Lys49 myotoxin from the Brazilian lancehead pit viper elicits pain through regulated ATP release.
Pain-producing animal venoms contain evolutionarily honed toxins that can be exploited to study and manipulate somatosensory and nociceptive signaling pathways. From a functional screen, we have identified a secreted phospholipase A2 (sPLA2)-like protein, BomoTx, from the Brazilian lancehead pit viper (Bothrops moojeni). BomoTx is closely related to a group of Lys49 myotoxins that have been shown to promote ATP release from myotubes through an unknown mechanism. Here we show that BomoTx excites a cohort of sensory neurons via ATP release and consequent activation of P2X2 and/or P2X3 purinergic receptors. We provide pharmacological and electrophysiological evidence to support pannexin hemichannels as downstream mediators of toxin-evoked ATP release. At the behavioral level, BomoTx elicits nonneurogenic inflammatory pain, thermal hyperalgesia, and mechanical allodynia, of which the latter is completely dependent on purinergic signaling. Thus, we reveal a role of regulated endogenous nucleotide release in nociception and provide a detailed mechanism of a pain-inducing Lys49 myotoxin from Bothrops species, which are responsible for the majority of snake-related deaths and injuries in Latin America.


Fidelity of the eukaryotic codon-anticodon interaction: interference by aminoglycoside antibiotics.
A homologous in vitro method was developed from Tetrahymena for ribosomal A-site binding of aminoacyl-tRNA to poly(uridylic acid)-programmed ribosomes with very low error frequency. The reaction mixture pH was the crucial factor in the stable A-site association of aminoacyl-tRNA with high fidelity. At a pH greater than 7.1, endogenous activity translocated A-site-bound aminoacyl-tRNA to the P site. If translocation was allowed to occur, a near-cognate amino-acyl-tRNA, Leu-tRNA, could stably bind to the ribosome by translocation to the ribosomal P site. Near-cognate aminoacyl-tRNA did not stably bind to either site when translocation was blocked. Misreading antibiotics stimulated the stable association of near-cognate aminoacyl-tRNA to the ribosomal A site, thereby increasing the error frequency by several orders of magnitude. Ribosome binding of total aminoacyl-tRNA near equilibrium was not inhibited by misreading antibiotics; however, initial rate kinetics of the binding reaction were dramatically altered such that a 6-fold rate increase was observed with paromomycin or hygromycin B. The rate increase was evident with both cognate and near-cognate aminoacyl-tRNAs. Several antibiotics were tested for misreading potency by the ribosome binding method. We found gentamicin G418 greater than paromomycin greater than neomycin greater than hygromycin B greater than streptomycin in the potentiation of misreading. Tetracycline group antibiotics effectively inhibited A-site aminoacyl-tRNA binding without promoting misreading.


Quenched disorder and the critical behavior of a partially frustrated system.
We report the direct observation of the effects of quenched disorder on the critical behavior of partially frustrated amorphous FeMnZr alloys by the systematic analysis of high-precision ac susceptibility data and dc magnetization data. Interestingly, the analysis reveals that the presence of short-range quenched disorder does not alter the actual critical behavior. However, it does affect quantities such as the Curie temperature, the peak value of effective exponent gamma, width of the peak, and crossover temperatures. The observed temperature dependence of the effective critical exponent can be understood in terms of the field-theoretical renormalization group approach. Also, the present results would help in identifying the main source of the spread in the exponent values reported in the literature.


Correlation between Epstein-Barr Virus Infection and Disease Activity of Systemic Lupus Erythematosus: a Cross-Sectional Study
Background: Systemic lupus erythematosus (SLE) is an autoimmune disease for whose pathogenesis viral infections are important. The Epstein-Barr virus (EBV) is the main infectious etiological agent. This study aimed to quantitative evaluation of EBV in SLE patients. Materials and Methods: In this cross-sectional study, 40 patients with SLE diagnosed based on American College of Rheumatology criteria were selected using purposive sampling. All were included in the study after obtaining informed consent for participation. Whole blood samples were taken and buffy coat preparations were isolated to determine viral load using the real-time polymerase chain reaction method and assessment with the SLE disease activity index (SLE-DAI). Results: From a total of 40 patients, 37 cases (92.5%) were women. The EBV test was positive in 67.5% and mean viral load was 5396 ± 1891.9 copy/ml. Twenty of forty patients had active and 50% inactive disease, mean EBV viral loads being 6798 and 28.25 copy/ml, respectively (P-value = 0.003). In terms of the severity of disease activity, 17.5 % of female patients had mild to moderate activity, whilst 32.5% of them had severe activity, with respective viral loads of 5,803.3 and 29.73 copy/ml (P-value = 0.003). Conclusion: The Epstein-Barr viral load in SLE patients with active disease was found to be markedly higher than in inactive cases. Thus, EBV may have an important role in the pathogenesis and activity of SLE.


Identification and sequencing of ESTs from the halophyte grass Aeluropus littoralis.
Aeluropus littoralis (Gouan) Parl. is a C4 perennial halophyte monocotyledonous plant belonging to the same family as wheat. Growing as weed in dry salty areas or marshes, it is salt-secreting, rhizomatous and is used as forage. It is diploid (2n=2X=14) and has a relative small genome of around 342 Mb. A. littoralis is highly salt-tolerant since this plant has the ability to secrete salt. Thus, A. littoralis has the potential to become an important genetic resource for biotechnological strategies to improve salt and drought tolerance in economically important crops such as wheat. We have constructed SSH (Suppression Subtractive Hybridization) cDNA libraries from root (RSD45) and leaf (LSD45) tissues of 45 days old plants grown in the presence of 300 mM NaCl. We have also constructed full-length cDNA library from 15 days old salt stressed (300 mM NaCl) roots (RSTL15). Sequencing revealed 25 and 42 independent transcripts from the RSD45 and LSD45 cDNA libraries respectively, in both cases this was less than 25% of the clones sequenced. In contrast, 425 (60%) of the clones from the RSTL15 library revealed independent transcripts. After comparison with protein databases using BlastX, 335 (68%) ESTs (Expressed Sequence Tag) were classified into putative known functions and unclassified proteins, 59 (12%) have homology only to unidentified homologous sequences. A total of 98 (20%) of the ESTs have no homologies to known sequences in the protein databases which can be considered as novel.


Network analysis of primary hepatocyte dedifferentiation using a shotgun proteomics approach.
The liver is the major site of xenobiotic metabolism and detoxification. Primary cultures of hepatocytes are a vital tool in the development of new therapeutic agents but their utility is hindered by the rapid loss of phenotype. Hepatocytes cultured in a sandwich of extracellular matrix protein maintain better hepatic function compared with cells cultured as a monolayer but a wide-ranging proteomics study of the differences in cultures has never been performed. We characterize the changing phenotype of rat hepatocytes in primary culture using iTRAQ proteomics and systems biology network analysis of the identified, significantly regulated, proteins. A total of 754 unique proteins were identified from 4 independent experiments. Of these, 413 proteins were common to at least 3 experiments and 328 proteins were identified in all experiments. Both culture systems displayed altered expression of many common proteins. Network analysis showed that the primary functions of these proteins were in metabolic pathways, immune responses and cytoskeleton remodelling. Monolayer cultures uniquely regulate proteins mapping to pathways of oxidative stress and cell migration, whereas sandwich culture affected translation regulation and apoptosis pathways. These experiments provide a detailed proteomics data set to direct further work into maintaining hepatic phenotype using cultured primary hepatocytes and stem cell derived hepatocyte-like cells.


Endobronchial Hamartoma Causing Postobstructive Pneumonia.
The current case highlights an uncommon cause of recurrent pneumonia. A 76 year old female presented to her primary care physician multiple times with recurrent left lower lobe pneumonia. After multiple chest radiographs continued to show no improvement despite antibiotics, a CT of the chest was performed and showed an intraluminal lesion in the left lower lobe bronchus. An endoscopic biopsy was performed and showed the lesion to be a hamartoma. The patient subsequently underwent resection of the lesion and her symptoms resolved. This case illustrates the need for further diagnostic testing when recurrent pneumonia presents with the same lobar distribution.


Complete mitochondrial genome of the kawakawa tuna Euthynnus affinis.
The present study represents the first report on the complete mitochondrial genome (mitogenome) of the kawakawa tuna Euthynnus affinis. Illumina next-generation sequencing generated a total of 38,428 reads with an average sequencing depth of 232.83×. The mitogenome of E. affinis was 16,513 bp in length and contains 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes and a 849 bp control region, as found in other typical teleosts. With the exception of ND6 and eight tRNA genes, all other mitochondrial genes were encoded on the heavy strand, whose nucleotide composition is 28.11% A, 24.77% T, 17.16% G and 29.96% C. All the protein-coding genes shared the start codon ATG except for COXI gene (start with GTG), and appeared to be terminated with four types of stop codons including two complete codons (TAA and TAG) and two incomplete codons (T and TA).


[Pyomyositis in an immunocompetent patient].
We report a case of pyomyositis in a 59 year-old immunocompetent male patient caused by methicillin-susceptible Staphylococcus aureus. A CT scan of the chest demonstrated a lesion of the pectoral muscles. The patient was treated with surgical drainage and cephalexin. Pyomyositis is a rare and potentially serious infection that requires early recognition and prompt treatment.


Damage to the inferior alveolar nerve as the result of root canal therapy.
Endodontic treatment of mandibular molar teeth has the potential to damage the inferior alveolar nerve via direct trauma, pressure or neurotoxicity. The author reviewed all cases of involvement of the inferior alveolar nerve resulting from root canal therapy in patients seen in a tertiary referral center during an eight-year period (1998 through 2005). The author had encouraged practitioners to refer patients immediately to a university clinic. The author saw 61 patients during the eight-year period. Eight patients were asymptomatic and received no treatment. Forty-two patients exhibited only mild symptoms or were seen more than three months after undergoing root canal therapy, and they received no surgical treatment. Only 10 percent of these patients experienced any resolution of symptoms. Eleven patients underwent surgical exploration. Five of these patients underwent exploration and received treatment within 48 hours, and all recovered completely. The remaining six patients underwent surgical exploration and received treatment between 10 days and three months after receiving endodontic therapy. Of these patients, four experienced partial recovery and two experienced no recovery at all. Early surgical exploration and débridement may reverse the side effects of endodontic treatment on the inferior alveolar nerve. If the radiograph obtained after endodontic therapy shows sealant in the inferior alveolar canal, then immediate referral to an oral and maxillofacial surgeon is indicated if the patient has continued symptoms of paresthesia or pain once the local anesthetic should have worn off. Immediate surgical exploration and débridement may provide satisfactory results.


Sexual offender recidivism among a population-based prison sample.
The present study examines recidivism rates in sexual offenders using officially registered reconvictions in a representative data set of N = 1,115 male sexual offenders from Austria. In general, results indicate that most sexual offenders do not reoffend sexually after release from prison. More detailed, within the first 5 years after release, the sexual recidivism rate was 6% for the total sample, 4% for the rapist subgroup, and 8% for the child molester subgroup. The findings confirmed previous studies about sex offender recidivism which have shown that first-time sexual offenders are significantly less likely to sexually reoffend than those with previous sexual convictions. With regard to the relationship between age and sexual recidivism, the results challenged the traditional assumption of a clear linear function between age and recidivism. Taken together, compared with previous studies, the recidivism rates found in the present investigation are substantially lower than previous research has indicated.


Oral Bacteria Combined with an Intranasal Vaccine Protect from Influenza A Virus and SARS-CoV-2 Infection.
The gut microbiota plays a critical role in the induction of adaptive immune responses to influenza virus infection. However, the role of nasal bacteria in the induction of the virus-specific adaptive immunity is less clear. Here, we found that disruption of nasal bacteria by intranasal application of antibiotics before influenza virus infection enhanced the virus-specific antibody response in a MyD88-dependent manner. Similarly, disruption of nasal bacteria by lysozyme enhanced antibody responses to intranasally administered influenza virus hemagglutinin (HA) vaccine in a MyD88-dependent manner, suggesting that intranasal application of antibiotics or lysozyme could release bacterial pathogen-associated molecular patterns (PAMPs) from disrupted nasal bacteria that act as mucosal adjuvants by activating the MyD88 signaling pathway. Since commensal bacteria in the nasal mucosal surface were significantly lower than those in the oral cavity, intranasal administration of HA vaccine alone was insufficient to induce the vaccine-specific antibody response. However, intranasal supplementation of cultured oral bacteria from a healthy human volunteer enhanced antibody responses to an intranasally administered HA vaccine. Finally, we demonstrated that oral bacteria combined with an intranasal vaccine protect from influenza virus and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. Our results reveal the role of nasal bacteria in the induction of the virus-specific adaptive immunity and provide clues for developing better intranasal vaccines. IMPORTANCE Intranasal vaccination induces the nasal IgA antibody which is protective against respiratory viruses, such as influenza virus and severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Therefore, understanding how mucosal immune responses are elicited following viral infection is important for developing better vaccines. Here, we focused on the role of nasal commensal bacteria in the induction of immune responses following influenza virus infection. To deplete nasal bacteria, we intranasally administered antibiotics to mice before influenza virus infection and found that antibiotic-induced disruption of nasal bacteria could release bacterial components which stimulate the virus-specific antibody responses. Since commensal bacteria in nasal mucosa were significantly lower than those in the oral cavity, intranasal administration of split virus vaccine alone was insufficient to induce the vaccine-specific antibody response. However, intranasal supplementation of cultured oral bacteria from a healthy human volunteer enhanced antibody responses to the intranasally administered vaccine. Therefore, both integrity and amounts of nasal bacteria may be critical for an effective intranasal vaccine.


Mast Cells in the Developing Brain Determine Adult Sexual Behavior.
Many sex differences in brain and behavior are programmed during development by gonadal hormones, but the cellular mechanisms are incompletely understood. We found that immune-system-derived mast cells are a primary target for the masculinizing hormone estradiol and that mast cells are in turn primary mediators of brain sexual differentiation. Newborn male rats had greater numbers and more activated mast cells in the preoptic area (POA), a brain region essential for male copulatory behavior, than female littermates during the critical period for sexual differentiation. Inhibiting mast cells with a stabilizing agent blunted the masculinization of both POA neuronal and microglial morphology and adult sex behavior, whereas activating mast cells in females, even though fewer in number, induced masculinization. Treatment of newborn females with a masculinizing dose of estradiol increased mast cell number and induced mast cells to release histamine, which then stimulated microglia to release prostaglandins and thereby induced male-typical synaptic patterning. These findings identify a novel non-neuronal origin of brain sex differences and resulting motivated behaviors.SIGNIFICANCE STATEMENT We found that immune-system-derived mast cells are a primary target for the masculinizing hormone estradiol and that mast cells are in turn primary mediators of brain sexual differentiation. These findings identify a novel non-neuronal origin of brain sex differences and resulting motivated behaviors.


Integration of Ag Plasmonic Metal and WO3/InGaN Heterostructure for Photoelectrochemical Water Splitting.
In this study, a Ag/WO3/InGaN hybrid heterostructure was successfully developed by sputtering and molecular beam epitaxy techniques, to obtain unique Ag nanospheres adorned with cauliflower-like WO3 nanostructure over the InGaN nanorods (NRs). Exploiting the localized surface plasmon resonance of Ag, the Ag/WO3/InGaN heterostructure exhibited superior photoabsorption ability in the visible region (400-700 nm) of the solar spectrum, with a surface plasmon resonance band centered around 440 nm. Comprehensive analysis through photoluminescence spectroscopy, photocurrent measurements, and electrochemical impedance spectroscopy revealed that the Ag/WO3/InGaN hybrid heterostructure significantly enhances the charge carrier separation and transfer kinetics leading to improved overall photoelectrochemical (PEC) performance. The photocurrent density of the Ag/WO3/InGaN photoanode is 1.17 mA/cm2, which is about 2.72 times higher than that of pure InGaN NRs under visible light irradiation. The photoanode exhibited excellent stability for about 12 h. From the study, it has been found that the maximum applied bias photon-to-current efficiency (ABPE) is ∼1.67% at the applied bias of 0.6 V. The improved PEC water splitting efficiency of the Ag/WO3/InGaN photoanode is attributed to the synergistic effects of localized surface plasmon resonance (LSPR), efficient charge carrier separation and transport, and the presence of a Schottky junction. Consequently, the plasmonic metal-assisted heterojunction-based semiconductor Ag/WO3/InGaN demonstrates immense potential for practical applications in photoelectrochemical water splitting.


Bivalent SARS-CoV-2 mRNA vaccines increase breadth of neutralization and protect against the BA.5 Omicron variant in mice.
The emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) variants in the Omicron lineage has resulted in diminished Coronavirus Disease 2019 (COVID-19) vaccine efficacy and persistent transmission. In this study, we evaluated the immunogenicity and protective efficacy of two, recently authorized, bivalent COVID-19 vaccines that contain two mRNAs encoding Wuhan-1 and either BA.1 (mRNA-1273.214) or BA.4/5 (mRNA-1273.222) spike proteins. As a primary two-dose immunization series in mice, both bivalent vaccines induced greater neutralizing antibody responses against Omicron variants than the parental, monovalent mRNA-1273 vaccine. When administered to mice as a booster at 7 months after the primary vaccination series with mRNA-1273, the bivalent vaccines induced broadly neutralizing antibody responses. Whereas most anti-Omicron receptor binding domain antibodies in serum induced by mRNA-1273, mRNA-1273.214 and mRNA-1273.222 boosters cross-reacted with the antecedent Wuhan-1 spike antigen, the mRNA-1273.214 and mRNA-1273.222 bivalent vaccine boosters also induced unique BA.1-specific and BA.4/5-specific responses, respectively. Although boosting with parental or bivalent mRNA vaccines substantially improved protection against BA.5 compared to mice receiving two vaccine doses, the levels of infection, inflammation and pathology in the lung were lowest in animals administered the bivalent mRNA vaccines. Thus, boosting with bivalent Omicron-based mRNA-1273.214 or mRNA-1273.222 vaccines enhances immunogenicity and confers protection in mice against a currently circulating SARS-CoV-2 strain.


A Novel Strategy to Reveal the Latent Abnormalities in Human Embryonic Stages from a Large Embryo Collection.
The cause of spontaneous abortion of normal conceptuses remains unknown in most cases. The study was aimed to reveal the latent abnormalities by using a large collection of embryo images from a magnetic resonance imaging (MRI) database and novel phase-contrast radiographic computed tomography (PXCT). MRI from 1,156 embryos between Carnegie stage (CS) 14 and CS23 from the Kyoto Collection were screened by using the volume of the liver as the target organ. Embryos with liver volumes ≥2 SD above or below the mean for the stage of development were screened and examined precisely on MRI. Embryos with potentially abnormal livers were further analyzed by using PXCT. Liver abnormality was detected in all 7 embryos in the extra-small liver group and in 2 of 8 embryos in the extra-large liver group. The abnormalities in the extra-small liver group consisted of hepatic agenesis (2 embryos), hepatic hypogenesis (4), and liver lobe defect (1). Among the 7 extra-small liver group, 2 had only liver abnormalities and 5 exhibited complications in other organs. Of the 2 embryos in the extra-large liver group, one had only a single liver abnormality and the other had a morphologically abnormal liver with complications in other organs. Most of such liver abnormality cases are not survive, as liver function becomes essential. The prevalence of liver malformations in CS18 and CS21 in the intrauterine population of externally normal embryos is approximately 1.7%. The present study is the first step toward the elucidation of the latent abnormalities resulting in spontaneous abortion in externally normal embryos.


HCCS loss-of-function missense mutation in a female with bilateral microphthalmia and sclerocornea: a novel gene for severe ocular malformations?
To analyze if mutations in HCCS, encoding the mitochondrial holocytochrome c-type synthase, are associated with phenotypes other than the microphthalmia with linear skin defects (MLS) syndrome, including severe eye malformations such as microphthalmia and/or anophthalmia. In addition, we investigated the impact of the p.E159K missense mutation on sorting of HCCS to mitochondria and its functional integrity. In a cohort of 27 females obtained from a population-based study on infants and fetuses with congenital eye malformations we performed mutation analysis of HCCS by PCR amplification of the coding exons and direct sequencing. The X-inactivation pattern was determined by analyzing the methylation pattern at the AR locus in one patient. For functional analysis of the identified missense mutation, we transfected CHO-K1 cells with wild-type HCCS or HCCS E159K mutant construct and analyzed subcellular localization of the expressed proteins by immunofluorescence analysis and confocal microscopy. Functional integrity of the mutated HCCS protein was investigated by complementation studies in yeast. Therefore, we ectopically expressed HCCS wild type and the E159K mutant in the S. cerevisiae strain B-8025, carrying a deletion of the HCCS ortholog CYC3, and analyzed the capacity of the yeast strain to grow on nonfermentable carbon sources. We detected the heterozygous c.475G>A mutation in exon 5 of HCCS, predicting an amino acid substitution of the highly conserved glutamate at position 159 by lysine, in a female presenting with bilateral microphthalmia and sclerocornea. This point mutation was not found on more than 460 X chromosomes. We identified a skewed X-inactivation in the patient's peripheral blood cells. Similar to HCCS wild type, ectopically expressed HCCS E159K was targeted to mitochondria in CHO-K1 cells. In contrast, expression of HCCS E159K did not complement respiratory growth of the CYC3-deficient yeast strain B-8025, while wild-type HCCS and the yeast heme lyase Cyc3p could rescue growth on nonfermentable carbon sources. Identification of the novel missense mutation p.E159K of HCCS, which leads to loss-of-function of the encoded holocytochrome c-type synthase, in a sporadic female patient with microphthalmia of both eyes and bilateral sclerocornea may suggest HCCS as candidate for severe ocular manifestations.


Pharmacodynamics and tolerance development during multiple intravenous bolus morphine administration in rats.
Limited information is available about how the time course of the development of tolerance to morphine-induced antinociception is related to the kinetics of drug administration and disposition. The objectives of the present experiment were to characterize the rate and extent of tolerance development during the administration of multiple increasing i.v. bolus doses of morphine to rats, and to construct a pharmacokinetic-pharmacodynamic model of morphine tolerance. Morphine was administered according to two different treatment (TXT) regimens: a 12-hr TXT, in which a total morphine exposure of 24 mg/kg was administered in seven escalating doses, and a 13-day TXT, in which escalating doses of morphine were administered daily up to a maximum of 6 mg/kg. Analgesic effect, expressed as the percent of maximum possible response, was assessed with the hot water-induced tail flick. Serum samples were collected for determination of morphine concentrations by HPLC. Concentration-normalized peak effects, measured after each morphine dose, remained constant throughout the 12-hr study period, which suggests that there was little or no tolerance development during the 12-hr TXT. In contrast, tolerance appeared more significant during administration of the 13-day TXT; a large decrease in normalized peak effect occurred between days 1 and 8. Effect remained constant thereafter, with administration of the maximum dose of morphine for the remainder of the treatment period. A pharmacokinetic-pharmacodynamic model describing the development of tolerance during the 13-day TXT was constructed. The applicability of this model of tolerance to morphine-induced antinociception with different modes of administration is discussed.


Protective effect of dietary phytochemicals against arsenite induced genotoxicity in mammalian V79 cells.
Chronic arsenic exposure causes skin diseases, gastrointestinal and neurological disorders, diabetes and cancer in various organs. Oxidative stress associated with arsenic exposure cause genetic instabilities and may initiate carcinogenesis. Phytochemicals present in vegetables, fruits, spices, tea, and medicinal plants, have shown to suppress experimental carcinogenesis in various organs. The aim of the present study was to elucidate the protective effect of some of the phytochemicals against the arsenite induced DNA damage in normal mammalian V79 cells. Comet assay was used for assessment of DNA damage and 2', 7'-dichlorofluorescein dihydroacetate for estimation of ROS generated by arsenite. The effect of the phytochemicals was observed during simultaneous treatment with arsenic, before arsenite exposure and during repair experiments. Of all the phytochemicals tested against arsenic, curcumin gave better protection during simultaneous treatment and resveratrol during pre treatment, which was evident both from comet assay and ROS generation experiments. During pre treatment a longer duration of treatment with lower dose of phytochemicals proved fruitful in reducing the genotoxicity. During repair experiments the phytochemicals enhanced recovery of DNA damage and ellagic acid gave promising results. The results indicated that natural phytochemicals may have the efficacy in reducing arsenic induced genotoxicity, in scavenging ROS and in enhancing the process of DNA repair in V79 cells.


Resection of urethral diverticulum in pregnancy.
Female urethral diverticulum is a rare finding, less often diagnosed during pregnancy. There are few reports in the literature on urethral diverticulum in pregnancy with most undergoing expectant management. A young woman presented during the third trimester of pregnancy with pelvic and vaginal pain and a vaginal bulge. On examination, she was noted to have a 5-cm periurethral mass consistent with a urethral diverticulum. We describe the successful excision of this diverticulum during pregnancy. The procedure was performed without adverse consequences to the pregnancy, and the patient had a normal course for the remainder of her pregnancy and an uneventful cesarean delivery.


Fully convolutional networks for automated segmentation of abdominal adipose tissue depots in multicenter water-fat MRI.
An approach for the automated segmentation of visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) in multicenter water-fat MRI scans of the abdomen was investigated, using 2 different neural network architectures. The 2 fully convolutional network architectures U-Net and V-Net were trained, evaluated, and compared using the water-fat MRI data. Data of the study Tellus with 90 scans from a single center was used for a 10-fold cross-validation in which the most successful configuration for both networks was determined. These configurations were then tested on 20 scans of the multicenter study beta-cell function in JUvenile Diabetes and Obesity (BetaJudo), which involved a different study population and scanning device. The U-Net outperformed the used implementation of the V-Net in both cross-validation and testing. In cross-validation, the U-Net reached average dice scores of 0.988 (VAT) and 0.992 (SAT). The average of the absolute quantification errors amount to 0.67% (VAT) and 0.39% (SAT). On the multicenter test data, the U-Net performs only slightly worse, with average dice scores of 0.970 (VAT) and 0.987 (SAT) and quantification errors of 2.80% (VAT) and 1.65% (SAT). The segmentations generated by the U-Net allow for reliable quantification and could therefore be viable for high-quality automated measurements of VAT and SAT in large-scale studies with minimal need for human intervention. The high performance on the multicenter test data furthermore shows the robustness of this approach for data of different patient demographics and imaging centers, as long as a consistent imaging protocol is used.


Two novel mutations in exons 19a and 20 and a BsaBI [correction of BsaI] polymorphism in a newly characterized intron of the neurofibromatosis type 1 gene.
Neurofibromatosis type 1 (NF1) is a common autosomal dominant disorder. It is caused by mutations in the NF1 gene, which comprises 60 exons and is located on chromosome 17q11.2. A total of 170 unrelated NF1 patients were screened for mutations in four exons by temperature-gradient gel electrophoresis. Preparatory work revealed the presence of a previously uncharacterized intron (19a) in what was previously designated exon 19; this allowed us to develop assays for genomic mutation screening in the newly defined exons 19a and 19b. Two novel NF1 mutations were detected: a single-base insertion in exon 19a creating a frameshift, and a second mutation affecting the splice donor site of intron 20 and leading to skipping of exon 20. A novel BsaBI polymorphism was identified in intron 19a.


Changes in levels of immune activation and reconstitution markers among HIV-1-infected Africans receiving antiretroviral therapy.
To describe changes in immune activation and reconstitution markers among HIV-1-infected patients receiving antiretroviral therapy (ART) in Abidjan, Côte d'Ivoire. Between November 1998 and February 2001, we analyzed changes in immune activation and reconstitution markers among 52 patients. Good virologic responders (n = 26) were defined as those who had suppressed and maintained plasma viral load (VL) below the detection limit of the assay for at least 12 months. Poor virologic responders (n = 26) were defined as those with a detectable VL at 6 and 12 months after beginning ART. Of the 26 good virologic responders, 20 (77%) were on highly active antiretroviral therapy (HAART) compared with one (4%) of the poor responders. Among the 26 good responders, baseline median levels of CD38+CD8+ T cells were elevated, but had decreased significantly at 6 months (P < 0.001) and at 12 months of therapy (P < 0.001). Median levels of HLA-DR+CD8+ T cells also decreased from baseline at 6 months (P < 0.001) and at 12 months of therapy (P < 0.001). Levels of CD62L+CD4+ T cells increased steadily during the 6 and 12 months of therapy and reached levels observed among HIV-negative blood donors (P = 0.07). Among the 26 poor responders, median levels of CD38+CD8+ T cells decreased significantly at 12 months of therapy (P = 0.006), but were higher than levels in blood donors (P = 0.005). Levels of HLA-DR+CD8+ T cells decreased significantly at 12 months of therapy (P < 0.001). Levels of CD62L+CD4+ decreased over time. Our results suggest that HAART can be successfully used in African populations with elevated baseline immune activation markers.


Superior mesenteric blood flow in man studied with a dye-dilution technique.
The superior mesenteric blood flow in the post-absorptive state was studied with a dye-dilution technique using indocyanine green in 37 patients with apparently normal small bowel function. All the patients except 2 had been, or were to be, treated because of malignant disease, mostly rectal carcinoma. The investigations were performed in connection with portography. The dye was injected into the superior mesenteric artery as a single injection during continuous sampling of blood from the superior mesenteric vein, catheterized through the reopened umbilical vein. The blood flow was calculated from the dye-dilution curves obtained with a modified Stewart-Hamilton formula. As recirculating dye appeared in the later part of the dye-dilution curves, the appearance of the recirculating dye was timed in 4 patients. The hepatic extraction of the amount of dye usually injected, 1.25 mg, was 60%. The superior mesenteric blood flow averaged 706 ml/min (397 ml/min/m-2 body surface) or 12% of the cardiac output. The appearance time, mean transit time, vascular resistance and vascular volume were also determined.


A rare melanoma feature with primary ovarian origin: a case report and the literature review.
Primary ovarian melanoma arising on a mature ovarian cystic teratoma is extremely rare. As best of our knowledge, to date, 49 cases have been reported in literature. Few information was reported about best management and therapy. We present a case occurred in a 69-year-old woman, without symptoms, who come to our unit for stress incontinence. A pelvic mass was detected and, after imaging evaluation, surgery was performed. The diagnosis was ovarian melanoma arose on a mature teratoma. No other adjuvant treatment was proposed after surgery. She died 9 months after the first diagnosis. Primary ovarian melanoma is a definite entity associated with a variable natural history and poor prognosis. Differential diagnosis is a challenge for the pathologist, because it must be differentiated by metastatic melanoma. The corner stone treatment of this disease is surgery; however, chemotherapy, immunotherapy, and target therapy seem to have a role.


A Case of Amoxicillin-Induced Acute Generalized Exanthematous Pustulosis Presenting as Septic Shock.
This case report demonstrates the challenges of diagnosing and managing acute generalized exanthematous pustulosis (AGEP) presenting as septic shock. The disseminated, erythematous, pustular rash is a common feature. However, extensive organ involvement and life-threatening hypotension are unusual. The constellation of signs has not previously been documented following amoxicillin therapy. Toxic epidermal necrolysis (TEN) and toxic shock syndrome (TSS) were considered in addition to AGEP because of the systemic presentation. AGEP was diagnosed following histopathology (TEN was ruled out based on limited necrotic keratinocytes and lack of epidermal necrosis) and a negative antistreptolysin O titer (eliminated TSS). Antibiotic therapy for septic shock was provided before the diagnosis was confirmed as AGEP. Upon confirmation of the AGEP diagnosis, antibiotics were discontinued and a 5-day course of oral prednisone (40 mg/d) was initiated in addition to topical half-strength (0.05%) betamethasone valerate. The patient rapidly improved and was discharged. Outpatient patch testing confirmed amoxicillin as the culprit drug. In conclusion, it is critical to realize that AGEP cannot be ruled out with a septic shock presentation. Recent drug history is critical in recognizing an adverse drug reaction, and patch testing is useful for determining the culpable drug when the diagnosis is AGEP.


Lactogen enhances Nb2 cell GTPase activity after 4 hours incubation.
The lactogen receptor has been suggested to associate with one or more G proteins despite the absence of a 7-transmembrane spanning sequence. These studies were designed to determine whether lactogens acutely increase GTP binding to or GTPase activity in Nb2 cell membrane. Incubation of Nb2 cell membrane with either ovine PRL (10 ng/ml) or diluent for 0-1 h resulted in a decrease in total(35)S-GTP binding to both with no difference in GTP binding between PRL- and diluent-treated membranes. There was also no change in(35)S-GTP binding to Nb2 cell membrane incubated with increasing oPRL concentrations (0.001-100 ng/ml) for 60 min. α-(32)P-GTP photoaffinity labelling was used to evaluate changes in GTP binding to specific G proteins. Photoaffinity labelling of α-(32)P-GTP to no G protein was changed after preincubation with oPRL (10 ng/ml) for 0-60 min or with oPRL (0.01-10 ng/ml) for 60 min. Finally, it was determined whether oPRL had any acute effect on GTPase activity, as determined by release of(32)Pi from γ-(32)P-GTP. When Nb2 cell membrane was preincubated for 0-60 min with oPRL (10 ng/ml) or a range of oPRL concentrations (0-10 ng/ml), no change in GTPase activity was observed. However, when Nb2 cells were incubated with lactogen for 0-7 h, GTPase activity in equal quantities of Nb2 cell membrane prepared from those cells increased over time. Increased GTPase activity (64.9-74.4%;P<0.03 compared to 0 h) was observed after 4-7 h incubation with lactogen.In summary, addition of lactogen to Nb2 cell membrane did not acutely increase either GTP binding or GTPase activity. Yet when Nb2 cells were incubated with lactogen for 4 h prior to preparation of membrane, GTPase activity was significantly increased. This evidence, in addition to our previous results showing that 4 h incubation with lactogen increased G protein β subunit concentration and pertussis toxin-stimulated ADP-ribosylation of Gi, support a role for delayed lactogen modulation of one or more G proteins in the Nb2 cell, requiring at least 4 h for maximal effect.


Color-coded digital subtraction angiography in the management of a rare case of middle cerebral artery pure arterial malformation. A technical and case report.
The advent of flow dynamics and the recent availability of perfusion analysis software have provided new diagnostic tools and management possibilities for cerebrovascular patients. To this end, we provide an example of the use of color-coded angiography and its application in a rare case of a patient with a pure middle cerebral artery (MCA) malformation. A 42-year-old male chronic smoker was evaluated in the emergency room due to sudden onset of severe headache, nausea, vomiting and left-sided weakness. Head computed tomography revealed a right basal ganglia hemorrhage. Cerebral digital subtraction angiography (DSA) showed a right middle cerebral artery malformation consisting of convoluted and ectatic collateral vessels supplying the distal middle cerebral artery territory-M1 proximally occluded. An associated medial lenticulostriate artery aneurysm was found. Brain single-photon emission computed tomography with and without acetazolamide failed to show problems in vascular reserve that would indicate the need for flow augmentation. Twelve months after discharge, the patient recovered from the left-sided weakness and did not present any similar events. A follow-up DSA and perfusion study using color-coded perfusion analysis showed perforator aneurysm resolution and adequate, albeit delayed perfusion in the involved vascular territory. We propose a combined congenital and acquired mechanism involving M1 occlusion with secondary dysplastic changes in collateral supply to the distal MCA territory. Angiographic and cerebral perfusion work-up was used to exclude the need for flow augmentation. Nevertheless, the natural course of this lesion remains unclear and long-term follow-up is warranted.


States of carbon-12 in the Skyrme model.
The Skyrme model has two Skyrmion solutions of baryon number 12, with D_{3h} and D_{4h} symmetries. The first has an equilateral triangular shape and the second an extended linear shape, analogous to the triangle and linear chain structures of three alpha particles. We recalculate the moments of inertia of these Skyrmions, and deduce the energies and spins of their quantized rotational excitations. There is a good match with the ground-state band of carbon-12, and with the recently established rotational band of the Hoyle state. The ratio of the root mean square matter radii also matches the experimental value.


A phase II study of interferon-alpha 2b with dacarbazine, carmustine, cisplatin and tamoxifen in metastatic melanoma.
A phase II trial was conducted to determine the efficacy and toxicity of the addition of interferon-alpha 2b (IFN-alpha) to the chemotherapy combination of dacarbazine (DTIC), carmustine (BCNU), cisplatin and tamoxifen (DBCT), in patients with stage III or IV melanoma. Treatment consisted of DTIC 220 mg/m2 and cisplatin 25 mg/m2 intravenously on days 1-3, BCNU 100 mg/m2 IV on day 1 only, tamoxifen 20 mg orally twice daily and IFN-alpha 5 x 10(6) units/m2 subcutaneously on days 1-5. Cycles were repeated every 4 weeks. All patients received a loading dose of tamoxifen 100 mg orally twice daily for 5 days before the first course of therapy. Of the 24 patients treated, three (13%) achieved a complete response (CR) and six (25%) a partial response (PR), for an overall response rate of 38% (95% confidence interval, 17-58%). Two patients, one who achieved a clinical CR and one a PR, had pathologically confirmed complete responses. Severe myelosuppression occurred in 47% of cycles and constitutional symptoms were common. Overall, the addition of IFN-alpha to the DBCT regimen did not appear to enhance the response rate and may have increased toxicity.


Mutator clones of Neisseria meningitidis in epidemic serogroup A disease.
Serogroup A Neisseria meningitidis has repeatedly caused widespread epidemics of meningitis and septicemia throughout the 20th century. Recently, in a limited collection of strains, epidemic serogroup A isolates were found to have elevated mutation rates that was caused by defects in mismatch repair pathways. To ascertain the role of these mutators in the epidemic spread of this serogroup, the prevalence of hypermutability in a collection of 95 serogroup A N. meningitidis invasive isolates was determined. Overall mutability in Neisseriae can be described by measuring both missense mutation rates as well as phase variation frequencies of "contingency loci." Fifty-seven percent of serogroup A isolates possessed elevated mutability, which could be divided into two classes: intermediate and high level. Eleven of 20 high-level mutators, with phase variation rates >100-fold higher than wild-type isolates, were defective in mismatch repair. Ten of the 34 intermediate mutators possessing >10-fold increases in phase variation rates could be partially complemented by a wild-type mutL allele. A high prevalence of mutators in epidemic isolates indicates that hypermutability may play a major role in the transmission of this pathogen. The added diversity derived from increased phase variation rates may allow fixation of mutator alleles more frequently during epidemic spread.


Clinical effect of different maintenance doses of caffeine citrate in the treatment of preterm infants requiring assisted ventilation: a pilot multicenter study.
To explore the optimal maintenance dose of caffeine citrate for preterm infants requiring assisted ventilation and caffeine citrate treatment. A retrospective analysis was performed on the medical data of 566 preterm infants (gestational age ≤34 weeks) who were treated and required assisted ventilation and caffeine citrate treatment in the neonatal intensive care unit of 30 tertiary hospitals in Jiangsu Province of China between January 1 and December 31, 2019. The 405 preterm infants receiving high-dose (10 mg/kg per day) caffeine citrate after a loading dose of 20 mg/kg within 24 hours after birth were enrolled as the high-dose group. The 161 preterm infants receiving low-dose (5 mg/kg per day) caffeine citrate were enrolled as the low-dose group. Compared with the low-dose group, the high-dose group had significant reductions in the need for high-concentration oxygen during assisted ventilation (P=0.044), the duration of oxygen inhalation after weaning from noninvasive ventilation (P<0.01), total oxygen inhalation time during hospitalization (P<0.01), the proportion of preterm infants requiring noninvasive ventilation again (P<0.01), the rate of use of pulmonary surfactant and budesonide (P<0.05), and the incidence rates of apnea and bronchopulmonary dysplasia (P<0.01), but the high-dose group had a significantly increased incidence rate of feeding intolerance (P=0.032). There were no significant differences between the two groups in the body weight change, the incidence rates of retinopathy of prematurity, intraventricular hemorrhage or necrotizing enterocolitis, the mortality rate, and the duration of caffeine use (P>0.05). This pilot multicenter study shows that the high maintenance dose (10 mg/kg per day) is generally beneficial to preterm infants in China and does not increase the incidence rate of common adverse reactions. For the risk of feeding intolerance, further research is needed to eliminate the interference of confounding factors as far as possible.


Characteristics of "Early Type-I Pattern" (0-6 years) Hodgkin's Disease in Turkish Children.
The clinico-epidemiologic characteristics of 54 children with HD in 0-6 years of age group were retrospectively analyzed. This group represented 27% of 200 HD cases observed in our center and was named as early type-I pattern HD. The association of EBV with HD was also shown by serologic and immunohistochemical methods (LMP1) in these very young Turkish patients. T-cell immune deficiency, cytokine imbalance and Zn deficiency were additional findings found in these patients. This series seems to be the largest one studying early type-I HD, by several aspects.


SORL1 is genetically associated with neuropathologically characterized late-onset Alzheimer's disease.
SORL1 was shown to be genetically associated with late-onset Alzheimer's disease (LOAD) in a large-scale genome-wide association study (GWAS) involving clinically verified subjects. Here, we attempted to replicate the association of SORL1 in Japanese neuropathologically characterized brain donor subjects (LOAD, 213; control, 370) through a single-nucleotide polymorphism (SNP)-based genetic study involving 19 SNPs: 11 SNPs were selected from the initial study reported by Rogaeva et al. (2007), and the other eight were from our GWAS. Among these SNPs, five exhibited a significant association with LOAD after multiple test correction (p < 2.63E-03 [ = 0.05/19]), which was supported by means of multiple logistic regression analysis with adjustment for age, gender, and carrier status of the APOE ε4 allele. Three of these SNPs (rs985421, rs12364988 [Rogaeva's SNP 7], and rs4598682) were encompassed by a 5' linkage disequilibrium (LD) region, and the remaining two (rs3781834 and rs3781836) by a 3' LD region. Strong LD among SNPs was observed within each LD region, implying that there are two genomic regions showing association with LOAD in SORL1. Case-control haplotype analysis demonstrated that some haplotypes are associated with LOAD in both LD regions. Our replication study strongly supports the preceding evidence that SORL1 is likely one of the genes associated with LOAD.


Grasp Detection for Assistive Robotic Manipulation.
In this paper, we present a novel grasp detection algorithm targeted towards assistive robotic manipulation systems. We consider the problem of detecting robotic grasps using only the raw point cloud depth data of a scene containing unknown objects, and apply a geometric approach that categorizes objects into geometric shape primitives based on an analysis of local surface properties. Grasps are detected without a priori models, and the approach can generalize to any number of novel objects that fall within the shape primitive categories. Our approach generates multiple candidate object grasps, which moreover are semantically meaningful and similar to what a human would generate when teleoperating the robot-and thus should be suitable manipulation goals for assistive robotic systems. An evaluation of our algorithm on 30 household objects includes a pilot user study, confirms the robustness of the detected grasps and was conducted in real-world experiments using an assistive robotic arm.


Hox proteins mediate developmental and environmental control of autophagy.
Hox genes encode evolutionarily conserved transcription factors, providing positional information used for differential morphogenesis along the anteroposterior axis. Here, we show that Drosophila Hox proteins are potent repressors of the autophagic process. In inhibiting autophagy, Hox proteins display no apparent paralog specificity and do not provide positional information. Instead, they impose temporality on developmental autophagy and act as effectors of environmental signals in starvation-induced autophagy. Further characterization establishes that temporality is controlled by Pontin, a facultative component of the Brahma chromatin remodeling complex, and that Hox proteins impact on autophagy by repressing the expression of core components of the autophagy machinery. Finally, the potential of central and posterior mouse Hox proteins to inhibit autophagy in Drosophila and in vertebrate COS-7 cells indicates that regulation of autophagy is an evolutionary conserved feature of Hox proteins.


Vapor-Liquid-Solid Growth and Optoelectronics of Gallium Sulfide van der Waals Nanowires.
Nanowires of layered van der Waals (vdW) crystals are of interest due to structural characteristics and emerging properties that have no equivalent in conventional 3D crystalline nanostructures. Here, vapor-liquid-solid growth, optoelectronics, and photonics of GaS vdW nanowires are studied. Electron microscopy and diffraction demonstrate the formation of high-quality layered nanostructures with different vdW layer orientation. GaS nanowires with vdW stacking perpendicular to the wire axis have ribbon-like morphologies with lengths up to 100 μm and uniform width. Wires with axial layer stacking show tapered morphologies and a corrugated surface due to twinning between successive few-layer GaS sheets. Layered GaS nanowires are excellent wide-bandgap optoelectronic materials with Eg = 2.65 eV determined by single-nanowire absorption measurements. Nanometer-scale spectroscopy on individual nanowires shows intense blue band-edge luminescence along with longer wavelength emissions due to transitions between gap states and photonic properties such as interference of confined waveguide modes propagating within the nanowires. The combined results show promise for applications in electronics, optoelectronics, and photonics, as well as photo- or electrocatalysis owing to a high density of reactive edge sites, and intercalation-type energy storage benefiting from facile access to the interlayer vdW gaps.


The association between respiratory syncytial virus infection and reactive airway disease.
Evidence has been accumulating that respiratory syncytial virus (RSV) lower respiratory tract infection (LRTI) in infants may be linked to subsequent development of reactive airway disease (RAD) in childhood, and therefore research into the prevention of RSV LRTI may have important implications for the prevention of RAD. This article reviews the epidemiological evidence linking RSVand RAD and some ofthe theories concerning cellular and molecular mechanisms of post-viral airway inflammation in order to understand how RSV prophylaxis may assist in reducing the occurrence of RSV LRTI and RAD.


Acclimization in a hot, humid environment: cardiovascular adjustments.
Four trained young men worked for 4 h/day at 40-50% of their maximum aerobic capacity first for 3 days at 25 degrees C db, 18 degrees C wb, and then for 10 consecutive days at 45 degrees C db, 32 degrees C wb. This portion of the study was mainly concerned with central circulatory changes during acclimatization. The central circulatory adaptation to work in heat could be divided into four distinct phases: phase I (day 1) was characterized by a progressive fall in stroke volume (SV) during heat exposure but cardiac output (CO) was maintained above control values by high heart rates. Phase II (days 2 and 3) was marked by increases in SV ande decreases in heart rate but with little change in CO from phase I. During phase III (days 4-8 of acclimatization), CO increased due to increases in SV. Phase IV (days 6-8) was associated with decreases in rectal and skin temperature towards control levels. SV and HR both decline in this phase so that CO was not elevated greatly above control levels. The results indicated that central circulatory and temperature regulating events are not casually associated in acclimatization.


Trends and factors associated with antipsychotic use among elderly patients with dementia in Taiwan from 2005 to 2013: a population-based study.
This study aimed to examine the trends and factors associated with antipsychotic prescriptions for elderly outpatients with dementia in Taiwan from 2005 to 2013. We assessed the annual prescription patterns of antipsychotic medications among elderly patients attending outpatient visits for dementia between 2005 and 2013 using the National Health Insurance Research Database in Taiwan. We also carried out logistic regression analyses to test the trends and associated factors. We found that any antipsychotic prescriptions for elderly patients making visits for dementia increased slightly, from 25.5 to 26.5%, over the 9-year period. From 2005 to 2013, prescriptions for first-generation antipsychotics only decreased from 7.8 to 3.3%, whereas second-generation antipsychotic prescriptions only increased from 17.0 to 22.2%. Elderly dementia patients who were female, older, concomitantly using other psychotropic drugs (antidepressants, benzodiazepines, and Z-drugs), and treated by psychiatrists and at regional/local hospitals were prescribed significantly more antipsychotics, whereas patients with comorbid hypertension, hyperlipidemia, diabetes, and stroke used antipsychotics significantly less. Although physicians seemed to avoid prescribing antipsychotics for elderly outpatients with dementia and certain comorbid physical disorders, second-generation antipsychotic use increased during the study period. Physicians should balance the benefits and risks of antipsychotic use to ensure the safety of dementia patients.


Al4(2-); the anion-π interactions and aromaticity in the presence of counter ions.
The influence of presence of counter ions and π-complexation with benzene on the bonding and magnetic properties of Al(4)(2-), the most studied all-metal cluster, is studied here. It is shown that complexation by either counter ions or benzene decreases the delocalization index between Al atoms and the magnitude of bond magnetizability, that is a Quantum Theory of Atoms in Molecules, QTAIM, -based magnetic index of aromaticity. Benzene forms two types of π-complexes with the Al(4) framework; CH-π (T-shaped) complexes and parallel π-π stacking (PPS) complexes. It is shown that variation in the π-charge of the Al(4) framework affects the relative stability of the T-shaped/PPS complexes. Free Al(4)(2-) forms a stable T-shaped anion-π complex with benzene but in the presence of cations, formation of PPS complexes is more favourable, energetically. It is suggested that this property could be used for designing molecular switches and tuneable anion sensors.


Regulation of T cell apoptosis during the immune response.
Apoptosis of T-lymphocytes is a fundamental process regulating antigen receptor repertoire selection during T cell maturation and homeostasis of the immune system. It also plays a key role in elimination of autoreactive lymphocytes. Resting mature T cells are activated by antigen to elicit an appropriate immune response. In contrast, preactivated T cells undergo activation-induced cell death (AICD) in response to TCR triggering alone. Thus, death by apoptosis is essential for function, growth and differentiation of T-lymphocytes. This review focuses on apoptosis mechanisms involved in T cell development and during the course of an immune response.


Can takotsubo cardiomyopathy be familial?
Takotsubo cardiomyopathy is a form of reversible left ventricular dysfunction that is presumably precipitated by stress. Our observations of the recurrence of takotsubo cardiomyopathy in an individual and the occurrence of a reversible cardiomyopathy in her daughter lead us to speculate on the possibility of a familial predisposition to this entity or its variants.


Site-specific mutagenesis of human chorionic gonadotrophin (hCG)-beta subunit: influence of mutation on hCG production.
The heterodimeric glycoprotein hormones, human chorionic gonadotrophin (hCG), LH, TSH and FSH, consist of two non-covalently linked subunits, the alpha and beta subunits. The beta subunit is specific for each hormone and is responsible for the biological specificity, but the beta subunits of different hormones show some degree of structural homology. The CAGY (cysteine-alanine-glycine-tyrosine) region is one of the amino acid sequences that is homologous in different beta subunits and is highly conserved between species. In the present study, site-specific in-vitro mutagenesis was used to change three individual nucleotides in the centre of the CAGY region of the hCG-beta subunit, and the effects of these mutations on hCG production was determined by in-vitro transcription and then translation in Xenopus laevis oocytes. The results indicate that the CAGY region, particularly the glycine residue at position 36 in the beta subunit, is essential for the production of hCG. This finding is consistent with previous studies showing that this region is necessary for the biological activity of human TSH.


Characterization of floating activity of indigenous diesel-assimilating bacterial isolates.
Six diesel-degrading bacterial strains were isolated from oil-polluted sites located in central Taiwan. The floating activity of the isolates in an oil-supplemented liquid medium was monitored. Cell-surface hydrophobicity as well as cell-free and cell-residue emulsification activities were also investigated. Three isolates, identified as Gordonia alkanivorans CC-JG 39, Rhodococcus erythropolis CC-BC 04, and R. erythropolis CC-BC 11, were found to float and grow near the diesel layer on the surface. The other three isolates (namely, Comamonas testosteroni CC-CF3, Acinetobacter sp. CC-CF 5, and Sphingomonas yanoikuyae CC-CG 22) did not display floating activity, as they distributed uniformly in the liquid medium. Isolated cell walls of the floating strains appeared to settle at a lower sucrose density than the non-floating strains. The floating strains were also characterized by a higher cell-surface hydrophobicity and a higher cell-residue emulsification activity than the non-floating strains. In fact, the floating strains were thought to produce extracellular emulsifiers due to their higher supernatant emulsification activity than the non-floating strains. The floating activity of G. alkanivorans CC-JG 39 may be associated with the production of extracellular polymeric substances that formed an "air-bag" structure facilitating cell floating. The floating ability may also correlate with a high cellular hydrophobicity arising from unique cell wall compositions or cell-wall-bound surface active products.


Hard gelatin and hypromellose (HPMC) capsules: estimation of rupture time by real-time dissolution spectroscopy.
The objective of this study was to measure rupture time of gelatin and hypromellose (hydroxypropyl methylcellulose or HPMC) capsules using a novel approach based on real-time dissolution spectroscopy. Rupture time was measured in standard dissolution apparatus at a constant temperature using a dip-type fiber-optic probe. Labrasol released from the capsules was treated as the marker of the rupture process. Light scatter generated by the emulsified labrasol was detected by an ultrafast monochromator at scan rates approximating 24,000 nm/min. This technique was validated by measuring the dissolution time of gelatin capsules. Rupture times of hypromellose capsules were studied as a function of capsule size, capsule grade, and dissolution medium. Statistical correlations were analyzed by ANOVA. Rupture time of hypromellose capsules was dependent on both the medium and the grade of the capsule, and was independent of capsule size. The composition of the dissolution medium contributes to the rupture time of the capsules and should be considered when fast release and quick biological response is desired. Release delay, however, may not manifest itself in vivo and the time to maximum plasma concentration may not be significant.


[Lupus vulgaris as the etiology of tuberculous mastitis].
We report the rare case of granulomatous tuberculous mastitis based on lupus vulgaris in a 73-year-old female patient. The most important diagnostic procedure was the detection of mycobacterial DNA-sequences in paraffin-embedded tissues using the polymerase-chain-reaction (PCR). A combined antituberculous therapy lead to complete remission within 14 months after the diagnosis was established. The incidence of tuberculous mastitis among surgically treated lesions of the breast is less than 0.025 percent. Because of the frequent localisation of this disease in the upper outer quadrant of the breast and suspicious findings in mammography, tuberculous mastitis is one, even if rare, differential diagnosis for breast cancer.


Cutaneous metastasis of renal cell carcinoma: Fine needle aspiration provides rapid diagnosis.
The diagnosis of cutaneous metastasis of renal cell carcinoma is challenging in a young person in absence of a prior history of cancer. In such situation, fine needle aspiration alone as a minimally invasive procedure can provide rapid, accurate and cost effective diagnosis, even in case of unknown primary.


Variation of kQclin,Qmsr (fclin,fmsr) for the small-field dosimetric parameters percentage depth dose, tissue-maximum ratio, and off-axis ratio.
Evaluate the ability of different dosimeters to correctly measure the dosimetric parameters percentage depth dose (PDD), tissue-maximum ratio (TMR), and off-axis ratio (OAR) in water for small fields. Monte Carlo (MC) simulations were used to estimate the variation of kQclin,Qmsr (fclin,fmsr) for several types of microdetectors as a function of depth and distance from the central axis for PDD, TMR, and OAR measurements. The variation of kQclin,Qmsr (fclin,fmsr) enables one to evaluate the ability of a detector to reproduce the PDD, TMR, and OAR in water and consequently determine whether it is necessary to apply correction factors. The correctness of the simulations was verified by assessing the ratios between the PDDs and OARs of 5- and 25-mm circular collimators used with a linear accelerator measured with two different types of dosimeters (the PTW 60012 diode and PTW PinPoint 31014 microchamber) and the PDDs and the OARs measured with the Exradin W1 plastic scintillator detector (PSD) and comparing those ratios with the corresponding ratios predicted by the MC simulations. MC simulations reproduced results with acceptable accuracy compared to the experimental results; therefore, MC simulations can be used to successfully predict the behavior of different dosimeters in small fields. The Exradin W1 PSD was the only dosimeter that reproduced the PDDs, TMRs, and OARs in water with high accuracy. With the exception of the EDGE diode, the stereotactic diodes reproduced the PDDs and the TMRs in water with a systematic error of less than 2% at depths of up to 25 cm; however, they produced OAR values that were significantly different from those in water, especially in the tail region (lower than 20% in some cases). The microchambers could be used for PDD measurements for fields greater than those produced using a 10-mm collimator. However, with the detector stem parallel to the beam axis, the microchambers could be used for TMR measurements for all field sizes. The microchambers could not be used for OAR measurements for small fields. Compared with MC simulation, the Exradin W1 PSD can reproduce the PDDs, TMRs, and OARs in water with a high degree of accuracy; thus, the correction used for converting dose is very close to unity. The stereotactic diode is a viable alternative because it shows an acceptable systematic error in the measurement of PDDs and TMRs and a significant underestimation in only the tail region of the OAR measurements, where the dose is low and differences in dose may not be therapeutically meaningful.


[Clinical and radiological studies of seizure in chronic subdural hematoma--case control study].
We studied the mechanism underlying seizure induction in patients with chronic subdural hematoma. In our study population of 1,009 patients with chronic subdural hematoma, 26 (2.6%) had seizure-related complications. Six of them had already been diagnosed with epilepsy (4 patients) or suspected of having secondary epilepsy (2 patients) after experiencing traffic accidents or cerebral bleeding. Twenty patients (seizure group) had been tentatively diagnosed as having hematoma-induced convulsion. Of the remaining 989 patients without convulsion, 40 randomly sampled patients were included in the non-seizure group by matching with clinical terms. Intergroup comparisons showed that patients with dementia were more common in the seizure group than in the non-seizure group; however, no intergroup differences were observed for other clinical parameters. Radiological examinations showed that bilateral hematomas were relatively more common and sulcal hyperintensity on FLAIR MR images was significantly more frequent in the seizure group than in the non-seizure group. Interestingly, many patients presenting with sulcal hyperintensity exhibited mixed-density hematomas on CT images. These findings suggest the mechanism by which hematoma content infiltrates into the brain parenchyma and the subsequent induction of convulsions by the stimulatory component.


Glucose-dependent activation of protein kinase A activity in Saccharomyces cerevisiae and phosphorylation of its TPK1 catalytic subunit.
Protein kinase A (PKA), in yeast, plays a major role in controlling metabolism and gene expression in connection with the available nutrient conditions. We here measure, for the first time, a transient change in the in vivo PKA activity, along a cAMP peak produced by 100 mM glucose addition to glycerol-growing cells as well as a change in the phosphorylation state of its catalytic subunit (Tpk1p) following PKA activation. PKA activity was measured in situ in permeabilized cells, preserving its intracellular localization. Comparison of total PKA activity, measured in situ in permeabilized cells with data obtained from in vitro assays in crude extracts, underscores the inhibitory potency of the regulatory subunit within the cell. Tpk1p phosphorylation was detected through non-denaturing gel electrophoresis. Phosphorylation of Tpk1p increases its specificity constant toward kemptide substrate. The use of mutants of the cAMP pathway showed that phosphorylation depends on the activation of PKA via the G-protein coupled receptor pathway triggered by glucose. The phosphorylation state of Tpk1p was followed during the diauxic shift. Tpk1p phosphorylation is dynamic and reversible: its up-regulation correlates with a fully fermentative metabolism, while its down-regulation with stationary phase or respiratory metabolism. Reversible phosphorylation can thus be considered a new control mechanism possibly pointing to a fine-tuning of PKA activity in response to environmental conditions.


Successful endovascular recanalization of a partially occluded basilar artery fenestration.
A 76-year-old man with a history of arterial hypertension, obstructive sleep apnea, dyslipidemia, family history of cardiovascular events, prestroke and overweight presented 90 minutes after acute onset of right-sided sensorimotor hemiparesis, hemiataxia and dysarthria (National Institutes of Health Stroke Scale (NIHSS) 9/42). Magnetic resonance imaging (MRI) revealed a pontine ischemia and MR angiography showed a thrombus in the middle to distal portion of the basilar artery. Owing to the location, an occlusion of one lumen of a fenestrated basilar artery was suspected. Fearing the risk of peripheral dislocation, intravenous thrombolysis was withheld after an interdisciplinary discussion and direct endovascular thrombectomy (Solitaire stent retriever) was successfully performed by passing the stent retriever specifically through the affected lumen and between the thrombus and the vascular wall, which would normally be avoided. Angiography after complete reperfusion (Thrombolysis in Cerebral Infarction grade 3) confirmed a fenestration in the middle to distal portion of the basilar artery where the thrombus was initially located (blue and green arrow). Follow-up MRI after 24 hours showed only minimal ischemic damage in the left pontine area, and the patient was discharged home with ambulatory physiotherapy for residual minimal gait disturbance (NIHSS 0).


A major class of L-selectin ligands is eliminated in mice deficient in two sulfotransferases expressed in high endothelial venules.
The interaction of L-selectin on lymphocytes with sulfated ligands on high endothelial venules leads to rolling and is critical for recruitment of lymphocytes into peripheral lymph nodes. Peripheral node addressin represents a class of L-selectin ligands recognized by the function-blocking monoclonal antibody MECA-79. Its epitope overlaps with sialyl 6-sulfo Lewis X, an L-selectin recognition determinant. Here, mice lacking two N-acetylglucosamine-6-O-sulfotransferases (GlcNAc6ST-1 and GlcNAc6ST-2) demonstrated elimination of both peripheral node addressin and sialyl 6-sulfo Lewis X in high endothelial venules, considerably reduced lymphocyte homing to peripheral lymph nodes and reduced sticking of lymphocytes along high endothelial venules. Our results establish an essential function for the sulfotransferases in L-selectin ligand synthesis and may have relevance for therapy of inflammatory diseases.


Detection of MAGE-1 tumor antigen in brain tumor.
The expression of MAGE tumor rejection antigens in brain tumors is unclear. We investigated the expression of MAGE-1 and -4 proteins in brain tumors with antibodies to recombinant MAGE-1 and -4 proteins. MAGE-1 or -4 protein was detected in a majority (12 of 14) or some (5 of 14) of gliomas, respectively, by the immunoblot analysis. In contrast, MAGE-1 or -4 protein was rarely detected in the non-glioma tumors (3 of 14 or 1 of 14, respectively). Cellular MAGE-1 protein was detected in the cytoplasma of the glioblastoma cells with immunohistochemical staining. The expression of the MAGE-4 protein was confirmed by enzyme-linked immunosorbent assay. The results suggest that the MAGE-1 protein is an appropriate target molecule for specific immunotherapy of glioma.


Why vocal production of atypical sounds in apes and its cerebral correlates have a lot to say about the origin of language.
Ackermann et al. mention the "acquisition of species-atypical sounds" in apes without any discussion. In our commentary, we demonstrate that these atypical sounds in chimpanzees not only include laryngeal sounds, but also have a major significance regarding the origins of language, if we consider looking at their context of use, their social properties, their relations with gestures, their lateralization, and their neurofunctional correlates as well.


CCR5 receptor expression is down-regulated in HIV type 2 infection: implication for viral control and protection.
HIV-2 is known to display an attenuated phenotype in vivo with prolonged time to disease and decreased rate of transmission. Observational studies in Senegal have demonstrated protection from HIV-1 infection, although the putative mechanism for immunoprotection remains undefined. We evaluated HIV-2-seropositive women from a cohort of commercial sex workers in Dakar, Senegal and identified individuals with very low surface CCR5 receptor expression on CD4+ T cells. In vitro up-regulation of the CCR5 receptor was readily achieved. Down-regulation of the CCR5 was not correlated with activation markers (HLA-DR), beta-chemokine levels, or plasma viral loads. A correlation was observed with HIV-2-specific CD8+ T cell activity as measured by intracellular cytokine production. We postulate that down-regulation of the CCR5 receptor in HIV-2 infection contributes to slower disease course and to the protective mechanism against HIV-1 superinfection, mediated in part by HIV-2-specific cellular immune responses.


General practitioners' perceptions of population based bowel screening and their influence on practice: a qualitative study.
Although largely preventable, Australia has one of the highest rates of bowel cancer in the world. General Practitioners (GPs) have an important role to play in prevention and early detection of bowel cancer, however in Australia this is yet to be optimised and participation remains low. This study sought to understand how GPs' perceptions of bowel screening influence their attitudes to, and promotion of the faecal occult blood test (FOBT), to identify opportunities to enhance their role. Interviews were conducted with 31 GPs from metropolitan and regional New South Wales (NSW), Australia. Discussions canvassed GPs' perceptions of their role in bowel screening and the national screening program; perceptions of screening tests; practices regarding discussing screening with patients; and views on opportunities to enhance their role. Transcripts were coded using Nvivo and thematically analysed. The study revealed GPs' perceptions of screening did not always align with broader public health definitions of 'population screening'. While many GPs reportedly understood the purpose of population screening, notions of the role of asymptomatic screening for bowel cancer prevention were more limited. Descriptions of screening centred on two major uses: the use of a screening 'process' to identify individual patients at higher risk; and the use of screening 'tools', including the FOBT, to aid diagnosis. While the FOBT was perceived as useful for identifying patients requiring follow up, GPs expressed concerns about its reliability. Colonoscopy by comparison, was considered by many as the gold standard for both screening and diagnosis. This perception reflects a conceptualisation of the screening process and associated tools as an individualised method for risk assessment and diagnosis, rather than a public health strategy for prevention of bowel cancer. The results show that GPs' perceptions of screening do not always align with broader public health definitions of 'population screening'. Furthermore, the way GPs understood screening was shown to impact their clinical practice, influencing their preferences for, and use of 'screening' tools such as FOBT. The findings suggest emphasising the preventative opportunity of FOBT screening would be beneficial, as would formally engaging GPs in the promotion of bowel screening.


Cardiac rehabilitation after heart transplantation in a patient with Becker's muscular dystrophy: a case report.
Becker's muscular dystrophy (BMD) is associated with abnormal cardiac findings in 75% of cases; up to one third will develop ventricular dilatation leading to congestive heart failure, at times necessitating cardiac transplant. Candidates are selected from a base of heart failure patients who are usually New York Heart Association (NYHA) class III or IV. Treatment in a phase II cardiac rehabilitation program after transplantation is associated with functional improvement in patients without BMD, but there are no reports of patients with this disorder. We present the case of a 38-year-old man diagnosed with BMD with associated dilated cardiomyopathy. The patient was a NYHA class IIIa and underwent orthotopic cardiac transplantation for intractable heart failure followed by treatment in a phase II cardiac rehabilitation program. At the end of cardiac rehabilitation, his 12-minute walking distance had improved from 716.28 to 929.64 m (30% improvement), he had increased his conditioning metabolic equivalent level from 3.5 to 5.5 (55% improvement), he had a weight loss from 81.65 to 78.93 kg, and his body mass index changed from 23 to 22 kg/m2. The patient now has returned to work, is using a stationary bicycle once a day for 30 minutes, and is walking 1 hour a day. This suggests that treatment in a cardiac rehabilitation program is effective in patients with BMD after cardiac transplant.


Secondary amenorrhea. A protocol for pinpointing the underlying cause.
Secondary amenorrhea presents the primary care physician with a diagnostic challenge because of the many disorders that can cause it. Thus, its evaluation warrants a systematic approach to avoid unnecessary and expensive diagnostic procedures. By using the step-by-step protocol outlined in this article, the clinician can isolate the dysfunctional compartment--outflow tract, ovary, anterior pituitary, CNS, or hypothalamus--and then pursue a more specific workup to pinpoint the cause. Treatment depends on the patient's desire for pregnancy, and the need for long-term follow-up is dictated by the specific diagnosis.


Patient centeredness, cultural competence and healthcare quality.
Cultural competence and patient centeredness are approaches to improving healthcare quality that have been promoted extensively in recent years. In this paper, we explore the historical evolution of both cultural competence and patient centeredness. In doing so, we demonstrate that early conceptual models of cultural competence and patient centeredness focused on how healthcare providers and patients might interact at the interpersonal level and that later conceptual models were expanded to consider how patients might be treated by the healthcare system as a whole. We then compare conceptual models for both cultural competence and patient centeredness at both the interpersonal and healthcare system levels to demonstrate similarities and differences. We conclude that, although the concepts have had different histories and foci, many of the core features of cultural competence and patient centeredness are the same. Each approach holds promise for improving the quality of healthcare for individual patients, communities and populations.


[Exploration on the common characters of sub-healthy people based on clinical epidemiology].
To study the common characters of sub-healthy people. Questionnaire of clinical epidemiological investigation was adopted in the sub-healthy people. The commonest symptoms and their frequency in sub-healthy people were fatigue 78.7%, poor quality of sleep 73.4%, amnesia 59.9%, fatigue irrelievable after rest 59.1%, dry throat 59.0%, dizziness 58.6%, dry and xerotic eyes 58.3%, ache with distension eyes 57.8%, ache 56.4%, early awakening 52.7%, difficulty in falling asleep 52.5%, irritability 51.8%, etc. The commonest risk factors were lack of relaxation 60.4%, lack of physical exercise 58.0%, working with frequent extra shifts 56.7%, air pollution 56.2%, noise pollution 51.3%, etc. The commonest syndrome types in sub-healthy people were insufficiency of Pi-qi type 10.2%, Gan-qi stagnation and Pi deficiency type 10.1%, deficiency of Xin and Pi type 9.7 %, retention of dampness due to Pi-deficiency type 7.4% and qi deficiency type 4.2%, et al. The commonest symptoms in sub-healthy people are fatigue, poor quality of sleep, amnesia, irrelievable fatigue after taking rest, dry throat, dizziness, dry and xeritic eyes, ache with distension in eyes, ache, early awakening, difficulty in falling asleep, irritability, etc. The commonest risk factors are lack of relaxation and physical exercise, working with frequent shift, air and noise pollution. The commonest syndromes of sub-healthy people are insufficiency of Pi-qi type, Gan qi stagnation and Pi deficiency type, deficiency of Xin and Pi type, retention of dampness due to Pi deficiency type, qi deficiency type. These provides evidence for prevention and treatment of sub-health with TCM.


Prognostic factors of primary gastrointestinal stromal tumors: a cohort study based on high-volume centers.
We aimed to evaluate the clinicopathologic characteristics, immunohistochemical expression and prognostic factors of patients with primary gastrointestinal stromal tumors (GISTs). Data from 2,570 consecutive GIST patients from four medical centers in China (January 2001-December 2015) were reviewed. Survival curves were constructed by the Kaplan-Meier method, and Cox regression models were used to identify independent prognostic factors. Of the included patients, 1,375 (53.5%) were male, and the patient age range was 18 to 95 (median, 58) years. The tumors were mostly found in the stomach (64.5%), small intestine (25.1%) and colorectal region (5.1%). At the time of diagnosis, the median tumor size was 4.0 (range: 0.1-55.0) cm, and the median mitotic index per 50 high power fields (HPFs) was 3 (range: 0-254). Of the 2,168 resected patients, 2,009 (92.7%) received curative resection. According to the modified National Institutes of Health (NIH) classification, 21.9%, 28.9%, 14.1% and 35.1% were very low-, low-, intermediate- and high-risk tumors, respectively. The rate of positivity was 96.4% for c-Kit, 87.1% for CD34, 96.9% for delay of germination 1 (DOG-1), 8.0% for S-100, 31.0% for smooth muscle actin (SMA) and 5.1% for desmin. However, the prognostic value of each was limited. Multivariate analysis showed that age, tumor size, mitotic index, tumor site, occurrence of curative resection and postoperative imatinib were independent prognostic factors. Furthermore, we found that high-risk patients benefited significantly from postoperative imatinib (P<0.001), whereas intermediate-risk patients did not (P=0.954). Age, tumor size, mitotic index, tumor site, occurrence of curative resection and postoperative imatinib were independent prognostic factors in patients with GISTs. Moreover, determining whether intermediate-risk patients can benefit from adjuvant imatinib would be of considerable interest in future studies.


Effect of age and inoculation route on the infection of duck Tembusu virus in Goslings.
Duck Tembusu virus (TMUV) is an emerging flavivirus that has caused variable levels of outbreaks in poultry in recent years. In order to study the effect of age and inoculation routes on the TMUV infection, one hundred healthy domestic 5-day-old and 20-day-old goslings were equally divided into five groups and four experimental groups of goslings were infected with the TMUV-SDSG strain by intravenous and intranasal routes, respectively. Severe clinical signs were observed in goslings infected at 5 days of age, including listlessness, growth retardation, severe neurological dysfunction and even death. However, goslings infected at 20 days of age showed mild symptoms and no mortality. The severity of gross lesions gradually reduced as goslings matured. The severe histopathological changes were observed in 5-day-old infected goslings, including cerebral edema, viral encephalitis, myocardial necrosis, hepatic steatosis, spleen lymphoid cell depletion, pancreatic epithelial cell shedding and interstitial hemorrhage. However, 20-day-old infected goslings showed mild histopathological changes. Viral loads in different tissues were detected by the SYBR Green I real-time PCR assay. The level of viral loads in most of tissues 5-day-old infected goslings was higher than that of 20-day-old infected goslings, correlating with the severity of clinical symptoms and lesions in these tissues. 20-day-old infected goslings developed significantly higher serum neutralizing antibody titers than 5-day-old infected goslings. Furthermore, goslings infected with TMUV intravenously demonstrated more severe clinical signs, lesions and higher viral loads in tissues than those of goslings infected with TMUV intranasally. Therefore, age and inoculation routes can affect the pathogenicity of TMUV in geese and younger geese are more susceptible to the virus. Age and inoculation route factors should be considered in study of the pathogenicity, pathogenesis, folumation of prevention and therapy strategies of TMUV infection in geese.


An optimized assay for transcription factor NF-kappaB with dsDNA-coupled microplate.
To develop an EMSA-free assay approach for analyzing the sequence-specific DNA-binding proteins (DBPs), an easy cost-effective dsDNA-coupled plate (dcPlate) was developed in our lab for this purpose. In this paper, the assay conditions of such dcPlate were fully optimized for detecting an important transcription factor, NF-kappaB. The optimized parameters of dcPlate for assay of NF-kappaB were as follows: immobilized DNA probe at the concentration of 25 pmol/100 microL-well, incubation time of 90 min for NF-kappaB binding to dcPlate, primary and secondary antibody concentration of 0.1 microL/100 microL dilution, incubation time of 90 min for primary antibody binding to NF-kappaB, temperature of 25 degrees C for the above process, colorimetric developing time for 30 min. After optimization, the signal was improved three times higher than that from not optimized conditions. The linear colorimetric detection ranges of the purified recombinant NF-kappaB p50 and the cell nuclear extract were from 0.59 to 75 ng/well and 0.313 to 10 microg/well, respectively.


Fate of Histoplasma capsulatum in guinea pig polymorphonuclear leukocytes.
Guinea pig polymorphonuclear leukocytes (PMN) were parasitized in vitro with yeast cells of Histoplasma capsulatum. Preparations stained after 3 h at 37 C with May Greenwald-Giemsa revealed that 87% of the yeasts were tinctorially altered. Such alterations corresponded to those displayed by fungus cells intentionally killed by heat or other means and thus the altered yeast were presumed to be dead. A combination of 10(-5) M H(2)O(2), 10(-5) M KI, and horseradish peroxidase killed H. capsulatum. Death was assessed by the eosin-y dye exclusion test. All of the listed components were required for death of the fungus. A granule lysate preparation derived from guinea pig PMN leukocytes could replace the horseradish peroxidase in the fungicidal system. The granule lysates behaved in keeping with the attributes of a myeloperoxidase. Thus, PMN leukocytes and certain extracellular peroxidase systems kill the yeast cell phase of the dimorphic fungus H. capsulatum.


Origin of human mast cells: development from transplanted hematopoietic stem cells after allogeneic bone marrow transplantation.
Although mast cells are hematopoietic cells, little is known about the origin of their precursors in vivo. In this study, the origin (donor v recipient genotype) of human mast cells (MCs) was analyzed in a patient who underwent allogeneic bone marrow transplantation (BMT). The patient presented with secondary acute myeloid leukemia (French-American-British classification, M2) arising from refractory anemia with excess of blast cells and bone marrow (BM) mastocytosis. Transplantation was performed in chemotherapy-induced complete remission. On days 88, 126, 198, and 494 after BMT, mast cells were enriched to homogeneity from bone marrow mononuclear cells (BM MNCs) by cell sorting for CD117+/CD34- cells. Purified mast cell populations were CD117(c-kit)+ (> 95%), CD34- (< 1%), CD3- (< 1%), CD14- (< 1%), and virtually free of contaminating cells as assessed by Giemsa staining. The genotype of MCs was analyzed after amplification by polymerase chain reaction (PCR) of a variable number tandem repeat (VNTR) region within intron 40 of the von Willebrand factor (vWF) gene. Unexpectedly, on days 88 and 126 after BMT, sorted MCs displayed recipient genotype as shown by vWF.VNTR-PCR. However, on days 198 and 494, PCR analysis showed a switch to donor genotype in isolated mast cells. Peripheral blood (PB) and BM MNC as well as highly enriched (sorted) CD3+ T cells (PB, BM), CD4+ helper T cells (PB), CD8+ T cells (PB), CD19+ B cells (PB), CD14+ monocytes (PB, BM), and CD34+ precursor cells (BM) showed donor genotype throughout the observation period. Together, these results provide evidence that human MCs developed in vivo from transplanted hematopoietic stem cells. Engraftment and in vivo differentiation of MCs from early hematopoietic progenitor cells may be a prolonged process.


Impaired hippocampal-prefrontal synchrony in a genetic mouse model of schizophrenia.
Abnormalities in functional connectivity between brain areas have been postulated as an important pathophysiological mechanism underlying schizophrenia. In particular, macroscopic measurements of brain activity in patients suggest that functional connectivity between the frontal and temporal lobes may be altered. However, it remains unclear whether such dysconnectivity relates to the aetiology of the illness, and how it is manifested in the activity of neural circuits. Because schizophrenia has a strong genetic component, animal models of genetic risk factors are likely to aid our understanding of the pathogenesis and pathophysiology of the disease. Here we study Df(16)A(+/-) mice, which model a microdeletion on human chromosome 22 (22q11.2) that constitutes one of the largest known genetic risk factors for schizophrenia. To examine functional connectivity in these mice, we measured the synchronization of neural activity between the hippocampus and the prefrontal cortex during the performance of a task requiring working memory, which is one of the cognitive functions disrupted in the disease. In wild-type mice, hippocampal-prefrontal synchrony increased during working memory performance, consistent with previous reports in rats. Df(16)A(+/-) mice, which are impaired in the acquisition of the task, showed drastically reduced synchrony, measured both by phase-locking of prefrontal cells to hippocampal theta oscillations and by coherence of prefrontal and hippocampal local field potentials. Furthermore, the magnitude of hippocampal-prefrontal coherence at the onset of training could be used to predict the time it took the Df(16)A(+/-) mice to learn the task and increased more slowly during task acquisition. These data suggest how the deficits in functional connectivity observed in patients with schizophrenia may be realized at the single-neuron level. Our findings further suggest that impaired long-range synchrony of neural activity is one consequence of the 22q11.2 deletion and may be a fundamental component of the pathophysiology underlying schizophrenia.


"Failure-to-Identify" Hunting Incidents: A Resilience Engineering Approach.
Objective The objective was to develop an understanding, using the Functional Resonance Analysis Method (FRAM), of the factors that could cause a deer hunter to misidentify their intended target. Background Hunting is a popular activity in many communities. However, hunters vary considerably based on training, experience, and expertise. Surprisingly, safety in hunting has not received much attention, especially failure-to-identify hunting incidents. These are incidents in which the hunter, after spotting and targeting their quarry, discharge their firearm only to discover they have been spotting and targeting another human, an inanimate object, or flora by mistake. The hunter must consider environment, target, time of day, weather, and many other factors-continuously evaluating whether the hunt should continue. To understand how these factors can relate to one another is fundamental to begin to understand how incidents happen. Method Workshops with highly experienced and active hunters led to the development of a FRAM model detailing the functions of a "Hunting FRAM." The model was evaluated for correctness based on confidential and anonymous near-miss event submissions by hunters. Results A FRAM model presenting the functions of a hunt was produced, evaluated, and accepted. Using the model, potential sources of incidents or other unintended outcomes were identified, which in turn helped to improve the model. Conclusion Utilizing principles of understanding and visualization tools of the FRAM, the findings create a foundation for safety improvements potentially through training or safety messages based on an increased understanding of the complexity of hunting.


DAGchainer: a tool for mining segmental genome duplications and synteny.
Given the positions of protein-coding genes along genomic sequence and probability values for protein alignments between genes, DAGchainer identifies chains of gene pairs sharing conserved order between genomic regions, by identifying paths through a directed acyclic graph (DAG). These chains of collinear gene pairs can represent segmentally duplicated regions and genes within a single genome or syntenic regions between related genomes. Automated mining of the Arabidopsis genome for segmental duplications illustrates the use of DAGchainer.


Displaced supracondylar humeral fractures in children: Comparison of three treatment approaches.
Closed reduction and percutaneous pinning are the most widely used treatment options for displaced supracondylar humerus fractures in children, but there is still no consensus concerning the most preferred technique in injuries of the extension type. The aim of this study was to compare three common orthopaedic procedures in the treatment of displaced extension type supracondylar humerus fractures in children. Total of 93 consecutive patients (66 boys and 27 girls) referred to our hospital with Gartland type II or III extension supracondylar humeral fractures were prospectively included in the study over a six-year period. At initial presentation 48 patients were classified as Gartland type II and 45 as Gartland type III fractures. The patients were subdivided into three groups based on the following treatment modality: closed reduction with percutaneous pinning, open reduction with Kirschner wires (K-wires) fixation, and closed reduction with cast immobilisation. The treatment outcome and clinical characteristics were compared among groups, as well as evaluated using Flynn's criteria. Excellent clinical outcome was reported in 70.3% of patients treated with closed reduction with percutaneous pinning and in 64.7% of patients treated with open reduction with K-wire fixation. The outcome was significantly worse in children treated with closed reduction and cast immobilisation alone, as excellent outcome is achieved in just 36.4% of cases (p = 0.011). Closed reduction with percutaneous pinning is the method of choice in the treatment of displaced pediatric supracondylar humeral fracture, while open reduction with K-wire fixation is as a good alternative in cases with clear indications.


New approaches on the study of the psychometric properties of the STAI.
The main purpose of this study was to analyze the psychometric properties of the State-Trait Anxiety Inventory (STAI1). Previous studies have indicated different factor solutions. Nevertheless, there is still a lack of consensus about the best dimensional model of STAI scores. The sample consisted of 417 participants, composed of 387 (29.71% male) healthy participants (comparison group: M=35.5 years; SD=8.40), and 30 (36.66% male) patient (clinical group M=35.8 years; SD=12.94). The internal consistency evaluated through Ordinal Alpha was good, 0.98 and 0.94 in the non-clinical and the clinical samples, respectively. Test-retest reliability (two weeks) for Total Score was 0.81 for the non-clinical subsample, and 0.93 for the clinical subsample. Confirmatory factor analyses supported both a four factor model and bifactor model. Also, STAI scores showed statistically significant correlations with Burns Anxiety Inventory (Burns-A) scores. Furthermore, results showed statistically significant differences in the mean scores of the STAI between the clinical and the non-clinical subsamples. The psychometric properties of the STAI were adequate. The present study contributes to better understand the STAI structure through the comparison of new approaches in the study of the STAI internal structure. The results found may contribute in the efforts to improve the evaluation and identification of anxiety symptoms and disorders.