residues affects only pro-OT/Np(7-15) flexibility. This has a measurable effect on the dynamics of the peptide. Since the same modifications have a major influence on the K(m) and V(max) values of the peptide cleavage, these kinetic parameters should depend on the peptide substrate motions. Therefore, the primary kinetic contribution of substrate cleavage should arise from substrate dynamics rather than from the enzyme.


Traceability of 'Limone di Siracusa PGI' by a multidisciplinary analytical and chemometric approach.
Food traceability is increasingly relevant with respect to safety, quality and typicality issues. Lemon fruits grown in a typical lemon-growing area of southern Italy (Siracusa), have been awarded the PGI (Protected Geographical Indication) recognition as 'Limone di Siracusa'. Due to its peculiarity, consumers have an increasing interest about this product. The detection of potential fraud could be improved by using the tools linking the composition of this production to its typical features. This study used a wide range of analytical techniques, including conventional techniques and analytical approaches, such as spectral (NIR spectra), multi-elemental (Fe, Zn, Mn, Cu, Li, Sr) and isotopic ((13)C/(12)C, (18)O/(16)O) marker investigations, joined with multivariate statistical analysis, such as PLS-DA (Partial Least Squares Discriminant Analysis) and LDA (Linear Discriminant Analysis), to implement a traceability system to verify the authenticity of 'Limone di Siracusa' production. The results demonstrated a very good geographical discrimination rate.


Polyacrylamide grafted cellulose as an eco-friendly flocculant: Key factors optimization of flocculation to surfactant effluent.
The discharge of effluents from surfactant manufacturers is giving rise to increasingly serious environmental problems. In order to develop the eco-friendly flocculation materials to achieve effective removal of pollutants from the surfactant effluents, the bamboo pulp cellulose from Phyllostachys heterocycla is employed as the skeleton material to synthesize an eco-friendly bamboo pulp cellulose-g-polyacrylamide (BPC-g-PAM) for flocculation. The BPC-g-PAM is used with the metal ions as the coagulant to treat the effluent from a surfactant manufacturer. The response surface methodology coupled with Box-behnken design is employed to optimize the key factors of coagulation-flocculation. The results show that the combination of Fe(3+) with BPC-g-PAM achieves the best coagulation-flocculation performance like, the fast treatment time, minimum coagulant and BPC-g-PAM dosages compared with the other two combinations of Al(3+) with BPC-g-PAM and Ca(2+) with BPC-g-PAM. Therefore, the combination of Fe(3+) with BPC-g-PAM is expected to promote its application for the pollution control in the surfactant manufacturers.


A hypothesis concerning the structural relationship between memory and emotion.
Limbic and temporal lobe structures have long been implicated as the anatomic substratum for emotion. Recent investigations have localized the processing of short term memory to a virtually identical anatomical locus. It would seem logical that the same structures subserve both functions if one assumes that the strength of the memory trace is dependent on the emotional value of the information for the organism. Under this hypothesis, the hippocampus emotionally weights sensory data as a preliminary to incorporation of such material into long term memory. It is not necessary, nor is it likely, that long term memory reside within the temporal lobe. Previous associations stored in remote memory can be accessed by the temporal lobe and these may serve to modulate the weighting of the present stimulus. Some of the functions of the temporal lobe can be modeled by a simple electronic filter; the addition of a single feedback loop to this system can account for other properties inherent in the hypothesis. The anatomical connections of the system would permit it to be an early and efficient repressor, in the pyschodynamic sense.


The corticotrophin releasing factor test: a valuable and rapid procedure for the differential diagnosis of hirsutism.
Plasma concentrations of adrenocorticotrophic hormone, cortisol, 17-hydroxyprogesterone, testosterone, delta 4-androstenedione and dehydroepiandrosterone sulphate were determined in 10 regularly menstruating hirsute women 0, 15, 30, 60, 90 and 120 min after intravenous injection of 100 micrograms corticotrophin releasing factor. The baseline concentrations of adrenocorticotrophic hormone, cortisol, progesterone and dehydroepiandrosterone sulphate were within the normal range, whereas the mean delta 4-androstenedione and testosterone concentrations were mildly elevated. The administration of corticotrophin releasing factor induced a rapid and significant increase in plasma concentrations of adrenocorticotrophic hormone, cortisol, 17-hydroxyprogesterone and delta 4-androstenedione after 60 min, whereas dehydroepiandrosterone sulphate and testosterone concentrations were not significantly elevated. These findings suggest that the corticotrophin releasing factor test can readily be used as an out-patient procedure to exclude adrenal hirsutism.


The mechanism of acentrosomal spindle assembly in human oocytes.
Meiotic spindle assembly ensures proper chromosome segregation in oocytes. However, the mechanisms behind spindle assembly in human oocytes remain largely unknown. We used three-dimensional high-resolution imaging of more than 2000 human oocytes to identify a structure that we named the human oocyte microtubule organizing center (huoMTOC). The proteins TACC3, CCP110, CKAP5, and DISC1 were found to be essential components of the huoMTOC. The huoMTOC arises beneath the oocyte cortex and migrates adjacent to the nuclear envelope before nuclear envelope breakdown (NEBD). After NEBD, the huoMTOC fragments and relocates on the kinetochores to initiate microtubule nucleation and spindle assembly. Disrupting the huoMTOC led to spindle assembly defects and oocyte maturation arrest. These results reveal a physiological mechanism of huoMTOC-regulated spindle assembly in human oocytes.


Oesophageal dysmotility, delayed gastric emptying and autonomic neuropathy correlate to disturbed glucose homeostasis.
Among diabetic patients, glucose homeostasis may be affected by abnormal gastrointestinal motility and autonomic neuropathy. This study analysed whether oesophageal dysmotility, delayed gastric emptying or autonomic neuropathy affect glucose homeostasis. Oesophageal manometry and gastric emptying scintigraphy were performed in 20 diabetic patients. Heart-rate variation during deep breathing (expiration/inspiration [E/I] ratio) and continuous subcutaneous glucose concentrations for a period of 72 h were also monitored in the same patients. Oesophageal dysmotility was found in eight of 14 patients. Eleven of 20 patients had delayed gastric emptying (abnormal gastric emptying half-time [T (50)]) and nine of 18 had an abnormal E/I ratio. Complaints of abdominal fullness were predictive of delayed gastric emptying. A low peristaltic speed of the oesophagus was associated with impaired T (50) (r ( s )=-0.67; p=0.02). One hour after breakfast, subcutaneous glucose levels decreased in patients with delayed gastric emptying but continued to rise in those with normal emptying. Consequently, the median glucose level 2.5 h after breakfast was lower in the former (9.1 [4.2-12.5] vs 14.3 [11.2-17.7] mmol/l; p<0.05). Glucose fluctuations during the 72 h were significantly higher in patients with an abnormal E/I ratio than in those with a normal E/I ratio (coefficient of variation: 41 [46-49] vs 28 [27-34]%; p=0.008). Abdominal fullness predicted delayed gastric emptying that was associated with diminished glucose uptake after breakfast. Low oesophageal peristaltic speed was associated with slow gastric emptying whereas parasympathetic neuropathy was associated with increased glucose variations.


Effects of prebiotics and precision biotics on performance, animal welfare and environmental impact. A review.
This review aims to analyze the recent studies about prebiotics and precision biotics, as alternatives to animal growth promoters. These substances improve intestinal health, growth performance and poultry environmental impact. Prebiotics are insoluble fibers, that have no nutritive value, but they promote the growth of positive bacteria, increase the nutrients absorption and modulate the immune response. Instead, precision biotics are carbohydrates with glycosidic linkages, which interact with gut bacteria metabolism, reducing the excretion of nitrogen and consequentially, the poultry environmental impact. In the last years, different studies were published in this field, and for this reason, it is necessary to organize the results found. It was shown that mannan-oligosaccharides and β-glucans increase ileal nutrient digestibility, nitrogen retention and antibodies titers. Inulin, arabinoxylans-derived oligosaccharides, and galacto-oligosaccharides improved intestinal morphology, arranging for a larger absorption surface area. It was reported that prebiotics enhance the colonization of positive bacteria and can reduce the count of Campylobacter colonies. Furthermore, xylo-oligosaccharides are often used in animal feed, due to their ability to form organic acids, which decompose noxious substances, improving litter quality, and consequentially, reducing the environmental impact. Litter quality is a relevant aspect for ammonia emissions and for animal welfare. Whether the litter quality is poor, footpad dermatitis increase, worsening animal welfare and increasing nitrogen emissions to air. Precision biotics select metabolic pathways to modulate amino acid degradation, reintegrating the nitrogen discarded, and reducing the ammonia level in litter. It was also reported an improvement of growth performance and a better animal welfare. In conclusion, prebiotics and precision biotics can have positive effects on animal performance and welfare, and they can be a new strategy to reduce the environmental impact of chickens' farms.


Independent effects of statistical learning and top-down attention.
It is well known that spatial attention can be directed in a top-down way to task-relevant locations in space. In addition, through visual statistical learning (VSL), attention can be biased towards relevant (target) locations and away from irrelevant (distractor) locations. The present study investigates the interaction between the explicit task-relevant, top-down attention and the lingering attentional biases due to VSL. We wanted to determine the contribution of each of these two processes to attentional selection. In the current study, participants performed a search task while keeping a location in spatial working memory. In Experiment 1, the target appeared more often in one location, and appeared less often in other location. In Experiment 2, a color singleton distractor was presented more often in location than in all other locations. The results show that when the search target matched the location that was kept in working memory, participants were much faster at responding to the search target than when it did not match, signifying top-down attentional selection. Independent of this top-down effect, we found a clear effect of VSL as responses were even faster when target (Experiment 1) or the distractor (Experiment 2) was presented at a more likely location in visual field. We conclude that attentional selection is driven by implicit biases due to statistical learning and by explicit top-down processing, each process individually and independently modulating the neural activity within the spatial priority map.


Is Twitter a forum for disseminating research to health policy makers?
Findings from scientific research largely remain inside the scientific community. Research scientists are being encouraged to use social media, and especially Twitter, for dissemination of evidence. The potential for Twitter to narrow the gap on evidence translated into policy presents new opportunities. We explored the innovative question of the feasibility of Twitter as a tool for the scientific community to disseminate to and engage with health policy makers for research impact. We created a list of federal "health policy makers." In December 2014, we identified members using several data sources, then collected and summarized their Twitter usage data. Nearly all health policy makers had Twitter accounts. Their communication volume varied broadly. Policy makers are more likely to push information via Twitter than engage with constituents, although usage varied broadly. Twitter has the potential to aid the scientific community in dissemination of health-related research to health policy makers, after understanding how to effectively (and selectively) use Twitter.


Effective reduction in inadequate Pap smears by using a saline-lubricated speculum and two glass slides.
The aim of the study was to propose a method using saline lubrication and two glass slides to reduce the proportion of inadequate Pap smears. This was a retrospective study of patients at a medical center in eastern Taiwan that performs 5000-6000 Pap smears annually. The extracted data only detailed the number and percentage of inadequate Pap smears. We applied two modifications to the conventional Pap smear technique. The first modification was lubricating the speculum with normal saline instead of jelly. The second modification was performing the smear on two glass slides instead of just one. We used the modified technique beginning in January 2017. Therefore, we collected data from 2016 (before the modified technique was employed) and 2018 (after the modified technique was employed). The categorical data are presented as numbers (percentages). The differences in the number and percentage of inadequate smears resulting from both techniques were assessed using the Chi-square test. During 2016 and 2018, 28 and 2 women received inadequate Pap smears among the total of 594 and 613 women who received Pap smears, respectively. The proportion of inadequate Pap smears was 4.71% and 0.33% in 2016 and 2018, respectively (P < 0.001). The use of this modified technique effectively reduced the percentage of inadequate Pap smears.


Development of chimeric candidate vaccine against HPV18: a proof of concept.
Human papillomaviruses (HPVs) are prerequisite for the development of cervical cancer, with HPV16 and HPV18 being the most prevalent. Despite the fact that two prophylactic vaccines against HPVs are in the market, wide-scale application of the vaccine in developing countries is a major problem as far as cost of the vaccine and lack of therapeutic efficacy are concerned. Hence, the aim of our study was to develop HPV18 L1E7 chimeric virus-like particles (CVLPs) vaccine candidate possessing both, prophylactic and therapeutic potential against HPV18-associated cervical cancer. In this study, we have developed a potential candidate vaccine against HPV18 involving HPV18 L1E7 CVLPs, which was expressed in E. coli and assembled in vitro. These CVLPs were able to induce a neutralizing antibody response as well as a cell-mediated immune response in mice.


Caffeic acid attenuates the decrease in cortical BDNF mRNA expression induced by exposure to forced swimming stress in mice.
We previously reported that caffeic acid produces antidepressive-like effects in the forced swimming test in mice, an animal model of depression. Increased evidence suggests that brain-derived neurotrophic factor (BDNF), a member of the neurotrophin family that has high affinity for the tyrosine kinase receptor B (TrkB), plays an important role in the pathophysiology and treatment of depression. The present study examined whether caffeic acid affects the expression levels of BDNF and TrkB mRNA in brain regions of mice subjected to a forced swimming test. Caffeic acid (4 mg/kg, i.p.) reduced the duration of immobility of mice in the forced swimming test. The levels of BDNF mRNA in the frontal cortex as well as TrkB mRNA in the amygdala were significantly decreased after the forced swimming test, and the former reduction was significantly inhibited by caffeic acid (4 mg/kg, i.p.). Caffeic acid (4 mg/kg, i.p.) did not modify the levels of BDNF and TrkB mRNA in brain regions of naive mice. These results suggest that caffeic acid can attenuate the down-regulation of BDNF transcription that results from stressful conditions.


Post-larvae movement network of marine shrimp during the 2013 outbreak of acute hepatopancreatic necrosis disease in Thailand.
This study aimed to assess the movement patterns of post-larvae (PL) of whiteleg shrimp and black tiger shrimp, which were linked to the epidemic of acute hepatopancreatic necrosis disease (AHPND) in Thailand in 2013, using social network analysis. Hatcheries or nurseries were assigned as origin nodes, whereas cultivated ponds were designated as destination nodes. From 376 farms, data of 1300 ponds were collected from network data using a questionnaire. We found that 1477 nodes with 1315 ties appeared in this study. These were divided into a network of cases and controls that presented 538 nodes with 436 ties and 1013 nodes with 879 ties, respectively. The top four out-degree centrality values in the entire partial network, cases, and controls were used to analyze ego-alter similarity. The results showed that each PL provider played a different role in the association with the disease sender. A simulated network using exponential random graph models presented edges as a significant microstructure. In addition, our analysis indicated that the hatchery or nursery (PL provider) is an important node for AHPND occurrence. In conclusion, this study led us to understand the source of PL, which plays a key role in the surveillance, prevention, and control of AHPND.


Staphylococcal protein A bound to Sepharose 4B is mitogenic for T cells but not B cells from rabbit tissues.
The mitogenicity of protein A from Staphylococcus aureus bound to Sepharose 4B (SpA-S) was tested against mononuclear cells from normal rabbit spleens and Peyer's patches after using Sephadex G-10 adherence chromatography to deplete macrophages and Sephadex G-200 anti-rabbit F(ab')2 immunoabsorbent columns to obtain B-enriched lymphocytes. Macrophage-depleted unseparated lymphocytes and the B-cell-depleted (T-cell-enriched) fractions from both tissues consistently showed good mitogenic responses to SpA-S. In contrast, the B-cell-enriched populations from either tissue did not respond to SpA-S. Supplementary experiments employing glass adherence and T-cell autorosetting did not support the possibility that the more efficient immunoabsorbent method of B-cell isolation had in some way caused the unresponsiveness to SpA-S. These results indicate that SpA-S is mitogenic for rabbit tissue T cells but will not serve as a T-independent mitogen for rabbit tissue B cells.


Three-dimensional ultrasound findings of the posterior iris region.
The aim of this study was to assess the benefit of the three-dimensional ultrasound biomicroscopy in examination of the posterior iris and ciliary body. Three-dimensional visualisation of the anterior eye section was achieved through extension of the existing ultrasound biomicroscope system (Humphrey Instruments). Visualisation of posterior iris and ciliary body pathologies in three patients was performed with a three-dimensional reconstruction technique of B-scans. The extended ultrasound system provided three-dimensional visualisation of alterations of the posterior iris region, i.e. iris cysts, ciliary body cysts and solid tumours of the ciliary body and iris. The three-dimensional ultrasound biomicroscopy yields extended diagnostic findings regarding iris and ciliary body pathology. This method offers an improved assessment of the posterior surface of the iris and the volume of the ciliary body. Furthermore, these data can be useful for procedures in computer simulation and calculation for a better understanding of the function of the ciliary body in the accommodation process.


Alterations on vitamin C synthesis and transportation and egg deposition induced by dietary vitamin C supplementation in Hy-Line Brown layer model.
In ovo feeding of vitamin C (VC) has positive effects on the growth performance, immune and antioxidant function in poultry, which indicates that increasing VC content in eggs may be of benefit. This study was to investigate the effects of dietary VC supplementation on VC synthesis and transportation and egg deposition. In Exp. 1, in order to select a suitable animal model, VC content was detected in different eggs from different layer species. Vitamin C content was lower in ISA Brown breeder eggs and Hy-Line Brown layer eggs (P < 0.05) then in Arbor Acres breeder eggs. In Exp. 2, a total of 24 Hy-Line Brown layers (42-week-old) were randomly divided into 3 treatments with 8 replicates and fed a basal diet with VC at 0, 200 and 400 mg/kg. Sodium-dependent VC transporter 1 and 2 (SVCT1 and SVCT2) expressions were higher in ileum than in duodenum and jejunum (P < 0.05). SVCT1 expression was higher but SVCT2 expression was lower in the magnum than in the ovary (P < 0.05). L-Gulonolactone oxidase (GLO) and SVCT1 expressions were higher but SVCT2 was lower in the kidney than in the liver (P < 0.05). Dietary VC supplementation at 400 mg/kg increased SVCT1 expression in duodenum, ovary and magnum, but decreased GLO and SVCT1 expression in liver (P < 0.05). Dietary VC supplementation at 200 and 400 mg/kg increased SVCT2 expression in duodenum, but decreased GLO and SVCT1 expression in kidney and SVCT2 expression in liver (P < 0.05). Dietary VC supplementation promoted VC absorption in duodenum and jejunum, but reduced endogenous VC synthesis in liver and kidney. Although dietary VC supplementation enhanced VC transportation in ovary and magnum, it did not increase VC deposition in produced eggs.


MIF/SCL3A2 depletion inhibits the proliferation and metastasis of colorectal cancer cells via the AKT/GSK-3β pathway and cell iron death.
This study investigated the mechanisms of migration inhibitory factor (MIF) and solute carrier family 3 member 2 (SLC3A2) in colorectal cancer progression. The levels of MIF and SLC3A2 expression in cells were measured by RT-qPCR. SW480 and SW620 cells were transfected with sh-MIF and sh-SLC3A2, respectively. MIF, SLC3A2, GPX4, E-cadherin and N-cadherin expression were detected by immunofluorescence (IF). CCK8 and Transwell assays were performed to detect cell proliferation and migration. Co-immunoprecipitation (CoIP) was used to measure the binding activity of MIF and SLC3A2. Finally, a nude mouse tumorigenicity assay was used to confirm the functions of MIF and SLC3A2 in colorectal cancer. Results showed that the levels of MIF and SLC3A2 expression were up-regulated in colorectal cancer cells. Inhibition of MIF or SLC3A2 expression prevented cell proliferation, migration, epithelial-mesenchymal transition (EMT) and invasion. In addition, knockdown of MIF and SLC3A2 promoted iron death in SW480 and SW620 cells. CoIP results showed that MIF and SLC3A2 directly interact with each other. Knockdown of both MIF and SLC3A2 inhibited tumour growth and metastasis via the AKT/GSK-3β pathway in vivo. The Akt/GSK-3β pathway was found to participate in regulating MIF and SLC3A2 both in vivo and in vitro. MIF and SLC3A2 might be potential biomarkers for monitoring the treatment of colorectal cancer.


Elastography: Imaging the elastic properties of soft tissues with ultrasound.
Elastography is a method that can ultimately generate several new kinds of images, called elastograms. As such, all the properties of elastograms are different from the familiar properties of sonograms. While sonograms convey information related to the local acoustic backscatter energy from tissue components, elastograms relate to its local strains, Young's moduli or Poisson's ratios. In general, these elasticity parameters are not directly correlated with sonographic parameters, i.e. elastography conveys new information about internal tissue structure and behavior under load that is not otherwise obtainable. In this paper we summarize our work in the field of elastography over the past decade. We present some relevant background material from the field of biomechanics. We then discuss the basic principles and limitations that are involved in the production of elastograms of biological tissues. Results from biological tissues in vitro and in vivo are shown to demonstrate this point. We conclude with some observations regarding the potential of elastography for medical diagnosis.


Impact of home monitoring for sudden infant death syndrome on family life. A controlled study.
As home monitoring has been advocated for the prevention of sudden infant death (SIDS) we investigated the influence of such a monitoring programme on family life. Twenty European middle and lower-class families with a child monitored at home after a near miss for SIDS event were investigated. Some psychological and social findings were compared with a group of 20 matched control families. It appears that home monitoring induces severe stress, mainly in the mothers. Fathers and siblings also manifested significant emotional stress. Poor schooling of the parents and previous marital discord seem to be predictors of bad adaptation to home monitoring.


GABA transport goes structural.
The γ-aminobutyric acid transporter 1 (GAT1) is a transporter which clears the inhibitory neurotransmitter γ-aminobutyric acid (GABA) from the synaptic cleft. The paper by Motiwala et al. documents a structure of GAT1 in complex with the antiepileptic drug tiagabine. This study will enable structure-based docking of large chemical libraries for the discovery of novel antiepileptics.


High-tech wound and ostomy care in the home setting.
The role of the home health care nurse has expanded as a result of a burgeoning client base. This client group is sicker and relies on technology at an ever increasing rate. As the critical care nurse transitions into the homecare setting, wound, ostomy, and fistula management will require a broader base of expertise. Adequate reimbursement is tied to accurate assessment and documentation. The clinician armed with these skills is better prepared to develop an effective plan of care for this group of clients.


Seed predation-induced Allee effects, seed dispersal and masting jointly drive the diversity of seed sources during population expansion.
The environmental factors affecting plant reproduction and effective dispersal, in particular biotic interactions, have a strong influence on plant expansion dynamics, but their demographic and genetic consequences remain an understudied body of theory. Here, we use a mathematical model in a one-dimensional space and on a single reproductive period to describe the joint effects of predispersal seed insect predators foraging strategy and plant reproduction strategy (masting) on the spatio-temporal dynamics of seed sources diversity in the colonisation front of expanding plant populations. We show that certain foraging strategies can result in a higher seed predation rate at the colonisation front compared to the core of the population, leading to an Allee effect. This effect promotes the contribution of seed sources from the core to the colonisation front, with long-distance dispersal further increasing this contribution. As a consequence, our study reveals a novel impact of the predispersal seed predation-induced Allee effect, which mitigates the erosion of diversity in expanding populations. We use rearrangement inequalities to show that masting has a buffering role: it mitigates this seed predation-induced Allee effect. This study shows that predispersal seed predation, plant reproductive strategies and seed dispersal patterns can be intermingled drivers of the diversity of seed sources in expanding plant populations, and opens new perspectives concerning the analysis of more complex models such as integro-difference or reaction-diffusion equations.


Use of surgical-site infection rates to rank hospital performance across several types of surgery.
Comparing and ranking hospitals based on health outcomes is becoming increasingly popular, although case-mix differences between hospitals and random variation are known to distort interpretation. The aim of this study was to explore whether surgical-site infection (SSI) rates are suitable for comparing hospitals, taking into account case-mix differences and random variation. Data from the national surveillance network in the Netherlands, on the eight most frequently registered types of surgery for the year 2009, were used to calculate SSI rates. The variation in SSI rate between hospitals was estimated with multivariable fixed- and random-effects logistic regression models to account for random variation and case mix. 'Rankability' (as the reliability of ranking) of the SSI rates was calculated by relating within-hospital variation to between-hospital variation. Thirty-four hospitals reported on 13 629 patients, with overall SSI rates per surgical procedure varying between 0 and 15·1 per cent. Statistically significant differences in SSI rate between hospitals were found for colonic resection, caesarean section and for all operations combined. Rankability was 80 per cent for colonic resection but 0 per cent for caesarean section. Rankability was 8 per cent in all operations combined, as the differences in SSI rates were explained mainly by case mix. When comparing SSI rates in all operations, differences between hospitals were explained by case mix. For individual types of surgery, case mix varied less between hospitals, and differences were explained largely by random variation. Although SSI rates may be used for monitoring quality improvement within hospitals, they should not be used for ranking hospitals.


Association between aortic calcification and the risk of osteoporosis in a chinese cohort: the Chongqing osteoporosis study.
The aim of this study was to examine the relationship between aortic calcification (AC) and the risk of osteoporosis for a community of residents in China. We conducted a 5-year prospective study on 1,729 Chinese elderly subjects. An AC scale (ACS) was determined semiquantitatively from baseline lateral radiographs of the lumbar spine. Dual-energy X-ray absorptiometry was used to determine bone mineral density (BMD). Only subjects who completed the study were included in the final analysis; 432 subjects (25.0 %) developed osteoporosis. The BMD of vertebrae, femoral neck, and the total hip in the Q4 quartile of the ACS (ACS > 6) was lower when compared with the Q1 quartile (ACS = 0, p < 0.05). Cox proportional hazards model analysis showed that ACS (Q4), BMD, CTX, current smoking, and daily drinking were associated with increased risk of osteoporosis. A higher ACS was associated with an increased risk of osteoporosis in both women and men. The present study found that AC carried a higher risk of osteoporosis.


Microsatellite and mitochondrial DNA analyses unveil the genetic structure of native sheep breeds from three major agro-ecological regions of India.
Sheep farming has been fundamental to many civilizations in the world and is practiced in India since antiquity. Several thousand years of adaptation to local environmental conditions and selective breeding have evolved 44 sheep breeds in India. They are paramount in terms of economic, scientific, and cultural heritage. Genetic characterization information is imperative for sustainable utilization and conservation of ovine heritage. In this study, the genetic diversity, differentiation, and structure of 11 indigenous sheep breeds from three different agro-ecological zones of India were explored with genomic microsatellite loci and mitochondrial DNA (D loop). The estimated diversity parameters indicated that populations retained high levels of genetic diversity (Na = 8.27 ± 0.17; Ho = 0.65 ± 0.01), which provides an optimistic viewpoint for their survival. However, significant inbreeding was also observed in the nine populations. Moderate genetic differentiation existed among the groups (FST = 0.129 ± 0.012), and most likely clusters existing in the dataset are seven. Phylogenetic clustering was in line with the geographical locations of sheep populations. Mitochondrial sequences revealed high haplotype diversity with the existence of maternal haplogroups A, B, and C, and signals of population expansion. Decreased genetic diversity and unique maternal lineage (C) in endangered Tibetan and Bonpala sheep breed, warrant their immediate scientific management. Overall, the quantitative data reported here on the extant variability, and genetic relationships among the Indian sheep breeds, provide critically important inputs that will be valuable for the decision-making process on their management, both for the conservation of endangered breeds, and formulation of breeding programs to check genetic erosion.


Autonomic responses to ethanol in adolescent and adult rats: a dose-response analysis.
Although the exact cause of the increase in ethanol consumption during adolescence is not known, age differences in sensitivity to some of ethanol's effects may play a contributory role. Prior research has shown little difference in the expression of ethanol-induced tachycardia between adolescents and adults following ethanol inhalation. In contrast, there is mounting evidence of ontogenetic differences in ethanol-induced hypothermia, although the nature of the ontogenetic effect observed has been found to vary across studies and even within labs. Relative ontogenetic differences in body temperature (BT) after ethanol administration appear to be driven in part by the amount of experimental perturbation associated with the test protocol, although differing ethanol exposure levels across studies may also have contributed to the variations in ontogenetic patterns that have been observed. To explore the latter possibility, the present study assessed ethanol-induced hypothermia and tachycardia in adolescent and adult male Sprague-Dawley rats examined in their home cages in the presence of their housing partner following intraperitoneal administration of 0.5, 1.5, or 3.0 g/kg ethanol. The results showed that, although adolescents did not show an adult-typical tachycardic effect at any dose, they proved more sensitive than adults to ethanol's hypothermic effects at the two highest doses. These findings suggest that not only the degree of experimental perturbation, but also the amount of ethanol exposure may differentially effect expression of age differences in ethanol-induced hypothermia, with adolescents showing greater hypothermia than adults at higher doses. Together with previous findings, these data contribute to the emerging picture that age differences in autonomic effects of ethanol appear to be particularly sensitive to dosing parameters and experimental protocols, unlike the generally more consistent ontogenetic findings observed across studies when using behavioral measures of ethanol sensitivity.


Preoperative Mechanical Bowel Preparation for Gynecologic Surgeries: A Systematic Review with Meta-analysis.
To assess the efficacy and safety of mechanical bowel preparation (MBP) before benign laparoscopic or vaginal gynecologic surgeries. Database searches of MEDLINE (PubMed), Embase (OVID), Cochrane Central Register of Controlled Trials, and Web of Sciences and citations and reference lists published up to December 2021. Randomized clinical trials in any language comparing MBP with no preparation were included. Two reviewers independently screened 925 records and extracted data from 12 selected articles and assessed the risk of bias with the Cochrane risk-of-bias tool for randomized trials tool. A random-effects model was used for the analysis. Surgeon findings (surgical field view, quality of bowel handling and bowel preparation), operative outcomes (blood loss, operative time, length of stay, surgical site infection), and patient's preoperative symptoms and satisfaction were collected. Thirteen studies (1715 patients) assessing oral and rectal preparations before laparoscopic and vaginal gynecologic surgeries were included. No significant differences were observed with or without MBP on surgical field view (primary outcome, risk ratio [RR] 1.01, 95% confidence interval [CI] 0.97-1.05, p = .66, I2 = 0%), bowel handling (RR 1.01, 95% CI 0.95-1.08, p = .78, I2 = 67%), or bowel preparation. In addition, there were no statistically significant differences in perioperative findings. MBP was associated with increased pain (mean difference [MD] 11.62[2.80-20.44], I2 = 76, p = .01), weakness (MD 10.73[0.60-20.87], I2 = 94, p = .04), hunger (MD 17.52 [8.04-27.00], I2 = 83, p = .0003), insomnia (MD 10.13[0.57-19.68], I2 = 82, p = .04), and lower satisfaction (RR 0.68, 95% CI 0.53-0.87, I2 = 76%, p = .002) compared with controls. MBP has not been associated with improved surgical field view, bowel handling, or operative outcome. However, in view of the adverse effects induced, its routine use before benign gynecologic surgeries should be abandoned.


HFE gene mutations in patients with primary iron overload: is there a significant improvement in molecular diagnosis yield with HFE sequencing?
Rare HFE variants have been shown to be associated with hereditary hemochromatosis (HH), an iron overload disease. The low frequency of the HFE p.C282Y mutation in HH-affected Brazilian patients may suggest that other HFE-related mutations may also be implicated in the pathogenesis of HH in this population. The main aim was to screen for new HFE mutations in Brazilian individuals with primary iron overload and to investigate their relationship with HH. Fifty Brazilian patients with primary iron overload (transferrin saturation>50% in females and 60% in males) were selected. Subsequent bidirectional sequencing for each HFE exon was performed. The effect of HFE mutations on protein structure were analyzed by molecular dynamics simulation and free binding energy calculations. p.C282Y in homozygosis or in heterozygosis with p.H63D were the most frequent genotypic combinations associated with HH in our sample population (present in 17 individuals, 34%). Thirty-six (72.0%) out of the 50 individuals presented at least one HFE mutation. The most frequent genotype associated with HH was the homozygous p.C282Y mutation (n=11, 22.0%). One novel mutation (p.V256I) was indentified in heterozygosis with the p.H63D mutation. In silico modeling analysis of protein behavior indicated that the p.V256I mutation does not reduce the binding affinity between HFE and β2-microglobulin (β2M) in the same way the p.C282Y mutation does compared with the native HFE protein. In conclusion, screening of HFE through direct sequencing, as compared to p.C282Y/p.H63D genotyping, was not able to increase the molecular diagnosis yield of HH. The novel p.V256I mutation could not be implicated in the molecular basis of the HH phenotype, although its role cannot be completely excluded in HH-phenotype development. Our molecular modeling analysis can help in the analysis of novel, previously undescribed, HFE mutations.


Characterization of cell lines derived from a multiply aneuploid human bladder transitional-cell carcinoma, UCRU-BL-13.
A series of cultured cell lines (designated UCRU-BL-13) has been established from different serial passages of a multiply aneuploid human bladder transitional-cell carcinoma xenografted in nude mice. Serial passage of the xenografts in vivo and of the cell lines in vitro was accompanied by shifts in the tumor ploidy, with dominance of different major peaks. Despite this, the expression of tumor markers remained constant, and consistent chromosomal markers were observed both in the 8th xenograft passage and in a subline in tissue culture established over a year apart. Chromosomal numbers reflected the predominant aneuploid peaks observed; consistent numerical and structural changes included a marker derived from chromosome 1, 8p-, -10, 11q+, and 17q+. The cell line derived from the initial xenograft comprised a mixture of transitional, adenocarcinoma and squamous carcinoma cells in early passage, but adenocarcinoma cells were absent from later passages. The lines expressed the B-blood-group antigen, histocompatibility antigens, receptors for transferrin and EGF, and reacted with a series of monoclonal antibodies (MAbs) directed to malignant human epithelial cell lines. These lines provide a model for studying the evolution of tumor heterogeneity and drug resistance in bladder carcinoma exhibiting multiple aneuploidy.


Systemic cancer therapy with engineered adenovirus that evades innate immunity.
Oncolytic virus therapy is a cancer treatment modality that has the potential to improve outcomes for patients with currently incurable malignancies. Although intravascular delivery of therapeutic viruses provides access to disseminated tumors, this delivery route exposes the virus to opsonizing and inactivating factors in the blood, which limit the effective therapeutic virus dose and contribute to activation of systemic toxicities. When human species C adenovirus HAdv-C5 is delivered intravenously, natural immunoglobulin M (IgM) antibodies and coagulation factor X rapidly opsonize HAdv-C5, leading to virus sequestration in tissue macrophages and promoting infection of liver cells, triggering hepatotoxicity. Here, we showed that natural IgM antibody binds to the hypervariable region 1 (HVR1) of the main HAdv-C5 capsid protein hexon. Using compound targeted mutagenesis of hexon HVR1 loop and other functional sites that mediate virus-host interactions, we engineered and obtained a high-resolution cryo-electron microscopy structure of an adenovirus vector, Ad5-3M, which resisted inactivation by blood factors, avoided sequestration in liver macrophages, and failed to trigger hepatotoxicity after intravenous delivery. Systemic delivery of Ad5-3M to mice with localized or disseminated lung cancer led to viral replication in tumor cells, suppression of tumor growth, and prolonged survival. Thus, compound targeted mutagenesis of functional sites in the virus capsid represents a generalizable approach to tailor virus interactions with the humoral and cellular arms of the immune system, enabling generation of "designer" viruses with improved therapeutic properties.


[Seminal plasma miR-122-3p and miR-141-5p stability and its diagnosis value for idiopathic asthenospermia].
To explore the stability of seminal plasma miR-122-3p and miR-141-5p and their diagnostic value for idiopathic asthenospermia. Seminal plasma miR-122-3p and miR-141-5p with various incubation time, freeze-thaw cycles and incubation time were analyzed. Real-time fluorescence quantitative polymerase chain reaction (RT-PCR) was used to detect the content of miR-122-3p and miR-141-5p, with U6 snRNA as the reference. The idiopathic asthenospermia group was further divided into grade I and II based on the progressive motility percent, and the content of miR-122-3p and miR-141-5p in the two grades were compared. No difference was detected after incubation at room temperature and 4 degrees Celsius. But seminal plasma miR-122-3p and miR-141-5p have decreased along with the increase in freeze-thaw cycles increases. RT-PCR assayed showed that the miR-122-3p content in the idiopathic asthenospermia group was significantly lower than the control group, while miR-141-5p was significantly higher. The difference in both miR-122-3p and miR-141-5p content was statistically significant (P < 0.05). A significant difference in miR-122-3p and miR-141-5p content between a and b groups was also detected (P < 0.05). The AUC-ROC of miR-122-3p was 0.88. At the cutoff value of 1.02, the sensitivity and specificity was 83% and 84%, respectively. At the cutoff of 2.95, the AUC-ROC of miR-141-5p was 0.88. The sensitivity and specificity was 84% and 70%, respectively. MiR-122-3p and miR-141-5p in seminal plasma are stable and have certain value for the diagnosis of idiopathic asthenospermia.


Negative correlation of urinary miR-199a-3p level with ameliorating effects of sarpogrelate and cilostazol in hypertensive diabetic nephropathy.
The prevalence of chronic kidney disease is increasing globally; however, effective therapeutic options are limited. In this study, we aimed to identify urinary miRNAs reflecting the effect of therapeutic intervention in rats with comorbid hypertension and diabetes. Additionally, the potential beneficial effects of anti-platelet sarpogrelate and cilostazol were investigated. Nephropathy progression in streptozotocin (STZ)-treated spontaneously hypertensive rats (SHRs), including albuminuria, collagen deposition, and histopathological changes, was alleviated by sarpogrelate and antihypertensive agent telmisartan. Global analysis of urinary miRNAs identified that miR-199a-3p was commonly reduced by sarpogrelate and telmisartan treatment. In vitro analysis suggested CD151 as a target gene of miR-199a-3p: miR-199a-3p overexpression repressed CD151 levels and miR-199a-3p interacted with the 3'-untranslated region of the CD151 gene. In addition, we demonstrated that the miR-199a-3p/CD151 axis is associated with the transforming growth factor-β1 (TGF-β1)-induced fibrogenic pathway. TGF-β1 treatment led to miR-199a-3p elevation and CD151 suppression, and miR-199a-3p overexpression or CD151-silencing enhanced TGF-β1-inducible collagen IV and α-smooth muscle actin (α-SMA) levels. In vivo analysis showed that the decrease in CD151 and the increase in collagen IV and α-SMA in the kidney from STZ-treated SHR were restored by sarpogrelate and telmisartan administration. In an additional animal experiment using cilostazol and telmisartan, there was a correlation between urinary miR-199a-3p reduction and the ameliorating effects of cilostazol or combination with telmisartan. Collectively, these results indicate that urinary miR-199a-3p might be utilized as a marker for nephropathy treatment. We also provide evidence of the benefits of antiplatelet sarpogrelate and cilostazol in nephropathy progression.