Bacterial communities of indoor surface of stingless bee nests.
Microbes have been identified as fundamental for the good health of bees, acting as pathogens, protective agent against infection/inorganic toxic compounds, degradation of recalcitrant secondary plant metabolites, definition of social group membership, carbohydrate metabolism, honey and bee pollen production. However, study of microbiota associated with bees have been largely confined to the honeybees and solitary bees. Here, I characterized the microbiota of indoor surface nest of four brazilian stingless bee species (Apidae: Meliponini) with different construction behaviors and populations. Bees that use predominantly plant material to build the nest (Frieseomelitta varia and Tetragonisca angustula) have a microbiome dominated by bacteria found in the phylloplane and flowers such as Pseudomonas sp. and Sphingomonas sp. Species that use mud and feces (Trigona spinipes) possess a microbiome dominated by coliforms such as Escherichia coli and Alcaligenes faecalis. Melipona quadrifasciata, which uses both mud / feces and plant resin, showed a hybrid microbiome with microbes found in soil, feces and plant material. These findings indicate that indoor surface microbiome varies widely among bees and reflects the materials used in the construction of the nests.


[Serum cystatin C concentration in children with urinary stones].
Stone formation precedes long period, when the crystals are accumulated in basement membranes of renal tubules and intestinal tissue. Accumulated, inside of renal tubules crystals and stones in urinary tract cause urinary tract obstruction, what may lead to impairment of renal function. The aim of work was the assessment of serum cystatin C (cys C) concentration in children with urolithiasis, confirmed by the presence of renal stones in renal pelvis in comparison to serum creatinine concentration and creatinine clearance (Cr cl). Examined group (B) consisted of 30 children aged (13.08 +/- 4.14 years) with urolithiasis, which was divided into 3 subgroups (I, II, Ill) in dependence on stones' diameter (0.35-1.6cm). Control group (C) consisted of 26 healthy children at the same age. Nephelometric method was used to determine serum cystatin C level, Jaffe method to assess serum creatinine and the Schwartz formula to estimate glomerular filtration rate. In control group (C) serum cys C did not exceed 0.95 mg/l. In group B serum cystatin C and serum creatinine concentration and Cr cl was similar to the results of control group (p > 0.05). However in 16% of children with urolithiasis, in whom the stones of 0.8-1.6cm diameter were found in both renal pelvis, the concentration of serum cys C exceed 1.2 mg/l, and the value differed significantly from the results of control group (p < 0.05). A weak positive correlation between cys C and creatinine concentration and also between cys C and Cr cl was found. The serum cys C concentration in children with single stones of 0.35-0.8 diameter was normal. Serum cystatin C increases with increased degrees of urolithiasis assessed by stone size and their number in kidney.


Involvement of sinoaortic afferents in renal sympathoinhibition and vasodilation induced by acute hypernatremia.
Despite the abundance of evidence that supports the important role of aortic and carotid afferents to short-term regulation of blood pressure and detection of variation in the arterial PO2 , PCO2 and pH, relatively little is known regarding the role of these afferents during changes in the volume and composition of extracellular compartments. The present study sought to determine the involvement of these afferents in the renal vasodilation and sympathoinhibition induced by hypertonic saline (HS) infusion. Sinoaortic-denervated and sham male Wistar rats were anaesthetised with intravenous (i.v.) urethane (1.2 g/kg body weight (bw)) prior to the measurement of the mean arterial pressure (MAP), renal vascular conductance (RVC) and renal sympathetic nerve activity (RSNA). In the sham group, the HS infusion (3 mol/L NaCl, 1.8 mL/kg bw, i.v.) induced transient hypertension (12 ± 4 mmHg from baseline, peak at 10 min; P < 0.05), an increase in RVC (127 ± 9% and 150 ± 13% from baseline, at 20 and 60 min respectively; P < 0.05) and a decrease in RSNA (-34 ± 10% and -29 ± 5% from baseline, at 10 and 60 min respectively; P < 0.05). In sinoaortic-denervated rats, HS infusion promoted a sustained pressor response (30 ± 5 and 17 ± 6 mmHg of baseline values, at 10 and 30 min respectively; P < 0.05) and abolished the increase in RVC (85 ± 8% from baseline, at 10 min) and decrease in RSNA (-4 ± 3% from baseline, at 10 min). These results suggest that aortic and carotid afferents are involved in cardiovascular and renal sympathoinhibition responses induced by acute hypernatremia.


Perspectives on leaf dorsoventral polarity.
Leaves occur in a vast array of shapes and sizes, with complex diversity contributing to optimization of the principal function of photosynthesis. The program of development from a self-renewing stem cell population to a mature leaf has been of interest to biologists for years. Many genes involved in this process have been identified, particularly in the model eudicot Arabidopsis, so that now we have a greater understanding of mechanisms of stem cell maintenance, cell differentiation and organogenesis. One aspect of leaf development that is of particular interest is the establishment of dorsoventral polarity: the distinct adaxial (upper) and abaxial (lower) sides of the leaf. Early studies postulated conceptual models of how establishment of polarity leads to the development of planar leaves. Studies over the past decade have defined genetic details of this model, and uncovered diverse mechanisms of gene regulation that facilitate development of leaf dorsoventral polarity, including transcriptional regulation, chromatin modification, DNA modification, regulation by short RNAs and translational and post-translational regulation. This review will discuss these regulatory mechanisms in the context of leaf dorsoventrality, and will conclude with unresolved questions and areas of future research.


Dietary chitosan enhances hepatic CYP7A1 activity and reduces plasma and liver cholesterol concentrations in diet-induced hypercholesterolemia in rats.
The present study was performed to elucidate the hypocholesterolemic action of chitosan on the diet-induced hypercholesterolemia in rats. Male Sprague-Dawley rats (n=24) were fed with chitosan-free diet (Control), diets containing 2% or 5% chitosan for 4 weeks. Hypercholesterolemia was induced by adding 1% cholesterol and 0.5% cholic acid to all diets. Body weight gain and food intake of rats did not differ among the groups. The chitosan treated groups showed significant improvement in the plasma concentration of total cholesterol and LDL-cholesterol compared to the control group (p<0.05). Also, the chitosan treated groups decreased the liver concentration of total lipid and total cholesterol compared to the control group (p<0.05). The activity of hepatic cholesterol 7alpha-hydroxylase (CYP7A1), the rate-limiting enzyme in the conversion of cholesterol to bile acids, was increased by 123% and 165% for the 2% or 5% chitosan diets, respectively. These findings suggest that enhancement of hepatic CYP7A1 activity may be a mechanism, which can partially account for the hypocholesterolemic effect of dietary chitosan in cholesterol metabolism.


Effect of Fiber Bundle Morphology on Fiber Dispersion for Long Fiber-Reinforced Thermoplastics.
Understanding the mechanics of fiber attrition during the extrusion process is highly important in predicting the strength of long fiber-reinforced thermoplastic composites. However, little work has been done to investigate the mechanics of fiber dispersion and its effects on fiber attrition. This study aims at investigating fiber dispersion in simple shear flows for long fiber-reinforced thermoplastic pellets. Depending on the fabrication process, fiber bundles display distinct levels of compaction within the pellets. Studies have shown that morphological differences can lead to differences in dispersion mechanics; therefore, using a Couette rheometer and a sliding plate rheometer, coated and pultruded pellets were subjected to simple shear deformation, and the amount of dispersion was quantified. Additionally, a new image-based analysis method is presented in this study to measure fiber dispersion for a multi-pellet-filled system. Results from the single-pellet dispersion study showed a small amount of correlation between the dimensionless morphological parameter and the dispersion measurement. Pultruded and coated pellets were both found to have similar dispersion rates in a multi-pellet system. However, pultruded pellets were found to have a higher dispersion value at all levels when compared with coated pellets in both dispersion studies.


Cryopreserved and frozen hyaline cartilage imaged by environmental scanning electron microscope. An experimental and prospective study.
To obtain images of the articular surface of osteochondral grafts (fresh, frozen, and cryopreserved in RPMI) using an environmental scanning electron microscope (ESEM). To evaluate and compare the main morphological aspects of the chondral surface of the fresh, frozen, and cryopreserved grafts as visualized via ESEM. The study was based on osteochondral fragments from the internal condyle of the knee joint of New Zealand rabbits, corresponding to the chondral surface from fresh, frozen, and cryopreserved samples. One hundred ESEM images were obtained from each group and then classified according to a validated system. The kappa index and the corresponding concordance index were calculated, and the groups were compared by Pearson's chi-squared test (p < 0.05). The articular surface of cryopreserved osteochondral grafts had fewer even surfaces and filled lacunae and a higher number of empty lacunae as compared to fresh samples; these differences correspond to images of cell membrane lesions that lead to destruction of the chondrocyte. Frozen grafts showed more hillocky and knobby surfaces than did fresh grafts; they also had a greater number of empty chondrocyte lacunae. ESEM is useful for obtaining images of the surface of osteochondral grafts. When compared to fresh samples, cryopreservation in RPMI medium produces changes in the surface of hyaline cartilage, but to a lesser extent than those produced by freezing.


Controlled study of lactoperoxidase gel on oral flora and saliva in irradiated patients with oral cancer.
The aim of this study was to determine if radiotherapy induces hyposalivation altering oral microbial flora. The purpose of this placebo-controlled, single-blind study was to determine beneficial effects of a saliva substitute and an oral hygiene product on irradiated patients with oropharyngeal cancer. Eighteen patients were assigned to the test group (Biotène Oral Balance gel [Lacléde Incorporated Healthcare Products, Gardena, CA] and toothpaste used daily), and another 18 were put on a conventional daily regimen (carboxymethylcellulose gel and Oral-B toothpaste [Laclede Pharmaceuticals, Gardena, CA]). Cultures for identifying and quantitating microorganisms, whole unstimulated saliva, and visual analog measurements for comfort were obtained before mucositis occurred and after treatment. Daily use of Biotène products enhanced control of microbial flora, improved salivary flow, and increased oral comfort as compared with control subjects. Four weeks after mucositis, some aerobic isolates disappeared in the test group; periodontal-associated bacteria were markedly decreased in the test group; and candidal species were significantly lowered in the test group. Although baseline saliva was lower in the test group (P = 0.001), after 4 weeks, no difference between groups existed; comfort was greater in the test group (P = 0.007). Use of enzyme-engineered Biotène products that assist in control of the oral microbial flora as well as supporting oral comfort through lubrication appear to be useful aids for irradiated patients with oropharyngeal cancer.


Three siblings with steroid-resistant nephrotic syndrome: new NPHS2 mutations in a Turkish family.
Steroid-resistant nephrotic syndromes often are resistant to additional immunosuppressive agents and tend to progress to end-stage renal disease. Genetic studies in children with familial nephrotic syndrome have identified mutations in genes that encode important podocyte proteins. NPHS2 mutations are responsible for autosomal recessive familial focal segmental glomerulosclerosis (FSGS), and these mutations were detected in both familial and sporadic forms of FSGS. Interethnic differences were suggested to play a role in the incidence of these mutations. In this report, the cases of 3 siblings with steroid-resistant nephrotic syndrome who carry NPHS2 mutations (R238S and P118L) are presented.


The role of formant amplitude in the perception of /i/ and /u/.
Although recent evidence reconfirmed the importance of spectral peak frequencies in vowel identification [Kiefte and Kluender (2005). J. Acoust. Soc. Am. 117, 1395-1404], the role of formant amplitude in perception remains somewhat controversial. Although several studies have demonstrated a relationship between vowel perception and formant amplitude, this effect may be a result of basic auditory phenomena such as decreased local spectral contrast and simultaneous masking. This study examines the roles that local spectral contrast and simultaneous masking play in the relationship between the amplitude of spectral peaks and the perception of vowel stimuli. Both full- and incomplete-spectrum stimuli were used in an attempt to separate the effects of local spectral contrast and simultaneous masking. A second experiment was conducted to measure the detectability of the presence/absence of a formant peak to determine to what extent identification data could be predicted from spectral peak audibility alone. Results from both experiments indicate that, while both masking and spectral contrast likely play important roles in vowel perception, additional factors must be considered in order to account for vowel identification data. Systematic differences between the audibility of spectral peaks and predictions of perceived vowel identity were observed.


Effector function does not contribute to protection from virus challenge by a highly potent HIV broadly neutralizing antibody in nonhuman primates.
Protection from immunodeficiency virus challenge in nonhuman primates (NHPs) by a first-generation HIV broadly neutralizing antibody (bnAb) b12 has previously been shown to benefit from interaction between the bnAb and Fcγ receptors (FcγRs) on immune cells. To investigate the mechanism of protection for a more potent second-generation bnAb currently in clinical trials, PGT121, we carried out a series of NHP studies. These studies included treating with PGT121 at a concentration at which only half of the animals were protected to avoid potential masking of FcγR effector function benefits by dominant neutralization and using a new variant that more completely eliminated all rhesus FcγR binding than earlier variants. In contrast to b12, which required FcγR binding for optimal protection, we concluded that PGT121-mediated protection is not augmented by FcγR interaction. Thus, for HIV-passive antibody prophylaxis, these results, together with existing literature, emphasize the importance of neutralization potency for clinical antibodies, with effector function requiring evaluation for individual antibodies.


Milwaukee shoulder syndrome.
Milwaukee shoulder syndrome or rapid destructive arthritis of the shoulder; is a particular form of arthritis which affects mainly elderly patients, predominantly women, with limited pain and sometimes neuropathic joints, rotator cuff tear, joint instability, mildly inflammatory or noninflammatory, blood-stained effusion, calcium apatite crystals, and marked joint and bone destruction. This is very uncommon condition and very sparsely reported and so far, there is no case report from India, hence, we report a case of Milwaukee shoulder syndrome from India.


Discovery of small molecule inhibitors for the C. elegans caspase CED-3 by high-throughput screening.
C. elegans has been widely used as a model organism for programmed cell death and apoptosis. Although the CED-3 caspase is the primary effector of cell death in C. elegans, no selective inhibitors have been identified. Utilizing high-throughput screening with recombinant C. elegans CED-3 protein, we have discovered and confirmed 21 novel small molecule inhibitors. Six compounds had IC50 values < 10 μM. From these, four distinct chemotypes were identified. The inhibitor scaffolds described here could lead to the development of selective molecular probes to facilitate our understanding of programmed cell death in this model organism.


Reconsidering Our Domestic Violence System.
Children's exposure to domestic violence is well established as an adverse childhood experience (ACE). Much is known about the impact of this exposure, but efforts to ameliorate its effects are too often unsuccessful. Reconsidering our response requires a candid assessment of whether convening large and disparate systems leads to the best outcome.


Sex differences in growth patterns of the airways and lung parenchyma in children.
Seventeen boys and 19 girls, 8-15 yr in age, were studied to ascertain, in the two sex groups, the predictors of airway size [assessed by measurement of tracheal cross-sectional area (CSA) and maximal expiratory flows (Vmax)] and the relative rates of growth of the major divisions of the airways and lung parenchyma. In boys, total lung capacity (TLC) accounted for 77% of the variance of CSA and for 66% of the variability of Vmax. In contrast, somatic growth and maturation in girls accounted for only 45% of the variance of CSA and for 64% of the variability of Vmax; TLC was relatively unimportant. In boys, but not in girls, TLC-corrected CSA was significantly and inversely related to height and to TLC. In girls, TLC-corrected Vmax at 50 and 75% of forced vital capacity were directly related to height. These observations suggest different patterns of airway-parenchymal-somatic growth relationships in the two sexes. Furthermore, parenchymal growth appears to be the best determinant of airway growth in boys. In girls, other factors, perhaps genetic in nature, besides growth of parenchyma, may help determine airway size.


An efficient convergent synthesis of GP1c ganglioside epitope.
In this report, we describe an efficient convergent synthesis of the GP1c glycolipid epitope, which is one of the most complex c-series gangliosides. The alpha(2,3) and alpha(2,8) sialylations were accomplished by use of 5N,4O-carbonyl and 7,8-O-isopropyliden as well as 5N,4O-carbonyl- and 7,8-di-O-chloroacetyl-protected sialyl donors in good yields with excellent alpha-selectivity, respectively. The two sialyl donors enable synthesis of the di- and trisialylgalactosides by simple glycosylation and deprotection. We synthesized the protected GP1c glycolipid epitope, which is a compact, rigid, branched structure, via direct coupling of tetarasaccharide and pentasaccharide units.


A human t-PA mutant cDNA cassette knocked in the murine fgfr-4 locus targeting for mammary gland expression.
The expression of foreign gene in transgenic animals produced by pronuclear microinjection is often confounded by the position effects caused by not only the nature of chromosomal integration site but also the number and arrangement of multiple transgene copies. Gene targeting provides a new way to overcome these inhibitions by introducing single-copy transgene into a chosen site. The choice of a good chromosomal site will favor transgene expression in a predictable fashion. In this study, we tested a new site (fgfr-4) for foreign gene integration and expression. A t-PA mutant (t-PAm) expression cassette under bovine alphas1-casein regulatory sequences was efficiently knocked-in fgfr-4 site through homologous recombination. The t-PAm was expressed in the milk of all targeted mice. Our experiment indicates that the fgfr-4 may be a candidate site for knocking foreign gene to make transgenic animals.


Comparability of cancer mortality data in different regions of Yugoslavia.
The standardized cancer mortality ratio based on well-defined causes of death (MSMR) was analysed. It was shown how the use of MSMR gave wrong results just because differences related to the age structure of compared populations were not eliminated. In this paper, the presented calculation pointed out that, actually, MSMR only standardizes age distribution of well-defined causes of death. The age standardization can be obtained with MSMR only if compared populations have the same age specific rates from all causes of death. At the same time standard population must also have the same age specific rates from all causes of death or have the same or similar ratios of age specific rates from all causes of death as populations under comparison.


Feeding the 1 to 7-year-old child. A support paper for the South African paediatric food-based dietary guidelines.
Young children embark on a transitional nutritional journey, progressing from total reliance on caregivers to independence, autonomy and self-determination. Appropriate nutritional intake in young children is a diverse concept, incorporating suitable nutritional choices and feeding behaviours. Lessons learnt in childhood will have long-term effects on the individual and society overall. Since South African children are raised in a country where under- and over-nutrition exist simultaneously, a careful balance should be achieved in any national public health message. The South African paediatric food-based dietary guidelines for children younger than 7 years strive to facilitate the education of carers of young children in the adoption of healthy eating practices. The guidelines address issues regarding variety in the diet that has been shown to improve both micro- and macronutrient intakes. Specific reference is made to starchy foods, vegetables, fruit and water along with protein sources, which should be consumed regularly. Milk, has been emphasised in these guidelines because of the poor calcium intake in South African children. The only guideline that limits intake is the sweet treats or drinks message, because of public health concerns such as obesity and dental caries. Other messages pertaining to eating habits take cognisance of the child's physiological limitations in gastric capacity and suggest small regular meals. Clean, safe drinking water is the beverage of choice. Non-food-based guidelines are also included, which recognize the importance of active play, for developmental and health purposes, as well as regular clinic attendance.


Diffusion-weighted MRI and quantitative biophysical modeling of hippocampal neurite loss in chronic stress.
Chronic stress has detrimental effects on physiology, learning and memory and is involved in the development of anxiety and depressive disorders. Besides changes in synaptic formation and neurogenesis, chronic stress also induces dendritic remodeling in the hippocampus, amygdala and the prefrontal cortex. Investigations of dendritic remodeling during development and treatment of stress are currently limited by the invasive nature of histological and stereological methods. Here we show that high field diffusion-weighted MRI combined with quantitative biophysical modeling of the hippocampal dendritic loss in 21 day restraint stressed rats highly correlates with former histological findings. Our study strongly indicates that diffusion-weighted MRI is sensitive to regional dendritic loss and thus a promising candidate for non-invasive studies of dendritic plasticity in chronic stress and stress-related disorders.


Solution-Processed Self-Assembled Nanodielectrics on Template-Stripped Metal Substrates.
The coupling of hybrid organic-inorganic gate dielectrics with emergent unconventional semiconductors has yielded transistor devices exhibiting record-setting transport properties. However, extensive electronic transport measurements on these high-capacitance systems are often convoluted with the electronic response of the semiconducting silicon substrate. In this report, we demonstrate the growth of solution-processed zirconia self-assembled nanodielectrics (Zr-SAND) on template-stripped aluminum substrates. The resulting Zr-SAND on Al structures leverage the ultrasmooth (r.m.s. roughness <0.4 nm), chemically uniform nature of template-stripped metal substrates to demonstrate the same exceptional electronic uniformity (capacitance ∼700 nF cm(-2), leakage current <1 μA cm(-2) at -2 MV cm(-1)) and multilayer growth of Zr-SAND on Si, while exhibiting superior temperature and voltage capacitance responses. These results are important to conduct detailed transport measurements in emergent transistor technologies featuring SAND as well as for future applications in integrated circuits or flexible electronics.


Analysis of boceprevir resistance associated amino acid variants (RAVs) in two phase 3 boceprevir clinical studies.
We investigated the frequency of RAVs among patients failing to achieve SVR in two clinical trials. We also investigated the impact of interferon responsiveness on RAVs and specific baseline RAVs relationship with boceprevir treatment failure. Data are from 1020 patients enrolled into either SPRINT-2 or RESPOND-2; patients received a 4-week PR lead-in prior to receiving boceprevir or placebo. RAVs were analyzed via population-based sequence analysis of the NS3 protease gene (success rate of >90% at a virus level of ≥ 10,000IU/mL) RESULTS: The high SVR rate in patients who received boceprevir resulted in a low rate of RAVs; 7% was detected at baseline in all patients, which rose to 15% after treatment. However, RAVs were detected in 53% of patients that failed to achieve SVR, which declined to 22.8% 6-14 months following cessation of boceprevir therapy. Baseline RAVs alone were not predictive of virologic outcome; poor interferon responsiveness was highly predictive of non-SVR. RAVs were more frequently detected in poor interferon responders. We detected no association between the presence of baseline amino acid variants at boceprevir resistance-associated loci and outcome in the context of good IFN response.


Sustained complete molecular remissions after treatment with imatinib-mesylate in patients with failure after allogeneic stem cell transplantation for chronic myelogenous leukemia: results of a prospective phase II open-label multicenter study.
In the era of molecular therapy of chronic myelogenous leukemia (CML) applying BCR-ABL tyrosine kinase inhibitors, the usefulness of molecular end points, in particular, quantitative polymerase chain reaction (PCR) for BCR-ABL in monitoring responses has been broadly accepted. Therefore, we have designed a prospective phase II trial in CML, which, for the first time, evaluated the feasibility and safety of molecular end points as surrogate markers to guide through a stratified treatment algorithm within a multicenter trial. As a clinical model, we adopted minimal residual disease (MRD) found in relapse after allogeneic stem cell transplantation (SCT) in CML. Forty-four patients were enrolled and received the BCR-ABL tyrosine kinase inhibitor imatinib (IM) at a starting dose of 400 mg/d. The quality of molecular responses achieved then decided on discontinuation of IM or dose escalation up to 800 mg/d, and finally, on application of donor lymphocyte infusions. Results Seventy percent of patients achieved a complete molecular response (CMR), defined as nested PCR-negativity for BCR-ABL in three consecutive samples. Interestingly, in four out of 10 patients who discontinued IM, CMR was durable even after cessation of IM with a median follow-up of 494 days. This suggests the possibility of long-term tumor control in a subset of patients. The treatment strategy showed that IM treatment was well-tolerated and highly efficacious in MRD after allogeneic SCT. Moreover, this study demonstrated that evaluation of a molecular end point within a multicenter trial can be a safe and effective tool for clinical decision making.


Association between Polymorphisms in MicroRNAs and Risk of Urological Cancer: A Meta-Analysis Based on 17,019 Subjects.
Accumulating evidence has demonstrated that some single nucleotide polymorphisms (SNPs) existing in miRNAs correlate with the susceptibility to urological cancers. However, a clear consensus still not reached due to the limited statistical power in individual study. Thus, we concluded a meta-analysis to systematically evaluate the association between microRNA SNPs and urological cancer risk. Eligible studies were collected from PubMed, Embase, Web of Science, and CNKI databases. Pooled odds ratio (OR) and corresponding 95% confidence interval (95% CI) were calculated to assess the strength of the relationships between three SNPs (miR-196a2, C>T rs11614913; miR-146a, G>C rs2910164; and miR-499, A>G rs3746444) and the risk of urological cancers. In addition, the stability of our analysis was evaluated by publication bias, sensitivity and heterogeneity analysis. Overall, a total of 17,019 subjects from 14 studies were included in this meta-analysis. We found that CT (miR-196a2, C>T rs11614913) was a risk factor for renal cell carcinoma (CT vs. CC: OR = 1.72, 95%CI = 1.05-2.80, P = 0.03, I2 = 66%), especially in Asian population (CT vs. CC: OR = 1.17, 95%CI = 1.04-1.32, P < 0.01, I2 = 0%). miR-146a G>C rs2910164 was a protective factor of urological cancers (C vs. G: OR = 0.87, 95%CI = 0.81-0.93, P < 0.01, I2 = 0%), especially for bladder cancer. miR-499 A>G rs3746444 was correlated with an increased risk of urological cancers, specifically in Asian population. In conclusion, our meta-analysis suggests that polymorphisms in microRNAs, miR-196a2, C>T rs11614913, miR-146a G>C rs2910164 and miR-499 A>G rs3746444, may be associated with the development of urological cancers and the risks mainly exist in Asian populations.


Drosophila melanogaster as a model host for Staphylococcus aureus infection.
Staphylococcus aureus is an important pathogen of humans, causing a range of superficial and potentially life-threatening diseases. Infection of the fruit fly Drosophila melanogaster with S. aureus results in systemic infection followed by death. Screening of defined S. aureus mutants for components important in pathogenesis identified perR and pheP, with fly death up to threefold slower after infection with the respective mutants compared to the wild-type. Infection of D. melanogaster with reporter gene fusion strains demonstrated the in vivo expression levels of the accessory gene regulator, agr, alpha-toxin, hla, and a manganese transporter, mntA. The use of the green fluorescent protein as a reporter under the control of the agr promoter (P3) showed S. aureus microcolony formation in vivo. The disease model also allowed the effect of antibiotic treatment on the flies to be determined. D. melanogaster is a genetically tractable model host for high-throughput analysis of S. aureus virulence determinants.


The effects of polyanions on NBT Reductions hexose monophosphate shunt activity, and ultrastructure of polymorphonuclear leukocytes.
Heparin causes enhanced nitroblue tetrazolium (NBT) reduction by polymorphonuclear leukocytes (PMN's). To determine the mechanism of this stimulation, samples of 1 to 3 x 10(7) PMN's were incubated with various concentrations of heparin, chondroitin sulfate A (CSA), and chondroitin sulfate B (CSB), with and without NBT. The effect of the polyanions (PA) on PMN hexose monophosphate shunt (HMPS) activity was determined by the production of 14CO2 from glucose-1-14C by the leukocytes. NBT reduction was evaluated histochemically and spectrophotometrically at 515 mmu. Samples of PMN's in heparin and heparin-NBT mixtures were examined by electron microscopy after various incubation periods. Increased NBT reductions by PMN's was found when leukocytes were incubated with heparin, CSA, and CSB, but these compounds had no effect on the HMPS activity of PMN's unless NBT was added. Electron microscopy of samples that contained heparin-NBT revealed an insoluble complex that was phagocytosed by the leukocytes. The stimulation of PMN oxidative metabolism and NBT reduction that follows incubation with PA-NBT appears to be directly related to ingestion of this particulate complex by the leukocytes.


Spatial anticorrelation between nonlinear white-light generation and single molecule surface-enhanced Raman scattering.
We investigate the correlation between plasmon-enhanced nonlinear white-light emission and single-molecule surface-enhanced Raman scattering (SERS) on fractal silver films using a conjugated polymer as a versatile analyte. Single molecule resonance SERS is preferentially observed from sample positions which do not exhibit nonlinear light emission under infrared excitation. The results suggest that the broad emission background often associated with single molecule SERS may not be intrinsic to the huge optical field amplifications characteristic of SERS. The two-photon imaging technique promises to offer a facile approach to prescreen substrates for their single molecule SERS capability.


Electrophysiologic effects of propranolol on sinus node function in patients with sinus node dysfunction.
The electrophysiologic effects of intravenously administered propranolol (0.1 mg/kg) on three parameters of sinus node function were examined in ten symptomatic patients with sinus node dysfunction. The patients ranged in age from 26 to 79 years. Symptoms ranged from fatigue to frank syncope. Sinoatrial (SA) block and sinus pauses were observed in one patient; sinus pauses alone were observed in three patients. Five (5/10) patients had intraatrial block; three (3/10) patients had atrioventricular block; four (4/10) patients had an intraventricular conduction disturbance. At the time of electrophysiologic study, two patients had a control spontaneous sinus cycle length that exceeded 1000 msec. Following propranolol, the mean spontaneous cycle length increased by 17.4% (924 to 1085 msec, P less than 0.005) and spontaneous second degree SA block reappeared in the one patient. The maximum escape cycle ranged from 116% to 229% of the prepacing spontaneous cycle length and was considered to be prolonged in two of ten patients. Propranolol had no significant effect on the maximum escape cycle/prepacing cycle length X 100 (%). The estimated sinoatrial conduction time (SACT) was determined in seven patients and ranged in value from 120 to 238 mes. Propranolol increased the mean value of the estimated SACT from 179 to 213 msec, P less than 0.025. Propranolol may cause marked bradyarrhythmias in some patients with sinus node dysfunction, and should be used with caution in these patients.


A step-by-step regressed pediatric kidney depth formula validated by a reasonable index.
In predicting pediatric kidney depth, we are especially interested in that the errors of most estimates are within a narrow range. Therefore, this study was intended to use the proportion of estimates within a range of -5 to 5 mm (P5 mm) to evaluate the formulas and tried to regress a kidney depth formula for children. The enrolled children aged from 1 to 19 years were randomly sampled into group A and group B (75% and 25% of all recruits, respectively). Using data of the group A, the test formula was regressed by nonlinear regression and subsequently Passing & Bablok regression, and validated in group B. The Raynaud, Gordon, Tonnesen, Taylor, and the test formulas were evaluated in the 2 groups. Accuracy was evaluated by bias, absolute bias, and P5 mm; and precision was evaluated by correlation coefficient. In addition, root-mean square error was used as a mixed index for both accuracy and precision. Body weight, height, and age did not have significant differences between the 2 groups. In the nonlinear regression, coefficients of the formula (kidney depth = a × weight/height + b × age) from group A were in narrower 95% confidence intervals. After the Passing & Bablok regression, biases of left and right kidney estimates were significantly decreased. In the evaluation of formulas, the test formula was obviously better than other formulas mentioned above, and P5 mm for left and right kidneys was about 60%. Among children younger than 10 years, P5 mm was even more than 70% for left and right kidney depths. To predict pediatric kidney depth, accuracy and precision of a step-by-step regressed formula were better than the 4 "standard" formulas.


Objective determination of claw pain and its relationship to limb locomotion score in dairy cattle.
We hypothesized that claw and foot pain could be objectively determined and have a strong effect on limb locomotion. Claw pain was measured using hoof testers equipped with a pressure gauge. Soft tissue pain was evaluated with an algometer. Pain was determined as the maximum pressure recorded at the time the limb was withdrawn following claw or soft tissue compression with the hoof tester or algometer. Locomotion scores and claw and soft tissue pain were determined on 263 Holstein cows from 2 commercial dairy farms. The frequency and the magnitude of pain had an effect on locomotion score in the ipsilateral limb for lateral, but not medial, claws. The magnitude of the lateral claw pain index for limbs with locomotion scores 1 to 5 was 0.95 +/- 0.01, 0.90 +/- 0.02, 0.67 +/- 0.04, 0.65 +/- 0.05, and 0.45 +/- 0.11, respectively. The magnitude of the medial claw pain index for limbs with locomotion scores 1 to 5 was 1.0 +/- 0.00, 0.99 +/- 0.01, 0.98 +/- 0.01, 1.0 +/- 0.00, and 1.0 +/- 0.00, respectively. The frequency of painful claws (n = 208) in limbs with locomotion scores 1, 2, and > or =3 was 0.529, 0.173, and 0.298, respectively. The frequency of painless claws (n = 318) in limbs with locomotion scores 1, 2, or > or =3 was 0.792, 0.088, and 0.120, respectively. The frequency of pain (27.1%) and total lesions (85.6%) was greater in lateral claws (n = 524) than that of pain (2.1%) and total lesions (14.4%) in medial claws (n = 524). Yet the magnitude of the pain index in sore claws was similar for medial (0.73 +/- 0.09) and lateral claws (0.64 +/- 0.02). The magnitude and frequency of claw pain in one hind limb was inconsistently and weakly affected by locomotion score or claw pain in the contralateral limb. The prevalence of unilateral (32.8%) and bilateral (23.3%) pain was similar and lower than the occurrence of bilaterally nonpainful claws (43.9%) in the study group. Painful claws (n = 78) occurred on sound limbs (n = 332) with a pain index (0.72 +/- 0.02) indicative of less pain than the pain index (0.61 +/- 0.02) of painful claws (n = 130) on lame limbs (n = 192). The results showed that lateral claw pain was related to ipsilateral limb locomotion score and subclinical pain was a relatively common occurrence. Objective measures of pain may provide a more reliable, continuous measure of clinical events used in modeling lameness.


The effect of left and right poses on the expression of facial emotion.
Research, using composite facial photographs has demonstrated that left-left composites are more emotionally expressive than are right-right composites. The present study investigated whether hemifacial asymmetries in expression are apparent in photographs, that feature one side of the face more than the other. Photographs were taken of the models who turned their heads: (a) 15 degrees to the left, (b) 15 degrees to the right or (c) faced directly towards the camera. It was predicted that left hemiface and midline photographs would be judged as more emotionally expressive than right hemiface photographs, where the left hemiface is less prominent. Three hundred and eighty-four participants viewed photographs of the three posing conditions, and rated each photograph along an emotional expressivity scale. Midline and left hemiface portraits were rated as more emotionally expressive than were right hemiface portraits. To investigate whether this effect was caused by observer's aesthetic/perceptual biases, mirror-reversed versions of the three posing conditions were included. Left hemiface and midline portraits were rated as more emotionally expressive, irrespective of whether they were mirror-reversed. It was concluded that head turns of just 15 degrees can bring about significant changes in the perceived emotionality. The relevance of these findings to painted portraits, which feature the left hemiface more than the right, is discussed.


Erythropoietin treatment in peritoneal dialysis patients.
The availability of recombinant EPO has greatly improved the lives of patients with end-stage renal disease. Knowledge is still accumulating regarding the use and effects of EPO in patients on PD, and several critical questions remain to be answered: 1. At what hematocrit level and when in the predialysis or dialysis course should EPO be started in PD patients? Recent studies by Golper suggest that the concomitant initiation of EPO and PD results in an increased hematocrit response compared to starting EPO after PD has been initiated for some time (63). 2. What is the best route of administration of EPO in PD patients? It is apparent that i.v., SC, and IP EPO can be effective in this population if utilized properly. The goal should be to tailor the route to the needs of the patient. Perhaps the daily SC route, for example, might be best for minimizing hypertension because of the slow, steady rise of hematocrit, while the IP route would be best tolerated by children (33,64). 3. What should the target hematocrit level be? This may vary depending on which organ function is being assessed. For the whole patient data are not currently available on appropriate hematocrit targets to maximize oxygen utilization, but near normal levels have recently been reported to be safe and beneficial (65-67). 4. What are the end-organ effects of anemia and its correction in PD patients? There is a dearth of information in this area for PD as well as HD patients. Additional research of this kind will increase our understanding of the pathophysiology of anemia as well as uremia.(ABSTRACT TRUNCATED AT 250 WORDS)


Online rheological investigation on ion-induced micelle transition for amphiphilic polystyrene-block-poly(acrylic acid) diblock copolymer in dilute solution.
The ion-induced micellar transition is online-investigated by the time dependence of the viscosity of the solution under shear flow for the first time. During the morphological transition, the change in the micellar structure can be tracked by the change in viscosity. Adding HCl or CaCl2 into pre-prepared spherical micelle solution from the self-assembly of polystyrene-block-poly(acrylic acid) (PS144-b-PAA22) in the N,N-dimethylformamide (DMF)/water mixture, the micellar structures change into short cylinders, long, entangled cylinders, and then lamellae or vesicles, corresponding to the viscosity increasing first and then declining. When HCl or CaCl2 is added to the pre-prepared spherical micelle solution formed by PS144-b-PAA50 in the dioxane/water mixture, the micellar structures are quickly transformed into cylinders or lamellae before carrying out the rheological measurement and then are turned to vesicles or spheres under the shearing, corresponding to a gradual decline in viscosity. This study shows that the rheology can be a very simple and effective online method on the investigation of the micellization, which plays an important role in understanding the micellization mechanism and micellar transition pathway of block copolymers in dilute solution.


Precarious Aging: Insecurity and Risk in Late Life.
Population aging and longevity in the context of declining social commitments raise concerns about disadvantage, inequality, and the well-being of older people. In this short piece, we use the concept of precarity as a lens to understand new and sustained forms of insecurity that affect late life, and we illustrate how these risks, when experienced over time and in relation to conditions such as austerity, can deepen disadvantage.


Trans-differentiation of iris pigmented epithelial cells of Euphlyctis cyanophlyctis tadpoles into lens in vitro.
Meshed pigmented iris epithelium along with neural retina of tadpoles of the frog Euphlyctis cyanophlyctis were found to undergo dedifferentiation and subsequently transdifferentiate into lens in culture medium. During lag period, depigmentation (dedifferentiation) occurred in many cells. When culture became confluent 3-4 weeks after seeding tiny lens like structures differentiated from foci of cultured pigmented iris epithelium cells. The percentage of lens formation was higher in vitamin A treated cases. The culture system appears to be a suitable for investigating the changes occurred during trans-differentiation of pigmented epithelial cells into lens.


The genetic basis of undiagnosed muscular dystrophies and myopathies: Results from 504 patients.
To apply next-generation sequencing (NGS) for the investigation of the genetic basis of undiagnosed muscular dystrophies and myopathies in a very large cohort of patients. We applied an NGS-based platform named MotorPlex to our diagnostic workflow to test muscle disease genes with a high sensitivity and specificity for small DNA variants. We analyzed 504 undiagnosed patients mostly referred as being affected by limb-girdle muscular dystrophy or congenital myopathy. MotorPlex provided a complete molecular diagnosis in 218 cases (43.3%). A further 160 patients (31.7%) showed as yet unproven candidate variants. Pathogenic variants were found in 47 of 93 genes, and in more than 30% of cases, the phenotype was nonconventional, broadening the spectrum of disease presentation in at least 10 genes. Our large DNA study of patients with undiagnosed myopathy is an example of the ongoing revolution in molecular diagnostics, highlighting the advantages in using NGS as a first-tier approach for heterogeneous genetic conditions.


Loss of exon identity is a common mechanism of human inherited disease.
It is widely accepted that at least 10% of all mutations causing human inherited disease disrupt splice-site consensus sequences. In contrast to splice-site mutations, the role of auxiliary cis-acting elements such as exonic splicing enhancers (ESE) and exonic splicing silencers (ESS) in human inherited disease is still poorly understood. Here we use a top-down approach to determine rates of loss or gain of known human exonic splicing regulatory (ESR) sequences associated with either disease-causing mutations or putatively neutral single nucleotide polymorphisms (SNPs). We observe significant enrichment toward loss of ESEs and gain of ESSs among inherited disease-causing variants relative to neutral polymorphisms, indicating that exon skipping may play a prominent role in aberrant gene regulation. Both computational and biochemical approaches underscore the relevance of exonic splicing enhancer loss and silencer gain in inherited disease. Additionally, we provide direct evidence that both SRp20 (SRSF3) and possibly PTB (PTBP1) are involved in the function of a splicing silencer that is created de novo by a total of 83 different inherited disease mutations in 67 different disease genes. Taken together, we find that ~25% (7154/27,681) of known mis-sense and nonsense disease-causing mutations alter functional splicing signals within exons, suggesting a much more widespread role for aberrant mRNA processing in causing human inherited disease than has hitherto been appreciated.


Rapid Bayesian optimisation for synthesis of short polymer fiber materials.
The discovery of processes for the synthesis of new materials involves many decisions about process design, operation, and material properties. Experimentation is crucial but as complexity increases, exploration of variables can become impractical using traditional combinatorial approaches. We describe an iterative method which uses machine learning to optimise process development, incorporating multiple qualitative and quantitative objectives. We demonstrate the method with a novel fluid processing platform for synthesis of short polymer fibers, and show how the synthesis process can be efficiently directed to achieve material and process objectives.


Bone assessment and reconstruction in revision hip surgery.
As a result of septic and aseptic complications, total hip replacements can fail and consequently have finite life expectancies. One of the most challenging aspects of revision surgery is the management of bone loss. We present a classification that maps the severity of bone loss with a variety of reconstructive options, which include cemented and uncemented fixations. This classification has been used successfully to guide preoperative planning, to help determine intraoperative needs (implants, specialized equipment, and bone grafts), and to help prepare the patient about the postoperative rehabilitation course.


New species of Chelonus (Microchelonus) Szépligeti, 1908 (Hymenoptera: Braconidae: Cheloninae) from Brazil.
Chelonus (Microchelonus) murici sp. nov. (Hymenoptera: Braconidae: Cheloninae) is described in this work. It was reared from an unknown host in murici fruits, Byrsonima verbascifolia (L.) Rich. ex A. L. Juss. (Malpighiaceae), a species from the Brazilian savannah whose fruits are widely consumed by the population in northern Brazil. The adult of this new species is illustrated.


Nanozyme Sensor Arrays Based on Heteroatom-Doped Graphene for Detecting Pesticides.
Pesticides, widely used for pest control and plant growth regulation, have posed a threat to the environment and human health. Conventional methods to analyze pesticide residues are not applied to resource-limited areas because of their high cost, complexity, and requirements for expensive instruments (such as GC/MS and LC/MS). To address these challenges, herein we fabricated colorimetric nanozyme sensor arrays based on heteroatom-doped graphene for detection of aromatic pesticides. The active sites of nanozymes could be differentially masked when different pesticides were adsorbed on the graphene, which in turn resulted in the decrease of their peroxidase-mimicking activities. On the basis of this principle, five pesticides (i.e., lactofen, fluoroxypyr-meptyl, bensulfuron-methyl, fomesafen, and diafenthiuron) from 5 to 500 μM were successfully discriminated by the sensor arrays. In addition, discrimination for different concentrations of each pesticide and different ratios of two mixed pesticides were also demonstrated. The practical application of the sensor arrays was further validated by successfully discriminating the pesticides in soil samples. This work not only provides a facile and cost-effective method to detect pesticides but also makes a positive contribution to food safety and environmental protection.


Queensland tick typhus in rural New South Wales.
We report five cases of Rickettsia australis infection from southern coastal New South Wales, Australia. All patients presented with a cutaneous eruption of erythematous papules and pustules and systemic features of malaise, headache, lymphadenopathy and myalgia. Acute kidney injury (AKI) was present in two of five cases and one of five cases had acute delirium. Improvement was only seen after treatment with doxycycline 100 mg b.i.d. Positive serology for R. australis was present in four of five cases and a positive polymerase chain reaction (PCR) was seen in one of five cases. Histology showed varying features, from neutrophilic vasculitis to Sweet's syndrome and lymphocytic vasculitis. Recent significant advances in the diagnosis of R. australis infection include an eschar swab or biopsy PCR and isolation of specific Rickettsia on serology. These investigations should be considered in the presence of any of the following features: eschar at site of a tick bite or lymphadenopathy and fever with an eruption of erythematous papules and pustules.


Molecular mechanism of hepcidin-mediated ferroportin internalization requires ferroportin lysines, not tyrosines or JAK-STAT.
Ferroportin is the primary means of cellular iron efflux and a key component of iron metabolism. Hepcidin regulates Fpn activity by inducing its internalization and degradation. The mechanism of internalization is reported to require JAK2 activation, phosphorylation of Fpn tyrosine residues 302 and 303, and initiation of transcription through STAT3 phosphorylation. These findings suggest Fpn may be a target for therapeutic intervention through JAK2 modulation. To evaluate the proposed mechanism, Fpn internalization was assessed using several techniques combined with reagents that specifically recognized cell-surface Fpn. In vitro results demonstrated that Hepc-induced Fpn internalization did not require JAK2 or phosphorylation of Fpn residues 302 and 303, nor did it induce JAK-STAT signaling. In vivo, inhibition of JAK2 had no effect on Hepc-induced hypoferremia. However, internalization was delayed by mutation of two Fpn lysine residues that may be targets of ubiquitination.


Quality of life in epilepsy: the clinician's view.
Quality of life in patients with epilepsy may be impaired by seizures, side effects of medication, and psychosocial problems. Doctors tend to focus their attention on managing the seizures and the side effects, but may not be meeting all of the patient's needs because of differences between their perception of these concerns and the patient's. Further, psychosocial problems often are addressed only superficially or not at all during office visits. The challenge is to integrate quality-of-life issues into clinical practice and to better assess the patient's perception of the disorder, the seizures, and the medications, as they impact on cognitive function, emotional well-being, and social and economic functioning.


Luminescent phosphine gold(I) thiolates: correlation between crystal structure and photoluminescent properties in [R3PAu{SC(OMe)=NC6H4NO2-4}] (R = Et, Cy, Ph) and [(Ph2P-R-PPh2){AuSC(OMe)=NC6H4NO2-4}2] (R = CH2, (CH2)2, (CH2)3, (CH2)4, Fc).
X-ray crystallography shows the gold atoms in [R3PAu{SC(OMe)=NC6H4NO2-4}] (R = Et, Cy, Ph; 1-3, respectively) and [(Ph2P-R-PPh2){AuSC(OMe)=NC6H4NO2-4}(2)] (R = CH2, (CH2)2, (CH2)3, (CH2)4, Fc; 4-8, respectively) are linearly coordinated by phosphorus and thiolate-sulfur; weak intramolecular Au...O interactions are featured in all structures. The smaller ethyl substituents in 1 allow for supramolecular association via Au...S and Au...Au interactions that are not found in 2 and 3, which contain larger phosphorus-bound Cy and Ph groups, respectively. Intramolecular Au...Au interactions are found in the dppm, dppe, dppp, and Fc structures but not in the dppp analogue, for which an anti conformation was found. The structures have been correlated with the results from photophysical study conducted in the solid state. Thus, photoexcitation of 1-7 with lambda > 350 in the solid state and in solution produces green and blue luminescence, respectively. The spectra in each medium are remarkably similar to each other, and so the emission energy and excitation maxima observed for 1-7 appear to be independent of the nature of the ancillary phosphines, as well as the presence or absence of Au...Au interactions, either intermolecularly or intramolecularly.


TSC1 regulates osteoclast podosome organization and bone resorption through mTORC1 and Rac1/Cdc42.
Reorganization of the podosome into the sealing zone is crucial for osteoclasts (OCLs) to resorb bone, but the underlying mechanisms are unclear. Here, we show that tuberous sclerosis complex 1 (TSC1) functions centrally in OCLs to promote podosome organization and bone resorption through mechanistic target of rapamycin complex 1 (mTORC1) and the small GTPases Rac1/Cdc42. During osteoclastogenesis, enhanced expression of TSC1 downregulates mTORC1 activity. TSC1 deletion in OCLs reduced podosome belt formation in vitro and sealing zone formation in vivo, leading to bone resorption deficiency and osteopetrosis. Mechanistically, TSC1 promoted podosome superstructure assembly by releasing mTORC1-dependent negative feedback inhibition of Rac1/Cdc42. Rapamycin and active Rac1/Cdc42 restore podosome organization and bone resorption and alleviate osteopetrotic phenotypes in mutant mice. Our findings reveal an essential role of TSC1 signaling in the regulation of bone resorption. Targeting TSC1 represents a novel strategy to inhibit bone resorption and prevent bone loss-related diseases.


Essential role of Skp2-mediated p27 degradation in growth and adaptive expansion of pancreatic beta cells.
Diabetes results from an inadequate mass of functional beta cells, due to either beta cell loss caused by immune assault or the lack of compensation to overcome insulin resistance. Elucidating the mechanisms that regulate beta cell mass has important ramifications for fostering beta cell regeneration and the treatment of diabetes. We report here that Skp2, a substrate recognition component of Skp1-Cul1-F-box (SCF) ubiquitin ligase, played an essential and specific role in regulating the cellular abundance of p27 and was a critical determinant of beta cell proliferation. In Skp2(-/-) mice, accumulation of p27 resulted in enlarged polyploid beta cells as a result of endoreduplication replacing proliferation. Despite beta cell hypertrophy, Skp2(-/-) mice exhibited diminished beta cell mass, hypoinsulinemia, and glucose intolerance. Increased insulin resistance resulting from diet-induced obesity caused Skp2(-/-) mice to become overtly diabetic, because beta cell growth in the absence of cell division was insufficient to compensate for increased metabolic demand. These results indicate that the Skp2-mediated degradation pathway regulating the cellular degradation of p27 is essential for establishing beta cell mass and to respond to increased metabolic demand associated with insulin resistance.


Two cases of early recurrence after transabdominal preperitoneal inguinal hernia repair.
We performed transabdominal preperitoneal inguinal hernia repair in 46 patients (58 diseases), two of whom experienced early recurrence after mesh repair. Case 1 was a 76-year-old man with a bilateral inguinal hernia (recurrence site, left indirect hernia) after appendectomy. The recurrence occurred 1 month after transabdominal preperitoneal inguinal hernia repair. The mesh was dislocated to the lateral side, and we repaired it using the direct Kugel® patch with an anterior technique. Case 2 was a 79-year-old man with a bilateral inguinal hernia (recurrence site, right direct hernia with an orifice >3 cm) after appendectomy. The recurrence occurred 3 months after transabdominal preperitoneal inguinal hernia repair. The mesh was dislocated to the lateral side, and we repaired it using an ULTRAPRO® Plug with an anterior technique under laparoscopic observation. We believe the recurrences resulted from insufficient internal exfoliation and fixation affected by complicated exfoliation of the preperitoneal space with omental adhesion after intraperitoneal surgery.


Intracorporeal anastomosis using a Lapra-ty clip in laparoscopic distal gastrectomy: initial clinical experiences.
The aim of this study was to assess the efficacy, reliability, and performance of the Lapra-Ty clip (LTc) during intracorporeal Roux-en-Y gastrojejunostomy in a totally laparoscopic distal gastrectomy (TLDG). Between May 2009 and October 2009, we performed TLDG on 25 consecutive early gastric cancer patients. Two-bowel anastomosis was done intracorporeally in the side-to-side fashion using Endo-GIA, and closure of the entry hole was done by LTc with a running suture. There were no postoperative leakages, bleeding, or strictures in the gastrojejunostomies and jejunojejunostomies. We had a single intraoperative complication where the Levin tube was jammed at the gastrojejunostomy site, so a revision was done. Total mean operation time was 254.4 minutes (range, 170-485) and mean anastomosis time was 49 minutes (range, 30-110). Mean hospital stay was 7.9 days (range, 5-17), and mean time to liquid diet was 2.6 days postoperatively (range, 2-5). The usage of LTc is safe and efficient for bowel anastomosis, especially TLDG in humans, and adoption of LTc may help to reduce overall operative times. Perhaps, most importantly, it can cover the disadvantages of TLDG by making TLDG more easily executable in gastric cancer.


Premenstrual affective syndrome and psychiatric disorder.
The relationship between premenstrual affective syndrome and psychiatric disorder was investigated, using 81 women presenting to a Neurology Clinic with functional headache. Premenstrual affective syndrome was significantly associated with a history of depressive syndrome in the population studied. Patients judged to have a non-affective psychiatric disorder reported no greater frequency of definite or probable premenstrual affective syndrome than patients considered psychiatrically normal. The premenstrual occurrence or exacerbation of affective symptoms has been noted. This symptom exacerbation maybe sufficient to require hospitalization. Data presented by Coppen indicate that women with affective disorder are more likely to report the premenstrual symptom of depression than women with other psychiatric disorders. These findings suggest that there may be some relationship between depressive disorder and premenstrual affective symptoms. As part of a larger study on the personality and psychiatric correlates of functional headache, data on the relationship between depressive syndrome and premenstrual affective symptoms were obtained.


Peeling off an adhesive layer with spatially varying modulus.
We analyze here displacement controlled peeling of a flexible adherent off a thin layer of elastic adhesive, the elastic modulus of which does not remain uniform but varies periodically along the direction of peeling. Calculation shows that with progressive peeling, the crack front does not propagate continuously at the interface but intermittently with crack arrests and subsequent initiations. The crack gets arrested close to the location of the minimum shear modulus of the layer and initiates again only at a sufficiently large peel off load. This effect is very similar to the peeling experiment off surface patterned and microchannel embedded adhesives which results in significant enhancement of fracture toughness of the interface over smooth adhesive layers. The fracture toughness of the interface increases with the increase in thickness of the layer and the amplitude of variation in modulus. Fracture toughness is calculated to be high also for the larger value of critical stress at the opening of the crack. With the wavelength of modulus variation, it varies nonmonotonically, maximizing at an intermediate value. These results define the criterion for designing adhesive layers with spatially modulated physical properties useful for variety of applications.


Intestinal and hepatobiliary disposition of drugs.
Biliary excretion of xenobiotics is a complex process involving uptake into liver, intracellular sequestration and/or biotransformation, and transport into bile. A description of liver morphology is included to aid in the understanding of how chemical and physiological factors affect hepatic uptake and biliary excretion. Data will be presented with morphine and nalorphine that illustrate how efficient the intestinal and hepatobiliary system is in clearing drugs from the body. However, enterohepatic circulation can interfere with these processes.


Hyperaccumulation of Gadolinium by Methylorubrum extorquens AM1 Reveals Impacts of Lanthanides on Cellular Processes Beyond Methylotrophy.
Lanthanides (Ln) are a new group of life metals, and many questions remain regarding how they are acquired and used in biology. Methylotrophic bacteria can acquire, transport, biomineralize, and use Ln as part of a cofactor complex with pyrroloquinoline quinone (PQQ) in alcohol dehydrogenases. For most methylotrophic bacteria use is restricted to the light Ln, which range from lanthanum to samarium (atomic numbers 57-62). Understanding how the cell differentiates between light and heavy Ln, and the impacts of these metals on the metabolic network, will advance the field of Ln biochemistry and give insights into enzyme catalysis, stress homeostasis, and metal biomineralization and compartmentalization. We report robust methanol growth with the heavy Ln gadolinium by a genetic variant of the model methylotrophic bacterium Methylorubrum extorquens AM1, named evo-HLn, for "evolved for Heavy Lanthanides." A non-synonymous single nucleotide polymorphism in a cytosolic hybrid histidine kinase/response regulator allowed for sweeping transcriptional alterations to heavy metal stress response, methanol oxidation, and central metabolism. Increased expression of genes for Ln acquisition and uptake, production of the Ln-chelating lanthanophore, PQQ biosynthesis, and phosphate transport and metabolism resulted in gadolinium hyperaccumulation of 36-fold with a trade-off for light Ln accumulation. Gadolinium was hyperaccumulated in an enlarged acidocalcisome-like compartment. This is the first evidence of a bacterial intracellular Ln-containing compartment that we name the "lanthasome." Carotenoid and toblerol biosynthesis were also upregulated. Due to its unique capabilities, evo-HLn can be used to further magnetic resonance imaging (MRI) and bioremediation technologies. In this regard, we show that gadolinium hyperaccumulation was sufficient to produce MRI contrast in whole cells, and that evo-HLn was able to readily acquire the metal from the MRI contrast agent gadopentetic acid. Finally, hyperaccumulation of gadolinium, differential uptake of light and heavy Ln, increased PQQ levels, and phosphate transport provide new insights into strategies for Ln recovery.


Heparan sulfate in perlecan promotes mouse atherosclerosis: roles in lipid permeability, lipid retention, and smooth muscle cell proliferation.
Heparan sulfate (HS) has been proposed to be antiatherogenic through inhibition of lipoprotein retention, inflammation, and smooth muscle cell proliferation. Perlecan is the predominant HS proteoglycan in the artery wall. Here, we investigated the role of perlecan HS chains using apoE null (ApoE0) mice that were cross-bred with mice expressing HS-deficient perlecan (Hspg2(Delta3/Delta3)). Morphometry of cross-sections from aortic roots and en face preparations of whole aortas revealed a significant decrease in lesion formation in ApoE0/Hspg2(Delta3/Delta3) mice at both 15 and 33 weeks. In vitro, binding of labeled mouse triglyceride-rich lipoproteins and human LDL to total extracellular matrix, as well as to purified proteoglycans, prepared from ApoE0/Hspg2(Delta3/Delta3) smooth muscle cells was reduced. In vivo, at 20 minutes influx of human (125)I-LDL or mouse triglyceride-rich lipoproteins into the aortic wall was increased in ApoE0/Hspg2(Delta3/Delta3) mice compared to ApoE0 mice. However, at 72 hours accumulation of (125)I-LDL was similar in ApoE0/Hspg2(Delta3/Delta3) and ApoE0 mice. Immunohistochemistry of lesions from ApoE0/Hspg2(Delta3/Delta3) mice showed decreased staining for apoB and increased smooth muscle alpha-actin content, whereas accumulation of CD68-positive inflammatory cells was unchanged. We conclude that the perlecan HS chains are proatherogenic in mice, possibly through increased lipoprotein retention, altered vascular permeability, or other mechanisms. The ability of HS to inhibit smooth muscle cell growth may also influence development as well as instability of lesions.


Molecular single cell studies of normal and transformed lymphocytes.
The polymerase chain reaction allows the characterization of RNA and DNA sequences from single cells. Methods were established to analyse single cells isolated from suspension or by multicolour flow cytometry. We established a method to isolate single immunostained cells from frozen tissue sections and to analyse those cells for immunoglobulin gene rearrangements. This method was first used to study B cell differentiation within human germinal centres. In another series of experiments, Hodgkin and Reed-Sternberg (HRS) cells from a total of 14 cases of HD were analysed for B lineage derivation and clonality. In 13 of the 14 cases, clonal V gene rearrangements were identified. This shows that HRS cells generally represent the outgrowth of a clonal population of B cells. The detection of somatic mutations in all VH gene rearrangements amplified from HRS cells and the nature of those mutations identifies a GC B cell as the HRS precursor.


Advances in Low-Lactose/Lactose-Free Dairy Products and Their Production.
With increasing health awareness worldwide, lactose intolerance has become a major concern of consumers, creating new market opportunities for low-lactose/lactose-free dairy foods. In recent years, through innovating processes and technologies, dairy manufacturers have significantly improved the variety, and functional and sensory qualities of low-lactose and lactose-free dairy products. Based on this, this paper first covers the pathology and epidemiology of lactose intolerance and market trends. Then, we focus on current advantages and disadvantages of different lactose hydrolysis technologies and improvements in these technologies to enhance nutritional value, and functional, sensory, and quality properties of lactose-free dairy products. We found that more and more cutting-edge technologies are being applied to the production of lactose-free dairy products, and that these technologies greatly improve the quality and production efficiency of lactose-free dairy products. Hopefully, our review can provide a theoretical basis for the marketing expansion and consumption guidance for low-lactose/lactose-free dairy products.


Lessons from managed care.
The infiltration of the health-care environment by health maintenance organizations has been promoted, in part, by instilling fear in physicians about their patient base and creating an illusion of free choice for patients. Within capitation arrangements, specialists are often underpaid for management of complex cases and uncompensated for investment of long hours in ancillary activities. Clinical endocrinologists have been expected to develop algorithms for conditions such as diabetic ketoacidosis, yet application of such guidelines by other clinicians may yield suboptimal results. The transformation of medicine to a business and the focus of insurance administrators on profit rather than high-quality care currently tend to eliminate specialists. Studies that show that high-quality specialty care improves outcomes clinically (and therefore financially) should curb this trend. Nevertheless, a new paradigm for medical care is evolving. Because capitation will not be the only model, clinical endocrinologists should not allow themselves to be exploited in such an arrangement.


Phycobiliproteins and phycobilisomes: the early observations.
The purpose of this minireview is to highlight the early observations that led to the discovery of the physico-chemical properties of the phycobiliproteins, their structure and function, and to their architectural organization in supramolecular complexes, the phycobilisomes. Generally attached on the stromal surface of the thylakoid membranes in both prokaryotic (cyanobacteria) and eukaryotic cells (cyanelles, red algae and cryptomonads), these complexes represent the most abundant soluble proteins and the major light-harvesting antennae for photosynthesis. This review mainly focuses on the years prior to the development of the molecular biology of cyanobacteria that flourished in the 1980s. We refer the reader to the comprehensive and excellent review by Sidler (1994) for more recent discoveries and more detailed literature on this topic. [-6pt]'It would be difficult to find another series of colouring matters of greater beauty or with such remarkable and instructive chemical and physical peculiarities.' -H. Sorby, 1877.


Effect of isoniazid preventive therapy on immune responses to mycobacterium tuberculosis: an open label randomised, controlled, exploratory study.
With the renewed emphasis to implement isoniazid preventive therapy (IPT) in Sub-Saharan Africa, we investigated the effect of IPT on immunological profiles among household contacts with latent tuberculosis. Household contacts of confirmed tuberculosis patients were tested for latent tuberculosis using the QuantiFERON®-TB Gold In-Tube (QFN) assay and tuberculin skin test (TST). HIV negative contacts aged above 5 years, positive to both QFN and TST, were randomly assigned to IPT and monthly visits or monthly visits only. QFN culture supernatants from enrolment and six months' follow-up were analysed for M.tb-specific Th1, Th2, Th17, and regulatory cytokines by Luminex assay, and for M.tb-specific IgG antibody concentrations by ELISA. Effects of IPT were assessed as the net cytokine and antibody production at the end of six months. Sixteen percent of contacts investigated (47/291) were randomised to IPT (n = 24) or no IPT (n = 23). After adjusting for baseline cytokine or antibody responses, and for presence of a BCG scar, IPT (compared to no IPT) resulted in a relative decline in M.tb-specific production of IFN gamma (adjusted mean difference at the end of six months (bootstrap 95% confidence interval (CI), p-value) -1488.6 pg/ml ((-2682.5, -294.8), p = 0.01), and IL- 2 (-213.1 pg/ml (-419.2, -7.0), p = 0.04). A similar decline was found in anti-CFP-10 antibody levels (adjusted geometric mean ratio (bootstrap 95% CI), p-value) 0.58 ((0.35, 0.98), p = 0.04). We found no effect on M.tb-specific Th2 or regulatory or Th17 cytokine responses, or on antibody concentrations to PPD and ESAT-6. IPT led to a decrease in Th1 cytokine production, and also in the anti CFP-10 antibody concentration. This could be secondary to a reduction in mycobacterial burden or as a possible direct effect of isoniazid induced T cell apoptosis, and may have implications for protective immunity following IPT in tuberculosis-endemic countries. ISRCTN registry, ISRCTN15705625. Registered on 30(th) September 2015.


Physiologically based pharmacokinetic models for everolimus and sorafenib in mice.
Everolimus is a mammalian target of rapamycin (mTOR) inhibitor approved as an immunosuppressant and for second-line therapy of hepatocellular carcinoma (HCC) and renal cell carcinoma (RCC). Sorafenib is a multikinase inhibitor used as first-line therapy in HCC and RCC. This study assessed the pharmacokinetics (PK) of everolimus and sorafenib alone and in combination in plasma and tissues, developed physiologically based pharmacokinetic (PBPK) models in mice, and assessed the possibility of PK drug interactions. Single and multiple oral doses of everolimus and sorafenib were administered alone and in combination in immunocompetent male mice and to severe combined immune-deficient (SCID) mice bearing low-passage, patient-derived pancreatic adenocarcinoma in seven different studies. Plasma and tissue samples including tumor were collected over a 24-h period and analyzed by liquid chromatography-tandem mass spectrometry (LC-MS/MS). Distribution of everolimus and sorafenib to the brain, muscle, adipose, lungs, kidneys, pancreas, spleen, liver, GI, and tumor was modeled as perfusion rate-limited, and all data from the diverse studies were fitted simultaneously using a population approach. PBPK models were developed for everolimus and sorafenib. PBPK analysis showed that the two drugs in combination had the same PK as each drug given alone. A twofold increase in sorafenib dose increased tumor exposure tenfold, thus suggesting involvement of transporters in tumor deposition of sorafenib. The developed PBPK models suggested the absence of PK interaction between the two drugs in mice. These studies provide the basis for pharmacodynamic evaluation of these drugs in patient-derived primary pancreatic adenocarcinomas explants.


Production performance of white Leghorn layers limited fed.
Three trials were conducted to determine if the age at start of limited feeding or the amount of time laying hens were given to consume feed could be used to reduce feed intake without affecting egg weight (EW) or egg production. The age at start of limiting feed did not affect hen-day egg production (HDEP) or EW in any trial. In Trial 2, a significant difference in HDEP due to feeding times at 26 weeks of age was observed. Laying hens fed 7 hr/day at 26 weeks of age had a significantly lower HDEP than any other feeding time. Limited feeding in Trials 1 and 2 significantly decreased feed consumption, body weight gain (WG), and EW when compared with laying hens fed ad libitum. No difference in HDEP or EW was observed due to limited feeding in Trial 3. Laying hens fed 9 or 10 hr/day gained significantly less weight than laying hens fed ad libitum (Trial 3). Limited feeding did not improve feed efficiency in any trial. No consistent improvement in mortality could be attributed to limited feeding.


Long non-coding RNA CCAT2 promotes cell proliferation and invasion through regulating Wnt/β-catenin signaling pathway in clear cell renal cell carcinoma.
Clear cell renal cell carcinoma (ccRCC) is a common urologic malignancy. Long non-coding RNA colon cancer-associated transcript 2 (CCAT2) has been suggested as serving pivotal roles in tumorigenesis. However, the clinical significance and biological role of CCAT2 in ccRCC remains elusive. The purpose of this study is to identify the function of CCAT2 in ccRCC and its possible molecular mechanism. Expression of CCAT2 was analyzed in 61 ccRCC tissues and two ccRCC cell lines (786-O and ACHN) by quantitative reverse transcription polymerase chain reaction. The functional roles of CCAT2 in ccRCC were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, colony formation assay, Transwell assay, and flow cytometric analysis. The influence of CCAT2 on tumorigenesis was monitored by in vivo mice xenograft model. The activation of Wnt/β-catenin signaling pathway was evaluated by the TOP/FOP Wnt luciferase reporter assay and western blot assay. CCAT2 expression was markedly higher in ccRCC cell lines and tissues, being positively associated with tumor size and tumor stage in ccRCC patients. Patients with higher CCAT2 expression had a markedly poorer overall survival than did patients with low CCAT2 expression. Knocking down CCAT2 expression led to reduced cell proliferation and increased apoptosis of ccRCC cells in vitro as well as the activation of Wnt/β-catenin signaling pathway, and CCAT2 overexpression remarkably enhanced these oncogenic properties. In vivo mice xenograft model also showed that knocking CCAT2 expression inhibited the growth of ccRCC xenografts. In conclusion, these results indicated that CCAT2 may play a critical role in ccRCC progression and will be further considered as a biomarker for predicting the survival of ccRCC patients and a potential therapeutic target for ccRCC intervention.


Engineering of a bioluminescent probe for imaging nitroxyl in live cells and mice.
A bioluminescent probe, BP-HNO, which exhibits a turn-on response to nitroxyl with high sensitivity and selectivity, is reported for the first time in this work. BP-HNO is free from the interference of biological autofluorescence to afford a high signal-to-noise ratio for bioimaging, and was successfully applied to imaging nitroxyl in live cells and mice.


Teaching evidence-based medicine in a managed care setting: from didactic exercise to pharmacopolicy development tool.
To implement a residency-based program for the teaching of evidence-based medicine in an Israeli HMO and to incorporate this effort into the HMO's routine drug policy formulation process. Residents and preceptors participating in the family practice residency program in The Leumit Health Fund, 1 of the 4 HMOs operating in Israel, were invited to participate in a workshop for the formulation of guidelines for antibiotic treatment of the common infectious diseases encountered in primary care. The participants were allocated to teams consisting of a preceptor (an attending physician) and a resident physician, with each team choosing a different disease to analyze. Upon completion of the program, a questionnaire was sent to all residents and preceptors who participated in the workshop to evaluate attitudes concerning the outcomes of the program. Guidelines for the treatment of 14 infectious diseases commonly seen in the primary care setting were formulated. The program was accepted by the participants, who ultimately cooperated with the relevant HMO stakeholders in the formulation of official HMO policies for drug prescribing. The utilization of family practice residents is a feasible method of formulating in-house clinical practice guidelines for a managed care setting. The program was mutually beneficial for both the residents and for the stakeholders in the HMO.


Mutation in the S gene a determinant of the hepatitis B virus associated with concomitant HBsAg and anti-HBs in a population in Northeastern Brazil.
Mutations in the a determinant of S gene may develop co-existence of hepatitis B surface antigen (HBsAg) and antibodies to HBsAg (anti-HBs) in the serum of infected hepatitis B virus (HBV) individuals. Mutations in this region may change the antigenicity of HBsAg, which in turn, lead to escape of neutralizing action of anti-HBs antibodies. This study identified individuals with concomitant HBsAg and anti-HBs serological markers in individuals of Maranhão, Northeastern Brazil. Samples from a population-based study were evaluated for HBsAg, anti-HBs, and anti-HBc, and those that tested positive for simultaneous HBsAg and anti-HBs were submitted to HBV DNA quantification and S gene characterization by Sanger sequencing. Mutations were investigated in the a determinant located in major hydrophilic region (MHR) of the S gene. Among 3,984 samples analyzed, 92 (2.3%) were positive for HBsAg and three had the atypical HBsAg and anti-HBs-positive profile (3.26%). The frequency of HBsAg and anti-HBs co-existence was similar to previous studies. Only one individual harbored mutation in the S gene a determinant associated with this profile. Little is known about this phenomenon; however, studies as ours may contribute for future enlightenment of this important issue. J. Med. Virol. 89:458-462, 2017. © 2016 Wiley Periodicals, Inc.


Development of miRNA-Based Approaches to Explore the Interruption of Mosquito-Borne Disease Transmission.
MicroRNA (miRNA or miR)-based approaches to interrupt the transmission of mosquito-borne diseases have been explored since 2005. A review of these studies and areas in which to proceed is needed. In this review, significant progress is reviewed at the level of individual miRNAs, and miRNA diversification and relevant confounders are described in detail. Current miRNA studies in mosquitoes include four steps, namely, identifying miRNAs, validating miRNA-pathogen interactions, exploring action mechanisms, and performing preapplication investigations. Notably, regarding the Plasmodium parasite, mosquito miRNAs generally bind to mosquito immunity- or development-related mRNAs, indirectly regulating Plasmodium infection; However, regarding arboviruses, mosquito miRNAs can bind to the viral genome, directly modifying viral replication. Thus, during explorations of miRNA-based approaches, researchers need select an ideal miRNA for investigation based on the mosquito species, tissue, and mosquito-borne pathogen of interest. Additionally, strategies for miRNA-based approaches differ for arboviruses and protozoan parasites.


Extended versus local fluctuations in quantum critical Ce(Ru1-xFex)2Ge2 (x=xc=0.76).
We present inelastic neutron scattering experiments, performed near the antiferromagnetic quantum critical point in Ce(Ru0.24Fe0.76)2Ge2. Both local and long-range fluctuations of the local moments are observed, but due to the Kondo effect only the latter are critical. We propose a phenomenological expression which fits the energy E, temperature T, and wave vector q dependences of the dynamic susceptibility, describing the non-Fermi liquid E/T scaling found at every q.


Corticospinal excitability changes following prolonged muscle tendon vibration.
The present experiment addressed the time course of corticospinal excitability changes following interventional muscle tendon vibration. Using transcranial magnetic stimulation, motor evoked potentials of the flexor carpi radialis and extensor carpi radialis brevis muscle were recorded for a period of 60 min after cessation of vibration (80 Hz, 0.5 mm, 30 min) to the distal wrist flexor tendons. A delayed corticospinal excitability increase in both the vibrated and non-vibrated antagonistic muscle was observed, with lasting levels of facilitation for the latter. No changes were observed following interventional cutaneous vibration. These results underscore a facilitatory influence of prolonged Ia-afferent activation on corticospinal excitability. Findings are discussed in light of recent advances in promoting motor recovery after brain injury by somatosensory stimulation.


Identification, Synthesis, and Characterization of Potential Oxidative Impurities of Venetoclax: Application of Meisenheimer Rearrangement.
Venetoclax is a potent BCL-2 inhibitor that is used for the treatment of several blood cancers. During the oxidative stress degradation of venetoclax, we observed the formation of two potential impurities at levels of about 8-10%, which have similar molecular weights. The two impurities were isolated and identified as 4-(3-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(((3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)sulfonyl)carbamoyl)phenyl)-1-((4'-chloro-5,5-dimethyl-3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-yl)methyl)piperazine 1-oxide (venetoclax N-oxide, VNO) and 2-((1H-pyrrolo[2,3-b]pyridin-5-yl)oxy)-4-(4-((4'-chloro-5,5-dimethyl-3,4,5,6-tetrahydro-[1,1'-biphenyl]-2-yl)methoxy)piperazin-1-yl)-N-((3-nitro-4-(((tetrahydro-2H-pyran-4-yl)methyl)amino)phenyl)sulfonyl)benzamide (venetoclax hydroxylamine impurity, VHA). To confirm these two compounds, we have synthesized each impurity individually and analyzed it by high-performance liquid chromatography, mass spectrometry, 1H NMR, 13C NMR, and 2D NMR. VNO was synthesized by the oxidation of venetoclax using m-CPBA in dichloromethane to get the required N-oxide impurity. After the confirmation of the VNO impurity, the VNO impurity was heated with water at reflux in a sealed tube for 36 h to get the VHA impurity of about 6-8% after 36 h. After thorough analysis, it was confirmed that venetoclax N-oxide undergoes [1,2] Meisenheimer rearrangement to form the venetoclax hydroxylamine impurity. These two impurities may be relevant reference standards in manufacturing venetoclax Active Pharmaceutical Ingredient (API) (or) tablets.


Localization of methionine-enkephalin, substance P, and somatostatin immunoreactivities in the main olfactory bulb of the hamster.
Methionine-enkephalin, substance P, and somatostatin immunoreactivities are associated with different classes of neurons in the main olfactory bulb of the hamster. In the glomerular layer, methionine-enkephalin immunoreactivity is observed in the somata and dendrites of periglomerular cells whereas substance P immunoreactivity is present in the somata and dendrites of external tufted cells. Substance P immunoreactivity may also be present in the superficial short axon cells. In the deeper layers, methionine-enkephalin immunoreactivity is observed in a mixed population of neurons which appear to be primarily granule cells, whereas somatostatin immunoreactivity is present only in deep short axon cells. Methionine-enkephalin-, substance P-, and somatostatin-positive fibers of central origin are also observed in the main olfactory bulb.


Comparison of the in vitro effects of the anti-CD20 antibodies rituximab and GA101 on chronic lymphocytic leukaemia cells.
The effects of two CD20 antibodies, namely rituximab, the current standard for treatment of chronic lymphocytic leukaemia (CLL) in combination with chemotherapy, and GA101, a glyco-engineered type II antibody were compared on CLL cells ex vivo. Antibody-induced phosphatidylserine exposure was examined in isolated CLL cells. For a more comprehensive assessment of antibody-mediated cell killing including Fc-mediated mechanisms, B cell depletion from whole blood samples was monitored. Treatment with rituximab or GA101 reduced the average viability of isolated CLL cells by 6% or 11%, and the ratio of B to T cells in whole blood samples by 12% or 33%, respectively. Combination with GA101 enhanced the cytotoxicity of the chemotherapeutic agent chlorambucil on isolated CLL cells. CD20 surface expression on CLL cells correlated with GA101-induced B cell depletion, but not with direct cell death induction. Treatment of whole blood samples from CLL patients with a CpG-containing oligonucleotide increased CD20 expression on CLL cells and GA101-dependent B cell depletion. Despite the variable responses of individual CLL samples, the CLL cell depletion from whole blood by GA101 was consistently much stronger than by rituximab, which argues for clinical investigation of GA101 in CLL patients.


Weight loss experiences and willingness to intervention with pharmacotherapy among obese and very obese Danish people.
Obesity is a growing issue with increasing impact on healthcare budgets, yet very little is known about weight loss experiences of people with body mass index (BMI)≥30kg/m(2) and their willingness to try and pay for weight loss interventions (WLI). The objective of this survey was to gather knowledge about weight loss experiences among obese and severely obese people. 1,003 Danish people >18 years of age with BMI≥30 who wanted to lose weight completed an online survey. Data included demographics, experience with WLI, awareness of anti-obesity medication (AOM), and willingness to try and pay for WLI including AOM. Respondents had been trying to lose weight for several years (medium [25% percentile;75% percentile]);5.1[2.0;10.3] years (BMI 30-35) and 10.0 [5.0;20.0] (very obese (BMI>35) with co-morbidities (OWC). The desired weight loss was 20.0 [15.0;25.0] kg (BMI 30-35) and 35.0 [28.0;47.5] kg (OWC). Independent of educational level and gender, health concern was the main incentive for weight loss. Several WLI had been tried repeatedly, yet 60% of respondents with BMI 30-35 and 50% of the OWC were unaware of AOM. Among those who had tried AOM, side effects and lack of effectiveness were the main reasons to stop. 50-73% were willing to try AOM dependent on expected weight loss. Willingness to try and pay for new AOM was strongest for the OWC. Respondents had made repeated attempts for up to a decade to lose weight, yet remained far from their ideal weight. They had spent a substantial amount of money on WLI, had limited information of AOM, and indicated a desire for increased professional support.


Novel mutations in the exon 5, intron 2 and 3' UTR regions of IL-12B gene were observed in clinically proven tuberculosis patients of south India.
Interleukin-12 (IL-12) is formed by the interaction of IL-12p35 and IL-12p40 expressed independently from IL-12A and IL-12B genes. This interleukin plays prominent role in the T-helper type-1 (Th1) response against intracellular pathogens. Variations in IL-12B gene causes disruption of various activities one of them is suppression of Th1 response and is one of the characteristic features observed in patients with active tuberculosis. Hence, in the present study IL-12B gene status was evaluated in 50 new sputum smear-positive pulmonary tuberculosis patients (NSP-PTB) as identified by Ziehl-Nielsen (ZN) staining and 50 apparently healthy control subjects (HCS) who were sputum smear-negative. The sequence analysis showed novel missense mutations p.Ser205Ile, p.Leu206Glu, p.Pro207Ser, p.Glu209Lys, p.Val210Ser, p.(Ser205_Cys327delinsIleGlu) and p.(Lys217_Leu218delinsIle) were found in exon 5 of the IL-12B gene in nine patients resulting formation of inactive IL-12 and three patients showed novel frame shift mutations p.(Asn222Leufs∗23) in exon 5 of causing the formation of truncated protein. Several mutations were noted in intron 2 of the IL-12B gene in 5 patients and in 13 patients mutations were observed in 3' UTR region. All together 30/50 patients (60%) showed mutations in IL-12B gene. Decreased levels of interferon-gamma (IFN-γ) and IL-12 as determined by ELISA and flow cytometry were observed in the peripheral blood mononuclear cell culture supernatants in TB patients having mutations compared with control subjects. Further, in silico analysis revealed due to frame shift mutations in exon 5 at Asn222 resulted in deletion of functional fibronectin type-III (FN3) domain which leads to formation of inactive IL-12 in these patients.


Cholecystokinin and D-fenfluramine inhibit food intake in oxytocin-deficient mice.
Results from previous studies indicate that oxytocin (OT)-containing neural pathways are activated in laboratory rats after systemic administration of CCK or d-fenfluramine and that centrally released OT may participate in the anorexigenic effects of these treatments. To explore the relationship between feeding behavior and OT function, the effects of CCK and d-fenfluramine on feeding and central c-Fos expression were compared in wild-type (OT+/+) and OT-deficient mice (OT-/-) of C57BL/6 background. Male OT+/+ and OT-/- mice were administered saline or CCK (1, 3, or 10 microg/kg ip) after overnight food deprivation. Saline-treated OT+/+ and OT-/- mice consumed equivalent amounts of food after an overnight fast. CCK inhibited deprivation-induced food intake in a dose-dependent manner to a similar extent in both genotypes. CCK treatment also induced similar hindbrain and forebrain patterns of increased c-Fos expression in mice of both genotypes. After treatment with d-fenfluramine (10 mg/kg ip), both OT+/+ and OT-/- mice consumed significantly less food than untreated controls, with no difference between genotypes. We conclude that OT signaling pathways are unnecessary for the anorexigenic effects of systemically administered CCK and d-fenfluramine in C57BL/6 mice.


Phase I clinical trial of drug-monoclonal antibody conjugates in patients with advanced colorectal carcinoma: a preliminary report.
Melphalan (MEL), an alkylating agent, has been modified to a derivative, N-acetylmelphalan (N-AcMEL), which can be conjugated to anticolon cancer monoclonal antibodies (MoAbs 30.6, I-1, and JGT) and used for immunochemotherapy. The final immunoconjugates possess potent cytotoxicity and specificity in preclinical studies. In a phase I clinical study, N-AcMEL-MoAb conjugates were administered via the hepatic artery to 10 patients, nine of whom had disseminated colorectal cancer (including the liver) and one of whom had Dukes' C colon cancer that had been resected. The selection of MoAb was based on the immunoperoxidase staining of the primary colon cancer tissue. Thus far doses of 1000 mg/m2 MoAb conjugated to 20 mg/m2 of N-AcMEL have been administered with no significant side effects, whereas MEL unconjugated to monoclonal antibodies would have caused myelosuppression in a proportion of patients at the same dosage. Serum antimouse antibody responses were noted in all of the patients; febrile reactions were noted with higher doses but were easily controlled with antipyretics, antihistamines and, if necessary, steroids. Serum sickness developed in one patient who was given a second course of treatment in the presence of human antimouse antibody, but the episode was self-limiting. Eight of the 10 patients had evaluable disease. Subjective improvement was noted in almost all of the patients examined, and 33%, or 3 of 9, of the treatments (nine courses of treatment in eight patients with evaluable disease; one of the patients had two courses of treatment) led to antitumor responses (minor response) by objective assessment with computed tomography of the liver. It is important to note that treatment with N-AcMEL-MoAb conjugates was safe at a dose of 20 mg/m2 of N-AcMEL, whereas the efficacy of such a form of treatment remains to be determined.


Lanthanide doped nanoparticles for reliable and precise luminescence nanothermometry in the third biological window.
In recent years, infrared emitting luminescent nanothermometers have attracted significant attention because their potential for the development of new diagnosis and therapy procedures. Despite their promising applications, concerns have been raised about their reliability due to the spectral distortions induced by tissues that are present even in the commonly used second biological window (1000-1370 nm). In this work, we present an innovative solution to this issue by demonstrating the effectiveness of shifting the operation range of these nanothermometers to the third biological window (1550-1850 nm). Through experimental evidence using ytterbium, erbium, and thulium tri-doped CaF2 nanoparticles, we demonstrate that luminescence spectra acquired in the third biological window are minimally distorted by the presence of tissue, opening the way to reliable luminescence thermometry. In addition, advanced analysis (singular value decomposition) of emission spectra allows sub-degree thermal uncertainties to be achieved.


Evaluation of the therapeutic community treatment model in Thailand: policy implications for compulsory and prison-based treatment.
This study, conducted in 2005 to 2007, presents results that are based on a proscriptive cohort design. The sample consisted of 769 residents in 22 drug user treatment programs who stayed in treatment for at least 30 days to one year; 510 former residents (66%) from 21 programs (95%) were interviewed again at a 6-month post-treatment follow-up assessment. A majority of the participants were male, lived with family or relatives, had completed only primary school, and had a full-time or a part-time job prior to entering treatment. The participating therapeutic community (TC) programs were a mixture of volunteer, compulsory-probation, and prison-based programs. In-person interview data and urine testing showed that the self-reported drug use prevalence rates are reliable. The results show large positive treatment effects on 30-day and 6-month illegal drug use and small to medium effects on the severity of alcohol use and related problems. A multilevel regression analysis suggests that residents' reduced stigma, adaptation of the TC model, and frequency of alcohol and drug use-related consequences partially predict treatment success. Study limitations and policy implications are discussed.