Germline mutation of the PTEN gene in a Japanese patient with Cowden's disease.
Cowden's disease (CD) is an autosomal dominant disorder which confers a high susceptibility to diverse benign and malignant tumors. The PTEN (phosphatase and tensin homologue deleted in chromosome ten) gene has been identified as a tumor suppressor gene responsible for cancers of the endometrium, ovary, prostate, and glioblastomas. Recently, germline mutations of this gene were also found in patients with CD, and it is now recognized as a gene responsible for this disease. We identified a germline nonsense mutation at codon 130 in exon 5 of PTEN in a 56-year-old Japanese woman with CD. The patient had adenoid facies and mucocutaneous lesions including multiple facial papules, acral keratoses on neck and shoulders, palmoplantar keratoses, multiple oral papillomas, scrotal tongue, mucosal and cutaneous hemangiomas, and a sclerotic fibroma on the arm. She also had benign and malignant polypoid neoplasms throughout the entire digestive tract, including adenocarcinoma of the colon and submucosal lipomas of the rectum, as well as bilateral breast carcinomas, multinodular goiters, an ovarian cyst with a fibroma-like nodule, hepatic hemangiomas, and abdominal hernia. We searched CD cases with the same genotypic PTEN mutation as the present case and compared their phenotypes. Further studies will disclose a better understanding of the role of mutation in the PTEN gene in the course of tumorigenesis of both benign and malignant tumors developed in patients with CD.


Using a concept map as a tool for strategic planning: The Healthy Brain Initiative.
Concept mapping is a tool to assist in strategic planning that allows planners to work through a sequence of phases to produce a conceptual framework. Although several studies describe how concept mapping is applied to various public health problems, the flexibility of the methods used in each phase of the process is often overlooked. If practitioners were more aware of the flexibility, more public health endeavors could benefit from using concept mapping as a tool for strategic planning. The objective of this article is to describe how the 6 concept-mapping phases originally outlined by William Trochim guided our strategic planning process and how we adjusted the specific methods in the first 2 phases to meet the specialized needs and requirements to create The Healthy Brain Initiative: A National Public Health Road Map to Maintaining Cognitive Health. In the first stage (phases 1 and 2 of concept mapping), we formed a steering committee, convened 4 work groups over a period of 3 months, and generated an initial set of 42 action items grounded in science. In the second stage (phases 3 and 4), we engaged stakeholders in sorting and rating the action items and constructed a series of concept maps. In the third and final stage (phases 5 and 6), we examined and refined the action items and generated a final concept map consisting of 44 action items. We then selected the top 10 action items, and in 2007, we published The Healthy Brain Initiative: A National Public Health Road Map to Maintaining Cognitive Health, which represents the strategic plan for The Healthy Brain Initiative.


PARP1 controls KLF4-mediated telomerase expression in stem cells and cancer cells.
Telomerase is highly expressed in cancer and embryonic stem cells (ESCs) and implicated in controlling genome integrity, cancer formation and stemness. Previous studies identified that Krüppel-like transcription factor 4 (KLF4) activates telomerase reverse transcriptase (TERT) expression and contributes to the maintenance of self-renewal in ESCs. However, little is known about how KLF4 regulates TERT expression. Here, we discover poly(ADP-ribose) polymerase 1 (PARP1) as a novel KLF4-interacting partner. Knockdown of PARP1 reduces TERT expression and telomerase activity not only in cancer cells, but also in human and mouse ESCs. Recruitment of KLF4 to TERT promoter is reduced in PARP1-suppressed cells. The poly(ADP-ribose) polymerase activity is dispensable, while the oligo(ADP-ribose) polymerase activity is required for the PARP1- and KLF4-mediated TERT activation. Repression of Parp1 in mouse ESCs decreases expression of pluripotent markers and induces differentiation. These results suggest that PARP1 recruits KLF4 to activate telomerase expression and stem cell pluripotency, indicating a positive regulatory role of the PARP1-KLF4 complex in telomerase expression in cancer and stem cells.


Manipulating recall vantage perspective of intrusive memories in dysphoria.
The current study attempted to experimentally manipulate mode of recall (field, observer perspective) in a sample of mildly dysphoric participants (N=134) who reported a distressing intrusive memory of negative autobiographical event. Specifically, the current study sought to ascertain whether shifting participants into a converse perspective would have differential effects on the reported experience of their memory. Results indicated that shifting participants from a field to an observer perspective resulted in decreased experiential ratings: specifically, reduced distress and vividness. Also, as anticipated, the converse shift in perspective (from observer to field) did not lead to a corresponding increase in experiential ratings, but did result in reduced ratings of observation and a trend was observed for decreased levels of detachment. The findings support the notion that recall perspective has a functional role in the regulation of intrusion-related distress and represents a cognitive avoidance mechanism.


Comparison of group B rotavirus genes 9 and 11.
Group B rotaviruses (GBRs) were recognized recently as causative agents of gastroenteritis. Investigations into the relatedness of various heterologous GBR strains have been hindered by the difficulty of growing these viruses in cell culture. Viral RNA extracted from experimentally infected rats used to prepare cDNA clones. From these, the nucleotide sequences of genes 9 and 11 of the IDIR strain of GBR were determined and compared with the corresponding sequences of the human ADRV strain of GBR. IDIR gene 11 is 643 bp in length with a single open reading frame (ORF) encoding 174 amino acids; IDIR gene 9 is 804 bp in length with a single ORF encoding 246 amino acids. Comparison of the IDIR sequences with those of ADRV showed that nucleotide sequence similarity was 60.6% and 71.9% for genes 9 and 11, respectively. The deduced amino acid sequence similarity was 51.2% for the gene 9 and 66.5% for the gene 11 product. This sequence diversity indicates that GBRs are more distantly related than strains of group A rotavirus.


Mobile phone use and risk of glioma: a case-control study in Korea for 2002-2007.
There has been a growing concern about the possible carcinogenic effects of the electromagnetic radiofrequency fields emitted from mobile phones. The purpose of this study was to investigate the association between mobile phone use and the development of gliomas in Korea. Our study methods were based on the International Interphone study that aimed to evaluate possible adverse effects of mobile phone use. This study included 285 histologically-confirmed Korean patients 15 to 69 years of age, with gliomas diagnosed between 2002 and 2007 in 9 hospitals. The 285 individually matched controls were healthy individuals that had their medical check-up in the same hospitals. Unconditional logistic regression was used to calculate the adjusted odds ratios (aORs) and 95% confidence intervals (CIs) for use of mobile phones. For the entire group, no significant relationship was investigated between gliomas and regular use of mobile phones, types of mobile phones, lifetime years of use, monthly service fee, and the other exposure indices. Analyses restricted to self-respondents showed similar results. For ipsilateral users, whose the body side for usual mobile phone use match the location of glioma, the aORs (95% CIs) for lifetime years of use and cumulative hours of use were 1.25 (0.55 to 2.88) and 1.77 (0.32 to 1.84), respectively. However, the contralateral users showed slightly lower risk than ipsilateral users. Our results do not support the hypothesis that the use of mobile phones increases the risk of glioma; however, we found a non-significant increase in risk among ipsilateral users. These findings suggest further evaluation for glioma risk among long-term mobile phone users.


Appetitive Motivation and Associated Neurobiology Change Differentially across the Life Course of Mouse Offspring Exposed to Peri- and Postnatal High Fat Feeding.
Alterations in neural pathways that regulate appetitive motivation may contribute to increased obesity risk in offspring born to mothers fed a high fat (HF) diet. However, current findings on the impact of maternal obesity on motivation in offspring are inconclusive, and there is no information about the long-lasting effects in aged animals. This study examined the longitudinal effect of perinatal and chronic postnatal HF intake on appetitive motivation in young and aged offspring. Female C57Bl/6 were fed either a control (C) or HF diet before mating through to lactation. At weaning, offspring were maintained on the C or HF diet, generating the following four diet groups: C/C, C/HF, HF/C, and HF/HF based on the pre/post weaning diet. At 6 months, motivation was higher in HF/C females, but lower in male and female C/HF and HF/HF mice. By 12 months, this difference was lost, as C-fed animals became less motivated, while motivation increased in HF-fed mice. The mRNA levels of dopamine receptor 1 and 2 increased with age, while cannabinoid receptor 1 and μ-opioid receptor expression remained stable or decreased in mesolimbic and mesocortical dopaminergic pathways. Results from this study suggest that perinatal and chronic postnatal HF feeding produced opposite effects on appetitive motivation in young adult offspring mice, which was also reflected in the shift in motivation over time. These results have significant implications for patterns of hedonic eating across the life course and the relative risk of obesity at different time points.


Reduction in injury rates in nursing personnel through introduction of mechanical lifts in the workplace.
Health care workers incur frequent injuries resulting from patient transfer and handling tasks. Few studies have evaluated the effectiveness of mechanical lifts in preventing injuries and time loss due to these injuries. We examined injury and lost workday rates before and after the introduction of mechanical lifts in acute care hospitals and long-term care (LTC) facilities, and surveyed workers regarding lift use. The post-intervention period showed decreased rates of musculoskeletal injuries (RR = 0.82, 95% CI: 0.68-1.00), lost workday injuries (RR = 0.56, 95% CI: 0.41-0.78), and total lost days due to injury (RR = 0.42). Larger reductions were seen in LTC facilities than in hospitals. Self-reported frequency of lift use by registered nurses and by nursing aides were higher in the LTC facilities than in acute care hospitals. Observed reductions in injury and lost day injury rates were greater on nursing units that reported greater use of the lifts. Implementation of patient lifts can be effective in reducing occupational musculoskeletal injuries to nursing personnel in both LTC and acute care settings. Strategies to facilitate greater use of mechanical lifting devices should be explored, as further reductions in injuries may be possible with increased use.


[Results of 769 stapedectomies performed between 1975 and 1993].
A retrospective review was made of 719 primary stapedectomies and 48 re-operations carried out between 1975 and 1993. Preoperative and postoperative audiograms (6-12 months) were studied, analyzing 250, 500, 1000, 2000 and 4000 Hz, auditory gain and closure of the air-bone gap < or = 10 dB. Ninety percent had an auditory gain of > or = 10 dB at conversational frequencies. Age, the surgeon, and average preoperative pure tone did not influence the final result. Polyethylene prostheses (70.9%) and steel and teflon pistons (21.1%) were the prostheses used most often. Neurosensorial hearing loss (> 10 dB) occurred in 24 cases (3.3%). In revision stapedectomy, 66.6% of patients had air-bone gap closure of < or = 20 dB and 71% had a hearing gain of < or = 10 dB. Prosthesis displacement (43.7%) and incus necrosis (18.7%) were the most common findings in revision stapedectomy.


Pyrosequencing Reveals a Core Community of Anodic Bacterial Biofilms in Bioelectrochemical Systems from China.
Bioelectrochemical systems (BESs) are promising technologies for energy and product recovery coupled with wastewater treatment, and the core microbial community in electrochemically active biofilm in BESs remains controversy. In the present study, 7 anodic communities from 6 bioelectrochemical systems in 4 labs in southeast, north and south-central of China are explored by 454 pyrosequencing. A total of 251,225 effective sequences are obtained for 7 electrochemically active biofilm samples at 3% cutoff level. While Alpha-, Beta-, and Gamma-proteobacteria are the most abundant classes (averaging 16.0-17.7%), Bacteroidia and Clostridia are the two sub-dominant and commonly shared classes. Six commonly shared genera i.e., Azospira, Azospirillum, Acinetobacter, Bacteroides, Geobacter, Pseudomonas, and Rhodopseudomonas dominate the electrochemically active communities and are defined as core genera. A total of 25 OTUs with average relative abundance >0.5% were selected and designated as core OTUs, and some species relating to these OTUs have been reported electrochemically active. Furthermore, cyclic voltammetry and chronoamperometry tests show that two strains from Acinetobacter guillouiae and Stappia indica, bacteria relate to two core OTUs, are electrochemically active. Using randomly selected bioelectrochemical systems, the study has presented extremely diverse bacterial communities in anodic biofilms, though, we still can suggest some potentially microbes for investigating the electrochemical mechanisms in bioelectrochemical systems.


Continuing Education for Prehospital Healthcare Providers in India - A Novel Course and Concept.
Emergency medical services (EMS) in India face enormous challenges in providing care to a geographically expansive and diverse patient population. Over the last decade, the public-private-partnership GVK EMRI (Emergency Management and Research Institute) has trained over 100,000 emergency medical technicians (EMTs), with greater than 21,000 currently practicing, to address this critical gap in the healthcare workforce. With the rapid development and expansion of EMS, certain aspects of specialty development have lagged behind, including continuing education requirements. To date, there have been no substantial continuing education EMT skills and training efforts. We report lessons learned during development and implementation of a continuing education course (CEC) for EMTs in India. From 2014 to 2017, we employed an iterative process to design and launch a novel CEC focused on five core emergency competency areas (medicine and cardiology, obstetrics, trauma, pediatrics, and leadership and communication). Indian EMT instructors and providers partnered in design and content, and instructors were trained to independently deliver the CEC. Many challenges had to be overcome: scale (>21,000 EMTs), standardization (highly variable skill levels among providers and instructors), culture (educational emphasis on rote memorization rather than practical application), and translation (22 major languages and a few hundred local dialects spoken nationwide). During the assessment and development phases, we identified five key strategies for success: (1) use icon-based video instruction to ensure consistent quality and allow voice-over for easy translation; (2) incorporate workbooks during didactic videos and (3) employ low-cost simulation and case discussions to emphasize active learning; (4) focus on non-technical skills; (5) integrate a formal training-of-trainers prior to delivery of materials. These key strategies can be combined with innovation and flexibility to address unique challenges of language, system resources, and cultural differences when developing impactful continuing educational initiatives in bourgeoning prehospital care systems in low- and middle-income countries.


[Advances in the management of neonatal hypoxia].
During the birth, physiological changes occur in virtually all organs of the child, including the central nervous system. In this transitional phase, it is possible some degree of hypoxemia, generally well tolerated by the newborn. But, if neonatal hypoxia is intense and continuous it can lead to neonatal encephalopathy, which characterizes a critical situation for the infant. The proper approach is essential to ensure a good long-term prognosis. We up-to-date information regarding hypoxia neonatal and review recent evidence-based medicine publications addressing its approach. Neonatal encephalopathy may be clinically classified into three levels of intensity. Mild cases usually have a good prognosis, moderate intensity cases have 30% chance of sequels, and severe intensity cases have more than 70% mortality and nearly all survivors have sequels. Recent advances occurred in two areas: in the diagnosis, with new EEG and MRI techniques, and in the treatment, with the advent of therapeutic hypothermia. There is the possibility of future use for stem cell therapy. The prognosis depends on the clinical classification, the neuroimaging data as well as the EEG.


Immunogenicity and Protection Efficacy of a Naked Self-Replicating mRNA-Based Zika Virus Vaccine.
To combat emerging infectious diseases like Zika virus (ZIKV), synthetic messenger RNAs (mRNAs) encoding viral antigens are very attractive as they allow a rapid, generic, and flexible production of vaccines. In this work, we engineered a self-replicating mRNA (sr-mRNA) vaccine encoding the pre-membrane and envelope (prM-E) glycoproteins of ZIKV. Intradermal electroporation of as few as 1 µg of this mRNA-based ZIKV vaccine induced potent humoral and cellular immune responses in BALB/c and especially IFNAR1-/- C57BL/6 mice, resulting in a complete protection of the latter mice against ZIKV infection. In wild-type C57BL/6 mice, the vaccine resulted in very low seroconversion rates and antibody titers. The potency of the vaccine was inversely related to the dose of mRNA used in wild-type BALB/c or C57BL/6 mice, as robust type I interferon (IFN) response was determined in a reporter mice model (IFN-β+/Δβ-luc). We further investigated the inability of the sr-prM-E-mRNA ZIKV vaccine to raise antibodies in wild-type C57BL/6 mice and found indications that type I IFNs elicited by this naked sr-mRNA vaccine might directly impede the induction of a robust humoral response. Therefore, we assume that the efficacy of sr-mRNA vaccines after intradermal electroporation might be increased by strategies that temper their inherent innate immunogenicity.


Structural organization of a filamentous influenza A virus.
Influenza is a lipid-enveloped, pleomorphic virus. We combine electron cryotomography and analysis of images of frozen-hydrated virions to determine the structural organization of filamentous influenza A virus. Influenza A/Udorn/72 virions are capsule-shaped or filamentous particles of highly uniform diameter. We show that the matrix layer adjacent to the membrane is an ordered helix of the M1 protein and its close interaction with the surrounding envelope determines virion morphology. The ribonucleoprotein particles (RNPs) that package the genome segments form a tapered assembly at one end of the virus interior. The neuraminidase, which is present in smaller numbers than the hemagglutinin, clusters in patches and are typically present at the end of the virion opposite to RNP attachment. Incubation of virus at low pH causes a loss of filamentous morphology, during which we observe a structural transition of the matrix layer from its helical, membrane-associated form to a multilayered coil structure inside the virus particle. The polar organization of the virus provides a model for assembly of the virion during budding at the host membrane. Images and tomograms of A/Aichi/68 X-31 virions show the generality of these conclusions to non-filamentous virions.


Trematode infection affects shell shape and size in Bulinus tropicus.
Trematodes can increase intraspecific variation in the phenotype of their intermediate snail host. However, the extent of such phenotypic changes remains unclear. We investigated the influence of trematode infection on the shell morphology of Bulinus tropicus, a common host of medically important trematodes. We focused on a snail population from crater lake Kasenda (Uganda). We sampled a single homogeneous littoral habitat to minimize the influence of environmental variation on shell phenotype, and barcoded snails to document snail genotypic variation. Among the 257 adult snails analysed, 99 tested positive for trematode infection using rapid-diagnostic PCRs. Subsequently we used high-throughput amplicon sequencing to identify the trematode (co-)infections. For 86 out of the 99 positive samples trematode species delineation could discriminate among combinations of (co-)infection by 11 trematode species. To avoid confounding effects, we focused on four prevalent trematode species. We performed landmark-based geometric morphometrics to characterize shell phenotype and used regressions to examine whether shell size and shape were affected by trematode infection and the developmental stage of infection (as inferred from read counts). Snails infected by Petasiger sp. 5, Echinoparyphium sp. or Austrodiplostomum sp. 2 had larger shells than uninfected snails or than those infected by Plagiorchiida sp. Moreover, the shell shape of snails infected solely by Petasiger sp. 5 differed significantly from that of uninfected snails and snails infected with other trematodes, except from Austrodiplostomum sp. 2. Shape changes included a more protuberant apex, an inward-folded outer apertural lip and a more adapically positioned umbilicus. Size differences were more pronounced in snails with 'late' infections (>25 days) compared to earlier-stage infections. No phenotypic differences were found between snails infected by a single trematode species and those harbouring co-infections. Further work is required to assess the complex causal links between trematode infections and shell morphological alterations of snail hosts.


The brain and intelligence.
A measure of brain development is proposed which might correlate with intelligence. The neocortex/medulla volume is given for many species of primate, and it is found that this measure correlates with a measure of responsiveness to novel objects, and with performance on visual discrimination learning set. It is shown that cranial capacity/foramen magnum area is closely related to brain/medulla volume, and may therefore be used as a related measure with fossil species.


Expression of mRNAs encoding for alpha and beta integrin subunits, MMPs, and TIMPs in stretched human periodontal ligament and gingival fibroblasts.
The biological mechanisms of tooth movement result from the cellular responses of connective tissues to exogenous mechanical forces. Among these responses, the degradation of the extracellular matrix takes place, but the identification of the molecular basis as well as the components implicated in this degradation are poorly understood. To contribute to this identification, we subjected human fibroblasts obtained from the periodontal ligament (PDLs) and from the gingiva (HGFs) to a continuous stretch to quantify the mRNAs encoding for various metalloproteinases (MMPs), their tissue inhibitors (TIMPs), and alpha and beta integrin subunits. Both cell lines reacted by inducing the expression of the mRNAs encoding for MMP-1, MMP-2, TIMP-1, and TIMP-2, while other mRNAs did not vary (MT1-MMP, TIMP-3) or were not expressed (MMP-9). PDLs expressed selectively the mRNAs encoding for alpha4 and alphav, with no difference measurable under stretching, while the mRNAs encoding for alpha6 and beta1 were increased and the one encoding for alpha5 was decreased. HGFs increased the mRNAs encoding for alpha2, alpha6, beta1, and beta3 and decreased the one encoding for alpha3. Analysis of our data indicated that stretched HGFs and PDLs induced the same pattern of mRNAs encoding for MMPs and TIMPs but differed for those encoding various integrin subunits, known to act as protein receptors in mechanotransduction.


Penicillin therapy of spontaneous streptococcal meningitis in pigs.
Oral prophylactic medication with either procaine penicillin G or a mixture of chlortetracycline, sulphadimidine and procaine penicillin G reduced the incidence of streptococcal meningitis in a herd of pigs with a high recorded prevalence of the disease, but to a significant extent (P less than 0.01) only in those pigs receiving procaine penicillin G. Subsequent studies showed that after oral administration of procaine penicillin G, benzylpenicillin was detectable in plasma only at very low concentration and similar results were obtained using the potassium salt of penicillin G. However, phenoxymethyl penicillin administered orally provided high plasma concentrations of this drug. A further investigation demonstrated that despite the low plasma concentrations of penicillin after oral administration of the procaine salt, gastrointestinal and urinary concentrations of the drug were relatively high for up to five hours.


Is the pathogenesis of hypertension different in black patients?
There is a higher prevalence of hypertension in the urban black population of South Africa and the USA in comparison to whites living in the same geographical areas. Several factors including age, urbanisation, socioeconomic status and acculturation contribute to the differences in the prevalence of hypertension between blacks and whites. In addition there may be a genetic element. Biochemical differences in the lipid profile of the blacks in Sub-Saharan Africa may play an Important role in the lower incidence of coronary heart disease. Blacks have an abnormal transport mechanism of sodium. There are varying responses to antihypertensive drugs in blacks compared to whites. It is probable that with a better understanding of the pathogenesis of hypertension in blacks, we will be able to reduce the high prevalence, prevent complications and institute more effective treatment and control of hypertension.


Assessment of Apical Pressures in Single and Joining Canals - An Ex Vivo Study Based on Computational Fluid Dynamic Analysis.
Computational fluid dynamic analysis (CFD) is claimed to be a reliable tool for analysing the fluid flow and the generated apical pressures in the simulated root canal. The current study aimed to analyse the apical pressures in extracted teeth with single and joining canals. Forty-six freshly extracted teeth were collected for the present study. The power was set at 95%, with an effect size of 0.55 (1-β=95%, α=0.05). Once the root canal anatomy was confirmed with cone-beam computed tomography (CBCT), they were divided into two groups: group I: mandibular second premolars with Vertucci type-I (n=23), and group II: maxillary second premolars with Vertucci type-II (n=23). The instrumentation of the specimens was carried out to a 0.04-taper using rotary instruments. A post-instrumentation CBCT was obtained, and computer-aided design models were obtained. The CFD simulations were then con- ducted with simulated 30-gauge side vented needles at 25, 50, and 75% short of the working length (WL). Group I recorded significantly (p<0.05) higher apical pressures at needle positions 25% short of the WL. However, no significant differences were elicited in the groups at other needle positions. Single canal specimens recorded higher apical pressures at needle positions 25% short of the WL. However, no differences were elicited between single and joining canals at higher needle positions.


Neurogenic Component of Chronic Renal Hypertension.
Infusion of angiotensin or renin in small quantities affects the sympathetic nervous system so that responses are increased to either drugs or reflexes that cause release of norepinephrine at nerve endings. Response to injected norepinephrine is relatively unchanged. This action of angiotensin is dependent upon an intact sympathetic nervous system. The direct vasoconstrictor action of angiotensin is not an essential part of the enhanced response. The phenomenon was shown to have relevance to acute and chronic experimental renal hypertension in dogs by the fact that in both the pressor response to tyramine was enhanced. We believe that the ability of angiotensin to intensify the effect of normal neurogenic vasomotor activity, along with an upward reset of the carotid sinus buffer mechanism, might account importantly for the neurogenic component of renal hypertension.


Occlusive peripheral arteriosclerosis: treatment by percutaneous transluminal recanalization--"the Dotter procedure".
We have performed 17 percutaneous transluminal dilatations in 15 patients with segmental stenoses of the femoral and popliteal arteries. Initial success was achieved in 14 instances. This technique must be considered a useful adjunct in the treatment of peripheral vascular disease in selected patients. Its use along with endarterectomy has not been previously reported and warrants further investigation.


Comparing the efficiencies of hydrazide labels in the study of protein carbonylation in human serum albumin.
In this work, we establish a methodology for comparing the efficiencies of different hydrazide labels for detecting protein carbonyls. We have chosen acrolein-modified human serum albumin as a model. This system provides a convenient means of reproducibly generating carbonylated protein. Five hydrazide-based labels were tested. Three carry a biotin affinity tag, and the others are simple fatty acid hydrazides. For the biotin-based labels, the yield of the labeling reaction varies considerably, and the most commonly used label, biotin hydrazide, gives the lowest yield. The total tandem mass spectrometry (MS/MS) spectrum counts of modified peptides are similar for all of the biotin-based tags, indicating that factors beyond the labeling efficiency are important in determining the effectiveness of the label. In addition, there is a large variation in the number of spectra obtained for specific, modified peptides depending on the nature of the labeling group. This variation implies that the relative detectability of a particular modification site is highly dependent on the tagging reagent, and more importantly, titration schemes aimed at identifying the most reactive site based on its threshold concentration will be biased by the choice of tagging reagent. The fatty acid hydrazides are somewhat more effective than the biotin-based hydrazides in generating identifiable MS/MS spectra but offer no opportunity for enrichment. For the biotin-based tags, avidin affinity chromatography was used with the tryptic digests, and each tag led to similar enrichment levels.


Encystation in Acanthamoeba castellanii: development of biocide resistance.
Since the early 1960s, axenic culture and the development of procedures for the induction of encystation have made Acanthamoeba spp. superb experimental systems for studies of cell biology and differentiation. More recently, since their roles as human pathogens causing keratitis and encephalitis have become widely recognized, it has become urgent to understand the parameters that determine differentiation, as cysts are much more resistant to biocides than are the trophozoites. Viability of trophozoites of the soil amoeba Acanthamoeba castellanii (Neff), is conveniently measured by its ability to form plaques on a lawn of Escherichia coli. Use of confocal laser scanning microscopy with Calcofluor white, Congo Red or the anionic oxonol dye, DiBAC4(3) or flow cytometry with propidium iodide diacetate and fluorescein or oxonol provides more rapid assessment. For cysts, the plaque method is still the best, because dye exclusion does not necessarily indicate viability and therefore the plate count method has been used to study the sequence of development of biocide resistance during the differentiation process. After two hours, resistance to HCl was apparent. Polyhexamethylene biguanide, benzalkonium chloride, propamidine isethionate, pentamidine isethionate, dibromopropamine isethionate, and H2O2 and moist heat, all lost effectiveness at between 14 and 24 h after trophozoites were inoculated into encystation media. Chlorhexidine diacetate resistance was observed at between 24 and 36 h. The molecular biology and biochemistry of the modifications that underlie these changes are now being investigated.


Interpersonal repression as a predictor of cancer.
This paper examines the possible relationship between two specific styles of interactive behavior which reflect active and passive coping with stressful experiences, and attendant illness susceptibility. In a longitudinal study of 1353 inhabitants of a Yugoslav town from 1965 to 1975 data show that being a passive receiver of repression is associated with subsequent incidence of cancer. Being an active emitter of interpersonal repression is found more commonly among subjects who experience cardiovascular and other circulatory diseases later on. Data are presented and discussed in the framework of recent findings in environmental physiology, neuroendocrinology and psychosomatics.


[Medicaments and oral healthcare. Adverse effects of medicines and self care products on the gingiva].
Medications and self care products may have adverse effects on the gingiva. These adverse effects can be divided into inflammation, intrinsic discolouration, irritation, trauma, cytotoxicity, lichenoid reaction, and proliferation. This article deals with the first 6 types of adverse effects mentioned: a subsequent article will deal with the last type mentioned. Since contraceptives were introduced, there have been indications that they cause or promote gingivitis, but with the current contraceptives this adverse effect is rarely seen. Intrinsic discolouration of the gingiva has been reported when using Staloral®, minocycline, contraceptives and hydroxychloroquine. Irritation and trauma of the gingiva are seen when self care products containing carbamide peroxide or hydrogen peroxide are used for external tooth whitening, or which have analgesic potential, such as acetylsalicylic acid and hydrogen peroxide or oral rinses containing alcohol. Several cytostatics may induce apoptosis of keratinocytes in the gingiva. Oral rinses with antibacterial ingredients have cytotoxic potential. Lichenoid reactions have been reported due to the use of several (groups of) medications.


Role of different voltage-gated Ca2+ channels in cortical spreading depression: specific requirement of P/Q-type Ca2+ channels.
Gain-of-function mutations in CaV 2.1 (P/Q-type) Ca2+ channels cause familial hemiplegic migraine type 1 (FHM1), a subtype of migraine with aura. Knockin (KI) mice carrying FHM1 mutations show increased neuronal P/Q-type current and facilitation of induction and propagation of cortical spreading depression (CSD), the phenomenon that underlies migraine aura and may activate migraine headache mechanisms. We recently studied cortical neurotransmission in neuronal microcultures and brain slices of FHM1 KI mice, and showed (1) gain-of-function of excitatory neurotransmission, due to increased action potential-evoked Ca2+ influx and increased probability of glutamate release at pyramidal cell synapses, but unaltered inhibitory neurotransmission at fast-spiking interneuron synapses, and (2) a causative link between enhanced glutamate release and facilitation of CSD induced by brief pulses of high K+ in cortical slices. Here, we show that after blockade of either the P/Q-type Ca2+ channels or the NMDA receptors, CSD cannot be induced in wild-type mouse cortical slices. In contrast, blockade of N- or R-type Ca2+ channels has only a small inhibitory effect on CSD threshold and velocity of propagation. Our findings support a model in which Ca2+ influx through presynaptic P/Q-type Ca2+ channels with consequent release of glutamate from recurrent cortical pyramidal cell synapses and activation of NMDA receptors are required for initiation and propagation of the CSD involved in migraine.


Delayed Maturation of Fast-Spiking Interneurons Is Rectified by Activation of the TrkB Receptor in the Mouse Model of Fragile X Syndrome.
Fragile X syndrome (FXS) is a neurodevelopmental disorder that is a leading cause of inherited intellectual disability, and the most common known cause of autism spectrum disorder. FXS is broadly characterized by sensory hypersensitivity and several developmental alterations in synaptic and circuit function have been uncovered in the sensory cortex of the mouse model of FXS (Fmr1 KO). GABA-mediated neurotransmission and fast-spiking (FS) GABAergic interneurons are central to cortical circuit development in the neonate. Here we demonstrate that there is a delay in the maturation of the intrinsic properties of FS interneurons in the sensory cortex, and a deficit in the formation of excitatory synaptic inputs on to these neurons in neonatal Fmr1 KO mice. Both these delays in neuronal and synaptic maturation were rectified by chronic administration of a TrkB receptor agonist. These results demonstrate that the maturation of the GABAergic circuit in the sensory cortex is altered during a critical developmental period due in part to a perturbation in BDNF-TrkB signaling, and could contribute to the alterations in cortical development underlying the sensory pathophysiology of FXS.SIGNIFICANCE STATEMENT Fragile X (FXS) individuals have a range of sensory related phenotypes, and there is growing evidence of alterations in neuronal circuits in the sensory cortex of the mouse model of FXS (Fmr1 KO). GABAergic interneurons are central to the correct formation of circuits during cortical critical periods. Here we demonstrate a delay in the maturation of the properties and synaptic connectivity of interneurons in Fmr1 KO mice during a critical period of cortical development. The delays both in cellular and synaptic maturation were rectified by administration of a TrkB receptor agonist, suggesting reduced BDNF-TrkB signaling as a contributing factor. These results provide evidence that the function of fast-spiking interneurons is disrupted due to a deficiency in neurotrophin signaling during early development in FXS.


Decisive role of water and protein dynamics in residence time of p38α MAP kinase inhibitors.
Target residence time plays a crucial role in the pharmacological activity of small molecule inhibitors. Little is known, however, about the underlying causes of inhibitor residence time at the molecular level, which complicates drug optimization processes. Here, we employ all-atom molecular dynamics simulations (~400 μs in total) to gain insight into the binding modes of two structurally similar p38α MAPK inhibitors (type I and type I½) with short and long residence times that otherwise show nearly identical inhibitory activities in the low nanomolar IC50 range. Our results highlight the importance of protein conformational stability and solvent exposure, buried surface area of the ligand and binding site resolvation energy for residence time. These findings are further confirmed by simulations with a structurally diverse short residence time inhibitor SB203580. In summary, our data provide guidance in compound design when aiming for inhibitors with improved target residence time.


Features and Outcomes of Methamphetamine-associated Pulmonary Arterial Hypertension.
Although amphetamines are recognized as "likely" agents to cause drug- and toxin-associated pulmonary arterial hypertension (PAH), (meth)amphetamine-associated PAH (Meth-APAH) has not been well described. To prospectively characterize the clinical presentation, histopathology, and outcomes of Meth-APAH compared with those of idiopathic PAH (iPAH). We performed a prospective cohort study of patients with Meth-APAH and iPAH presenting to the Stanford University Pulmonary Hypertension Program between 2003 and 2015. Clinical, pulmonary angiography, histopathology, and outcomes data were compared. We used data from the Healthcare Cost and Utilization Project to estimate the epidemiology of PAH in (meth)amphetamine users hospitalized in California. The study sample included 90 patients with Meth-APAH and 97 patients with iPAH. Patients with Meth-APAH were less likely to be female, but similar in age, body mass index, and 6-minute-walk distance to patients with iPAH. Patients with Meth-APAH reported more advanced heart failure symptoms, had significantly higher right atrial pressure (12.7 ± 6.8 vs. 9.8 ± 5.1 mm Hg; P = 0.001), and had lower stroke volume index (22.2 ± 7.1 vs. 25.5 ± 8.7 ml/m2; P = 0.01). Event-free survival in Meth-APAH was 64.2%, 47.2%, and 25% at 2.5, 5, and 10 years, respectively, representing more than double the risk of clinical worsening or death compared with iPAH (hazard ratio, 2.04; 95% confidence interval, 1.28-3.25; P = 0.003) independent of confounders. California data demonstrated a 2.6-fold increase in risk of PAH diagnosis in hospitalized (meth)amphetamine users. Meth-APAH is a severe and progressive form of PAH with poor outcomes. Future studies should focus on mechanisms of disease and potential therapeutic considerations.


Is ultrasound combined with computed tomography useful for distinguishing between primary thyroid lymphoma and Hashimoto's thyroiditis?
The aim of the study is to investigate the usefulness of ultrasound combined with computed tomography (CT) for distinguishing between primary thyroid lymphoma (PTL) and Hashimoto's thyroiditis (HT). The investigation was conducted retrospectively in 80 patients from January 2000 to July 2018. All patients underwent pathological tests to be classified into one of two groups: PTL group and HT group. The cut-off value of CT density was determined using receiver-operating characteristic (ROC) curve analysis. The accuracy, sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) of diagnosis for thyroid by CT alone, ultrasound alone, and the combination of CT plus ultrasound were calculated. Of the 80 study patients, 27 patients were PTL and 53 patients were HT. Mean CT density had a sensitivity of 90.6% and a specificity of 88.9% at a cut-off value of 53.5 HU, with area under the curve (AUC) 0.88. Ultrasound combined with CT had the highest specificity, accuracy, and PPV compared with CT alone and ultrasound alone (p value < 0.05). Features such as extremely hypoechogenicity, enhanced posterior echo, cervical lymphadenopathy in ultrasound image, and linear high-density strand signs, and very low density in CT imaging have high sensitivity and specificity in thyroid lymphoma. Therefore, ultrasound combined with CT may be useful for distinguishing between PTL and HT.


Clinical significance of CXCL16/CXCR6 expression in patients with prostate cancer.
We hypothesized that the CXCL16-CXCR6 ligand-receptor system may play an important role in prostate cancer progression. Levels of CXCL16 and CXCR6 expression were evaluated in prostate cancer cell lines (PC-3 and LNCaP) and normal prostate epithelial cells (PrEC), as well as in tissues from 354 patients. The immunohistochemical expression of CXCL16/CXCR6 was greater in the PC-3/LNCaP cells than in the PrEC cell line. The expression of CXCL16/CXCR6 was significantly higher in prostate cancer than in benign prostatic hypertrophy. Using RT-PCR, the expression of CXCL16/CXCR6 was found to be greater in the PC-3/LNCaP cells than in the PrEC cell line. CXCL16/CXCR6 was weakly detected in lung and liver tissues, whereas CXCL16 was highly expressed in specimens of bone metastasis. CXCL16 immunostaining was related to Gleason score, T stage, tumor volume, perineural invasion and lymph node metastasis. However, biochemical PSA recurrence was not related to the expression of CXCL16/CXCR6. High CXCL16/CXCR6 expression may be related to aggressive cancer behavior, and high CXCL16 expression to bone metastases.


Toxicity and Binding Studies of Bacillus thuringiensis Cry1Ac, Cry1F, Cry1C, and Cry2A Proteins in the Soybean Pests Anticarsia gemmatalis and Chrysodeixis (Pseudoplusia) includens.
Anticarsia gemmatalis (velvetbean caterpillar) and Chrysodeixis includens (soybean looper, formerly named Pseudoplusia includens) are two important defoliating insects of soybeans. Both lepidopteran pests are controlled mainly with synthetic insecticides. Alternative control strategies, such as biopesticides based on the Bacillus thuringiensis (Bt) toxins or transgenic plants expressing Bt toxins, can be used and are increasingly being adopted. Studies on the insect susceptibilities and modes of action of the different Bt toxins are crucial to determine management strategies to control the pests and to delay outbreaks of insect resistance. In the present study, the susceptibilities of both soybean pests to the Bt toxins Cry1Ac, Cry1Fa, Cry1Ca, and Cry2Aa have been investigated. Bioassays performed in first-instar larvae showed that both insects are susceptible to all these toxins. Competition-binding studies carried out with Cry1Ac and Cry1Fa 125-iodine labeled proteins demonstrated the presence of specific binding sites for both of them on the midgut brush border membrane vesicles (BBMVs) of both A. gemmatalis and C. includens Competition-binding experiments and specific-binding inhibition studies performed with selected sugars and lectins indicated that Cry1Ac and Cry1Fa share some, but not all, binding sites in the midguts of both insects. Also, the Cry1Ac- or Cry1Fa-binding sites were not shared with Cry1Ca or Cry2Aa in either soybean pest. This study contributes to the knowledge of Bt toxicity and midgut toxin binding sites in A. gemmatalis and C. includens and sheds light on the cross-resistance potential of Cry1Ac, Cry1Fa, Cry1Ca, and Cry2Aa Bt proteins as candidate proteins for Bt-pyramided crops.IMPORTANCE In the present study, the toxicity and the mode of action of the Bacillus thuringiensis (Bt) toxins Cry1Ac, Cry1Fa, Cry1Ca, and Cry2Aa in Anticarsia gemmatalis and Chrysodeixis includens (important defoliating pests of soybeans) have been investigated. These studies are crucial for determining management strategies for pest control. Bioassays showed that both insects were susceptible to the toxins. Competition-binding studies demonstrated the presence of Cry1Fa- and Cry1Ac-specific binding sites in the midguts of both pests. These results, together with the results from binding inhibition studies performed with sugars and lectins, indicated that Cry1Ac and Cry1Fa share some, but not all, binding sites, and that they were not shared with Cry1Ca or Cry2Aa in either soybean pest. This study contributes to the knowledge of Bt toxicity in A. gemmatalis and C. includens and sheds light on the cross-resistance potential of Cry1Ac, Cry1Fa, Cry1Ca, and Cry2Aa Bt proteins as candidate proteins for Bt-pyramided crops.


COVID-19 vaccine equity: a retrospective population-based cohort study examining primary series and first booster coverage among persons with a history of immigration and other residents of Ontario, Canada.
Immigrants were disproportionately impacted by COVID-19 and experience unique vaccination barriers. In Canada (37 million people), 23% of the population is foreign-born. Immigrants constitute 60% of the country's racialized (non-white) population and over half of immigrants reside in Ontario, the country's most populous province. Ontario had several strategies aimed at improving vaccine equity including geographic targeting of vaccine supply and clinics, as well as numerous community-led efforts. Our objectives were to (1) compare primary series vaccine coverage after it was widely available, and first booster coverage 6 months after its availability, between immigrants and other Ontario residents and (2) identify subgroups experiencing low coverage. Using linked immigration and health administrative data, we conducted a retrospective population-based cohort study including all community-dwelling adults in Ontario, Canada as of January 1, 2021. We compared primary series (two-dose) vaccine coverage by September 2021, and first booster (three-dose) coverage by March 2022 among immigrants and other Ontarians, and across sociodemographic and immigration characteristics. We used multivariable log-binomial regression to estimate adjusted risk ratios (aRR). Of 11,844,221 adults, 22% were immigrants. By September 2021, 72.6% of immigrants received two doses (vs. 76.4%, other Ontarians) and by March 2022 46.1% received three doses (vs. 58.2%). Across characteristics, two-dose coverage was similar or slightly lower, while three-dose coverage was much lower, among immigrants compared to other Ontarians. Across neighborhood SARS-CoV-2 risk deciles, differences in two-dose coverage were smaller in higher risk deciles and larger in the lower risk deciles; with larger differences across all deciles for three-dose coverage. Compared to other Ontarians, immigrants from Central Africa had the lowest two-dose (aRR = 0.60 [95% CI 0.58-0.61]) and three-dose coverage (aRR = 0.36 [95% CI 0.34-0.37]) followed by Eastern Europeans and Caribbeans, while Southeast Asians were more likely to receive both doses. Compared to economic immigrants, resettled refugees and successful asylum-claimants had the lowest three-dose coverage (aRR = 0.68 [95% CI 0.68-0.68] and aRR = 0.78 [95% CI 0.77-0.78], respectively). Two dose coverage was more equitable than 3. Differences by immigrant region of birth were substantial. Community-engaged approaches should be re-invigorated to close gaps and promote the bivalent booster.


Stress Symptoms Among Children and Their Parents After ICU Hospitalization.
Objective: Child survival after intensive care unit (ICU) hospitalization has increased, yet many children experience acute stress that may precipitate mental/behavioral health comorbidities. Parents report stress after their child's hospitalization. Little is known about the individual and family characteristics that may moderate intergenerational relationships of acute stress. Design: Following ICU admission at a large academic medical center, a prospective cross-sectional cohort study assessed the associations between intergenerational characteristics and acute stress among children and families. Patients: Parent-child dyads (N = 88) were recruited from the pediatric ICU and pediatric cardiovascular ICU (CVICU) following ICU discharge. Eligible children were between 8 and 18 years old with ICU stays longer than 24 hours. Children with developmental delays were excluded. Caregivers were proficient in English or Spanish. Surveys were collected before hospital discharge. Measurements/Main Results: The primary outcome was "child stress" defined as a score≥17, measured by the Children's Revised Impact of Events Scale (CRIES-8). "Parent stress" was defined as an elevated composite score on the Stanford Acute Stress Reaction Questionnaire. We used validated scales to assess the child's clinical and family social characteristics. Acute stress was identified in 34 (39.8%) children and 50 (56.8%) parents. In multivariate linear regression analyses adjusting for social characteristics, parent stress was associated with increased risk of child stress (adjusted odds ratio 2.58, 95% confidence interval 0.69, 4.46, p < .01). In unadjusted analyses, Hispanic ethnicity was associated with greater child stress. In adjusted analyses, race, income, ICU length of stay, and language were not associated with child stress and did not moderate the parent-child stress relationship. Conclusions: Parent stress is closely correlated with child stress during ICU hospitalization. Hispanic ethnicity may be associated with increased risk for child stress, but further studies are required to define the roles of other social and clinical measures.


Associations of Body Mass Index, Motor Performance, and Perceived Athletic Competence with Physical Activity in Normal Weight and Overweight Children.
Children who are overweight and obese display lower physical activity levels than normal weight peers. Measures of weight status, perceived motor competence, and motor skill performance have been identified as potential correlates explaining this discrepancy. 1881 children (955 males; 926 females; 9.9 years) were assessed as part of the Physical Health Activity Study Team project. The age, habitual physical activity participation (PAP), body mass index (BMI), socioeconomic status (SES), motor performance (MP), and perceived athletic competence (PAC) of each child included were assessed. Gender-specific linear regression analyses (main effects model) were conducted to identify the percent variance in PAP explained by the following variables: BMI, MP, and PAC. For males, 18.3% of the variance in PAP was explained by BMI, MP, and PAC. PAC explained 17% of the variance, while MP, BMI, and SES only accounted for 0.6%, 0.7%, and 0.5%, respectively. PAC explained 17.5% of PAP variance in females; MP explained 0.8%. BMI, SES, and chronological age were not significant correlates of PAP in girls. An established repertoire of motor skill performance has been seen as a vehicle to PAP in children; however, this study indicates that PAC should not be overlooked in intervention strategies to promote increased PAP.


miR-149-3p reverses CD8+ T-cell exhaustion by reducing inhibitory receptors and promoting cytokine secretion in breast cancer cells.
Blockade of inhibitory receptors (IRs) is one of the most effective immunotherapeutic approaches to treat cancer. Dysfunction of miRNAs is a major cause of aberrant expression of IRs and contributes to the immune escape of cancer cells. How miRNAs regulate immune checkpoint proteins in breast cancer remains largely unknown. In this study, downregulation of miRNAs was observed in PD-1-overexpressing CD8+ T cells using miRNA array analysis of mouse breast cancer homografts. The data reveal that miR-149-3p was predicted to bind the 3'UTRs of mRNAs encoding T-cell inhibitor receptors PD-1, TIM-3, BTLA and Foxp1. Treatment of CD8+ T cells with an miR-149-3p mimic reduced apoptosis, attenuated changes in mRNA markers of T-cell exhaustion and downregulated mRNAs encoding PD-1, TIM-3, BTLA and Foxp1. On the other hand, T-cell proliferation and secretion of effector cytokines indicative of increased T-cell activation (IL-2, TNF-α, IFN-γ) were upregulated after miR-149-3p mimic treatment. Moreover, the treatment with a miR-149-3p mimic promoted the capacity of CD8+ T cells to kill targeted 4T1 mouse breast tumour cells. Collectively, these data show that miR-149-3p can reverse CD8+ T-cell exhaustion and reveal it to be a potential antitumour immunotherapeutic agent in breast cancer.


Patterned Carboxylation of Graphene Using Scanning Electrochemical Microscopy.
A simple, direct, and versatile scanning electrochemical microscopy (SECM) approach for local carboxylation of multilayered graphene on nickel is demonstrated, in which carbon dioxide serves as the carboxylation agent under reductive conditions in N,N-dimethylformamide. The use of SECM gives control over both the spatial dimensions and the degree of carboxylation. While the pattern size, in general, is governed by the dimension of the SECM tip, the degree of modification, expressed as the surface coverage of carboxylate groups introduced at the graphene substrate, is found to be controlled by the electrolysis time. This is supported by electrochemical measurements, two-dimensional X-ray photoelectron spectroscopy, Raman spectroscopy mapping, and He ion microscopy. Surprisingly, intercalation of the supporting electrolyte in the multilayered graphene on nickel occurs to a relatively small extent when compared to corresponding results obtained in previously described carboxylations of this kind of multilayered graphene.


Association of stomatin (band 7.2b) with Glut1 glucose transporter.
Employing a monoclonal antibody directed against the C-terminal peptide of glucose transporter molecule 1 (Glut1), we identified a approximately 30-kDa polypeptide which coimmunoprecipitated with Glut1 from sample of human red blood cells (RBC) membranes. The approximately 30-kDa polypeptide reacted with an antibody directed against stomatin, an integral plasma membrane protein which is also present at a high abundance in the human RBC plasma membrane. Likewise, employing anti-stomatin antibody, we found that Glut1 coimmunoprecipitated with stomatin from samples of RBC membranes. However, neither band 3, which is the most abundant integral membrane protein in the RBC, nor actin coimmunoprecipitated with Glut1, indicating a specific interaction between Glut1 and stomatin. Similar to the results obtained in the RBC, Glut1 and stomatin immunoprecipitated with each other in lysates of Clone 9 cells, a rat liver cell line in which Glut1 is expressed at approximately 1/200 the level present in RBC. Employing conditions that resulted in immunoprecipitation of approximately 10% of Glut1 in RBC membranes led to a approximately 3% coimmunoprecipitation of stomatin. A mixed population of Clone 9 cells stably transfected with a plasmid overexpressing the mouse stomatin exhibited 30 +/- 3% reduction in the basal rate of glucose transport compared to control cells or cells stably transfected with the empty vector. The above results suggest that stomatin is closely associated with Glut1 in the plasma membrane and that overexpression of stomatin results in a depression in the basal rate of glucose transport.


Effective virtual patient simulators for medical communication training: A systematic review.
Despite the growing use of virtual patients (VPs) in medical education, few studies have explored the features and effectiveness of VP-based medical communication skills training. We undertook a systematic review to summarise the design and evaluation of VP-based medical communication skills training systems in order to identify features of successful cases. Following PRISMA guidelines, we searched four databases for studies published between 2006 and 2018. Using a refined classification scheme, we extracted data on instructional design (scenario and instructional intervention), technological design (modality and interaction), and evaluation (user experience, learning effectiveness and evaluator). We assessed the quality of studies using the Medical Education Research Study Quality Instrument (MERSQI) and the QualSyst standard assessment criteria. A total of 14 studies were included for review. Of these, 85.7% (n = 12) were quantitative and 71.4% (n = 10) involved undergraduate students. The most common VP training scenario was history taking followed by the delivery of bad news. Diverse instructional interventions, including tutorials, learning activities, and feedback, were embedded in the VPs. The first-person perspective animated within-screen size VP was a popular technological feature. Most evaluations concerned the reality of simulation (for user experience) and skill in expressing empathy (as a learning outcome). Of the eight comparative studies, half reported significant attitude or skill improvements in the VP group. The distinct features of VPs shown to be effective were well-designed instructional interventions (eg, a pre-activity with a protocol-informed tutorial), and post-activity (eg, debrief or reflection), scaffolding and human feedback, but not system feedback. Evidence-based VP training can enable students to gain communication skills in a safe and affordable learning environment. Elaborate technology alone cannot guarantee effective learning, but evidence-based instructional interventions can facilitate its optimal use and bring about better learning outcomes.


An evaluation of process and protocols for planned home birth attended by regulated midwives in British Columbia.
Midwifery emerged as a self-regulated profession in British Columbia in the context of a 2-year demonstration project beginning in 1998. The project evaluated accountability among midwives, defined as the provision of safe and appropriate care and maintenance of standards of communication set by the College of Midwives of British Columbia. Adherence to protocols was measured by using documentation designed specifically for the Home Birth Demonstration Project. Hospital and transport records for selected clients were reviewed by an expert committee. Outcomes among Home Birth Demonstration Project clients were compared with outcomes among women eligible for home birth but planning to deliver in hospital. Adherence to clinical and communication protocols was 96% or higher. Planned home birth was not associated with an increase in risk but prevalence of adverse outcomes was too low to be studied with precision. Recommendations of an expert review committee have been implemented or are under review. Midwives have demonstrated a high degree of compliance with reporting requirements and protocols. Comparisons of birth outcomes of planned home versus hospital births, while supporting home birth as a choice for women, were limited in scope and require ongoing study. Integration of home birth has been a dynamic process with guidelines and policy continuing to evolve.


Calcium mobilization, water balance, and growth in embryos of the agamid lizard Amphibolurus barbatus.
Embryos of the agamid lizard Amphibolurus barbatus are at developmental stage 29 of Dufaure and Hubert at the time of oviposition. Mobilization of calcium and other nutrients from the yolk proceeds slowly for the first half of incubation, during which time embryonic growth also proceeds slowly. During the second half of incubation, however, embryos withdraw calcium and other nutrients from the yolk very rapidly, and growth rates are correspondingly high. Approximately 60% of the calcium used by developing embryos is obtained from the yolk, but fully 40% of their requirement is met by calcium mobilized from the eggshell. Very little calcium remains in residual yolk of hatchlings, so this yolk must be used in maintenance metabolism rather than in growth of neonates. No dichotomy exists among oviparous, amniotic vertebrates with respect to sources of calcium used by developing embryos, but one does exist with respect to patterns of mobilization of this element. Whereas calcium is extracted from yolk of embryonic reptiles throughout incubation, it actually is deposited in yolk of embryonic birds after the midpoint in development.


Influence of Age and Phylogenetic Background on Blood Parameters Associated With Bone Metabolism in Laying Hens.
The high laying performance of today's laying hens places enormous demands on their mineral metabolism. While up-to-date data are rare, the present study aimed to describe blood parameters associated with egg laying and bone metabolism during the pre-laying period, in the course of the laying period and the daily egg laying cycle. Ten to 15 laying hens of two high-performing, phylogenetically divergent lines (BLA: brown-egg layer; WLA: white-egg layer), kept in single cages were blood sampled at 17, 25, 29, 49, and 69 weeks of age. Sampling was made at 6 a.m., 10 a.m., 2 p.m. and, with the exception of week 17, 6 p.m. Blood samples were analyzed for concentrations of total and ionized calcium, inorganic phosphate (PO4), markers of bone formation (osteocalcin) and resorption [carboxyterminal crosslinked telopeptide of type I collagen (CTX-I)], 25-hydroxycholecalciferol (25(OH)D3) and estradiol-17β. In the pre-laying period (17 week), the estradiol-17β level calculated for WLA was more than twice as high as the level calculated for BLA, while no significant difference could be observed in the laying period (25 to 69 weeks). BLA hens had significantly higher total calcium concentrations at 49 weeks of age as well as up to twice as high levels of osteocalcin and 25(OH)D3 than WLA at any time of the day from 25 to 69 weeks of age. While osteocalcin, CTX-I and 25(OH)D3 concentrations were significantly higher before the onset of lay, total calcium and estradiol-17β levels significantly increased from 17 to 69 weeks of age. In contrast, PO4 values varied only slightly during the experimental period and ionized calcium was highest at 17 and 49 weeks of age and lowest around peak production (29 week). In the course of the daily egg laying cycle blood concentrations clearly reflected the stage of egg formation. Our results provide up-to-date data of bone- and egg laying-associated blood parameters of two contemporary purebred layer lines over the course of the pre- and egg-laying period and the daily egg laying cycle. Differences between brown- and white-egg layers raise questions, whether phylogenetic background determines their efficiency to cope with high calcium demands relating to egg production.


Kinetics of oxygen evolution at α-Fe2O3 photoanodes: a study by photoelectrochemical impedance spectroscopy.
Photoelectrochemical Impedance Spectroscopy (PEIS) has been used to characterize the kinetics of electron transfer and recombination taking place during oxygen evolution at illuminated polycrystalline α-Fe(2)O(3) electrodes prepared by aerosol-assisted chemical vapour deposition from a ferrocene precursor. The PEIS results were analysed using a phenomenological approach since the mechanism of the oxygen evolution reaction is not known a priori. The results indicate that the photocurrent onset potential is strongly affected by Fermi level pinning since the rate constant for surface recombination is almost constant in this potential region. The phenomenological rate constant for electron transfer was found to increase with potential, suggesting that the potential drop in the Helmholtz layer influences the activation energy for the oxygen evolution process. The PEIS analysis also shows that the limiting factor determining the performance of the α-Fe(2)O(3) photoanode is electron-hole recombination in the bulk of the oxide.


Airborne transmission of respiratory viruses.
The COVID-19 pandemic has revealed critical knowledge gaps in our understanding of and a need to update the traditional view of transmission pathways for respiratory viruses. The long-standing definitions of droplet and airborne transmission do not account for the mechanisms by which virus-laden respiratory droplets and aerosols travel through the air and lead to infection. In this Review, we discuss current evidence regarding the transmission of respiratory viruses by aerosols-how they are generated, transported, and deposited, as well as the factors affecting the relative contributions of droplet-spray deposition versus aerosol inhalation as modes of transmission. Improved understanding of aerosol transmission brought about by studies of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection requires a reevaluation of the major transmission pathways for other respiratory viruses, which will allow better-informed controls to reduce airborne transmission.


Fragmentation of carbohydrate anomeric alkoxyl radicals: new synthesis of chiral 1-fluoro-1-halo-1-iodoalditols.
A new general methodology for the synthesis of 1,1,1-trihaloalditols by starting from 1,5-anhydro-2-deoxy-hex-1-enitol derivatives (glycals) is described. The halogens are introduced sequentially in each of the three different steps of the process. The fluorine is introduced in the first step by electrophilic fluorination of the starting glycal; next, hydroxyhalogenation of the resulting vinyl fluoride allows the addition of any halogen (F, Cl, Br or I) at will, and finally, an iodine atom is inserted through an alkoxyl radical fragmentation reaction. This methodology allows the preparation of diverse types of 1,1,1-trihalogenated compounds (R--CF(2)I, R--CFI(2), R--CFClI and R--CFBrI) under mild conditions compatible with sensitive substituents. In some cases, the diastereomeric mixtures generated from R--CFClI and R--CFBrI can be chromatographically separated, and their configuration determined by X-ray crystallographic analysis. The synthetic usefulness of these compounds has been preliminarily assessed by examining the reactivity of the fluorinated radical generated by rupture of the C--I bond.


Critical concepts in adaptive clinical trials.
Adaptive clinical trials are an innovative trial design aimed at reducing resources, decreasing time to completion and number of patients exposed to inferior interventions, and improving the likelihood of detecting treatment effects. The last decade has seen an increasing use of adaptive designs, particularly in drug development. They frequently differ importantly from conventional clinical trials as they allow modifications to key trial design components during the trial, as data is being collected, using preplanned decision rules. Adaptive designs have increased likelihood of complexity and also potential bias, so it is important to understand the common types of adaptive designs. Many clinicians and investigators may be unfamiliar with the design considerations for adaptive designs. Given their complexities, adaptive trials require an understanding of design features and sources of bias. Herein, we introduce some common adaptive design elements and biases and specifically address response adaptive randomization, sample size reassessment, Bayesian methods for adaptive trials, seamless trials, and adaptive enrichment using real examples.


Novel Formulation of Adaptive MPC as EKF Using ANN Model: Multiproduct Semibatch Polymerization Reactor Case Study.
In this paper, a novel formulation for nonlinear model predictive control (MPC) has been proposed incorporating the extended Kalman filter (EKF) control concept using a purely data-driven artificial neural network (ANN) model based on measurements for supervisory control. The proposed scheme consists of two modules focusing on online parameter estimation based on past measurements and control estimation over control horizon based on minimizing the deviation of model output predictions from set points along the prediction horizon. An industrial case study for temperature control of a multiproduct semibatch polymerization reactor posed as a challenge problem has been considered as a test bed to apply the proposed ANN-EKFMPC strategy at supervisory level as a cascade control configuration along with proportional integral controller [ANN-EKFMPC with PI (ANN-EKFMPC-PI)]. The proposed approach is formulated incorporating all aspects of MPC including move suppression factor for control effort minimization and constraint-handling capability including terminal constraints. The nominal stability analysis and offset-free tracking capabilities of the proposed controller are proved. Its performance is evaluated by comparison with a standard MPC-based cascade control approach using the same adaptive ANN model. The ANN-EKFMPC-PI control configuration has shown better controller performance in terms of temperature tracking, smoother input profiles, as well as constraint-handling ability compared with the ANN-MPC with PI approach for two products in summer and winter. The proposed scheme is found to be versatile although it is based on a purely data-driven model with online parameter estimation.


A comparison of costs of Medicare Part D prescriptions dispensed at retail and mail order pharmacies.
Plan sponsors encourage the use of mail order pharmacies because they believe mail order dispensing will lower their prescription drug costs. Health plans and pharmacy benefit management companies (PBMs) usually offer patients substantially lower copayments to incentivize them to use mail order pharmacies. A number of health plans and PBMs now require patients to use these pharmacies for maintenance prescriptions. To (a) compare costs for prescriptions dispensed through mail order and retail pharmacies in Medicare Part D plans and (b) examine whether mail order or retail pharmacies provided lower all third-party costs for each of the top 300 products; the relationship between whether a product was available generically and whether mail order or retail pharmacies provided lower prices; and the generic substitution rates at mail order and retail pharmacies. The sample for this study consisted of 2010 Medicare Part D prescription drug data for the 300 products with the highest sales at mail order pharmacies. The prescriptions included in the study were dispensed in the initial coverage limit phase of Part D by retail or mail order pharmacies to patients who were insured by Part D for 12 months in 2010, who received no Part D subsidies, and who were alive for the full year in 2010. Mean-per-unit costs were calculated for both mail order and retail prescriptions for each of the top 300 products. Products were defined by Medi-Span Generic Product Indentifier. Summary statistics for the overall costs of mail order and retail prescriptions were calculated as the weighted mean-per-unit costs of the top 300 products. The weighting factor for both mail order and retail prescriptions included both the mail order quantity dispensed per prescription and the number of prescriptions dispensed. Weighting both mail and retail prescriptions by mail order quantities dispensed and numbers of prescriptions ensured that the results reflected actual cost differences rather than differences in the mix or quantities of prescriptions dispensed. These calculations were made for total costs, costs covered by the Medicare Standard Benefit (MSB), costs paid by all third-party payers (including Medicare), and patient costs. The top 300 products accounted for 84.8% of mail order costs. Among all prescriptions in the sample--both mail order and retail--mail order prescriptions accounted for 7.8% of prescriptions dispensed and 14.1% of total spending. Comparison of 90-day or greater supplies indicated that costs per unit of medication for retail pharmacies were lower for total costs ($0.94 for retail pharmacies vs. $0.96 for mail order pharmacies), MSB costs ($0.59 for retail pharmacies vs. $0.63 for mail order pharmacies), and all third-party payer costs ($0.64 for retail pharmacies vs. $0.72 for mail order pharmacies), but higher for patient costs ($0.31 for retail pharmacies vs. $0.24 for mail order pharmacies). Retail pharmacies had lower all third-party payer costs for 244 products, while mail pharmacies had lower costs for 56 products. Retail pharmacies were more likely to have lower costs for products that included generic alternatives, while mail order pharmacies were more likely to have lower costs for products that included only branded drugs. Generic substitution rates were 91.4% for retail pharmacies versus 88.8% for mail order pharmacies. Results from secondary analyses that compared all prescriptions which met the inclusion criteria, regardless of days supply, and that compared exactly 90-day supplies, yielded similar results. Third-party payers, including Medicare, paid more for prescriptions dispensed at mail order pharmacies than for those dispensed at retail pharmacies in the Medicare Part D program. The higher payments appeared to result, for the most part, because of higher patient cost sharing at retail pharmacies. Further, total costs--including both third-party payer and patient payments--for 90-day and 90-day or greater supplies were lower at retail pharmacies than at mail pharmacies. These results suggest that, all other things being equal, Medicare Part D plan sponsors do not realize savings when patients use mail order pharmacies.


Sleep Mediates the Association Between PTSD Symptoms and Chronic Pain in Youth.
Symptoms of post-traumatic stress disorder (PTSS) and chronic pain have been shown to co-occur at high rates in adolescents and this co-occurrence is linked to worse pain and quality of life. Sleep disturbance has been posited as a mechanism underlying this co-occurrence in conceptual models of mutual maintenance. This study examined the mediating role of sleep in the relationship between PTSS and pain in youth (aged 10-17 years) with chronic pain. Ninety-seven participants completed measures of PTSS, pain (intensity and interference), anxiety symptoms, and sleep quality, in addition to demographic characteristics. Mediation models were conducted. Findings revealed that, over and above the influence of associated demographic characteristics (age, race) and anxiety symptoms, sleep quality partially mediated the relationships between PTSS and pain intensity and interference for youth with chronic pain. Specifically, higher levels of PTSS was linked to higher levels of pain intensity and pain interference, and these relationships were partially explained by poor sleep quality. Findings highlight the potential mechanistic role of sleep in explaining the co-occurrence of chronic pain and PTSS and suggest sleep might be an important target in future interventions. Consistent with the pediatric model of mutual maintenance in PTSS and chronic pain, poor sleep quality was found to underlie this co-occurrence in youth.


Reduced impact of alcohol use on next-day tiredness in older relative to younger adults: A role for sleep duration.
Recent work has suggested that older adults may be less susceptible to the next-day effects of alcohol relative to younger adults. The effects of alcohol in younger adults may be mediated by sleep duration, but due to age differences in the contexts of alcohol use, this mediation process may not generalize to older adults. The present study examined age-group (younger vs. older adults) differences in how alcohol use influenced next-day tiredness during daily life. Reports of alcohol use, sleep duration, and next-day tiredness obtained on ∼101 days from 91 younger adults (ages 20-31 years) and 75 older adults (ages 65-80 years) were modeled using a multilevel, moderated mediation framework. Findings indicated that (a) greater-than-usual alcohol use was associated with greater-than-usual tiredness in younger adults only, (b) greater-than-usual alcohol use was associated with shorter-than-usual sleep duration in younger adults only, and (c) shorter-than-usual sleep duration was associated with greater tiredness in both younger and older adults. For the prototypical younger adult, a significant portion (43%) of the association between alcohol use and next-day tiredness could be explained assuming mediation through sleep duration, whereas there was no evidence of mediation for the prototypical older adult. Findings of age differences in the mediation process underlying associations among alcohol use, sleep, and tiredness provide insight into the mechanisms driving recent observations of reduced next-day effects of alcohol in older relative to younger adults. (PsycINFO Database Record


Human cytomegalovirus infection and disorders of the nervous system.
Infection with human cytomegalovirus (CMV) occurs in nearly 1% of live-born infants, and from 60% to 80% of the inhabitants of the United States acquire CMV infection by mid adulthood. While neurologic disorders do not develop in the majority of congenitally infected infants, congenital CMV infection can severely damage the developing nervous system, causing microcephaly, psychomotor retardation, seizures, and deafness. Furthermore, approximately 10% of infants who are asymptomatic at birth subsequently exhibit sensorineural hearing loss. In the adult, CMV infection has been associated with the Guillain-Barré syndrome, meningoencephalitis, and retinitis. Although Guillain-Barré syndrome can accompany CMV infections in previously healthy adults, meningoencephalitis and retinitis occur more commonly in immunosuppressed patients, particularly among organ transplant recipients. In total, at least 7,000 persons in the United States each year have neurologic disorders attributable to CMV infection.


Asymmetric syntheses of α-methyl γ-amino acid derivatives.
This project was undertaken to demonstrate the potential of asymmetric hydrogenations mediated by the chiral, carbene-oxazoline analogue of Crabtree's catalyst "cat" in asymmetric hydrogenations of allylic amine derivatives of amino acids. Peripheral features of the substrates (protecting groups, functional groups related by redox processes, and alkene geometries) were varied to optimize the stereochemical vectors exerted by the substrate and align them with the catalyst vector. N-Acetyl-protected, O-TBDPS-protected allylic substrates 9a-e emerged as the best for this reaction; syn-products were formed from the E-alkenes, while the Z-isomers gave anti-target materials, both with high diastereoselectivities. This study featured asymmetric catalysis to elaborate optically active substrates into more stereochemically complex chirons; we suggest that the approach used, optimization of stereocontrol by varying peripheral aspects of the substrate, tends to be easier than de novo catalyst design for each substrate type. In other words, optimization of the substrate vector is likely to be more facile than enhancement of the catalyst vector via ligand modifications.


Prospects for biological control of nematode parasites of ruminants.
Control of nematode parasites in ruminants is directed almost exclusively at the parasitic stage in the host using conventional anthelmintic therapy. However, to complete their life cycle, parasites have to develop through a series of free-living stages on pasture. It is within this environment that there is a vast range of cohabiting micro-organisms, some of which are known to infect or prey on nematodes. These include bacteria, protozoa, viruses and fungi. To date, most interest has been focused on those organisms producing chemical toxins which have been developed as anthelmintics, e.g. Streptomyces/avermectins. Yet the potential of predacious and nematoxic fungi and bacteria for the biological control of nematode parasites of livestock remains largely untapped. These organisms may well offer a cheaper and more sustainable approach to reducing the impact of parasitism in livestock.


The use of lithium in childhood and adolescence: an overview.
Identifying those children and adolescents with lithium-responsive behavior disorders is often difficult. Work done in those areas is reviewed with consideration given to diagnostic issues, outcome studies, and the immediate or long-term risks of lithium use in this younger age group. A balance should be struck between potential side effects and the need for proper treatment of potentially devastating illness.


A rapid fluorescence assay for danofloxacin in beef muscle: effect of muscle type on limit of quantitation.
A simple, rapid fluorescence screening assay was applied to the analysis of beef muscle for danofloxacin at the U.S. tolerance level of 200 ng/g. Muscle samples were homogenized in acetic acid-acetonitrile, the resultant mixture centrifuged, and fluorescence of the supernatants was then measured. The significant difference between the fluorescence of control muscle sample extracts and extracts of samples fortified at 200 ng/g allowed for successful discrimination between the samples. Setting a threshold level at the average 200 ng/g fortified sample extract fluorescence -3sigma allowed for identification of potentially violative samples. Successful analysis of a group of blind fortified samples over a range of concentrations was accomplished in this manner, without any false-negative results. The limits of quantitation for danofloxacin, as well as enrofloxacin, using this assay were determined in three types of beef muscle (hanging tenderloin, neck, and eye round steak), as well as in serum. Significant differences in limits of quantitation were found among the three different muscle types examined, with hanging tenderloin muscle providing the lowest value. This work not only shows the potential for use of the fluorescence screening assay as an alternative to currently used microbial or antibody-based assays for the analysis of danofloxacin in beef muscle, but also suggests that assays using beef muscle may vary in performance depending on the specific muscle selected for analysis.


Healthcare utilization and direct costs of non-infectious comorbidities in HIV-infected patients in the USA.
To estimate the incremental healthcare utilization and costs associated with common non-infectious comorbid conditions among commercially and Medicaid-insured HIV-infected patients in the US. US administrative claims were used to select adult HIV patients with chronic kidney disease (CKD), cardiovascular disease (CVD) events, or fracture/osteoporosis, three common comorbidities that have been associated with HIV and HIV treatment, between 1 January 2004 and 30 June 2013. Propensity score matched controls with no CKD, no CVD events, and no fracture/osteoporosis were identified for comparison. All-cause healthcare utilization and costs were reported as per patient per month (PPPM). The commercial cohort comprised 381 CKD patients, 624 patients with CVD events, and 774 fracture/osteoporosis patients, and 1013, 1710, and 2081 matched controls, respectively; while the Medicaid HIV cohort comprised 207 CKD and 271 CVD cases, and 516 and 735 matched controls, respectively. There was insufficient Medicaid data for fracture analyses. Across both payers, HIV patients with CKD or CVD events had significantly higher healthcare utilization and costs than controls. The average incremental PPPM costs in HIV patients with CKD were $1403 in the commercial cohort and $3051 in the Medicaid cohort. In those with CVD events, the incremental costs were $2655 (commercial) and $4959 (Medicaid) for HIV patients compared to controls (p < .001). The results suggested a considerable increase in healthcare utilization and costs associated with CKD, CVD and fracture/osteoporosis comorbidities among HIV patients in the past decade. Because these conditions have been associated with treatment, it is critical to consider their impact on costs and outcomes when optimizing patient care.


Health-risk factors and the prevalence of hypertension: cross-sectional findings from a national cohort of 87,143 Thai Open University students.
Thailand is undergoing a health-risk transition which increases chronic diseases, particularly hypertension, as a result of a rapid transition from a developing to a developed country. This study analyzes the effect of health-risk factors such as demography, socioeconomic status (SES) and body mass index (BMI) on the prevalence of hypertension. This was a cross-sectional analysis using data obtained in 2005 from 87,143 Sukhothai Thammathirat Open University (STOU) students participating in the Thai Cohort Study (mean age 30.5 years, 54.7% female). Adjusted odds ratios of the association between risk factors and hypertension were analysed across two age groups by sex, after controlling for the confounding factors such as SES and BMI. The prevalence of hypertension in men was approximately twice as high as that in women (6.9% vs 2.6%). Hypertension was associated with ageing, a lower education attainment, a higher BMI and having underlying diseases in both sexes. In men, hypertension was associated with being single, having a high income, spending more time on screens (TV & PC), cigarette smoking and drinking alcohol. In women, it was directly correlated with instant and roasted or smoked food consumption. Hypertension was highly associated with obesity and having underlying disease. The Thai health-risk transition is in a later stage. Thais should now be educated about the danger of high blood pressure and the protective power of a low fat and low salt diet, and a normal BMI. Cessation of smoking and moderation in alcohol intake should be promoted.


The effect of pathophysiological changes in the vaginal milieu on the signs and symptoms of genitourinary syndrome of menopause (GSM).
The aim of this study was to provide an overview of the most recent literature on genitourinary syndrome of menopause (GSM), to explore the key elements of GSM diagnosis, and the potential impact of pathophysiological changes in the vaginal milieu on vulvovaginal symptoms. The MEDLINE database was searched, and only articles written in English were considered. Additional references were identified by hand searching the bibliographies of the included articles. The vaginal milieu plays important roles in producing bothersome symptoms in the host. In women with GSM, low hormone states can result in pathophysiological changes in the vaginal milieu, including the vaginal microbiome and the mucosal immunity. Hormone-associated disruption of the balance of the indigenous microbiota and the dysregulation of these immune responses are the pathophysiological basis of GSM symptoms. However, whether the microbiome and mucosal immunity are markers of vulvovaginal disorder or agents actively promoting a healthy vagina are still not fully understood. It is an important area of focus.


The impact of childhood obesity on inflammation, innate immune cell frequency, and metabolic microRNA expression.
Obesity is characterized by chronic inflammation, immune dysregulation, and alteration of gene expression, associated with type 2 diabetes mellitus and cardiovascular disease. The degree to which these changes occur in childhood obesity is not fully defined. The aim was to investigate the effect of childhood obesity on immune cell frequency, macrophage activation, cytokine production, and specific regulators of metabolic gene expression. Profiling was performed on peripheral blood from 29 obese and 20 nonobese children using real-time PCR, ELISA, and flow cytometry. Fasting glucose was similar in both groups, but there was a higher degree of insulin resistance in obese subjects (homeostasis model of assessment for insulin resistance, 4.8 vs 0.84; P < .001). Soluble CD163, a marker of macrophage polarization to a proinflammatory profile, was elevated in the obese compared to nonobese children (135 vs 105 ng/mL; P = .03). Invariant natural killer T cells were reduced in the obese children (CD3 T cells, 0.31 vs 0.53%; P = .001). Cytokine profiling revealed significantly elevated TNF-α (6.7 vs 5.1 pg/mL; P = .01) and leptin (1186 vs 432 pg/mL; P < .001) and reduced adiponectin (884 vs 1321 pg/mL; P = .001) in obese compared to nonobese children. Stimulation of peripheral blood mononuclear cells from obese children resulted in higher levels of IL-1β (2100 vs 1500 pg/mL; P = .018). There was a 4-fold increase in expression of microRNA33a (P = .001) and a 3-fold increase in microRNA33b (P = .017) in obese children. Childhood obesity is associated with changes in immune cell frequency, inflammatory environment, and regulation of metabolic gene expression. These changes have been causally linked to the onset of metabolic disease in adulthood and suggest the future trajectory of obese children to the development of type 2 diabetes mellitus and premature cardiovascular disease.


Significant high expression of cytokeratins 7, 8, 18, 19 in pulmonary large cell neuroendocrine carcinomas, compared to small cell lung carcinomas.
The aim of the present study was to clarify protein profiling in small cell lung carcinoma (SCLC) and pulmonary large cell neuroendocrine carcinoma (LCNEC). The proteomic approach was used, and involved cell lysate from two cell lines (N231 derived from SCLC and LCN1 derived from LCNEC), with 2-D gel electrophoresis (2-DE). In the present study, 25 protein spots with greater than twofold quantitative differences between LCN1 and N231 cells on 2-DE gels were confirmed. Within the 25 identified proteins, cytokeratins (CK) 7, 8, 18 and 19 were upregulated in LCN1 cells compared with N231 cells. The expression of CK7, 8, 18, and 19 was further studied on immunohistochemistry with 81 formalin-fixed and paraffin-embedded pulmonary carcinomas, which included 27 SCLC, 30 LCNEC, 14 adenocarcinomas, and 10 squamous cell carcinomas. Although the expression of CK7, 8, 18, and 19 was observed in all histological types, the mean immunostaining scores of CK7, 8, 18, and 19 were significantly higher in LCNEC than in SCLC (P < 0.001, P < 0.001, P < 0.01 and P < 0.001, respectively). These data suggest that the biological characteristics of LCNEC and SCLC may be different and the expression of CK may serve as differential diagnostic markers.


Soft tissue digital procedures.
The purpose of this article is to discuss the common soft tissue procedures used to correct the reducible digital deformities. The reader should keep in mind that often these procedures are used in combination with osseous techniques for semi- or nonreducible digital deformities.


Postmenopausal estrogen replacement and risk for venous thromboembolism: a systematic review and meta-analysis for the U.S. Preventive Services Task Force.
Postmenopausal estrogen replacement is widely used in the United States but poses important health risks. To assess the risk for venous thromboembolism with postmenopausal estrogen replacement by using literature review and meta-analysis. All relevant English-language studies identified in searches of the MEDLINE (1966 to December 2000), HealthSTAR (1975 to December 2000), and Cochrane Library databases, and references lists of key articles. All published studies of postmenopausal estrogen replacement reporting venous thromboembolism as an outcome or adverse event. 12 studies of estrogen were identified (3 randomized, controlled trials; 8 case-control studies; and 1 cohort study). Data were extracted on participants, interventions, event rates, and confounders. Two reviewers independently rated study quality on the basis of established criteria. A Bayesian meta-analysis was conducted. When data from all studies were pooled, current estrogen use was associated with an increased risk for venous thromboembolism (relative risk, 2.14 [95% credible interval, 1.64 to 2.81]). Estimates did not significantly change when studies were pooled according to study design, quality score, or whether participants had preexisting coronary artery disease. The absolute rate increase was 1.5 venous thromboembolic events per 10 000 women in 1 year. Six case-control studies that reported risk according to duration of use found that risk was highest in the first year of use (relative risk, 3.49 [credible interval, 2.33 to 5.59]). Postmenopausal estrogen replacement is associated with an increased risk for venous thromboembolism, and this risk may be highest in the first year of use.


Chest CT findings in patients with dysphagia and aspiration: a systematic review.
The objective of this systematic review was to characterize chest CT findings in patients with dysphagia and pulmonary aspiration, identifying the characteristics and the methods used. The studies were selected from among those indexed in the Brazilian Virtual Library of Health, LILACS, Indice Bibliográfico Español de Ciencias de la Salud, Medline, Cochrane Library, SciELO, and PubMed databases. The search was carried out between June and July of 2016. Five articles were included and reviewed, all of them carried out in the last five years, published in English, and coming from different countries. The sample size in the selected studies ranged from 43 to 56 patients, with a predominance of adult and elderly subjects. The tomographic findings in patients with dysphagia-related aspiration were varied, including bronchiectasis, bronchial wall thickening, pulmonary nodules, consolidations, pleural effusion, ground-glass attenuation, atelectasis, septal thickening, fibrosis, and air trapping. Evidence suggests that chest CT findings in patients with aspiration are diverse. In this review, it was not possible to establish a consensus that could characterize a pattern of pulmonary aspiration in patients with dysphagia, further studies of the topic being needed.


Enhanced Excito-Repellency of Binary Mixtures of Plant-Based Mosquito Repellents Against Culex quinquefasciatus Say (Diptera: Culicidae), a Night Biting Mosquito Species.
Efficacies of essential oils (EOs) of Vetiveria zizanioides (L.) Nash. (Poales: Poaceae) (VZ EO), Cananga odorata (Lam) Hook. F. & Thomson (Magnoliales: Annonaceae) (CO EO), and crude extract (CE) of Andrographis paniculata (Burm.F.) Wall ex. Nees (Lamiales: Acanthaceae) (AP CE), against laboratory (lab) and field strains of Culex quinquefasciatus Say were investigated. Irritant and repellent activities of individual and binary mixtures of plant extracts were compared with N,N-diethyl-m-toluamide (DEET) using an excito-repellency system. The irritant activity (direct tarsal contact), the mean percent escape response of VZ EO (91.67%, 83.33%), and CO EO (80%, 88.33%) were not significantly different compared with DEET (88.33%, 95%) against lab and field strains, respectively. Similarly, irritant responses in combinations (1:1 and 1:2, v:v) of either VZ EO or CO EO with AP CE were not significantly different from DEET against both strains (P > 0.001). The repellent activity (no tarsal contact), the mean percent escape response of VZ EO (68.33%), CO EO (61.67%), and VZ EO+AP CE (1:1, v:v) (81.67%) against lab strain and CO EO (85%) against field strain were not significantly different from that of DEET (P > 0.001). Interestingly, the greatest contact irritancy of VZ EO+AP CE (1:1, v:v) (96.67%) (P = 0.0026) and a stronger repellency response of CO EO (85%) (P = 0.0055) produced significantly different patterns of escape response compared with DEET against both lab and field strains, respectively. The EOs of VZ EO and CO EO or their mixture with AP CE showed potential as plant-based active ingredients for mosquito repellents. In addition, the major chemical constituents of VZ EO were β-vetivone (6.4%), khusimol (2.96%), and α-vetivone (2.94%) by gas chromatograpy-mass spectrometry.


[Detection of 29 types of fusion gene in leukemia by multiplex RT-PCR].
To analyze the fusion genes derived from 29 types of chromosome structural aberrations in children with leukemia. Bone marrow samples from 191 children with leukemia were analyzed with a novel multiplex nested RT-PCR. Of the 191 leukemic samples, 86 (45.0%) carried 14 types of fusion genes including SIL/TAL1, MLL/AF1q, E2A/PBX1, MLL/AF6, AML1/ETO, MLL/AF9, TEL/ABL, BCR/ABL, MLL/AF10, dupMLL, MLL/ENL, TEL/AML1, PML/RARalpha and CBFbeta/MYH11. The activation of oncogene HOX11 was detected in 31 cases, with or without other chromosome aberrations in 15 (7.8%) and 16 cases (8.4%), respectively. This multiplex nested RT-PCR reaction could screen 29 types of chromosome structural aberrations at the same time. It may be helpful for the detection of minimal residual diseases after chemotherapy and bone marrow transplantation.