Disruption of NAD(P)H:quinone oxidoreductase 1 gene in mice leads to radiation-induced myeloproliferative disease.
NAD(P)H:quinone oxidoreductase 1 null (NQO1(-/-)) mice exposed to 3 Gy of gamma-radiation showed an increase in neutrophils, bone marrow hypercellularity, and enlarged lymph nodes and spleen. The spleen showed disrupted follicular structure, loss of red pulp, and granulocyte and megakarocyte invasion. Blood and histologic analysis did not show any sign of infection in mice. These results suggested that exposure of NQO1(-/-) mice to gamma-radiation led to myeloproliferative disease. Radiation-induced myeloproliferative disease was observed in 74% of NQO1(-/-) mice as compared with none in wild-type (WT) mice. NQO1(-/-) mice exposed to gamma-radiation also showed lymphoma tissues (32%) and lung adenocarcinoma (84%). In contrast, only 11% WT mice showed lymphoma and none showed lung adenocarcinoma. Exposure of NQO1(-/-) mice to gamma-radiation resulted in reduced apoptosis in granulocytes and lack of induction of p53, p21, and Bax. NQO1(-/-) mice also showed increased expression of myeloid differentiation factors CCAAT/enhancer binding protein alpha (C/EBPalpha) and Pu.1. Intriguingly, exposure of NQO1(-/-) mice to gamma-radiation failed to induce C/EBPalpha and Pu.1, as was observed in WT mice. These results suggest that decreased p53/apoptosis and increased Pu.1 and C/EBPalpha led to myeloid hyperplasia in NQO1(-/-) mice. The lack of induction of apoptosis and differentiation contributed to radiation-induced myeloproliferative disease in NQO1(-/-) mice.


A Rare Case of Isolated Vesical Mucormycosis in a Patient with COVID-19 Pneumonitis.
We present the case of a 60-year-old diabetic female who was diagnosed with coronavirus disease 2019 (COVID-19) pneumonitis. After her recovery during follow-up, she presented with recurrent hematuria, burning of micturition and, occasional lower abdominal discomfort, with unsatisfactory response to oral antibiotics. On imaging evaluation, there was mild right hydronephrosis and hydroureter with urothelial thickening involving the right lower ureter and a filling defect in the urinary bladder close to the vesicoureteric junction seen on excretory phase images. Cystoscopy revealed a whitish friable mass-like lesion that was retrieved, histopathology of which revealed fungal elements, and Rhizopus was isolated in culture. Thus, the diagnosis of urinary bladder fungal ball due to mucormycosis infection of the urinary tract was reached. The patient was asymptomatic after 10 weeks of antifungal treatment. The treating physicians, urologist, and radiologists need to have a high index of suspicion of urinary mucormycosis in patients with COVID-19 pneumonia.


Role of hepatic enzymes in the biochemical assessment of the severity of sickle cell anemia.
Various clinical and hematological indices have been used to assess the severity of Sickle Cell Anemia (SCA), however biochemical indices are lacking. Hepatomegaly has been a frequent finding in SCA and its persistence has been associated with increased disease severity. The association between hepatic enzymes and disease severity in SCA is undefined. This study was therefore designed to look at the association between clinical severity and hepatic enzymes in SCA subjects with persistent hepatomegaly (that is, lasting more than six months) in order to determine a possible role for hepatic enzymes as a biochemical index of severity. Serum levels of aspartate transaminase (AST), alanine transaminase (ALT), alkaline phosphatase (SAP) and gamma-glutamyl-transferase (GGT) were determined in two groups of SCA subjects and in hemoglobin genotype AA (HbAA) controls. SCA group comprised of 37 subjects with persistent hepatomegaly equal to or greater than 10 cm (below right coastal margin) while the second group comprised another 38 SCA subjects without palpable hepatomegaly. 40 individuals with hemoglobin genotype AA served as control for both groups. Clinical and hematological parameters of severity which included steady state haematocrit, number of transfusions per year, number of crises per year and percentage HbF level were determined and scored in a manner similar to the Glasgow coma scale. Results obtained were analyzed with the aid of statistical package on EPI-INFO version 6.02. There was a significant increase in serum ALT, ALP and GGT levels in SCA with persistent hepatomegaly over those without hepatomegaly (p < 0.05, p < 0.05 and p < 0.01 respectively). All the index scores and the final aggregate severity scores were also significantly higher in SCA subjects with persistent hepatomegaly. Only GGT demonstrated a fairly positive and significant correlation (r = 0.46, P < 0.05) with increased clinical severity among the hepatic enzymes. Elevated serum level of GGT in SCA during steady state is suggestive of increased disease severity.


Women's perception of bone-mineral density testing and osteoporosis education in the community pharmacy.
The objective of this study was to assess women's level of acceptance and attitudes toward bone-mineral density (BMD) testing and osteoporosis education in the community pharmacy. The study design was a questionnaire with 13 statements using a five-point scale anchored from "strongly disagree" to "strongly agree." The statements focused on the value and comfort level of BMD testing in a pharmacy, confidence in pharmacists, osteoporosis education in the pharmacy, and payment for such services. One-sample t-tests were used to assess whether the responses statistically differed from the midpoint scale value. Comparisons also were made between demographic characteristics (education level, family history of osteoporosis, and daily calcium intake). An independently owned grocery store chain (Bashas') in the Phoenix, and Tucson, Arizona, metropolitan areas. The survey was administered to women upon completion of a BMD screening. Responses to 13 statements and demographic questions. Eighty women completed the survey. Women strongly agreed that BMD testing in community pharmacies was valuable and that it would increase the public's awareness of osteoporosis. For all 13 statements, respondents strongly agreed or agreed with the importance and role of BMD testing. All responses were statistically significantly greater than the midpoint scale value (P<0.001). There were no statistically significant differences in responses among women with higher education, with a family history of osteoporosis, and who take daily calcium (P>0.05). These women's opinions suggest that BMD testing has a useful role in community pharmacies.


Risk factors for early use of e-cigarettes and alcohol: Dimensions and profiles of temperament.
Adolescent e-cigarette use has been labeled an epidemic and alcohol use during this developmental period is associated with deleterious outcomes. Though specific temperamental dimensions have been shown to predict substance use, profiles of temperament have rarely been examined as predictors. This study examines dimensions and profiles of adolescent temperament as predictors of early use of e-cigarettes and alcohol. The sample was comprised of adolescent (62.07% female, 87.59% White, 82.76% Hispanic/Latinx)/caregiver dyads (N = 146) who completed the first two timepoints (M age at second timepoint = 16.16, SD = 0.68) of a longitudinal adolescent substance use study. Models showed parent-reported effortful control predicted protection against adolescent use of e-cigarettes, whereas adolescent report of effortful control predicted protection against alcohol use. Though dissimilar in temperamental pattern, three profiles emerged from both parent- and adolescent-report-based latent profile analysis models. Adolescents characterized by parents as displaying a Resilient profile had greater odds of e-cigarette use than those characterized by a Reserved profile, whereas adolescents who self-characterized as Mixed-type had markedly greater odds of alcohol use than those who self-characterized as Resilient. Utilization of temperamental profiles may aid in identification of particularly vulnerable subgroups of adolescents who may benefit from relevant preventative programing.


Drivers of Infectious Disease Seasonality: Potential Implications for COVID-19.
Not 1 year has passed since the emergence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the causative agent of coronavirus disease 2019 (COVID-19). Since its emergence, great uncertainty has surrounded the potential for COVID-19 to establish as a seasonally recurrent disease. Many infectious diseases, including endemic human coronaviruses, vary across the year. They show a wide range of seasonal waveforms, timing (phase), and amplitudes, which differ depending on the geographical region. Drivers of such patterns are predominantly studied from an epidemiological perspective with a focus on weather and behavior, but complementary insights emerge from physiological studies of seasonality in animals, including humans. Thus, we take a multidisciplinary approach to integrate knowledge from usually distinct fields. First, we review epidemiological evidence of environmental and behavioral drivers of infectious disease seasonality. Subsequently, we take a chronobiological perspective and discuss within-host changes that may affect susceptibility, morbidity, and mortality from infectious diseases. Based on photoperiodic, circannual, and comparative human data, we not only identify promising future avenues but also highlight the need for further studies in animal models. Our preliminary assessment is that host immune seasonality warrants evaluation alongside weather and human behavior as factors that may contribute to COVID-19 seasonality, and that the relative importance of these drivers requires further investigation. A major challenge to predicting seasonality of infectious diseases are rapid, human-induced changes in the hitherto predictable seasonality of our planet, whose influence we review in a final outlook section. We conclude that a proactive multidisciplinary approach is warranted to predict, mitigate, and prevent seasonal infectious diseases in our complex, changing human-earth system.


Prospective comparison of rectal dose reduction during intracavitary brachytherapy for cervical cancer using three rectal retraction techniques.
To compare three rectal retraction methods on dose to organs at risk, focusing on rectal dose, in cervix cancer patients treated with high-dose-rate intracavitary brachytherapy. A prospective study was conducted on patients with cervical carcinoma treated with chemoradiotherapy, including external beam radiation and four fractions of high-dose-rate intracavitary brachytherapy prescribed to Point A using a ring and tandem applicator under conscious sedation. Rectal retraction methods included: a rectal retractor blade (RR), vaginal gauze packing (VP), and a tandem Foley balloon (FB). All three methods were used in all patients. The RR was used first, and the following applications were randomly assigned to VP or FB. CT planning was used to calculate D2cc for rectum, sigmoid, small bowel, and bladder. The Wilcoxon signed rank test was used to determine if the median dose differences between methods were statistically significant. In these 11 patients, median dose (min, max) in cGy to the rectum using RR, FB, and VP was 131 (102, 165), 199 (124, 243), and 218 (149, 299), respectively. The RR demonstrated lower median intrapatient doses to rectum compared with FB and VP (-55 cGy; p = 0.014 and -76 cGy; p = 0.004, respectively). The RR also resulted in lower sigmoid doses. No differences in dose were observed between the VP and FB methods. The rectal retractor significantly reduced the dose to rectum and sigmoid compared with FP and VP. In patients treated under conscious sedation, the RR method provides the best rectal sparing. There were no significant differences in dose observed between the FB and VP techniques.


Patients with multiple unexplained symptoms. Their characteristics, functional health, and health care utilization.
To define groups of patients with inappropriately high health care utilization better, we have studied a group of 41 chronically ill patients to determine their characteristics, symptoms, functional health status, and amount of health care utilization. This group had somatization disorder diagnosed by specific criteria. They can be easily recognized by primary care physicians by their multiple complaints, negative physical examination results, and a history of multiple medical examinations. We found them to be functionally disabled, spending an average of seven days ill in bed each month. They perceived themselves as severely ill and were willing to undergo multiple hospitalizations, diagnostic studies, and operations. Their health care charges were extraordinary, averaging $4,700 annually. The physician can most effectively treat these patients by scheduling regular visits, relying on physical examinations rather than diagnostic studies, and avoiding costly hospitalizations.


Assessment of trace metals using lichen transplant from automobile mechanic workshop in Ile-Ife metropolis, Nigeria.
The level of air pollution around the automobile mechanic workshops has been generally overlooked. This study, examined the level of trace metals in automobile mechanic workshops and the suitability of using transplanted lichen thalli of Lepraria incana for measuring air pollution in such areas. Samples of the lichen thalli were transplanted into seven different sites and were attached to the bark of trees at each site. The samples were harvested from the sites after 3-month exposure. Concentrations of Pb, Cu, Cd, Fe, Zn, and S content were determined using an atomic absorption spectrophotometer. Results showed that there was a significant difference in the trace metals concentrations across the sites (p < 0.05). The analyzed lichen samples showed a range of 91.26-119.35 ppm for Fe, 30.23-61.32 ppm for Zn, 1.25-2.45 ppm for Cu, 0.017-0.043 ppm for Cd, 0.018-0.051 ppm, and 0.37-0.42 ppm for S. From the study, sites 6 and 7 presented higher concentrations of Cd, Pb, and Zn than other sites. The enrichment factor calculated showed that Zn, Cd, and Pb were greatly enriched from the workshops. The trend in the concentration of these heavy metals suggests that activities in these workshops might become a major source of certain heavy metals in the environment and if the pollution activities persist, it might become worrisome over time.


Tuberculosis is associated with a down-modulatory lung immune response that impairs Th1-type immunity.
Immune mediators associated with human tuberculosis (TB) remain poorly defined. This study quantified levels of lung immune mediator gene expression at the time of diagnosis and during anti-TB treatment using cells obtained by induced sputum. Upon comparison to patients with other infectious lung diseases and volunteers, active pulmonary TB cases expressed significantly higher levels of mediators that counteract Th1-type and innate immunity. Despite the concomitant heightened levels of Th1-type mediators, immune activation may be rendered ineffectual by high levels of intracellular (SOCS and IRAK-M) and extracellular (IL-10 and TGF-betaRII, IL-1Rn, and IDO) immune suppressive mediators. These modulators are a direct response to Mycobacterium tuberculosis as, by day 30 of anti-TB treatment, many suppressive factors declined to that of controls whereas most Th1-type and innate immune mediators rose above pretreatment levels. Challenge of human immune cells with M. tuberculosis in vitro up-regulated these immune modulators as well. The observed low levels of NO synthase-2 produced by alveolar macrophages at TB diagnosis, along with the heightened amounts of suppressive mediators, support the conclusion that M. tuberculosis actively promotes down-modulatory mediators to counteract Th1-type and innate immunity as an immunopathological strategy. Our data highlight the potential application of immune mediators as surrogate markers for TB diagnosis or treatment response.


Quantitative determination of Na+-K+-ATPase and other sarcolemmal components in muscle cells.
A recurring problem in the characterization of plasma membrane enzymes in tissues and cells is whether the samples tested are representative for the entire population of enzyme molecules present in the starting material. Measurements of [3H]-ouabain binding, enzyme activity, and maximum transport capacity all indicate that the concentration of Na+-K+ pumps in mammalian skeletal muscle is high (300-800 pmol/g wet wt). Studies on Na+-K+-ATPase activity in isolated sarcolemma, however, generally give little or no information on total cellular enzyme concentration. Due to the low and variable enzyme recovery (0.2-8.9%), such subcellular preparations may, therefore, give misleading data on factors regulating Na+-K+-ATPase in heart and skeletal muscle cells. As the same isolation and purification procedures are used for the study of other sarcolemmal components (lipids, hormone receptors, enzymes, and other transport systems), this inadequate recovery has general implications for statements on regulatory changes in the sarcolemmal composition of muscle cells. On the other hand, complete quantification of Na+-K+-ATPase in muscle tissue can now be achieved using simple procedures and the entire material (intact muscle fibers, biopsies, and whole homogenates). Recent studies have shown that regulatory changes in the entire population of Na+-K+ pumps in muscle can be quantified in measurements of [3H]-ouabain binding, K+-activated 3-O-methylfluorescein phosphatase activity, as well as maximum ouabain suppressible Na+-K+ transport capacity.


Expression of heat-shock proteins in Streptococcus pyogenes and their immunoreactivity with sera from patients with streptococcal diseases.
The heat-shock response of Streptococcus pyogenes following exposure to elevated growth temperatures, and the immunological reactivity of heat-shock proteins (HSPs) in streptococcal infections were studied. Two major proteins of 65 and 75 kDa were expressed when a S. pyogenes strain was shifted from 37 degrees C to heat-shock temperatures of 40, 42 and 45 degrees C. Such proteins are members of the GroEL and DnaK families recognised in a Western blot assay with polyclonal antibodies against Escherichia coli GroEL and E. coli DnaK, respectively. Two-dimensional autoradiograms of polypeptides labelled at 37 or 42 degrees C showed an increased intensity of three spots at 42 degrees C. A monoclonal antibody (MAb) against HSP 63 of Bordetella pertussis also recognised the 65-kDa inducible protein, although MAbs against Mycobacterium tuberculosis HSP 65 failed to recognise this protein. Immunoblot analysis of sera from individuals with rheumatic fever or uncomplicated streptococcal diseases revealed seven major immunogenic protein bands, two of which also reacted with anti-E. coli GroEL and DnaK polyclonal antibodies. Furthermore, antibodies to the GroEL and DnaK proteins were also detected in sera from patients with either rheumatoid arthritis or systemic lupus erythematosus. These results demonstrated a heat-shock response of S. pyogenes, and indicated the presence of an immune response against HSPs in streptococcal diseases.


POU4F3 Acts as a Tumor Suppressor in Lung Adenocarcinoma via the Endoplasmic Reticulum Stress Signaling Pathway.
Lung cancer is the most common malignancy worldwide, and LUAD is the primary type of lung cancer. Recently, the POU transcription factor family has been associated with the development of multicancer, especially lung cancer. However, the relationship between POU domain class Ⅳ transcription factor Ⅲ (POU4F3) and lung cancer remains unknown. We detected the expression of POU4F3 in human LUAD and adjacent tissues with immunohistochemical staining, and we found that POU4F3 was expressed less in LUAD tissues compared with adjacent tissues. Patients with higher POU4F3 expression have more prolonged overall survival. We then constructed SPCA1 and A549 cells with stable overexpression or inhibition of POU4F3. We found that overexpressed POU4F3 suppressed LUAD cell proliferation in vitro and in vivo, according to CCK-8, colony formation, and xenograft assays. LUAD cell apoptosis was suppressed by POU4F3 overexpression based on Flow cytometry. The downregulation of POU4F3 yielded the opposite patterns. Next, we explored the possible mechanisms through which POU4F3 promoted the apoptosis of LUAD cells. Western blotting suggested that overexpression of POU4F3 significantly increased protein expression levels of the PERK/eIF2α/ATF4/CHOP and IRE1α/XBP1s pathways of ERS, while POU4F3 absence reversed the expressions of the above essential proteins in ERS pathways in SPCA1 and A549 cells. However, we found that PERK inhibitor but not IRE1 inhibitor can reverse the effect of POU4F3 overexpression on apoptosis. This study indicated that POU4F3 may work as a tumor suppressor in LUAD via regulating the PERK/eIF2α/ATF4/CHOP pathway. We made it possible to develop POU4F3 as a diagnostic, therapeutic, and prognostic target of LUAD.


Partial anomalous pulmonary venous drainage. A novel approach to repair.
Isolated partial anomalous pulmonary venous drainage with an intact atrial septum is a rare finding. The authors describe their experience with three patients (ages 9, 37, and 54 years), with partial anomalous pulmonary venous connection to the superior vena cava, right atrium, and inferior vena cava, who underwent extracardiac conduit repair of this anomaly. In all three patients, a synthetic Gortex graft was used for reconstruction of the venous pathways to the left atrium. The follow-up period ranged from 10 to 82 months (mean, 42 months). All three patients were evaluated with intravenous digital angiography, transesophageal echocardiography, or both at 10, 33, and 82 months postoperatively. Patency of the grafts with no evidence of obstruction and excellent pulmonary venous flow was shown. This surgical technique is an excellent option for correction of this anomaly, and intravenous digital subtraction angiography is a useful diagnostic tool during the postoperative period to evaluate patency of the repair.


Lipid peroxidation as a possible mechanism for the neurotoxic and nephrotoxic effects of a combination of lithium carbonate and haloperidol.
Lithium carbonate as well as haloperidol increased lipid peroxidation in rat cerebral cortex synaptosomes and rat kidney homogenates while sulpiride had no significant effects. A combination of lithium carbonate and haloperidol caused a greater increase in lipid peroxidation than did either lithium carbonate or haloperidol alone. It is concluded that the neurotoxic and nephrotoxic effects of lithium carbonate and haloperidol might be a consequence of increased lipid peroxidation in the cerebral cortex and the kidney, respectively.


The role of lex2 in lipopolysaccharide biosynthesis in Haemophilus influenzae strains RM7004 and RM153.
The locus lex2, comprising lex2A and lex2B, contributes to the phase-variable expression of lipopolysaccharide (LPS) of Haemophilus influenzae and was found to be present in 74 % of strains investigated. lex2A contains 5'-GCAA repeats which vary in number from 4 to 46 copies between strains. The locus was cloned from the serotype b strains RM7004 and RM153 and showed >99 % nucleotide sequence identity between these strains and the published lex2 sequence. Disruption of the lex2B gene in strain RM7004 resulted in truncation of some LPS glycoforms, shown by gel fractionation, with only one glycoform reacting with a digalactoside-specific monoclonal antibody, 4C4, compared with four LPS glycoforms in the more elongated LPS of the parent strain. Mass spectrometry and NMR analyses of LPS from the lex2B mutant revealed loss of the terminal digalactoside as well as the second beta-glucose extending from the first heptose of the inner core. The authors conclude that Lex2B is the beta-(1-4)-glucosyltransferase that adds the second beta-glucose to the first beta-glucose as part of the oligosaccharide extension from the first heptose of the LPS of strain RM7004. Investigation of the expression of the lex2 locus indicated that the genes are co-transcribed and that both reading frames are required for addition of this second beta-glucose in a phase-variable manner.


Optimized delignification of wood-derived lignocellulosics for improved enzymatic hydrolysis.
One of the major bottlenecks in the bioconversion of lignocelluosic feedstocks to liquid ethanol is the recalcitrance of residue following pretreatment, specifically softwood derived residues. Peroxide delignification has previously been shown to effectively aid in the removal of condensed lignaceous moieties from substrates following pretreatment, and thereby improve the hydrolyzability of the polymeric carbohydrates to their monomeric constituents. Despite the effectiveness of peroxide, drawbacks in this system still remain, as the concentration of peroxide required for adequate hydrolysis performance is currently uneconomical. In an attempt to improve the efficacy of the delignification process, we evaluated other post-treatment operations and concurrently attempted to limit the decomposition of peroxide loading; with the over arching aim to improve the efficiency of the bioconversion process. By employing several peroxide stabilizers and pre-chelating the steam exploded recalcitrant substrates, we were able to substantially improve the delignification treatment of Douglas-fir wood chips, and to reduce peroxide loading by more than 50% without negative effects on the hydrolysis rates and yield.


Organotypic Culture of Bone-Like Structures Using Composite Ceramic-Fibrin Scaffolds.
We have developed an organotypic culture system that allows the production of bone tissue features on a centimeter scale. A composite, calcium phosphate-strained fibrin gel system is able to organize itself in the presence of osteoblastic cells, creating basic hierarchical units as seen in vivo, and can be modified to produce a range of other tissues that require such directional structuring. Constructs evolve over time into multi-compositional structures containing a high mineral content and terminally differentiated, osteocyte-like cells. These tissues can be cultured over extended durations (exceeding 1 year) and are responsive to a variety of chemical and biological agents. The platform can reduce the number of animals used in experimentation by acting as an intermediate stage in which more personalized research conditions can be generated. We provide a thorough description of the protocol used to successfully culture and modify this system, as well as guidance on compositional characterization. © 2019 by John Wiley & Sons, Inc.


Hierarchical constraint processes for shape analysis.
A major application of syntactic pattern recognition is the analysis of two-dimensional shape. This paper describes a new syntactic shape analysis technique which combines the constraint propagation techniques which have been so successful in computer vision with the syntactic representation techniques which have been successfully applied to a wide variety of shape analysis problems. Shapes are modeled by stratified shape grammars. These grammars are designed so that local constraints can be compiled from the grammar describing the appearance of pieces of shape at various levels of description. Applications to the analysis of airplane shapes are presented.


Study modality and false recall.
False memories occur when individuals mistakenly report an event as having taken place when that event did not in fact occur. The DRM (Deese, 1959; Roediger & McDermott, 1995) paradigm provides an effective technique for creating and investigating false memories. In this paradigm participants study a list of words (e.g., SOUR, CANDY,…) that are highly associated to a non-presented critical item (e.g., SWEET). The study phase is followed by a test of memory for the study list words. Researchers typically find very high levels of false recall of the critical non-presented item. However, the likelihood of falsely remembering the non-presented critical items can be reduced by presenting studied associates visually rather than auditorally (e.g., Smith & Hunt, 1998). This is referred to as the modality effect in false memory. The current study investigated the role of resource availability in the expression of this modality effect in false recall. In Experiment 1 false recall was reduced in the visual study presentation condition relative to the auditory condition for participants with higher working memory capacity, but not for participants with lower working memory capacity. In Experiment 2 the effect of study modality on false recall was eliminated by the addition of a divided attention task at encoding. Both studies support the proposal that resource availability plays a role in the expression of the modality effect in the DRM paradigm (Smith, Lozito, & Bayen, 2005).


Serological findings in leprosy and tuberculosis with the Wassermann, Meinicke, and VDRL tests.
In the course of a venereal disease survey in Egypt, 820 cases of leprosy and 720 cases of tuberculosis were serologically examined with the Wassermann, Meinicke (MKR II), and VDRL tests; the results are reported in this paper.On serological and anamnestic evidence, 31 cases of syphilis were discovered among the leprosy cases and 37 among the tuberculosis cases. Apparently false positive reactions were seen in 203 cases of leprosy (25%) and in 38 cases of tuberculosis (5%). The author discusses the probability that a fairly high proportion of these reactions were in fact caused by otherwise undetected syphilis or were non-specific.The Meinicke test proved the most specific of the three, followed, in that order, by the Wassermann and the VDRL tests.It was found that syphilis was more frequent among males with tuberculosis than among those with leprosy; this is attributed to the fact that leprosy patients are kept in greater isolation. Less easily explicable is the fact that more females than males with leprosy were found to have syphilis, whereas in tuberculous persons the difference in syphilis incidence between male and female patients was not very great.


Amelioration of 5-Fluorouracil Induced Nephrotoxicity by Acacia catechu through Overcoming Oxidative Damage and Inflammation in Wistar Rats.
The research intended to explore the possible nephroprotective potential of the ethyl acetate fraction derived from Acacia catechu leaves against nephrotoxicity brought about by 5-fluorouracil (5-FU) in Wistar rats. While possessing strong anticancer properties, 5-FU is hindered in its therapeutic application due to significant organ toxicity linked to elevated oxidative stress and inflammation. The study is undertaken to conduct an analysis of the ethyl acetate fraction of A. catechu leaves both in terms of quality and quantity, examining its impact on different biochemical and histopathological parameters within the context of 5-FU-induced renal damage in rats and elucidation of the mechanism behind the observed outcomes. Intraperitoneal injection of 5-FU at a dosage of 20 mg/kg/day over 5 days was given to induce nephrotoxicity in rats. The evaluation of nephrotoxicity involved quantifying serum creatinine, urea, uric acid, and electrolyte concentrations. Furthermore, superoxide dismutase, catalase antioxidant enzymes, and TNF-α concentration in serum were also measured. 5-FU injection led to the initiation of oxidative stress within the kidneys, leading to modifications in renal biomarkers (including serum creatinine, urea, uric acid, and Na+ , K+ levels), and a reduction in antioxidant enzymes namely superoxide dismutase and catalase. Notably, the presence of the inflammatory cytokine TNF-α was significantly elevated due to 5-FU. Microscopic examination of renal tissue revealed tubular degeneration and congestion. However, treatment involving the ethyl acetate fraction derived from A. catechu leaves effectively and dose-dependently reversed the changes observed in renal biomarkers, renal antioxidant enzymes, inflammatory mediators, and histopathological features, bringing them closer to normal conditions. The observed recuperative impact was mainly attributed to the antioxidant and antiinflammatory properties of the fraction. The ethyl acetate fraction of A. catechu leaves exhibited a mitigating influence on the renal impairment caused by 5-FU, showcasing its potential as a nephroprotective agent capable of preventing and ameliorating 5-FU-induced nephrotoxicity.


LIQUID NITROGEN FREEZING IN MICROBIOLOGICAL ASSAY SYSTEMS. I. PRESERVATION OF LACTOBACILLUS LEICHMANNII FOR DIRECT USE IN THE VITAMIN B12 ASSAY.
Suspensions of Lactobacillus leichmannii were stored in liquid nitrogen and were used as direct inocula in vitamin B(12) assays. Complete recovery of viable cells was obtained when the suspensions in basal B(12) medium were rapidly frozen by direct immersion into liquid nitrogen and rapidly thawed by agitating the suspensions in a water bath at 40 C. Greater than 90% destruction occurred when the suspensions were in saline. However, both suspensions were usable in the B(12) assay system. Assay results on a number of test materials indicated good correlation between freshly prepared suspensions and frozen suspensions in basal medium stored 3 months. Suspensions in saline stored for 1 year in liquid nitrogen showed no detectable difference from the first day after freezing. Suspensions frozen slowly at the rate of 1 degree per min from 4 to -40 C and subsequently immersed in liquid nitrogen had a longer lag period of growth and were not usable in the 18-hr assay incubation system. A major advantage of a stored inoculum for direct use in a microbiological assay is the reduced day-to-day variation in the inoculum.


"To be a phenomenal doctor you have to be the whole package": physicians' interpersonal behaviors during difficult conversations in pediatrics.
Delivery of bad news is a challenging task for physicians and other health care professionals. Several studies have assessed parental perceptions of the delivery of bad news, but none have focused on the role of physicians' interpersonal behaviors in the communication process. The study's objective was to assess parental perceptions of physicians' interpersonal behaviors and their role in communication of bad news. The design was a cross-sectional qualitative interview study of 13 parents of patients hospitalized or previously hospitalized in the pediatric intensive care unit or oncology/bone marrow transplant unit at an academic children's hospital. Eleven interpersonal behaviors were identified as important by parents. The majority of parents identified empathy in physicians as critical. Availability, treating the child as an individual, and respecting the parent's knowledge of the child were mentioned by almost half of parents. Themes also considered important but by a smaller number of parents were allowing room for hope, the importance of body language, thoroughness, going beyond the call of duty, accountability, willingness to accept being questioned, and attention to the suffering of the child. To increase parental satisfaction and enhance the parent-physician therapeutic partnership, we recommend that physicians consider attending to the 11 interpersonal behaviors described in this manuscript, and that educational programs pay particular attention to these behaviors when training health care providers in the communication of bad news.


Molecular and pharmacological characterization of zebrafish 'contractile' and 'inhibitory' prostanoid receptors.
Prostanoids comprising prostaglandins (PGs) and thromboxanes (TXs) have been shown to play physiological and pathological roles in zebrafish. However, the molecular basis of zebrafish prostanoid receptors has not been established. Here, we demonstrate that there exist at least five 'contractile' (Ca(2+)-mobilizing) and one 'inhibitory' (Gi-coupled) prostanoid receptors in zebrafish; five 'contractile' receptors consisting of two PGE2 receptors (EP1a and EP1b), two PGF2α receptors (FP1 and FP2), and one TXA2 receptor TP, and one 'inhibitory' receptor, the PGE2 receptor EP3. [(3)H]PGE2 specifically bound to the membranes of cells expressing zebrafish EP1a, EP1b and EP3 with a Kd of 4.8, 1.8 and 13.6nM, respectively, and [(3)H]PGF2α specifically bound to the membranes of cells expressing zebrafish FP1 and FP2, with a Kd of 6.5 and 1.6nM, respectively. U-46619, a stable agonist for human and mouse TP receptors, significantly increased the specific binding of [(35)S]GTPγS to membranes expressing the zebrafish TP receptor. Upon agonist stimulation, all six receptors showed an increase in intracellular Ca(2+) levels, although the increase was very weak in EP1b, and pertussis toxin abolished only the EP3-mediated response. Zebrafish EP3 receptor also suppressed forskolin-induced cAMP formation in a pertussis toxin-sensitive manner. In association with the low structural conservation with mammalian receptors, most agonists and antagonists specific for mammalian EP1, EP3 and TP failed to work on each corresponding zebrafish receptor. This work provides further insights into the diverse prostanoid actions mediated by their receptors in zebrafish.


Dengue, chikungunya and Zika in GeoSentinel surveillance of international travellers: a literature review from 1995 to 2020.
GeoSentinel is a global surveillance network of travel medicine providers seeing ill-returned travellers. Much of our knowledge on health problems and infectious encountered by international travellers has evolved as a result of GeoSentinel surveillance, providing geographic and temporal trends in morbidity among travellers while contributing to improved pre-travel advice. We set out to synthesize epidemiological information, clinical manifestations and time trends for dengue, chikungunya and Zika in travellers as captured by GeoSentinel. We conducted a systematic literature search in PubMed on international travellers who presented with dengue, chikungunya or Zika virus infections to GeoSentinel sites around the world from 1995 until 2020. Of 107 GeoSentinel publications, 42 articles were related to dengue, chikungunya and/or Zika. The final analyses and synthesis of and results presented here are based on the findings from 27 original articles covering the three arboviral diseases. Dengue is the most frequent arboviral disease encountered in travellers presenting to GeoSentinel sites, with increasing trends over the past two decades. In Southeast Asia, annual proportionate morbidity increased from 50 dengue cases per 1000 ill returned travellers in non-epidemic years to an average of 159 cases per 1000 travellers during epidemic years. The highest number of travellers with chikungunya virus infections was reported during the chikungunya outbreak in the Americas and the Caribbean in the years 2013-16. Zika was first reported by GeoSentinel already in 2012, but notifications peaked in the years 2016-17 reflecting the public health emergency in the Americas at the time.


Invasive congeners differ in successional impacts across space and time.
Invasive species can alter the succession of ecological communities because they are often adapted to the disturbed conditions that initiate succession. The extent to which this occurs may depend on how widely they are distributed across environmental gradients and how long they persist over the course of succession. We focus on plant communities of the USA Pacific Northwest coastal dunes, where disturbance is characterized by changes in sediment supply, and the plant community is dominated by two introduced grasses--the long-established Ammophila arenaria and the currently invading A. breviligulata. Previous studies showed that A. breviligulata has replaced A. arenaria and reduced community diversity. We hypothesize that this is largely due to A. breviligulata occupying a wider distribution across spatial environmental gradients and persisting in later-successional habitat than A. arenaria. We used multi-decadal chronosequences and a resurvey study spanning 2 decades to characterize distributions of both species across space and time, and investigated how these distributions were associated with changes in the plant community. The invading A. breviligulata persisted longer and occupied a wider spatial distribution across the dune, and this corresponded with a reduction in plant species richness and native cover. Furthermore, backdunes previously dominated by A. arenaria switched to being dominated by A. breviligulata, forest, or developed land over a 23-yr period. Ammophila breviligulata likely invades by displacing A. arenaria, and reduces plant diversity by maintaining its dominance into later successional backdunes. Our results suggest distinct roles in succession, with A. arenaria playing a more classically facilitative role and A. breviligulata a more inhibitory role. Differential abilities of closely-related invasive species to persist through time and occupy heterogeneous environments allows for distinct impacts on communities during succession.


Machine Learning Based Risk Prediction for Major Adverse Cardiovascular Events.
Patients with major adverse cardiovascular events (MACE) such as myocardial infarction or stroke suffer from frequent hospitalizations and have high mortality rates. By identifying patients at risk at an early stage, MACE can be prevented with the right interventions. The aim of this study was to develop machine learning-based models for the 5-year risk prediction of MACE. The data used for modelling included electronic medical records of more than 128,000 patients including 29,262 patients with MACE. A feature selection based on filter and embedded methods resulted in 826 features for modelling. Different machine learning methods were used for modelling on the training data. A random forest model achieved the best calibration and discriminative performance on a separate test data set with an AUROC of 0.88. The developed risk prediction models achieved an excellent performance in the test data. Future research is needed to determine the performance of these models and their clinical benefit in prospective settings.


Bone mineral density is linked to 1p36 and 7p15-13 in a southern Chinese population.
Genome-wide linkage scans have identified a number of quantitative trait loci (QTLs) affecting bone mineral density (BMD), mainly in the Caucasian population. In this study, we aim to determine whether seven well-replicated QTLs also contribute to BMD variation in the southern Han Chinese population. Thirty-three microsatellite markers in the proximity of seven QTLs were genotyped in 1,459 subjects from 306 families ascertained through a proband with BMD Z-score equal to or less than -1.3 at either the lumbar spine or hip. Regression-based multipoint linkage analysis was performed. In the entire study population, good linkage evidence of total hip BMD to 7p14 [maximum log of odds (LOD) score (MLS) = 2.75; nominal P = 0.0002] and 1p36 (MLS = 1.6, P = 0.003) was revealed. In the subgroup analysis of 1,166 female subjects, MLS of 3.42, 2.65, 2.42, and 1.54 were obtained on 7p12 for total hip, lumbar spine, trochanter, and femoral neck BMD, respectively. A suggestive linkage signal was achieved at 7p14-15 with a MLS of 3.38 and 3.15 for trochanter and total hip BMD in the 678 premenopausal women, and at 7p12 for femoral neck and total hip BMD with MLS of 2.22 and 3.04 in postmenopausal women. Subgroup analysis of premenopausal women also provided additional evidence of suggestive linkage of total hip BMD to 1p36, with a MLS of 2.84 at 17.07 cM. Thus, linkage of BMD to 1p36 and 7p15-13 is confirmed in southern Chinese. Computational prioritization strategy and published genome-wide association studies suggested RERE and SFRP4 as two promising candidate genes in which variants responsible for the linkage signal may be identified by follow-up gene-wide association studies.


The Effect of Triage Assessments on Identifying Inflammatory Arthritis and Reducing Rheumatology Wait Times in Ontario.
We evaluated the influence of triage assessments by extended role practitioners (ERP) on improving timeliness of rheumatology consultations for patients with suspected inflammatory arthritis (IA) or systemic autoimmune rheumatic diseases (SARD). Rheumatologists reviewed primary care providers' referrals and identified patients with inadequate referral information, so that a decision about priority could not be made. Patients were assessed by an ERP to identify those with IA/SARD requiring an expedited rheumatologist consult. The time from referral to the first consultation was determined comparing patients who were expedited to those who were not, and to similar patients in a usual care control group identified through retrospective chart review. Seven rheumatologists from 5 communities participated in the study. Among 177 patients who received an ERP triage assessment, 75 patients were expedited and 102 were not. Expedited patients had a significantly shorter median (interquartile range) wait time to rheumatologist consult: 37.0 (24.5-55.5) days compared to non-expedited patients [105 (71.0-135.0) days] and controls [58.0 (24.0-104.0) days]. Accuracy comparing the ERP identification of IA/SARD to that of the rheumatologists was fair (κ 0.39, 95% CI 0.25-0.53). Patients triaged and expedited by ERP experienced shorter wait times compared to usual care; however, some patients with IA/SARD were missed and waited longer. Our findings suggest that ERP working in a triage role can improve access to care for those patients correctly identified with IA/SARD. Further research needs to identify an ongoing ERP educational process to ensure the success of the model.


Molecular and clinical evidence for an ARMC5 tumor syndrome: concurrent inactivating germline and somatic mutations are associated with both primary macronodular adrenal hyperplasia and meningioma.
Primary macronodular adrenal hyperplasia (PMAH) is a rare cause of Cushing's syndrome, which may present in the context of different familial multitumor syndromes. Heterozygous inactivating germline mutations of armadillo repeat containing 5 (ARMC5) have very recently been described as cause for sporadic PMAH. Whether this genetic condition also causes familial PMAH in association with other neoplasias is unclear. The aim of the present study was to delineate the molecular cause in a large family with PMAH and other neoplasias. Whole-genome sequencing and comprehensive clinical and biochemical phenotyping was performed in members of a PMAH affected family. Nodules derived from adrenal surgery and pancreatic and meningeal tumor tissue were analyzed for accompanying somatic mutations in the identified target genes. PMAH presenting either as overt or subclinical Cushing's syndrome was accompanied by a heterozygous germline mutation in ARMC5 (p.A110fs*9) located on chromosome 16. Analysis of tumor tissue showed different somatic ARMC5 mutations in adrenal nodules supporting a second hit hypothesis with inactivation of a tumor suppressor gene. A damaging somatic ARMC5 mutation was also found in a concomitant meningioma (p.R502fs) but not in a pancreatic tumor, suggesting biallelic inactivation of ARMC5 as causal also for the intracranial meningioma. Our analysis further confirms inherited inactivating ARMC5 mutations as a cause of familial PMAH and suggests an additional role for the development of concomitant intracranial meningiomas.


Assessment of Academic-Community Partnerships in Translational Research.
The Community Engagement Core (CEC) of the Center of Collaborative Research in Health Disparities focuses on developing and implementing strategies to increase academic-community collaborations and partnerships, enhance the recruitment and retention of study participants, disseminate research findings to a broader audience, and mitigate health disparities in Puerto Rico. In order to assess the current state of academic-community relationships and also collect ideas for their improvement, a strengths, weaknesses, opportunities, and threats (SWOT) analysis was conducted. Participants for the SWOT analysis activity were drawn from a pool of stakeholders at the University of Puerto Rico Medical Sciences Campus and a group of community representatives. The areas identified by the group for the CEC to focus on were weaknesses such as 1) a lack of interaction and involvement with community leaders, 2) limited numbers of advocacy groups in several health areas, and 3) few research consortia. Opportunities identified included the possibilities of 1) creating alliances between academia and industry, municipalities, and community-based organizations, 2) advocating integration in research proposals, and 3) establishing a network of researchers and community leaders. The SWOT analysis activity served to foster relationships with diverse community stakeholders and select members for a community advisory board to collaborate in developing educational activities for our researchers and communities. These findings will also help the CEC establish a strategic plan that should be able to supply a strong community-based participatory research approach that would help mitigate health disparities in Puerto Rico, as well as define the strategies to implement such recommendations.


Collaborative involvement of woody plant roots and rhizosphere microorganisms in the formation of pedogenetic clays.
Previous studies have described the laying down of specific B horizons in south-western Australian ecosystems. This paper presents biomolecular, morphological and physicochemical analyses elucidating the roles of specific woody plant taxa and rhizosphere bacteria in producing these phenomena. Clayey deposits within lateral root systems of eucalypts and appropriate background soil samples were collected aseptically at multiple locations on sand dunes flanking Lake Chillinup. Bacterial communities were profiled using tagged next-generation sequencing (Miseq) of the 16S rRNA gene and assigned to operational taxonomic units. Sedimentation, selective dissolution and X-ray diffraction analyses quantitatively identified clay mineral components. Comparisons were made of pedological features between the above eucalypt systems, giant podzols under proteaceous woodland on sand dunes at the study site of Jandakot and apparently similar systems observed elsewhere in the world. Bacterial communities in clay pods are highly diverse, resolving into 569 operational taxonomic units dominated by Actinobacteria at 38.0-87.4 % of the total reads. Multivariate statistical analyses of community fingerprints demonstrated substrate specificity. Differently coloured pods on the same host taxon carry distinctive microfloras correlated to diversities and abundances of Actinobacteria, Acidobacteria, Firmicutes and Proteobacteria. A number of these microbes are known to form biominerals, such as phyllosilicates, carbonates and Fe-oxides. A biogenic origin is suggested for the dominant identified mineral precipitates, namely illite and kaolinite. Comparisons of morphogenetic features of B horizons under eucalypts, tree banksias and other vegetation types show remarkably similar developmental trajectories involving pods of precipitation surrounding specialized fine rootlets and their orderly growth to form a continuous B horizon. The paper strongly supports the hypothesis that B-horizon development is mediated by highly sophisticated interactions of host plant and rhizosphere organisms in which woody plant taxa govern overall morphogenesis and supply of mineral elements for precipitation, while rhizosphere microorganisms execute biomineralization processes.


A competitive labelling method for determining the ionization constants and reactivity of individual histidine residues in proteins. The histidines of -chymotrypsin.
A competitive labelling method (Kaplan et al., 1971), using tritiated 1-fluoro-2,4-dinitrobenzene as the labelling reagent, is described for determining the ionization constants and reactivities of individual histidine residues in proteins. When this method was applied to the two histidines of alpha-chymotrypsin, histidine-57 was found to have pK(a) 6.8 and a reactivity ten times that of alpha-N-acetyl-l-histidine. Histidine-40 had pK(a) 6.7 and a reactivity approximately six times that of alpha-N-acetyl-l-histidine. Between pH7.5 and 8 the reactivities of both histidines decrease simultaneously to approximately that of alpha-N-acetyl-l-histidine. The high reactivities of the histidines are attributed to hydrogen bonding, which increases the nucleophilicity of the imidazole ring. The sharp decrease in reactivity between pH7.5 and 8 is attributed to a conformational change that disrupts the hydrogen bonding by these residues. The reactivity data support the proposal of a charge-relay mechanism involving histidine-57 (Blow et al., 1969), which makes serine-195 more nucleophilic but indicates that this system is fully operative only in the enzyme-substate complex.


Gas gangrene secondary to Clostridium perfringens in pediatric oncology patients.
To report 2 cases of severe gas gangrene secondary to Clostridium perfringens in pediatric oncology patients. We describe 2 children with acute presentations of gas gangrene secondary to C. perfringens. Both children were initially seen and treated in a community hospital emergency department and subsequently were cared for in a pediatric intensive care unit in a tertiary care, university-based children's hospital. Both children demonstrated severe and unrelenting decompensation and required operative intervention within the first hospital day, which included amputation of the infected limb. One child survived and one child expired despite heroic measures. Gas gangrene secondary to C. perfringens is an uncommon but life-threatening and limb-threatening condition in pediatric cancer patients. A high index of suspicion in a immunocompromised child with cancer who presents with extremity pain in combination with neutropenia is the key to early diagnosis and may lead to improved survival. This disease requires prompt recognition and aggressive treatment to allow any hope of recovery. Emergency medicine physicians who treat these children should be aware of this severe and potentially fatal infectious process and should not delay treatment or prompt orthopedic surgery consultation.


Subaxolemmal cytoskeleton in squid giant axon. I. Biochemical analysis of microtubules, microfilaments, and their associated high-molecular-weight proteins.
Using the squid giant axon, we analyzed biochemically the molecular organization of the axonal cytoskeleton underlying the axolemma (subaxolemmal cytoskeleton). The preparation enriched in the subaxolemmal cytoskeleton was obtained by squeezing out the central part of the axoplasm using a roller. The electrophoretic banding pattern of the subaxolemmal cytoskeleton was characterized by large amounts of two high-molecular-weight (HMW) proteins (260 and 255 kD). The alpha, beta-tubulin, actin, and some other proteins were also its major constituents. The 260-kD protein is known to play an important role in maintaining the excitability of the axolemma (Matsumoto, G., M. Ichikawa, A. Tasaki, H. Murofushi, and H. Sakai, 1983, J. Membr. Biol., 77:77-91) and was recently designated "axolinin" (Sakai, H., G. Matsumoto, and H. Murofushi, 1985, Adv. Biophys., 19:43-89). We purified axolinin and the 255-kD protein in their native forms and further characterized their biochemical properties. The purified axolinin was soluble in 0.6 M NaCl solution but insoluble in 0.1 M NaCl solution. It co-sedimented with microtubules but not with actin filaments. In low-angle rotary-shadowing electron microscopy, the axolinin molecule in 0.6 M NaCl solution looked like a straight rod approximately 105 nm in length with a globular head at one end. On the other hand, the purified 255-kD protein was soluble in both 0.1 and 0.6 M NaCl solution and co-sedimented with actin filaments but not with microtubules. The 255-kD protein molecule appeared as a characteristic horseshoe-shaped structure approximately 35 nm in diameter. Furthermore, the 255-kD protein showed no cross-reactivity to the anti-axolinin antibody. Taken together, these characteristics lead us to conclude that the subaxolemmal cytoskeleton in the squid giant axon is highly specialized, and is mainly composed of microtubules and a microtubule-associated HMW protein (axolinin), and actin filaments and an actin filament-associated HMW protein (255-kD protein).


High performance computing environment for multidimensional image analysis.
The processing of images acquired through microscopy is a challenging task due to the large size of datasets (several gigabytes) and the fast turnaround time required. If the throughput of the image processing stage is significantly increased, it can have a major impact in microscopy applications. We present a high performance computing (HPC) solution to this problem. This involves decomposing the spatial 3D image into segments that are assigned to unique processors, and matched to the 3D torus architecture of the IBM Blue Gene/L machine. Communication between segments is restricted to the nearest neighbors. When running on a 2 Ghz Intel CPU, the task of 3D median filtering on a typical 256 megabyte dataset takes two and a half hours, whereas by using 1024 nodes of Blue Gene, this task can be performed in 18.8 seconds, a 478x speedup. Our parallel solution dramatically improves the performance of image processing, feature extraction and 3D reconstruction tasks. This increased throughput permits biologists to conduct unprecedented large scale experiments with massive datasets.


Application of cell culture technology and genetic engineering for production of future foods and crop improvement to strengthen food security.
The growing population and the climate changes put a pressure on food production globally, therefore a fundamental transformation of food production is required. One approach to accelerate food production is application of modern biotechnology such as cell culture, marker assisted selection, and genetic engineering. Cell culture technology reduces the usage of arable land, while marker-assisted selection increases the genetic gain of crop breeding and genetic engineering enable to introduce a desired traits to crop. The cell culture technology has resulted in development of cultured meat, fungal biomass food (mycoprotein), and bioactive compounds from plant cell culture. Except cultured meat which recently begin to penetrate the market, the other products have been in the market for years. The marker-assisted selection and genetic engineering have contributed significantly to increase the resiliency against emerging pests and abiotic stresses. This review addresses diverse techniques of cell culture technology as well as advanced genetic engineering technology CRISPR Cas-9 and its application for crop improvement. The pros and cons of different techniques as well as the challenges and future perspective of application of modern biotechnology for strengthening food security are also discussed.


Apropos of a case of pancreatic lymphoepithelial cyst: importance of the preoperative diagnosis by ecoendoscopy.
The case was a 75-year-old female with dyspeptic symptoms of a 2 month duration. There was a polycystic lesion at the level of the uncinate pancreatic process of 44 x 42 x 34 mm on abdominal ultrasound and MRI, which caused a slight dilation of the main pancreatic duct. EUS was performed that identified a multicystic formation of 25x36 mm in the pancreatic body and FNA was performed of the lesion. The cytological analysis identified squamous epithelial cells with lymphoid tissue, which was compatible with a lymphoepithelial cyst, with no evidence of cell atypia or other signs of malignancy. Radiological control and monitoring of the lesion was performed and it has remained stable for 2 years.


Integrative analysis of circadian transcriptome and metabolic network reveals the role of de novo purine synthesis in circadian control of cell cycle.
Metabolism is the major output of the circadian clock in many organisms. We developed a computational method to integrate both circadian gene expression and metabolic network. Applying this method to zebrafish circadian transcriptome, we have identified large clusters of metabolic genes containing mostly genes in purine and pyrimidine metabolism in the metabolic network showing similar circadian phases. Our metabolomics analysis found that the level of inosine 5'-monophosphate (IMP), an intermediate metabolite in de novo purine synthesis, showed significant circadian oscillation in larval zebrafish. We focused on IMP dehydrogenase (impdh), a rate-limiting enzyme in de novo purine synthesis, with three circadian oscillating gene homologs: impdh1a, impdh1b and impdh2. Functional analysis revealed that impdh2 contributes to the daily rhythm of S phase in the cell cycle while impdh1a contributes to ocular development and pigment synthesis. The three zebrafish homologs of impdh are likely regulated by different circadian transcription factors. We propose that the circadian regulation of de novo purine synthesis that supplies crucial building blocks for DNA replication is an important mechanism conferring circadian rhythmicity on the cell cycle. Our method is widely applicable to study the impact of circadian transcriptome on metabolism in complex organisms.


Meiotic recombination at the ends of chromosomes in Saccharomyces cerevisiae.
Meiotic reciprocal recombination (crossing over) was examined in the outermost 60-80 kb of almost all Saccharomyces cerevisiae chromosomes. These sequences included both repetitive gene-poor subtelomeric heterochromatin-like regions and their adjacent unique gene-rich euchromatin-like regions. Subtelomeric sequences underwent very little crossing over, exhibiting approximately two- to threefold fewer crossovers per kilobase of DNA than the genomic average. Surprisingly, the adjacent euchromatic regions underwent crossing over at twice the average genomic rate and contained at least nine new recombination "hot spots." These results prompted an analysis of existing genetic mapping data, which showed that meiotic reciprocal recombination rates were on average greater near chromosome ends exclusive of the subtelomeres. Thus, the distribution of crossovers in S. cerevisiae appears to resemble that found in several higher eukaryotes where the outermost chromosomal regions show increased crossing over.


Contrasting microbial assemblages in adjacent water masses associated with the East Australian Current.
Different oceanographic provinces host discrete microbial assemblages that are adapted to local physicochemical conditions. We sequenced and compared the metagenomes of two microbial communities inhabiting adjacent water masses in the Tasman Sea, where the recent strengthening of the East Australian Current (EAC) has altered the ecology of coastal environments. Despite the comparable latitude of the samples, significant phylogenetic differences were apparent, including shifts in the relative frequency of matches to Cyanobacteria, Crenarchaeota and Euryarchaeota. Fine-scale variability in the structure of SAR11, Prochlorococcus and Synechococcus populations, with more matches to 'warm-water' ecotypes observed in the EAC, indicates the EAC may drive an intrusion of tropical microbes into temperate regions of the Tasman Sea. Furthermore, significant shifts in the relative importance of 17 metabolic categories indicate that the EAC prokaryotic community has different physiological properties than surrounding waters.


Expression of the laminin gamma 2 chain in pancreatic adenocarcinoma.
Forty-two pancreatic adenocarcinomas were investigated immunohistochemically and by in situ hybridization for the expression of the laminin gamma 2 chain. In 41 cases, intracytoplasmic immunoreactivity for the gamma 2 chain was seen. Positive tumour cells were located especially at the epithelial-stromal interface of the tumour cell islands. In 22 cases, diffuse laminin gamma 2 chain immunoreactivity could also be seen in stroma and in seven cases, occasional positivity was detected in the neoplastic basement membranes. Signals for laminin gamma 2 chain mRNA in tumour cells displayed a distribution similar to that observed on immunohistochemistry. There were significantly more cases with less than 20 per cent of laminin gamma 2 chain-positive tumour cells in tumours extending to peripancreatic tissues and/or tumours with regional or distant metastases (p = 0.029). A corresponding statistical significance could also be noted in the mRNA level (P = 0.025). The results show that pancreatic adenocarcinomas display a high activity of laminin gamma 2 chain synthesis. Tumours with a strong laminin gamma 2 chain synthesis show a lower invasive and metastatic potential than tumours with a weak or moderate laminin gamma 2 chain expression.


Salvianolic Acid A Protects H9c2 Cells from Arsenic Trioxide-Induced Injury via Inhibition of the MAPK Signaling Pathway.
This study aimed to investigate whether Salvianolic acid A (Sal A) conferred cardiac protection against Arsenic trioxide (ATO)-induced cardiotoxicity in H9c2 cells by inhibiting MAPK pathways activation. H9c2 cardiac cells were exposed to 10 µM ATO for 24 h to induce cytotoxicity. The cells were pretreated with Sal A for 4 h before exposure to ATO. Cell viability was determined utilizing the MTT assay. The percentage of apoptosis was measured by a FITC-Annexin V/PI apoptosis kit for flow cytometry. Mitochondrial membrane potential (∆Ψm) was detected by JC-1. The intracellular ROS levels were measured using an Image-iTTM LIVE Green Reactive Oxygen Species Detection Kit. The apoptosis-related proteins and the MAPK signaling pathways proteins expression were quantified by Western blotting. Sal A pretreatment increased cell viability, suppressed ATO-induced mitochondrial membrane depolarization, and significantly altered the apoptotic rate by enhancing endogenous antioxidative enzyme activity and ROS generation. Signal transduction studies indicated that Sal A suppressed the ATO-induced activation of the MAPK pathway. More importantly, JNK, ERK, and p38 inhibitors mimicked the cytoprotective activity of Sal A against ATO-induced injury in H9c2 cells by increasing cell viability, up-regulating Bcl-2 protein expression, and down-regulating both Bax and caspase-3 protein expression. Sal A decreases the ATO-induced apoptosis and necrosis of H9c2 cells, and the underlying mechanisms of this protective effect of Sal A may be connected with the MAPK pathways.


Molecular profiling and utility of cell-free DNA in nonsmall carcinoma of the lung: Study in a tertiary care hospital.
Lung carcinoma accounts to the most common cause of cancer globally. Optimal management of nonsmall cell lung carcinoma (NSCLC) requires prognostic biomarkers that help in targeted therapy and identification of tumor subsets with a distinctive molecular profile that can foretell response to therapy. Quantitative analysis of circulating cell-free DNA is considered as a possible aid for lung cancer screening. The main aim of our study was detection of the clinicopathological spectrum of NSCLC, immunohistochemical (IHC) study of lung adenocarcinoma with epidermal growth factor receptor (EGFR), anaplastic lymphoma kinase (ALK), and molecular expression of EGFR mutation using Formalin fixed paraffin embedded tissue (FFPE) and cell-free DNA (cfDNA) from blood samples. It was a prospective and observational study conducted in the Department of Pathology in association with the Department of Chest Medicine in a tertiary care hospital for 18 months, done on 50 patients. Histological subtyping of lung carcinomas was done, followed by IHC analysis using P40, thyroid transcription factor (TTF1), EGFR, and ALK. Molecular analysis for EGFR mutation was done using FFPE and cfDNA from the patient's blood samples. On histological subtyping, majority (66%) of the cases were found to be adenocarcinoma. All adenocarcinoma (66%) cases show TTF1 positivity and all squamous cell carcinoma (32%) cases show P40 positivity. All the ALK-positive (6%) cases were never smokers and histologically diagnosed as adenocarcinoma. About 58% of the NSCLC cases were found to be EGFR IHC positive. Formalin-fixed paraffin tissue (FFPE) showed EGFR mutation in 32% cases, of which majority were deletion (19, 28%) and rest (4%) of the cases involving mutation in exon 21. From cfDNA, mutations were noticed in 16% of the cases where majority involved deletion 19 (12%), whereas the rest of the cases were positive for missense mutation in exon 21 of the EGFR gene (2%) and compound heterozygous mutation involving deletion 19 and missense mutation for exon 21 (2%). On correlation of EGFR mutation studies from FFPE with that of cfDNA analysis, the study was statistically significant (P = 0.000). This study reports clinicopathological, immunochemical, and molecular analysis of EGFR among NSCLC cases. EGFR mutation detection from cfDNA has its advantage of being a noninvasive technique to avoid rebiopsy in cases of the progressive disease to detect resistance to a drug and emergence of a newer mutation. Mutation detection from FFPE samples still remains the gold standard for targeted therapy using EGFR tyrosine kinase inhibitors. ALK rearrangement detection using IHC serves as an adjunct to EGFR diagnosis.


Nerve growth factor activation of the TrkA receptor induces cell death, by macropinocytosis, in medulloblastoma Daoy cells.
Ectopic expression of the TrkA receptor tyrosine kinase in tumors of the nervous system can mediate nerve growth factor (NGF)-dependent cell death by apoptosis and /or autophagy. Herein, we demonstrate that TrkA can also induce cell death in medulloblastoma Daoy cells by a caspase-independent mechanism that involves the hyperstimulation of macropinocytosis. Specifically, NGF-stimulates the uptake of AlexaFluor546-dextran into lysosome-associated membrane protein-1 positive vacuoles which fuse with microtubule associated protein light chain 3 (LC3) positive autophagosomes, to form large intracellular vacuoles (> 1 mum), which then fuse with lysotracker positive lysosomes. While LC3 cleavage and the appearance of LC3 positive vacuoles suggest the induction of autophagy, siRNA reduced expression of four proteins essential to autophagy (beclin-1, Atg5, LC3 and Atg9) neither blocks NGF-induced vacuole formation nor cell death. TrkA activated cell death does not require p38, JNK or Erk1/2 kinases but does require activation of class III PI-3 kinase and is blocked by the casein kinase 1 (CK1) inhibitor, D4476. This inhibitor does not interfere with TrkA activation but does block NGF-dependent AlexaFluor546-dextran uptake and CK1 dependent phosphorylation of beta-catenin. Collectively, these data demonstrate that TrkA stimulates cell death by a novel mechanism involving CK1-dependent hyperstimulation of macropinocytosis.


Isolation and partial characterization of Bermuda grass-pollen allergen: BG-60b.
Our earlier studies have shown that the pollen of Bermuda grass (Cynodon dactylon)-pollen contain at least 12 IgE-binding proteins which can be analyzed by the immunoblot technique. One of the highly active components was found to be a basic protein with a molecular weight of 60,000 daltons, designated as BG-60. This component was showed to consist of a group of proteins. One of them, BG-60a (pI 9.7), has been isolated and characterized. In this study, we have further isolated and characterized the second component of the antigen, designated as BG-60b. Its purity was demonstrated by gel electrophoresis experiment and antigen-antibody precipitation studies. The antigen is of glycoprotein nature with a pI of 10.0. It exhibits IgE-binding activity and shows cross-reactivity to antigen BG-60a in double diffusion. Its chemical and physical properties are similar to antigen BG-60a.


Acute Effects of Walking on the Deformation of Femoral Articular Cartilage in Older Adults.
Although discomfort during walking is a common complaint in individuals with knee osteoarthritis (OA), how an acute bout of walking affects femoral cartilage remains unclear. Current literature has suggested that frontal plane knee malalignment (ie, varus and valgus) is associated with the initiation and/or progression of knee OA. However, the association between knee alignment and femoral cartilage deformation after an acute bout of loading has not yet been investigated. This study was aimed to compare the acute effects of walking on femoral cartilage deformation between older adults with and without knee OA. We also examined the association between frontal plane knee alignment and loading-induced femoral cartilage deformation. Ten persons without OA (Kellgren Lawrence grading = 0 or 1; 5 females and 5 males; 55.0 [1.8] years of age; 78.8 [14.1] kg; 1.8 [0.2] m) and 9 persons with OA (Kellgren Lawrence grading ≥2; 4 females and 5 males; 55.6 [4.5] years of age; 97.4 [15.0] kg; 1.7 [0.1] m) participated. Each participant underwent magnetic resonance imaging before and immediately after 30 minutes of fast walking at 3 to 4 miles per hour. To obtain cartilage deformation postwalking, the medial and lateral femoral cartilage of the weight-bearing areas was segmented on participants' magnetic resonance imaging. Cartilage thickness was quantified by computing the average perpendicular distance between opposing voxels defining the edges of the femoral cartilage. Cartilage deformation of the medial and lateral femurs was defined as the percent changes in cartilage thickness after walking. Frontal plane knee alignment was obtained by measuring the angle between the long axes of femur and tibia. Independent t tests were used to compare cartilage deformation between the 2 groups. Pearson correlation coefficients were used to assess the association between cartilage deformation and knee alignment. There was no significant difference in cartilage deformation between the OA and control groups in lateral (P = .69) or medial (P = .87) femur. A significant correlation was found between lateral femoral cartilage deformation and increased knee valgus alignment (r = 0.497; P = .03). No difference was found between medial femoral cartilage deformation and frontal plane knee alignment (r = 273; P = .26). This is the first study comparing the acute effects of walking on femoral cartilage deformation between older adults with and without knee OA. Although there was not a difference in walking-induced femoral cartilage deformation between the OA and control groups, knee valgus was related to lateral femoral cartilage deformation after walking. Our findings suggested that walking exercises may be used safely in older adults without knee malalignment.


Archaeal nucleosome positioning in vivo and in vitro is directed by primary sequence motifs.
Histone wrapping of DNA into nucleosomes almost certainly evolved in the Archaea, and predates Eukaryotes. In Eukaryotes, nucleosome positioning plays a central role in regulating gene expression and is directed by primary sequence motifs that together form a nucleosome positioning code. The experiments reported were undertaken to determine if archaeal histone assembly conforms to the nucleosome positioning code. Eukaryotic nucleosome positioning is favored and directed by phased helical repeats of AA/TT/AT/TA and CC/GG/CG/GC dinucleotides, and disfavored by longer AT-rich oligonucleotides. Deep sequencing of genomic DNA protected from micrococcal nuclease digestion by assembly into archaeal nucleosomes has established that archaeal nucleosome assembly is also directed and positioned by these sequence motifs, both in vivo in Methanothermobacter thermautotrophicus and Thermococcus kodakarensis and in vitro in reaction mixtures containing only one purified archaeal histone and genomic DNA. Archaeal nucleosomes assembled at the same locations in vivo and in vitro, with much reduced assembly immediately upstream of open reading frames and throughout the ribosomal rDNA operons. Providing further support for a common positioning code, archaeal histones assembled into nucleosomes on eukaryotic DNA and eukaryotic histones into nucleosomes on archaeal DNA at the same locations. T. kodakarensis has two histones, designated HTkA and HTkB, and strains with either but not both histones deleted grow normally but do exhibit transcriptome differences. Comparisons of the archaeal nucleosome profiles in the intergenic regions immediately upstream of genes that exhibited increased or decreased transcription in the absence of HTkA or HTkB revealed substantial differences but no consistent pattern of changes that would correlate directly with archaeal nucleosome positioning inhibiting or stimulating transcription. The results obtained establish that an archaeal histone and a genome sequence together are sufficient to determine where archaeal nucleosomes preferentially assemble and where they avoid assembly. We confirm that the same nucleosome positioning code operates in Archaea as in Eukaryotes and presumably therefore evolved with the histone-fold mechanism of DNA binding and compaction early in the archaeal lineage, before the divergence of Eukaryotes.


Parenteral iron use: possible contribution to exceeding target hemoglobin in hemodialysis patients.
Use of parenteral iron for anemia management in dialysis patients has greatly increased. Exceeding hemoglobin target levels is not without risk, and whether parenteral iron administration contributes to exceeding targets has not been tested. The authors aimed to determine prevalence of parenteral iron administration and its contribution to exceeding hemoglobin target levels. The authors performed a retrospective observational study of 149,292 hemodialysis patients using Centers for Medicaid & Medicare Services data. All patients were point prevalent on January 1, 2004; survived through June 30, 2004; had Medicare as primary payer; were treated with erythropoiesis stimulating agents (ESAs); and had valid hemoglobin values in April, May, and June, 2004. Of the cohort, 58% received parenteral iron; use was more likely among men, whites, younger patients, and patients with end-stage renal disease as a result of diabetes. Age > 75 yr, African American and other races, baseline hemoglobin > 12 g/dl, higher ESA dose, and iron use in months 1 to 4 of the study period were independently associated with the risk of exceeding hemoglobin levels of 12, 13, and 14 g/dl. Receiving iron in month 4 of the study period showed the highest probability of exceeding targets (odds ratios 1.49, 1.43, 1.50 for hemoglobin levels 12, 13, 14 g/dl, respectively). Parenteral iron use is prevalent, and although adequate iron stores are central to ESA response, iron use may contribute to exceeding recommended hemoglobin levels. Only data from a prospective trial can confirm this association.


KIT Suppresses BRAFV600E-Mutant Melanoma by Attenuating Oncogenic RAS/MAPK Signaling.
The receptor tyrosine kinase KIT promotes survival and migration of melanocytes during development, and excessive KIT activity hyperactivates the RAS/MAPK pathway and can drive formation of melanomas, most notably of rare melanomas that occur on volar and mucosal surfaces of the skin. The much larger fraction of melanomas that occur on sun-exposed skin is driven primarily by BRAF- or NRAS-activating mutations, but these melanomas exhibit a surprising loss of KIT expression, which raises the question of whether loss of KIT in these tumors facilitates tumorigenesis. To address this question, we introduced a kit(lf) mutation into a strain of Tg(mitfa:BRAFV600E); p53(lf) melanoma-prone zebrafish. Melanoma onset was accelerated in kit(lf); Tg(mitfa:BRAFV600E); p53(lf) fish. Tumors from kit(lf) animals were more invasive and had higher RAS/MAPK pathway activation. KIT knockdown also increased RAS/MAPK pathway activation in a BRAFV600E-mutant human melanoma cell line. We found that pathway stimulation upstream of BRAFV600E could paradoxically reduce signaling downstream of BRAFV600E, and wild-type BRAF was necessary for this effect, suggesting that its activation can dampen oncogenic BRAFV600E signaling. In vivo, expression of wild-type BRAF delayed melanoma onset, but only in a kit-dependent manner. Together, these results suggest that KIT can activate signaling through wild-type RAF proteins, thus interfering with oncogenic BRAFV600E-driven melanoma formation. Cancer Res; 77(21); 5820-30. ©2017 AACR.


Cloning and nucleotide analysis of segment A gene of infectious bursal disease virus detected in Korea.
A strain of infectious bursal disease virus (IBDV) was detected from bursal tissues of chicks which suffered from infectious bursal disease (IBD) in Chinju, Korea and provisionally named as Chinju strain. A full-length cDNA clone for segment A gene of the virus was constructed, and complete nucleotide sequence of the gene including noncoding region was determined and analyzed by comparison with that of other IBDV strains. The segment A gene of Chinju strain consisted of 3,269 nucleotides including 862 adenine (26.4%), 917 cytosine (28.0%), 854 guanine (26.1%) and 636 thymine (19.5%). There were regions for two open reading frames (ORFs), ORF1 encoding the VP5 with ATG codon at nucleotides 98-100 and ORF2 encoding the polyprotein of VP2, VP4 and VP3 in the nucleotides 132-3,170. In deduced translation the ORF2 encoded 1,012 amino acids. The full nucleotide sequence of segment A gene and amino acid sequence of ORF2 of the Chinju strain showed 98-99% homology with those of the very virulent IBDVs (vvIBDVs) such as HK46, OKYM, UK661, UPM97/ 61, D6948 and BD3/99. In phylogenetic analysis of nucleotide and amino acid sequences, the Chinju strain was also related closely to the vvIBDVs. Hence, it was suggested that the Chinju strain is a vvIBDV. The nucleotide and amino acid sequences of the Chinju strain with pertinent information can be useful for the development of genetically engineered vaccines and diagnostic reagents against vvIBDV.


BRCA1 functions independently of homologous recombination in DNA interstrand crosslink repair.
Brca1 is required for DNA repair by homologous recombination (HR) and normal embryonic development. Here we report that deletion of the DNA damage response factor 53BP1 overcomes embryonic lethality in Brca1-nullizygous mice and rescues HR deficiency, as measured by hypersensitivity to polyADP-ribose polymerase (PARP) inhibition. However, Brca1,53BP1 double-deficient cells are hypersensitive to DNA interstrand crosslinks (ICLs), indicating that BRCA1 has an additional role in DNA crosslink repair that is distinct from HR. Disruption of the nonhomologous end-joining (NHEJ) factor, Ku, promotes DNA repair in Brca1-deficient cells; however deletion of either Ku or 53BP1 exacerbates genomic instability in cells lacking FANCD2, a mediator of the Fanconi anemia pathway for ICL repair. BRCA1 therefore has two separate roles in ICL repair that can be modulated by manipulating NHEJ, whereas FANCD2 provides a key activity that cannot be bypassed by ablation of 53BP1 or Ku.


Enhanced permeability responses to inflammation in streptozotocin-induced diabetic rat venules: Rho-mediated alterations of actin cytoskeleton and VE-cadherin.
Diabetes is a progressive disease that often leads to microvascular complications. This study investigates the impact of diabetes on microvessel permeability under basal and inflammatory conditions. Streptozotocin-induced diabetic rats were used to mimic type 1 diabetes. Parallel experiments were conducted in intact mesenteric venules in normal rats and diabetic rats experiencing hyperglycemia for 2-3 wk. Microvessel permeability was determined by measuring hydraulic conductivity (Lp). The correlated changes in endothelial intracellular Ca(2+) concentration ([Ca(2+)]i), adherens junctions, and cytoskeleton F-actin were examined with fluorescence imaging. Diabetic vessels showed moderately increased basal Lp, but upon platelet-activating factor (PAF) exposure, these vessels showed an ~10-fold higher Lp increase than the normal vessels. Concomitantly, we observed higher increases in endothelial [Ca(2+)]i, enhanced stress fiber formation, vascular endothelial-cadherin separation, and larger gap formation between endothelial cells than those occurring in normal vessels. PAF receptor staining showed no significant difference between normal and diabetic vessels. The application of Rho kinase inhibitor Y27632 did not affect PAF-induced increases in endothelial [Ca(2+)]i but significantly reduced PAF-induced Lp increases by 90% in diabetic vessels. The application of both Y27632 and nitric oxide (NO) synthase inhibitor attenuated PAF-induced Lp increases more than using one inhibitor alone. Our studies indicate that diabetic conditions prime endothelial cells into a phenotype with increased susceptibility to inflammation without altering receptor expression and that the increased Rho activation and NO production play important roles in exaggerated permeability increases when diabetic vessels were exposed to inflammatory mediators, which may account for the exacerbated vascular dysfunction when diabetic patients are exposed to additional inflammation.


Structural basis of the stereospecificity of bacterial B12-dependent 2-hydroxyisobutyryl-CoA mutase.
Bacterial coenzyme B12-dependent 2-hydroxyisobutyryl-CoA mutase (HCM) is a radical enzyme catalyzing the stereospecific interconversion of (S)-3-hydroxybutyryl- and 2-hydroxyisobutyryl-CoA. It consists of two subunits, HcmA and HcmB. To characterize the determinants of substrate specificity, we have analyzed the crystal structure of HCM from Aquincola tertiaricarbonis in complex with coenzyme B12 and the substrates (S)-3-hydroxybutyryl- and 2-hydroxyisobutyryl-CoA in alternative binding. When compared with the well studied structure of bacterial and mitochondrial B12-dependent methylmalonyl-CoA mutase (MCM), HCM has a highly conserved domain architecture. However, inspection of the substrate binding site identified amino acid residues not present in MCM, namely HcmA Ile(A90) and Asp(A117). Asp(A117) determines the orientation of the hydroxyl group of the acyl-CoA esters by H-bond formation, thus determining stereospecificity of catalysis. Accordingly, HcmA D117A and D117V mutations resulted in significantly increased activity toward (R)-3-hydroxybutyryl-CoA. Besides interconversion of hydroxylated acyl-CoA esters, wild-type HCM as well as HcmA I90V and I90A mutant enzymes could also isomerize pivalyl- and isovaleryl-CoA, albeit at >10 times lower rates than the favorite substrate (S)-3-hydroxybutyryl-CoA. The nonconservative mutation HcmA D117V, however, resulted in an enzyme showing high activity toward pivalyl-CoA. Structural requirements for binding and isomerization of highly branched acyl-CoA substrates such as 2-hydroxyisobutyryl- and pivalyl-CoA, possessing tertiary and quaternary carbon atoms, respectively, are discussed.


Synthesis of new chlorin e6 trimethyl and protoporphyrin IX dimethyl ester derivatives and their photophysical and electrochemical characterizations.
In view of increasing demands for efficient photosensitizers for photodynamic therapy (PDT), we herein report the synthesis and photophysical characterizations of new chlorin e6 trimethyl ester and protoporphyrin IX dimethyl ester dyads as free bases and Zn(II) complexes. The synthesis of these molecules linked at the β-pyrrolic positions to pyrano[3,2-c]coumarin, pyrano[3,2-c]quinolinone, and pyrano[3,2-c]naphthoquinone moieties was performed by using the domino Knoevenagel hetero Diels-Alder reaction. The α-methylenechromanes, α-methylenequinoline, and ortho-quinone methides were generated in situ from a Knoevenagel reaction of 4-hydroxycoumarin, 4-hydroxy-6-methylcoumarin, 4-hydroxy-N-methylquinolinone, and 2-hydroxy-1,4-naphthoquinone, respectively, with paraformaldehyde in dioxane. All the dyads as free bases and as Zn(II) complexes were obtained in high yields. All new compounds were fully characterized by 1D and 2D NMR techniques, UV/Vis spectroscopy, and HRMS. Their photophysical properties were evaluated by measuring the fluorescence quantum yield, the singlet oxygen quantum yield by luminescence detection, and also the triplet lifetimes were correlated by flash photolysis and intersystem crossing (ISC) rates. The fluorescence lifetimes were measured by a time-correlated single photon count (TCSPC) method, fluorescence decay associated spectra (FDAS), and anisotropy measurements. Magnetic circular dichroism (MCD) and circular dichroism (CD) spectra were recorded for one Zn(II) complex in order to obtain information, respectively, on the electronic and conformational states, and interpretation of these spectra was enhanced by molecular orbital (MO) calculations. Electrochemical studies of the Zn(II) complexes were also carried out to gain insights into their behavior for such applications.


Exploring Challenges in Conducting E-Mental Health Research Among Asian American Women.
In this discussion paper, we explore the challenges of conducting e-mental health intervention research among Asian American women and propose a model for addressing these barriers. Based on an extensive literature review, we identify two main types of barriers to conducting e-mental health intervention research among Asian American women: recruitment barriers and adherence barriers. Recruitment barriers are further subcategorized into those related to (1) stigmatized cultural beliefs about mental illness and mental health services; (2) lack of awareness about mental health services; and (3) language barrier. As to adherence barriers, the two identified subtypes concern (1) acuity and severity of mental health condition; and (2) lack of time. In order to enhance recruitment and adherence in e-mental health intervention research among the studied population, we formulate the following three main research strategies, namely: (1) considering the cultural and social contexts of Asian American women in the development of e-mental health interventions; (2) determining appropriate program length; and (3) conducting feasibility studies to test e-mental health interventions. We suggest that nurse researchers integrate our proposed model in conducting e-mental health interventions among Asian American women. Our proposed model also implies that nurses play an important role in encouraging Asian American women's acceptance of and adherence to e-mental health interventions. In order to overcome the obstacles to conducting e-mental health research among Asian American women, we recommend that nurses familiarize themselves with credible, relevant, and evidence-based e-mental health resources and integrate online mental health services and information within their nursing practice.


Zanubrutinib for treatment-naïve and relapsed/refractory chronic lymphocytic leukaemia: long-term follow-up of the phase I/II AU-003 study.
The phase I/II AU-003 study in patients with treatment-naïve (TN) or relapsed/refractory (R/R) chronic lymphocytic leukaemia/small lymphocytic lymphoma demonstrated that zanubrutinib therapy results in clinically meaningful and durable responses with acceptable safety and tolerability. We report updated safety and efficacy data for 123 patients with a median follow-up of 47·2 months. Patients received zanubrutinib 160 mg twice daily (81 patients), 320 mg once daily (40), or 160 mg once daily (two). Discontinuations due to adverse events or disease progression were uncommon. The overall response rate (ORR) was 95·9% (TN, 100%; R/R, 95%) with 18·7% achieving complete response (CR). Ongoing response at 3 years was reported in 85·7%. The ORR in patients with del(17p)/tumour protein p53 mutation was 87·5% (CR 16·7%). The 2- and 3-year progression-free survival estimates were 90% (TN, 90%; R/R, 91%) and 83% (TN, 81%; R/R, 83%) respectively. The most reported Grade ≥3 adverse events were neutropenia (15·4%), pneumonia (9·8%), hypertension (8·9%) and anaemia (6·5%). The annual incidence of atrial fibrillation, major haemorrhage, Grade ≥3 neutropenia and Grade ≥3 infection decreased over time. With a median follow-up of ~4 years, responses remain clinically meaningful and durable and long-term tolerability to zanubrutinib therapy continues.


[Marked endocrine abnormalities in polyneuritis with skin hyperpigmentation, hypertrichosis, edema and increased lambda type immunoglobulin A: a case report].
A syndrome which is known as plasma cell dyscrasia with polyneuropathy and various endocrine manifestations or plasma cell dyscrasia with polyneuropathy, organomegaly, endocrinopathy, abnormal M protein and skin changes is very interesting because this syndrome has miscellaneous manifestations such as skin hyperpigmentation, hypertrichosis, polyneuropathy, M protein abnormality, plasma cell dyscrasia and endocrine disturbances. Miscellaneous endocrine abnormalities which have not been described so far are reported here. A 47 year old female was admitted with the chief complaints of edema and gait disturbance. Past and family histories were noncontributory. In April 1981, edema appeared in her face and legs. In June she noticed paresthesia in her legs. Edema increased gradually and she had difficulty walking. Her skin became pigmented and hairy. In October she was admitted because of polyneuropathy with increased cerebrospinal fluid protein without pleocytosis. Prednisolone was started. Walking improved slightly, but edema and paresthesia remained unchanged. Prednisolone was stopped at the end of the following March. In May 1982, she was admitted for further evaluation of edema and polyneuropathy. The patient was alert and cooperative. On standing the skin of her legs became cyanotic. There was hypertrichosis on the arms and legs. Her fingers were clubbed. A moderate swelling of the cervical lymph nodes was noted. There was mild hepatomegaly without splenomegaly. All tendon reflexes were lost. Plantar response was flexor. Muscular strength diminished mildly. She complained of paresthesia on the soles. Superficial sensation was normal. Vibratory sense decreased mildly. Cerebellar function and cranial nerves were normal. There was no sphincter disturbance. The examination of urine, stool and peripheral blood was normal.(ABSTRACT TRUNCATED AT 250 WORDS)


HOXA10 deteriorates gastric cancer through activating JAK1/STAT3 signaling pathway.
Background: HOXA10 has been reported to be deregulated in many kinds of cancers including gastric cancer. But its role in gastric cancer progression is controversial. Therefore, the current study was performed to explore the role and mechanism of HOXA10 in gastric cancer. Materials and methods: IHC and Western blotting assays were used to assess HOXA10 expression in gastric cancer tissues and cells. Lentivirus infection was used to alter HOXA10, STAT3 and JAK1 expression in gastric cancer NCI-N87 and MKN28 cells. MTT, cloning formation, flow cytometry and in vivo xenotransplantation experiments were carried out to assess cell proliferation, cloning formation, apoptosis and tumorigenesis. Results: HOXA10 expression was obviously increased in gastric cancer tissues and cells when compared with the normal gastric tissue samples and cells. Upregulation of HOXA10 significantly enhanced cell proliferation, cloning formation and tumorigenesis abilities and reduced cell apoptosis in gastric cancer, and promoted the activation of JAK1/STAT3 signaling. In addition, we showed that the effects of HOXA10 on the promotion of cell viability and tumorigenesis and cell apoptosis repression were all weakened when JAK1 or STAT3 was downregulated. Conclusion: This study demonstrates that HOXA10 functions as an oncogene in gastric cancer through activating JAK1/STAT3 signaling.


Preservation of protein expression systems at elevated temperatures for portable therapeutic production.
Many biotechnology capabilities are limited by stringent storage needs of reagents, largely prohibiting use outside of specialized laboratories. Focusing on a large class of protein-based biotechnology applications, we address this issue by developing a method for preserving cell-free protein expression systems for months above room temperature. Our approach realizes unprecedented long-term stability at elevated temperatures by leveraging the sugar alcohol trehalose, a simple, low-cost, open-air drying step, and strategic separation of reaction components during drying. The resulting preservation capacity enables efficient production of a wide range of on-demand proteins under adverse conditions, for instance during emergency outbreaks or in remote locations. To demonstrate application potential, we use cell-free reagents subjected to months of exposure at 37°C and atmospheric conditions to produce sufficient concentrations of a pyocin protein to kill Pseudomonas aeruginosa, a troublesome pathogen for traumatic and burn wound injuries. Our work makes possible new biotechnology applications that demand ruggedness and scalability.


Understanding mental illness: the experience of families of psychiatric patients.
This qualitative study was designed to elicit the family's perspective of their experience of the first hospitalization of a relative. A major focus of the study was the process by which families developed an understanding of mental illness and its treatment. During unstructured interviews, seven families explained how they interpreted unfamiliar events and surroundings. They identified their needs and concerns related to their interpretation of 'mental illness', and they described their efforts to understand the patient from this perspective. The study identified processes by which families attached meaning to events: they reviewed past experiences and beliefs to make sense of the present experience; they examined their own behaviour as a contributor to the development of the illness; they observed and evaluated the hospitalization as treatment; and they asked questions about the future. The direction for nursing practice was identified in terms of providing support and information to families, as well as being aware of the emotional responses of families at all stages of the patient's illness.