A quantitative survey of intern's knowledge of communication skills: an Iranian exploration.
It is a high priority that health care providers have effective communication skills. It has been well documented that the doctor-patient relationship is central to the delivery of high quality medical care, and it has been shown to affect patient satisfaction, to decrease the use of pain killers, to shorten hospital stays, to improve recovery from surgery and a variety of other biological, psychological and social outcomes. This study sought to quantify the current knowledge of interns in Iran about communication skills. A cross-sectional study using a self-report questionnaire was conducted among interns. Data analysis was based on 223 questionnaires. The internal consistency of the items was 0.8979. Overall, knowledge levels were unsatisfactory. Results indicated that interns had a limited knowledge of communication skills, including identification of communication skills. In addition, there was a significant difference between the mean scores of interns on breaking bad news and sex education. The confidence of males about their communication skills was significantly higher than for females. Analysis of the total scores by age and sex showed that there was a statistically significant main effect for sex and the interaction with age was statistically significant. Free response comments of the interns are also discussed. It is argued that there is a real need for integrating a communication skills course, which is linked to the various different ethnic and religious backgrounds of interns, into Iranian medical curricula. Some recommendations are made and the limitations of the study are discussed.


Bile acids and biliary carcinoma in pancreaticobiliary maljunction.
Pancreaticobiliary maljunction is frequently associated with biliary carcinoma, whether or not there is choledochal dilatation. In this anomalous condition, pancreatic juice regurgitates and the mixture of bile and pancreatic juice stagnates in the biliary tree. In cystic choledochal dilatation, cancers arise, mainly in the dilated bile ducts, while in patients not having cystic dilatation, tumors arise in the gallbladder. Gallbladder bile and/or bile duct bile from fifteen cases of pancreaticobiliary maljunction, including five cancer patients, was analysed biochemically and compared with control bile from 6 patients with a normal pancreaticobiliary junction. Bile levels of pancreatic enzymes were extremely high in the anomalous junction group. In the bile duct bile from patients with cystic choledochal dilatation with pancreaticobiliary maljunction, the concentrations of deoxycholic acid (DCA), lithocholic acid (LCA) and unconjugated bile acid fractions were increased regardless of the presence of cancer. Increases of these bile acid fractions, which are known to have a cancer-promoting effect, were also seen in gallbladder bile from the cancer patients without cystic dilatation.


Perinatal malpractice. Risks and prevention.
An analysis of 25 perinatal malpractice cases filed in Wisconsin between 1978 and 1984 clearly revealed that the unfavorable outcomes were preventable and caused by professional negligence. Each case was settled or adjudicated in favor of the plaintiff, and the awards totaled $25.1 million. Although cases with a total recovery of over $1 million are relatively infrequent, they are not so rare as to be considered extra-ordinary occurrences. Fifty-six percent of physicians involved failed to recognize a high-risk pregnancy or fetal distress, while 44% failed to render proper care. The most common errors involved inadequate fetal monitoring, the injudicious use of oxytocin and the failure to recognize a high-risk pregnancy, such as prematurity or postterm or multiple gestation. The purpose of this study was to examine the medicolegal issues operating in such cases in order to develop guidelines for reducing professional negligence, thereby preventing injury to women and their newborns.


Development of a Nanoemulgel for the Topical Application of Mupirocin.
Mupirocin (MUP) is an effective topical antibiotic with poor skin permeability; however, its skin permeability can be improved by a nanoemulsion formulation based on eucalyptus oil or eucalyptol. Despite this improvement, the nanoemulsion has limitations, such as low viscosity, low spreadability, and poor retention on the skin. To overcome these limitations, the aim of this study was to develop a nanoemulgel formulation that would enhance its rheological behaviour and physicochemical properties. The MUP nanoemulgel was prepared by incorporating a preprepared MUP nanoemulsion into Carbopol gel at a concentration of 0.75% in a 1:1 ratio. The nanoemulgel formulations were characterised and evaluated for their physicochemical and mechanical strength properties, rheological behaviour, and in vitro skin permeation and deposition, as well as antibacterial studies. Both nanoemulgels exhibited stability at temperatures of 4 and 25 °C for a period of 3 months. They had a smooth, homogenous, and consistent appearance and displayed non-Newtonian pseudoplastic behaviour, with differences in their viscosity and spreadability. However, both nanoemulgels exhibited lower skin permeability compared to the marketed control. The local accumulation efficiency of MUP from nanoemulgel after 8 h was significantly higher than that of the control, although there was no significant difference after 24 h. Micro-CT scan imaging allowed visualisation of these findings and interpretation of the deposited drug spots within the layers of treated skin. While there were no significant differences in the antibacterial activities between the nanoemulgels and the control, the nanoemulgels demonstrated superiority over the control due to their lower content of MUP. These findings support the potential use of the nanoemulgel for targeting skin lesions where high skin deposition and low permeability are required, such as in the case of topical antibacterial agents.


Isolation of a novel cutinase homolog with polyethylene terephthalate-degrading activity from leaf-branch compost by using a metagenomic approach.
The gene encoding a cutinase homolog, LC-cutinase, was cloned from a fosmid library of a leaf-branch compost metagenome by functional screening using tributyrin agar plates. LC-cutinase shows the highest amino acid sequence identity of 59.7% to Thermomonospora curvata lipase. It also shows the 57.4% identity to Thermobifida fusca cutinase. When LC-cutinase without a putative signal peptide was secreted to the periplasm of Escherichia coli cells with the assistance of the pelB leader sequence, more than 50% of the recombinant protein, termed LC-cutinase*, was excreted into the extracellular medium. It was purified and characterized. LC-cutinase* hydrolyzed various fatty acid monoesters with acyl chain lengths of 2 to 18, with a preference for short-chain substrates (C(4) substrate at most) most optimally at pH 8.5 and 50°C, but could not hydrolyze olive oil. It lost activity with half-lives of 40 min at 70°C and 7 min at 80°C. LC-cutinase* had an ability to degrade poly(ε-caprolactone) and polyethylene terephthalate (PET). The specific PET-degrading activity of LC-cutinase* was determined to be 12 mg/h/mg of enzyme (2.7 mg/h/μkat of pNP-butyrate-degrading activity) at pH 8.0 and 50°C. This activity is higher than those of the bacterial and fungal cutinases reported thus far, suggesting that LC-cutinase* not only serves as a good model for understanding the molecular mechanism of PET-degrading enzyme but also is potentially applicable for surface modification and degradation of PET.


Ginsenoside Rb1 ameliorates CKD-associated vascular calcification by inhibiting the Wnt/β-catenin pathway.
Vascular calcification (VC) is a pathological process underpinning major cardiovascular conditions and has attracted public attention due to its high morbidity and mortality. Chronic kidney disease (CKD) is a common disease related to VC. Ginsenoside Rb1 (Rb1) has been reported to protect the cardiovascular system against vascular diseases, yet its role in VC and the underlying mechanisms remain unclear. In this study, we established a CKD-associated VC rat model and a β-glycerophosphate (β-GP)-induced vascular smooth muscle cell (VSMC) calcification model to investigate the effects of Rb1 on VC. Our results demonstrated that Rb1 ameliorated calcium deposition and VSMC osteogenic transdifferentiation both in vivo and in vitro. Rb1 treatment inhibited the Wnt/β-catenin pathway by activating peroxisome proliferator-activated receptor-γ (PPAR-γ), and confocal microscopy was used to show that Rb1 inhibited β-catenin nuclear translocation in VSMCs. Furthermore, SKL2001, an agonist of the Wnt/β-catenin pathway, compromised the vascular protective effect of Rb1. GW9662, a PPAR-γ antagonist, reversed Rb1's inhibitory effect on β-catenin. These results indicate that Rb1 exerted anticalcific properties through PPAR-γ/Wnt/β-catenin axis, which provides new insights into the potential theraputics of VC.


Gene-gene interactions in the folate metabolic pathway and the risk of conotruncal heart defects.
Conotruncal and related heart defects (CTRD) are common, complex malformations. Although there are few established risk factors, there is evidence that genetic variation in the folate metabolic pathway influences CTRD risk. This study was undertaken to assess the association between inherited (i.e., case) and maternal gene-gene interactions in this pathway and the risk of CTRD. Case-parent triads (n = 727), ascertained from the Children's Hospital of Philadelphia, were genotyped for ten functional variants of nine folate metabolic genes. Analyses of inherited genotypes were consistent with the previously reported association between MTHFR A1298C and CTRD (adjusted P = .02), but provided no evidence that CTRD was associated with inherited gene-gene interactions. Analyses of the maternal genotypes provided evidence of a MTHFR C677T/CBS 844ins68 interaction and CTRD risk (unadjusted P = .02). This association is consistent with the effects of this genotype combination on folate-homocysteine biochemistry but remains to be confirmed in independent study populations.


Human c-Ki-ras2 proto-oncogene on chromosome 12.
A human colonic adenocarcinoma transforming gene, recently identified as a cellular homolog of the Kirsten sarcoma gene (v-ras), was used to assign the human cellular Kirsten ras2 gene to chromosome 12 by the Southern hybridization method. A single 640 base-pair Eco RI--Hind III fragment of the transforming gene, isolated by DNA transfection and molecular cloning, can detect a single Eco RI fragment (2.9 kilobase pairs) of DNA from phenotypically normal cells. The data suggest a constant chromosomal location of c-Ki-ras2.


Superconductivity in the PbO-type structure alpha-FeSe.
The recent discovery of superconductivity with relatively high transition temperature (Tc) in the layered iron-based quaternary oxypnictides La[O(1-x)F(x)] FeAs by Kamihara et al. [Kamihara Y, Watanabe T, Hirano M, Hosono H (2008) Iron-based layered superconductor La[O1-xFx] FeAs (x = 0.05-0.12) with Tc = 26 K. J Am Chem Soc 130:3296-3297.] was a real surprise and has generated tremendous interest. Although superconductivity exists in alloy that contains the element Fe, LaOMPn (with M = Fe, Ni; and Pn = P and As) is the first system where Fe plays the key role to the occurrence of superconductivity. LaOMPn has a layered crystal structure with an Fe-based plane. It is quite natural to search whether there exists other Fe based planar compounds that exhibit superconductivity. Here, we report the observation of superconductivity with zero-resistance transition temperature at 8 K in the PbO-type alpha-FeSe compound. A key observation is that the clean superconducting phase exists only in those samples prepared with intentional Se deficiency. FeSe, compared with LaOFeAs, is less toxic and much easier to handle. What is truly striking is that this compound has the same, perhaps simpler, planar crystal sublattice as the layered oxypnictides. Therefore, this result provides an opportunity to better understand the underlying mechanism of superconductivity in this class of unconventional superconductors.


Clinical observation, pharmacotherapy and referral on discharge of patients with anxiety disorder in a psychiatric emergency service.
To analyze factors associated with clinical observation, pharmacotherapy and referral on discharge of patients with anxiety disorder (AD) seeking care at a psychiatric emergency unit. A total of 5003 consecutive visits were reviewed over a three-year period at a psychiatric emergency service in a tertiary university hospital. Data collected included sociodemographic and clinical information as well as the Global Assessment of Functioning (GAF) and the Severity Psychiatric Illness (SPI) scale scores. Of all the visits, 992 (19.8%) were diagnosed of AD. Of these, 19.6% required clinical observation and 72.2% were referred to a psychiatrist at discharge. Regression analysis showed that referral to psychiatry was associated with being male, native, psychiatric background, greater severity, lower global functioning, and behavioral disorders. Clinical observation (in a box) was associated with being female, greater severity, and psychotic or behavioral symptoms. Prescription of benzodiazepines was associated with anxiety, no history of addiction, and lower global functioning. Antidepressants were associated with being a native, anxiety with no history of addiction, and lower functioning. Antipsychotics were associated with being native, psychiatric background (not addiction), anxiety, and lower functioning. Behavior, psychiatric background and illness severity were determinants of referral to a specialist. Besides these, psychotic symptoms and non-specific clinical symptoms were determinants of observation. Drug prescription in AD is less frequent if the main complaint is not anxiety and depends more on the level of functioning than on that of severity.


Heme Oxygenase-1 Expression Affects Murine Abdominal Aortic Aneurysm Progression.
Heme oxygenase-1 (HO-1), the rate-limiting enzyme in heme degradation, is a cytoprotective enzyme upregulated in the vasculature by increased flow and inflammatory stimuli. Human genetic data suggest that a diminished HO-1 expression may predispose one to abdominal aortic aneurysm (AAA) development. In addition, heme is known to strongly induce HO-1 expression. Utilizing the porcine pancreatic elastase (PPE) model of AAA induction in HO-1 heterozygous (HO-1+/-, HO-1 Het) mice, we found that a deficiency in HO-1 leads to augmented AAA development. Peritoneal macrophages from HO-1+/- mice showed increased gene expression of pro-inflammatory cytokines, including MCP-1, TNF-alpha, IL-1-beta, and IL-6, but decreased expression of anti-inflammatory cytokines IL-10 and TGF-beta. Furthermore, treatment with heme returned AAA progression in HO-1 Het mice to a wild-type profile. Using a second murine AAA model (Ang II-ApoE-/-), we showed that low doses of the HMG-CoA reductase inhibitor rosuvastatin can induce HO-1 expression in aortic tissue and suppress AAA progression in the absence of lipid lowering. Our results support those studies that suggest that pleiotropic statin effects might be beneficial in AAA, possibly through the upregulation of HO-1. Specific targeted therapies designed to induce HO-1 could become an adjunctive therapeutic strategy for the prevention of AAA disease.


Association of Rgs7/Gβ5 complexes with Girk channels and GABAB receptors in hippocampal CA1 pyramidal neurons.
In the hippocampus, signaling through G protein-coupled receptors is modulated by Regulators of G protein signaling (Rgs) proteins, which act to stimulate the rate of GTP hydrolysis, and consequently, G protein inactivation. The R7-Rgs subfamily selectively deactivates the G(i/o)-class of Gα subunits that mediate the action of several GPCRs. Here, we used co-immunoprecipitation, electrophysiology and immunoelectron microscopy techniques to investigate the formation of macromolecular complexes and spatial relationship of Rgs7/Gβ5 complexes and its prototypical signaling partners, the GABAB receptor and Girk channel. Co-expression of recombinant GABAB receptors and Girk channels in combination with co-immunoprecipitation experiments established that the Rgs7/Gβ5 forms complexes with GABAB receptors or Girk channels. Using electrophysiological experiments, we found that GABAB -Girk current deactivation kinetics was markedly faster in cells coexpressing Rgs7/Gβ5. At the electron microscopic level, immunolabeling for Rgs7 and Gβ5 proteins was found primarily in the dendritic layers of the hippocampus and showed similar distribution patterns. Immunoreactivity was mostly localized along the extrasynaptic plasma membrane of dendritic shafts and spines of pyramidal cells and, to a lesser extent, to that of presynaptic terminals. Quantitative analysis of immunogold particles for Rgs7 and Gβ5 revealed an enrichment of the two proteins around excitatory synapses on dendritic spines, virtually identical to that of Girk2 and GABAB1 . These data support the existence of macromolecular complexes composed of GABAB receptor-G protein-Rgs7-Girk channels in which Rgs7 and Gβ5 proteins may preferentialy modulate GABAB receptor signaling through the deactivation of Girk channels on dendritic spines. In contrast, Rgs7 and Girk2 were associated but mainly segregated from GABAB1 in dendritic shafts, where Rgs7/Gβ5 signaling complexes might modulate Girk-dependent signaling via a different metabotropic receptor(s).


Exclusive Breastfeeding and Risk of Dental Malocclusion.
The distinct effect of exclusive and predominant breastfeeding on primary dentition malocclusions is still unclear. We hypothesized that exclusive breastfeeding presents a higher protective effect against malocclusions than predominant breastfeeding and that the use of a pacifier modifies the association between breastfeeding and primary dentition malocclusions. An oral health study nested in a birth cohort study was conducted at age 5 years (N = 1303). The type of breastfeeding was recorded at birth and at 3, 12, and 24 months of age. Open bite (OB), crossbite, overjet (OJ), and moderate/severe malocclusion (MSM) were assessed. Poisson regression analyses were conducted by controlling for sociodemographic and anthropometric characteristics, sucking habits along the life course, dental caries, and dental treatment. Predominant breastfeeding was associated with a lower prevalence of OB, OJ, and MSM, but pacifier use modified these associations. The same findings were noted between exclusive breastfeeding and OJ and between exclusive breastfeeding and crossbite. A lower prevalence of OB was found among children exposed to exclusive breastfeeding from 3 to 5.9 months (33%) and up to 6 months (44%) of age. Those who were exclusively breastfed from 3 to 5.9 months and up to 6 months of age exhibited 41% and 72% lower prevalence of MSM, respectively, than those who were never breastfed. A common risk approach, promoting exclusive breastfeeding up to 6 months of age to prevent childhood diseases and disorders, should be an effective population strategy to prevent malocclusion.


Primary lymphomas of the small intestine: east-west contrast.
Primary small intestinal lymphoma (PSIL) represents a heterogenous group of disorders with variable clinical and pathologic features and a characteristic age, socioeconomic, and geographic distribution. In developed countries, PSIL usually occurs as a localized ileal tumor, shows a bimodal age distribution, and most frequently presents with abdominal pain and obstructive symptoms. Histologically, most of these tumors are diffuse histiocytic, lymphocytic, or undifferentiated lymphomas. Other variants of PSIL, collectively referred to as immunoproliferative small intestinal disease, occur most often among young patients of poor socioeconomic status in Third World countries, mostly in the Middle East and Mediterranean area. They are characterized by involvement of long loops of the upper small intestine and commonly present with abdominal pain, diarrhea, malabsorption, and clubbing of the fingers. A subgroup of these patients shows a serological abnormality with the appearance of part of the alpha heavy chain of IgA in the serum. Histologically, the lesion appears as a dense diffuse lymphoplasmacytic infiltrate of the mucosa of the upper jejenum or duodenum. A form of malignant lymphoma of true histiocytic origin complicates long-standing celiac disease. The contrasting clinical, epidemiological, histopathological, and immunological features of these variants of PSIL raise interesting questions about the pathogenesis of small bowel lymphoma.


The effect of smoking on serum IgG2 reactive with Actinobacillus actinomycetemcomitans in early-onset periodontitis patients.
High titers of serum IgG2 reactive with Actinobacillus actinomycetemcomitans are present in early-onset periodontitis (EOP) patients and it appears that anti-A. actinomycetemcomitans may be protective. Smoking is associated with increased periodontal disease severity in generalized early-onset periodontitis (G-EOP) patients, but is not associated with periodontal disease severity in patients with localized juvenile periodontitis (LJP). Furthermore, smoking is associated with reduced serum IgG2 levels in black patients with G-EOP but not in those with LJP. Based on this selective effect of smoking, we hypothesized that smoking would be associated with a reduction of specific IgG2 reactive with A. actinomycetemcomitans in black G-EOP patients but not black LJP patients. In addition, we examined IgG2 responses to carbohydrate antigens from non-periodontal pathogens including Haemophilus influenzae b oligosaccharide antigen (Hib) and the Streptococcus pneumoniae antigen phosphocholine (PC). Smoking status was assessed from serum cotinine levels, and IgG2 specific for A. actinomycetemcomitans, Hib, and PC was assessed by ELISA. Our study revealed that smoking was correlated with a dramatic reduction in serum IgG2 anti-A. actinomycetemcomitans in G-EOP smokers but not in LJP smokers. In contrast, anti-Hib IgG2 and anti-PC IgG2 were not affected in either G-EOP or LJP patients. In short, these results indicate that smoking is associated with a reduction in serum IgG2 anti-A. actinomycetemcomitans in black G-EOP subjects, but IgG2 reactive with other antigens may not be reduced in G-EOP smokers.


Eccrine Syringofibroadenoma in Association With Acquired Epidermodysplasia Verruciformis.
A 75-year-old man with human immunodeficiency virus infection and numerous biopsy-proven warts for 10 years, refractory to cryosurgery, cimetidine, and topical imiquimod, presented with numerous pink to hypopigmented verrucous papules and plaques involving the face, trunk, buttocks, and groin. Laboratory evaluation revealed a CD4 T-cell count of 62 cells per microliter and human immunodeficiency virus viral load of <117 copies per milliliter. Biopsy of a plaque groin lesion was performed. Histopathology revealed vertically oriented anastomosing strands of basaloid epithelium arising from multiple points along the epidermis in a background fibrovascular stroma. Ductal differentiation was identified. Areas of epidermis showed compact orthokeratosis, coarse hypergranulosis, and keratinocytes with abundant steel-blue-gray cytoplasm, indicative of viral cytopathic changes. Cytologic atypia was not identified. Human papillomavirus (HPV) genotyping of this lesion was positive for types 5 and 14. Overall, the findings were consistent with epidermodysplasia verruciformis in association with eccrine syringofibroadenoma (ESFA). The patient was subsequently treated with acitretin and showed clinical improvement. ESFA is an uncommon benign adnexal tumor with unknown pathogenesis. Although its association with HPV has rarely been reported, ESFA in the setting of acquired epidermodysplasia verruciformis has not been described. The development of ESFA in this case may be the result of HPV-induced cellular transformation.


Intestinal immune responses in patients infected with enterotoxigenic Escherichia coli and in vaccinees.
Immune responses against enterotoxigenic Escherichia coli (ETEC) were examined in Bangladeshi adults with naturally acquired disease and compared to responses in age-matched Bangladeshi volunteers who had been orally immunized with a vaccine consisting of inactivated ETEC bacteria expressing different colonization factor antigens (CFs) and the B subunit of cholera toxin. B-cell responses in duodenal biopsy samples, feces, intestinal washings, and blood were determined. Because most of the patients included in the study were infected with ETEC expressing CS5, immune responses to this CF were studied most extensively. Vaccinees and patients had comparable B-cell responses against this antigen in the duodenum: the median numbers of antibody-secreting cells (ASC) were 3,300 immunoglobulin A (IgA) ASC/10(7) mononuclear cells (MNC) in the patient group (n = 8) and 1,200 IgA ASC/10(7) MNC in the vaccinees (n = 13) (not a significant difference). Similarly, no statistically significant differences were seen in the levels of duodenal B cells directed against enterotoxin among vaccinees and patients. A comparison of the capacities of the various methods used to assess mucosal immune responses revealed a correlation between numbers of circulating B cells and antibody levels in saponin extracts of duodenal biopsy samples (r = 0.58; n = 13; P = 0.04) after vaccination. However, no correlation was seen between blood IgA ASC and duodenal IgA ASC after two doses of vaccine. Still, a correlation between numbers of CF-specific B cells in blood sampled from patients early during infection and numbers of duodenal B cells collected 1 week later was apparent (r = 0.70; n = 10; P = 0.03).


Enhanced podocalyxin expression alters the structure of podocyte basal surface.
Glomerular basement membrane (GBM) and podocalyxin are essential for podocyte morphology. We provide evidence of functional interconnections between basement membrane components (collagen IV and laminin), the expression of podocalyxin and the morphology of human glomerular epithelial cells (podocytes). We demonstrated that GBM and laminin, but not collagen IV, up-regulated the expression of podocalyxin. Scanning electron microscopy revealed that laminin induced a modified morphology of podocytes with process formation, which was more extensive in the presence of GBM. Under high magnification, podocytes appeared ruffled. Using transmission electron microscopy we observed that raised areas occurred in the basal cell surface. Furthermore, the presence of anti-podocalyxin antibody increased the extent of adhesion and spreading of podocytes to both collagen IV and laminin, thus podocalyxin apparently inhibits cell-matrix interactions. We also performed adhesion and spreading assays on podocytes grown under increased glucose concentration (25 mM). Under these conditions, the expression of podocalyxin was almost totally suppressed. The cells adhered and spread to basement membrane components but there was no increase in the extent of adhesion and spreading in the presence of anti-podocalyxin antibody, or ruffling of the cell edges. Additionally, in podocytes expressing podocalyxin, the presence of anti-podocalyxin antibody partially reversed the inhibition of adhesion to collagen IV provoked by anti-beta1 integrin antibody, thus podocalyxin should compete with beta1-related cell adhesion. We suggest that the observed podocalyxin-mediated inhibition of binding to the matrix could be in part responsible for the specialized conformation of the basal surface of podocytes.


Treatment of acute focal cerebral ischemia with propranolol.
Propranolol has been found to have a protective effect in experimental myocardial ischemia. Protection of ischemic kidneys was subsequently demonstrated following treatment with propranolol and its weaker beta blocking isomer, d-propranolol. The objective of the present investigation was to study the effects of propranolol (i.e., racemic d,1 mixture) and d-propranolol upon regional cerebral blood flow (rCBF) and early ischemic changes following experimental middle cerebral artery (MCA) occlusion. Thirty adult cats, lightly anesthetized with ketamine hydrochloride, underwent 3 hours or right MCA occlusion. Ten cats were untreated. Ten cats were given a continuous infusion of propranolol (1 mg/kg/hr) for 4 hours beginning 1 hour before MCA occlusion and a 4 mg/kg bolus immediately before occlusion. Ten cats were given a continuous infusion of d-propranolol (0.5 mg/kg/hr) for 4 hours beginning 1 hour before MCA occlusion and a 2 mg/kg bolus immediately before occlusion. The therapeutic agents were injected directly into the right carotid artery. The rCBF in the right Sylvian region was not significantly different in the 3 groups. EEG changes also were similar. Carbon filling defects were found to be smallest in the d-propranolol-treated group. Light microscopic studies demonstrated a reduction in infarct size in the propranolol and d-propranolol groups. The findings of the investigation indicated that propranolol and d-propranolol do not have a deleterious effect on rCBF after MCA occlusion and suggested that these agents have a protective effect upon ischemic cerebral tissue.


Second capsule endoscopy for patients with severe iron deficiency anemia.
Patients with iron deficiency anemia are subjected to multiple endoscopic and radiologic examinations of the GI tract. If negative, some of the examinations are repeated, occasionally with positive findings. The diagnostic yield of a second capsule endoscopy in such patients is unknown. The aim of the current study was to assess the diagnostic yield of a second capsule endoscopy in patients with significant iron deficiency anemia and a previous negative evaluation. Twenty patients with iron deficiency anemia (Hb < 10 g/dL) were enrolled. All had at least one normal evaluation of the GI tract, including capsule endoscopy. A second capsule endoscopy examination was offered to all patients. The time between the first and the second capsule endoscopy ranged from 2 months to 1 year. Depending on the nature of an abnormality and its relevance to blood loss, the findings were classified as positive, suspicious, clinically irrelevant, or negative. In 7 patients, the second capsule endoscopy disclosed findings that were classified as either positive or suspicious findings, including arteriovenous malformations (2), flat polypoid lesion (1), edematous inflamed mucosa (1), erosions (1), and hemorrhagic gastritis (1). Lesions were located in the small intestine (5), the stomach (1), and the cecum (1). Based on the findings of the second capsule endoscopy, therapy was changed in two patients (10%). A second capsule endoscopy should be considered for patients with severe iron deficiency anemia and negative initial evaluation.


Structural aspects of the extracellular matrix of the tendon: an atomic force and scanning electron microscopy study.
The mutual interactions of small proteoglycans with collagen fibrils in the extracellular matrix remain to be completely understood. The present research investigated the extracellular matrix of the rat tail tendon by atomic force microscopy (AFM) as well as by scanning electron microscopy (SEM). Observations showed simply dehydrated specimens made of large heterogeneous fibrils, tightly packed in mutual contact with no visible interfibrillar spaces. Proteoglycans usually extended onto neighboring fibrils, forming an intricate interfibrillar weaving highly sensitive to chondroitinase digestion. Pre-treatment with cupromeronic blue only affected the proteoglycans side chains, which appeared better preserved but somewhat thickened. Observation of hydrated specimens by AFM confirmed the close packing of collagen fibrils and the abundance of collagen-bound proteoglycans. Interfibrillar bridges were only occasionally observed in this tissue, whose fibrils are instead tightly bound together by proteoglycans in a structure quite consistent with its functional requirements. The molecular machinery responsible for these interactions is the subject of ongoing research.


Therapeutic trends in pancreatic ductal adenocarcinoma (PDAC).
Prognosis remains dismal for pancreatic ductal adenocarcinoma (PDAC). Genomics and proteomics have depicted heterogeneity in PDAC. Collectively, this information could be useful in improving diagnosis, prognosis, modalities of therapy, treatment responses, deciphering drug resistance and new drug development. We describe major advances in the cellular and molecular subtypes based on next-generation sequencing and their predictive and prognostic value in PDAC patients. We review aberrant genes involving in defined cellular processes in PDAC. Finally, the current state of drug development with novel investigational agents targeting cell fate, cell survival, genomic instability, tumor-stroma, and immune checkpoints are discussed. Molecular techniques have revealed distinct driver mutations in PDAC. Common genes and cellular processes are dysregulated in the pathogenesis of PDAC. These cellular processes categorized by aberrant pathways include control cell fate, genome maintenance, and cell survival. Dysregulation of the tumor microenvironment promotes an intense fibrosis and immune suppression that play a major role in drug resistance. New information on tumor biology has led to the development of targeted/stromal therapies, immunotherapies or combinations with current chemotherapy in PDAC. New drug development targeting multiple hallmarks of PDAC we hope will positively impact the quality and survival of PDAC patients.


Affect dysregulation in borderline personality disorder and somatoform disorder: differentiating under- and over-regulation.
Although affect dysregulation is considered a core component of borderline personality disorder (BPD) and somatoform disorders (SoD), remarkably little research has focused on prevalence and nature of affect dysregulation in these disorders. BPD and SoD diagnoses were confirmed or ruled out in 472 psychiatric inpatients using clinical interviews. Three qualitatively different forms of affect dysregulation were identified: under-regulation, over-regulation of affect and combined under- and over-regulation of affect. BPD was associated with under-regulation of affect, and SoD was associated with over-regulation of affect. However, one in five patients with BPD also reported substantial over-regulation, and one in six patients with SoD reported clinically significant under-regulation, and the comorbid BPD and SoD group reported more frequently both over- and under-regulation than patients diagnosed with BPD or SoD alone or those with other psychiatric disorders.


Modelling retailer-based exemptions in flavoured tobacco sales restrictions: national estimates on the impact of product availability.
More than 250 US localities restrict sales of flavoured tobacco products (FTPs), but comprehensiveness varies, and many include retailer-based exemptions. The purpose of this study is to examine resulting changes in the US retail environment for FTPs if there was a hypothetical national tobacco control policy that would prohibit FTP sales in all retailers except (1) tobacco specialty stores or (2) tobacco specialty stores and alcohol outlets. A cross-sectional analysis of the FTP retail environment in every US Census tract (n=74 133). FTP retailers (n=3 10 090) were enumerated using nine unique codes from a national business directory (n=296 716) and a national vape shop directory (n=13 374). We assessed FTP availability using static-bandwidth and adaptive-bandwidth kernel density estimation. We then calculated the proportion of FTP stores remaining and the mean density of FTP retailers under each policy scenario for the overall population, as well as across populations vulnerable to FTP use. Exempting tobacco specialty stores alone would leave 25 276 (8.2%) FTP retailers nationwide, while exempting both tobacco specialty stores and alcohol outlets would leave 54 091 (17.4%) retailers. On average, the per cent remaining FTP availability per 100 000 total population was 7.1% for a tobacco specialty store exemption and 18.1% for a tobacco specialty store and alcohol outlet exemption. Overall, density estimate trends for remaining FTP availability among racial/ethnic populations averaged across Census tracts mirrored total population density. However, estimates varied when stratified by metropolitan status. Compared with the national average, FTP availability would remain 47%-49% higher for all racial/ethnic groups in large metropolitan areas. Retailer-based exemptions allow greater FTP availability compared with comprehensive policies which would reduce FTP availability to zero. Strong public policies have the greatest potential impact on reducing FTP availability, particularly among urban, and racial/ethnic minority populations.


Effects of simultaneous intakes of fish oil and green tea extracts on plasma, glucose, insulin, C-peptide, and adiponectin and on liver lipid concentrations in mice fed low- and high-fat diets.
The purpose of this study was to clarify the effects of a simultaneous intake of fish oil and green tea extracts (GTE) on plasma lipids, glucose (Glu), insulin, C-peptide, and adiponectin concentrations and on liver lipids in mice fed low- and high-fat diets. Male mice (5 months old) were fed low-fat (5%) or high-fat (20%) diets containing 0% (5 or 20% lard), 1% (4 or 19% lard) or 5% (0 or 15% lard) fish oil with or without 1% GTE for 3 months. Single or combined intakes of fish oil and GTE intakes significantly lowered plasma and liver total cholesterol (T-chol) and triacylglycerol (TG) concentrations in mice on high-fat diets. However, a positive effect of GTE alone was not observed in the plasma T-chol and TG concentrations of mice on low-fat diets. Plasma Glu concentrations were significantly lowered by dietary fish oil in mice on the low- and high-fat diets. A tendency of GTE intake to decrease plasma Glu in mice on both the low- and high-fat diets was not significant. An intake of GTE only minimally influenced plasma insulin, C-peptide, and adiponectin concentrations, but fish oil supplementation increased the adiponectin concentration in mice on the low- and high-fat diets. GTE addition to the diets had no adverse effects on reductions by fish oil. These results suggest that a simultaneous intake of fish oil and GTE can improve plasma and liver lipid and plasma Glu profiles in mice fed a high-fat diet without any negative interactions of fish oil and GTE.


Flowering as metamorphosis: two sequential signals regulate floral initiation in Lolium temulentum.
We investigated floral initiation in the long-day monocot Lolium temulentum, strain Ceres, by culturing apices explanted from photoperiodically induced plants at various times after one inductive long day onto medium with, and without, gibberellin. Apices cultured on the first day after the inductive long day usually required gibberellin in the medium to initiate floral morphogenesis while apices explanted on the second day after induction did not require gibberellin. Apices explanted on the first day after induction onto medium without gibberellin grew vegetatively for many days but a several-day exposure to culture medium with gibberellin at any time caused most apices to initiate floral morphogenesis. The gibberellin synthesis inhibitor, ancymidol, when applied to plants before apex excision and when present in the culture medium reduced floral initiation by more than 50% in the absence of added gibberellin in the medium, but it was ineffective in the presence of gibberellin. These results indicated that floral initiation in photoperiodically induced plants resulted from two signals acting at the apex. The first signal induced the apex into a florally determined state and then the second signal, gibberellin, elicited expression of the florally determined state. Leaf removal and culture of apices from plants previously treated with gibberellin provided evidence that the leaf-applied gibberellin did not itself act on the apex to cause floral determination or initiation. Rather, the exogenous gibberellin appeared to stimulate the production of a signal in the leaves that then led to floral initiation.


Repeated imipramine treatment enhances the 7-OH-DPAT-induced hyperactivity in rats: the role of dopamine D2 and D3 receptors.
Previous studies have shown that antidepressant drugs with different pharmacological profiles, administered repeatedly, increase the locomotor hyperactivity induced by various dopaminomimetics, among others by (+/-)7-hydroxydipropylaminotetralin (7-OH-DPAT). Since, according to a recent study, this drug shows a high affinity for not only dopamine D3 but also dopamine D2 receptors, a question arises whether dopamine D3 receptors are involved in the increase in 7-OH-DPAT-elicited locomotor hyperactivity induced by repeated treatment with antidepressant drugs. The aim of the present study was to investigate the effect of imipramine (IMI), administered repeatedly, on the hyperactivity induced by 7-OH-DPAT, a dopamine D3 receptor-preferring agonist. Male Wistar rats were treated with IMI (10 mg/kg po) either acutely (single dose) or repeatedly (twice daily for 14 days). The locomotor hyperactivity induced by 7-OH-DPAT (3 mg/kg sc) was measured in photoresistor actometers. The influence of nafadotride (0.2 and 0.4 mg/kg ip), a dopamine D3-preferring antagonist or sulpiride (10 and 25 mg/kg ip), a dopamine D2/D3 antagonist, on the 7-OH-DPAT-induced locomotor hyperactivity was studied. Nafadotride (in both doses used) or sulpiride (in the higher dose only) reduced (by about 50%) the hyperactivity induced by 7-OH-DPAT. Combined treatment with nafadotride (0.2 mg/kg) and sulpiride (25 mg/kg) completely abolished the effect of 7-OH-DPAT. IMI administered repeatedly (but not acutely) enhanced the 7-OH-DPAT-induced hyperactivity. Neither nafadotride, 0.2 mg/kg (or sulpiride, 10 mg/kg), given alone nor combined treatment with both these substances changed the hyperactivity induced by repeated treatment with IMI and 7-OH-DPAT (given 2 h after the last dose of IMI). Joint treatment with nafadotride, 0.2 mg/kg, and sulpiride, 25 mg/kg, completely abolished the enhancing effect of repeated treatment with IMI and 7-OH-DPAT. The above results indicate that both types of dopamine receptors, D3 and D2, may play a substantial role in the mechanism of the 7-OH-DPAT-induced hyperactivity, as well as in the increase evoked by repeated treatment with IMI in rats.


[Flowmetric assessment of coronary bypass grafts in the conditions of artificial circulation and on the beating heart].
Advantages and shortcomings of aortocoronary bypass grafting on the beating heart and in the conditions of artificial circulation (AC) have long been discussed. The data on patency of bypass grafts in the remote period are indicative of comparable results of operations with and without AC or advantages of using AC. In order to determine benefits of each method it is necessary to reveal intraoperative predictors of bypass grafts occlusion in the remote period. We analyzed the results of ultrasound flowmetry of the blood flow through the left internal thoracic artery during bypass grafting of the anterior descending artery with the use of AC and on the beating heart. A retrospective study included a total of 352 patients subdivided into 2 groups: Group One was composed of 120 patients undergoing surgery in the conditions of AC and Group Two comprised 232 patients subjected to similar operations on the beating heart. Blood flow was measured with the help of flowmeter VeryQ MediStim® after termination of AC and inactivation of heparin by protamine, with systolic pressure of 100-110 mm Hg. There were no statistically significant differences between the groups by the diameter and degree of stenosis of the anterior descending artery, diameter of the left internal thoracic artery. The mean volumetric blood flow velocity (Qmean) along the shunts in Group One was higher (p=0.01). No statistically significant differences by the pulsatility index (PI) between the groups were revealed (p=0.2). A conclusion was drawn that coronary bypass grafting of the anterior descending artery by the left internal thoracic artery in the conditions of artificial circulation made it possible to achieve higher volumetric velocity of blood flow through the conduit as compared with operations on the beating heart, with similar resistance index. The immediate results of the operations with the use of the both techniques did not differ.


Overexpression of p34cdc2 protein kinase in epithelial ovarian carcinoma.
To investigate the role of expression of p34cdc2 protein kinase in normal, benign, and malignant ovarian epithelium. Tissue sections from 24 patients with epithelial ovarian carcinoma (EOC) along with 6 normal ovarian specimens and 12 benign cystadenomas were incubated with mouse IgG monoclonal antibody to human p34cdc2 protein kinase, followed by detection with use of a standard peroxidase-labeled streptavidin-biotin technique. Immunohistochemical staining was graded and compared. Clinical data were also reviewed. Normal surface epithelium and 10 of 12 benign cystadenomas failed to stain for p34cdc2 protein kinase. Of the 24 EOC specimens, however, 19 (79%) stained positively. The staining pattern or intensity was not associated with the histologic grade or surgical stage. Expression of p34cdc2 protein kinase is strongly up-regulated in most cases of EOC but not in normal epithelial ovarian tissue or in most cases of benign epithelial tumors evaluated. Therefore, it may be associated with early events in carcinogenesis. Redundant overexpression of cyclin-dependent kinases such as p34cdc2 may contribute to deranged cell cycle progression and proliferation of EOC. Observation of overexpression of p34cdc2 protein kinase in other malignant lesions suggests a common mechanism.


Resistance to apoptosis: mechanism for the development of HIV reservoirs.
New insights into the physiological cell death program in mammalian cells have further confounded the central issue of T lymphocyte destruction during HIV infection. Although infected and uninfected cells die following infection, recent evidence indicates that infected and uninfected cells may have unique pathways controlling death. Two widely accepted models for apoptosis have been described, namely the intrinsic and extrinsic apoptotic pathway. In the extrinsic pathway, ligation of TNF death ligands to their receptors causes oligomerization of the death receptors and recruitment of adaptor proteins typically involving caspase 8 activation. In the intrinsic pathway, apoptotic signals converge on mitochondria to cause activation and subsequent release of cytochrome c, which leads to formation of a multiprotein complex containing Apaf-1, cytochrome c, dATP and procaspase 9. Expression of HIV proteins alters these pathways resulting in enhanced levels of apoptosis. Although HIV infection results in T cell apoptosis, under some circumstances a small fraction of CD4+ T cells and macrophages do not die following infection, indicating that this may be a critical step in the development of viral reservoirs. In addition, monocyte differentiation into macrophages leads to an apoptosis resistant phenotype characterized by upregulation of antiapoptotic molecules and lower levels of proapoptotic molecules. The development of these stable antiretroviral resistant cells represents the major obstacle in achieving a complete sterile cure. However, this provides a unique opportunity to further understand the regulation of apoptosis and may facilitate novel immune based therapies aimed at modifying apoptosis in HIV disease.


A cross-sectional study on selected child health outcomes in India: Quantifying the spatial variations and identification of the parental risk factors.
This study examined association between selected child health indicators- anaemia, stunting and no/incomplete immunization by inter-linking maternal characteristics at district level and parental characteristics at individual level. A spatial analysis and a binary logit model estimation were employed to draw inferences using the data from the fourth round of National Family Health Survey, 2015-16 of India. Significant spatial clustering of the selected child health outcomes was observed in the country. Mother's educational attainment explained significant district level differential in the selected child health outcomes. At the individual level, parents who are very young, not-educated, socially excluded, belong to poor class were found to be significantly associated with the poor child health outcomes. This study indicates that parental characteristics, such as age, educational attainment and employment substantially determine child health in India, suggesting that an intervention by targeting the households where children are vulnerable is important to improve child health in the country.


Policy statement--Pediatrician-family-patient relationships: managing the boundaries.
All professionals are concerned about maintaining the appropriate limits in their relationships with those they serve. Pediatricians should be aware that, under normal circumstances, caring for one's own children presents significant ethical issues. Pediatricians also must strive to maintain appropriate professional boundaries in their relationships with the family members of their patients. Pediatricians should avoid behavior that patients and parents might misunderstand as having sexual or inappropriate social meaning. Romantic and sexual involvement between physicians and patients is unacceptable. The acceptance of gifts or nonmonetary compensation for medical services has the potential to affect the professional relationship adversely.


Induction of apoptosis and reversal of permeability glycoprotein-mediated multidrug resistance of MCF-7/ADM by ginsenoside Rh2.
Multidrug resistance is a phenomenon that cancer cells develop a cross-resistant phenotype against several unrelated drugs, and permeability glycoprotein derived from the overexpression of multidrug resistance gene 1 has been taken as the most significant cause of multidrug resistance. In the present study, ginsenoside Rh2 was used to reverse permeability glycoprotein-mediated multidrug resistance of MCF-7/ADM cell line. Effects of ginsenoside Rh2 on the apoptotic process and caspase-3 activity of MCF-7 and MCF-7/ADM cell lines were determined using flow cytometry and microplate reader. Methyl thiazolyl tetrazolium test was conducted to assess the IC50 values of ginsenoside Rh2 and adriamycin on MCF-7 and MCF-7/ADM cultures; Rhodamin 123 assay was used to assess the retention of permeability glycoprotein after ginsenoside Rh2 treatment; flow cytometry and real time polymerase chain reaction were used to determine the expression levels of permeability glycoprotein and multidrug resistance gene 1 in drug-resistant cells and their parental cells after exposure to ginsenoside Rh2. The results showed that ginsenoside Rh2, except for inducing apoptosis, had the ability to reverse multidrug resistance in MCF-7/ADM cell line without changing the expression levels of permeability glycoprotein and multidrug resistance gene 1. Our findings provided some valuable information for the application of ginsenoside Rh2 in cancer therapy, especially for multidrug resistance reversal in clinic.


Publish or perish, information overload, and journal impact factors - A conflicting tripod of forces.
The academic mantra, to the point of cliché, is "publish or perish." Academia is generally too preoccupied with research and publishing to stand back and consider the driving forces behind the actual processes and systems involved. Indeed, academics are generally unaware of the factors that influence one's ability to publish: The drive to publish itself, readers' information overload, and editors' desire to increase journals' impact factors. This paper will detail these forces, and it behooves potential researchers to keep this veritable tripod of forces in mind since understanding the tripod may facilitate publication chances through the invocation and active implementation of news media theory. Media writers' remuneration is dependent on readers clicking on their articles. The media reel in readers by displaying an intriguing/bold/provocative headline and then keep the readers interested and hooked with initial sentences that not only give information but also tantalize with the promise of more to come. A paper's title and abstract should adhere to these precepts so as to increase the chances of avoiding immediate rejection at editorial or initial reviewer level.


A Laboratory Exercise Demonstrating the Limited Circumstances in which the Cerebral Cortex is Engaged in Over Ground Locomotion.
For neuroscience, memorable demonstrations of principles in action are crucial. Neural control of walking is particularly difficult to understand because the interaction of the cerebral cortex with a central pattern generator (CPG) makes the mode of control context-dependent. Beginning students tend to consider corticospinal control the basis of all movement, so they may not distinguish the limited circumstances in which the cerebral cortex bypasses the CPG to control leg movements directly for walking. The demonstration described here is designed to show that cortical involvement in normal walking is minimal unless visual control of foot placement is required. Cortical involvement in motor control is assessed by probing for spare attention while a student volunteer performs three different tasks: sitting, walking down a hallway, and walking through an obstacle course. Simple math quizzes with 20 oral questions are the probes. The class observes the demonstration and discusses the results. To evaluate learning, a multiple-choice question was administered two months after the demonstration, as well as 14 months later to cohorts from the previous year's class. The demonstration succeeded: quiz scores were similar for sitting and level walking, but lower for the obstacle course. Two months later, 86% of students correctly answered the multiple choice question; 42% of the previous year's cohorts answered correctly after 14 months. The demonstration shows that the cortex is engaged by walking through an obstacle course, not walking on a flat indoor surface. Initially, most students learned this distinction well, but after a year, many reverted to the idea that the corticospinal tract controls details of leg movements during walking. Thus this result emphasizes the need for review of advanced concepts. Overall, the experience was fun and could easily fit into basic or clinical neuroscience courses.


Thymol as a reciprocal regulator of T cell differentiation: Promotion of regulatory T cells and suppression of Th1/Th17 cells.
Regulatory T cells (Tregs) are critical for maintaining immune response and enhancing their differentiation has therapeutic implications for autoimmune diseases. In this study, we investigated the effects of thymol a well-known monoterpene from Thyme on differentiation and function of Tregs. In vitro generation of Tregs from purified naïve CD4+CD25- T cells in the presence of thymol was carried out. Suppressor activity of generated Tregs was examined by changes in the proliferation of CFSE-labeled conventional T cells. Thymol promotes differentiation of naïve CD4+CD25- T cells to CD4+CD25+Foxp3+ Tregs [66.9-71.8% vs. control (47%)] and increased intensity of Foxp3 expression on Tregs (p < 0.01). In functional assay, an increased immune suppression by thymol-induced Tregs (≈2.5 times of untreated Tregs) was detected. For in vivo study, thymol was intraperitoneally administered to ovalbumin (Ova)-immunized mice. Flow cytometry assessment of spleens from thymol-treated Ova-immunized mice showed increased number of CD4+ Foxp3+ Tregs (>8%, p < 0.01(and decreased levels of CD4+T-bet+ Th1 and CD4+RORγt+ Th17 cells resulted in significant decreased Th1/Treg and Th17/Treg ratios. In ex vivo Ova challenge of splenocytes from thymol-treated Ova-immunized mice, similarly higher levels of CD4+ Foxp3+ Tregs, and also elevated TGF-β expression in CD4+Foxp3+ population (48.1% vs. 18.9% in untreated Ova-immunized group) and reduced IFN-γ-producing CD4+T-bet+ T cells and IL-17-producing CD4+RORγt+ T cells were detected. This led to marked decreased ratios of IFNγ/TGF-β and IL-17/TGF-β expressions. In conclusion, this study revealed thymol as a compound with enhancing effects on Treg differentiation and function, which may have potential benefits in treatment of immune-mediated diseases with Th1/Th17 over-activation.


Association of the positive T wave in lead aVR with short-term mortality in patients with acute pulmonary embolism.
Background: Lead aVR provides prognostic information in various settings for patients with cardiovascular diseases. The present study aimed to investigate the prognostic value of a positive T wave in lead aVR (TaVR) for patients with acute pulmonary embolism (APE).Methods: We screened a total of 412 consecutive patients who were hospitalised with a diagnosis of APE between 2008 and 2018. We investigated electrocardiograms (ECGs) for the presence of a positive TaVR and classified other abnormal ECG findings. Additionally, clinical data, such as echocardiographic findings were recorded, and pulmonary embolism severity index (PESI) scores were calculated. The predictors of mortality at 30 days were investigated as the clinical outcome by logistic regression analysis.Results: In our study population, 54 patients (13.1%) died within 30 days. The prevalence of female gender, congestive heart failure, chronic obstructive pulmonary disease, and mean PESI scores were significantly higher in patients with a positive TaVR compared to those without it. Systolic blood pressure of patients with positive TaVR was significantly lower than that of patients without positive TaVR. In multivariate regression analysis; PESI scores (OR: 1.03; 95% CI: 1.01-1.04, p < .01), Right ventricular end-diastolic diameter (RVEDD) (OR: 1.07 95% CI: 1.01-1.13, p = .02), and a positive TaVR (OR: 4.41; 95% CI: 1.63-11.96, p < .01) were independently correlated with mortality.Conclusion: Positive TaVR, PESI scores, and RVEDD at hospital admission may have prognostic value in patients with APE. Positive T wave in lead aVR could be a useful marker in early risk stratification of pulmonary embolism.


Comparative efficacy and safety of long-acting oral opioids for chronic non-cancer pain: a systematic review.
Opioids have been endorsed as appropriate treatment for refractory chronic non-cancer pain when used according to published guidelines. They are widely used for this indication. However, there appear to be gaps in our understanding of the efficacy and safety of individual long-acting opioids compared to each other or as a class compared to short-acting opioids. This systematic review summarizes and assesses the evidence for the comparative efficacy and safety of long-acting opioids in the management of chronic non-cancer pain. Randomized trials (for comparative efficacy and adverse events) and observational studies (for adverse events only) that included non-parenteral long-acting opioids were sought using electronic databases, handsearching reference lists, and soliciting pharmaceutical company submissions. Searches were performed through October 2002. The validity of each included study was assessed using a data abstraction form and predefined criteria. An overall grade was allocated for the body of evidence for each key question. A total of 16 randomized trials (comparative efficacy and adverse events), enrolling 1427 patients, and 8 observational studies (adverse events) of 1190 patients were included in this review. No randomized trial was rated good quality; observational studies were generally of poorer quality than the trials. There was insufficient evidence to prove that different long-acting opioids are associated with different efficacy or safety profiles. There was also insufficient evidence to determine whether long-acting opioids as a class are more effective or safer than short-acting opioids. A subgroup of three studies on long-acting versus short-acting oxycodone was more homogeneous and provided fair evidence that these formulations are equally effective for pain control.


Click-Chemistry-Mediated Cell Membrane Glycopolymer Engineering to Potentiate Dendritic Cell Vaccines.
Dendritic cell vaccine (DCV) holds great potential in tumor immunotherapy owing to its potent ability in eliciting tumor-specific immune responses. Aiming at engineering enhanced DCV, we report the first effort to construct a glycopolymer-engineered DC vaccine (G-DCV) via metabolicglycoengineering and copper-free click-chemistry. Model G-DCV was prepared by firstly delivering tumor antigens, ovalbumin (OVA) into dendritic cells (DC) with fluoroalkane-grafted polyethyleneimines, followed by conjugating glycopolymers with a terminal group of dibenzocyclooctyne (DBCO) onto dendritic cells. Compared to unmodified DCV, our G-DCV could induce stronger T cell activation due to the enhanced adhesion between DCs and T cells. Notably, such G-DCV could more effectively inhibit the growth of the mouse B16-OVA (expressing OVA antigen) tumor model after adoptive transfer. Moreover, by combination with an immune checkpoint inhibitor, G-DCV showed further increased anti-tumor effects in treating different tumor models. Thus, our work provides a novel strategy to enhance the therapeutic effectiveness of DC vaccines.


Association of LRP1B Mutation With Tumor Mutation Burden and Outcomes in Melanoma and Non-small Cell Lung Cancer Patients Treated With Immune Check-Point Blockades.
Background: Tumor mutation burden (TMB) have been served as the most prevalent biomarkers to predict immunotherapy response. LRP1B (low-density lipoprotein receptor-related protein 1B) is frequently mutated in melanoma, non-small cell lung cancer (NSCLC) and other tumors; however, its association with TMB and survival in patients with immunotherapy remains unknown. Methods: We curated somatic mutation data and clinicopathologic information from 332 melanoma immunotherapy samples for discovery and 113 NSCLC samples for further corroboration. Bayesian variants non-negative matrix factorization was used to extract tumor mutational signatures. Multivariate Cox and logistic regression models were applied to adjust confounding factors. The CIBERSORT and GSEA algorithm were separately used to infer leukocyte relative abundance and significantly enriched pathways. Results: Patients with LRP1B mutation were identified to be associated with prolonged survival in both immunotherapy cohort. Higher tumor mutation burden was found in LRP1B mutated patients, and the association remained significant after controlling for age, gender, stage, mutations in TP53 and ATR, and mutational signatures. Immune response and cell cycle regulation circuits were among the top enriched pathways in samples with LRP1B mutations. Conclusion: Our studies suggested sequencing even a single, frequently mutated gene may provide insight into genome-wide mutational burden, and may serve as a biomarker to predict immune response.


Protective effect of cyclo(his-pro) on streptozotocin-induced cytotoxicity and apoptosis in vitro.
Cyclo(His-Pro) (CHP) is a naturally occurring, cyclic dipeptide structurally related to thyrotropin-releasing hormone (TRH). CHP was efficiently obtained from soybean meal by hydrolysis with flavourzyme and alcalase. In this study, the effects of CHP on streptozotocin (STZ)-induced beta-cell dysfunction and apoptosis were investigated in rat insulinoma cells (RINm5F) secreting insulin. When the RINm5F cells were treated with 2mM STZ, insulin secretion decreased to approximately 54% that of control cells. However, CHP treatment restored the insulin-secreting activity of RINm5F cells to approximately 71% that of the untreated control cells. Moreover, CHP significantly protected the cells from STZ-mediated cytotoxicity via reduction of nitric oxide (NO) production (2.3-fold) and lipid peroxidation (1.9-fold), which were induced by STZ. Moreover, CHP treatment also attenuated STZ-induced apoptotic events, such as activation of caspase-3, poly(ADP-ribose) polymerase (PARP) cleavage, and DNA fragmentation in RINm5F cells, indicating that CHP could protect the cells from apoptotic cell death induced by oxidative stress of STZ by increasing the expression of an anti-apoptotic protein, Bcl-2. These results suggest that CHP could be a candidate material for a protective and therapeutic agent against STZ-mediated cytotoxicity and apoptosis.


The role of nelarabine in the treatment of T-cell acute lymphoblastic leukemia/lymphoma: challenges, opportunities, and future directions.
Nelarabine is a guanine nucleoside analog and functions to terminate DNA synthesis in dividing cells. Pre-clinical and clinical studies have shown that it preferentially accumulates in T-cells where it exerts its cytotoxic effects. After generations of treatment protocol advances, it has been incorporated into numerous treatment regimens against T-lineage acute lymphoblastic leukemia/lymphoma (T-ALL/LLy). On 8 March 2023, the FDA approved the use of nelarabine for its use in T-ALL due to clear evidence of clinical benefits. This announcement concludes a nearly 6-decade period of evaluation for nelarabine and its role in the management of high-grade, aggressive T-cell malignancies. We review the medicinal biology of nelarabine, its evaluation through decades of clinical studies, its dose-limited adverse effects, and its areas of highest impact in the treatment of T-ALL/LLy. We provide a context of when nelarabine might be considered in treatments against T-ALL/LLy, and also alternative strategies when it has or has not been used in therapies prior to relapse. We anticipate that an increasing number of treatment regimens will include nelarabine as a part of front-line therapy.


Molecular dynamics study of surfactant-like peptide based nanostructures.
Surfactant-like peptide (SLP) based nanostructures are investigated using all-atomistic molecular dynamics (MD) simulations. We report structure properties of nanostructures belonging to the ANK peptide group. In particular, the mathematical models for the two A3K membranes, A6K nanotube, and A9K nanorod were developed. Our MD simulation results are consistent with the experimental data, indicating that A3K membranes are stable in two different configurations: (1) SLPs are tilted relative to the normal membrane plane; (2) SLPs are interdigitated. The former configuration is energetically more stable. The cylindrical nanostructures feature a certain order of the A6K peptides. In turn, the A9K nanorod does not exhibit any long-range ordering. Both nanotube and nanorod structure contain large amounts of water inside. Consequently, these nanostructures behave similar to hydrogels. This property may be important in the context of biotechnology. Binding energy analysis-in terms of Coulomb and van der Waals contributions-unveils an increase as the peptide size increases. The electrostatic interaction constitutes 70-75% of the noncovalent attraction energy between SLPs. The nanotubular structures are notably stable, confirming that A6K peptides preferentially form nanotubes and A9K peptides preferentially form nanorods.


Hydatidosis in the province of La Pampa, Argentina, 1998.
Echinococcus granulosus is the agent that causes the classical hydatidosis or cystic echinococcosis. The most spread cycle in South America is the sheep-dog cycle, and offers favorable conditions for the development of the parasite. As the province of La Pampa shows an endemic presence with notification of cases and infected dogs, the present work has the aim of contributing to the study of the distribution and prevalence of the disease in the district of Maracó, La Pampa, Argentina. Dogs of 36 farms that had ovines, were dosed with arecoline hydrobromide at a dose of 3.5 mg/kg. Dogs from 27 (75%) farms resulted positive to Tenia sp. and 3 (8.3%) to E. granulosus. Blood samples were taken to all the people living in all the inquired farm and were processed by the ELISA test for hydatidosis. A retrospective survey on cases of human hydatidosis attended in the Hospital of General Pico, head if the sanitary region, and in two private clinics was performed. Analysis of clinical records indicated 11 cases operated during the period 1996/1997 (annual incidence rate of 6.8 per 100,000 inhabitants). It is of public health interest to fulfill epidemiologic investigations in other districts of the Province of La Pampa with the aim of delimitating the endemic distribution within the province and also organizing surveillance systems on human hydatidosis.


Organ Donation From Patients on Extracorporeal Membrane Oxygenation at the Time of Death.
To describe the clinical characteristics and organ donation rate of patients supported by extracorporeal membrane oxygenation (ECMO) at the time of death. Retrospective observational study. Pearson chi-square and Fisher exact tests were used in statistical analyses. One hundred twenty-seven acute care hospitals in New Jersey, Pennsylvania, and Delaware. Adult and pediatric patients who were on ECMO at the time of referral to a large organ procurement organization (OPO) between 2016 and 2020. None. Nineteen thousand nine hundred thirty patients were referred to the OPO between November 2016 and September 2020, of which 5,034 were medically suitable potential donors. Of this cohort, 143 patients were supported on ECMO at the time of OPO referral and 141 were included in analyses (median age 47 yr, 60% male). Thirty-three percent (46/141, median age 48 yr, 52% male) donated organs, compared with 50% of non-ECMO patients (p ≤ 0.0005). ECMO and non-ECMO patients had organs recovered but not transplanted at similar rates (11% vs 10%, p = 0.8). There were no significant differences in sex (p = 0.16) or ethnicity (p = 0.50) between organ donor and nondonor groups. Fifty-one percent (21/41) of organ donors donated after circulatory death and 49% (20/41) after brain death. Patients declared dead by neurologic criteria were more likely to donate (51%) than those declared dead by circulatory criteria (21%, p < 0.001). Frequency of cardiac arrest prior to ECMO was similar between donors and nondonors (p = 0.68). Thirty-nine percent (16/41) of donors had an out-of-hospital cardiac arrest (OHCA) and 51% (21/41) were cannulated via extracorporeal cardiopulmonary resuscitation (ECPR). The most common reason patients were not donors was that family declined (57%). One-third of patients referred to the OPO on ECMO at the time of death donated organs. While donation occurred less frequently after ECMO, ECMO and non-ECMO patients had organs used rather than discarded at a similar rate. Patients successfully donated following OHCA and/or ECPR. Clinicians should not consider ECMO a barrier to organ donation.


Photodynamic therapy for hypoxic solid tumors via Mn-MOF as a photosensitizer.
Hypoxia, as an important feature in tumor sites, greatly hinders the performance of photosensitizers, thus affecting the efficacy of photodynamic therapy (PDT). Therefore, designing and preparing new photosensitizer with high photosensitivity under hypoxic condition presents a great challenge that urgently needs to be solved. In this work, a new nano-MOF material using Mn(ii) as the active center can catalytically decompose high concentrations of H2O2 in tumor cells to generate O2, thereby improving the PDT efficacy in hypoxic tumors. The Mn-MOF also produces 1O2 under light irradiation, which finally induces cancer cell apoptosis. This work offers a new strategy for the design and discovery of effective photosensitizers for PDT.


Transcranial Doppler Ultrasonography Provides Insights into Neurovascular Changes in Children with Cerebral Malaria.
To evaluate neurovascular changes in pediatric patients with cerebral malaria. African children with cerebral malaria were enrolled and underwent daily transcranial Doppler ultrasound (TCD) examinations through hospital day 8, discharge, or death. Neurologic outcomes were assessed 2 weeks after enrollment. In total, 160 children with cerebral malaria and 155 comparison patients were included. In patients with cerebral malaria, TCD flow changes characterized as hyperemia were seen in 42 (26%), low flow in 46 (28%), microvascular obstruction in 35 (22%), cerebral vasospasm in 21 (13%), and isolated posterior hyperemia in 7 (4%). Most had a single neurovascular phenotype observed throughout participation. Among comparison patients, 76% had normal TCD findings (P < .001). Impaired autoregulation was present in 80% of cases (transient hyperemic response ratio 1.01 ± 0.03) but improved through day 4 (1.1 ± 0.02, P = .014). Overall mortality was 24% (n = 39). Neurologic deficits were evident in 21% of survivors. Children meeting criteria for vasospasm were most likely to survive with sequelae, and children meeting criteria for low flow were most likely to die. Autoregulation was better in children with a normal neurologic outcome (1.09, 95% CI 1.06-1.12) than in others (0.98, 95% CI 0.95-1) (P ≤ .001). Several distinct changes in TCD measurements were identified in children with cerebral malaria that permitted phenotypic grouping. Groups had distinct associations with neurologic outcomes. Validation of pathogenic mechanisms associated with each phenotype may aid in developing TCD as a portable, easy-to-use tool to help guide targeted adjunctive therapy in cerebral malaria aimed at causative mechanisms of injury on an individual level.


Effect of wind on the chemical uptake kinetics of a passive air sampler.
Passive air samplers (PASs) operate in different types of environment under various wind conditions, which may affect sampling rates and thus introduce uncertainty to PAS-derived air concentrations. To quantify the effect of wind speed and angle on the uptake in cylindrical PASs using XAD-resin as the sampling medium, we measured the uptake kinetics of polychlorinated biphenyls (PCBs) in XAD and of water in silica-gel, both under quasi wind-still condition and with lab-generated wind blowing toward the PASs at various speeds and angles. Passive sampling rates (PSRs) of PCBs under laboratory generated windy conditions were approximately 3-4 times higher than under wind-still indoor conditions. The rate of water uptake by silica-gel increased with wind speed, following a logarithmic function so that PSRs are more strongly influenced at lower wind speed. PSRs of both PCBs and water varied little with wind angle, which is consistent with computational fluid dynamic simulations showing that different angles of wind incidence cause only minor variations of air velocities within the cylindrical sampler housing. Because modifications of the design of the cylindrical PAS were not successful in eliminating the wind speed dependence of PSRs at low wind levels, indoor and outdoor deployments require different sets of PSRs. The effect of wind speed and angle on the PSRs of the cylindrical PAS are much smaller than what has been reported for the double-bowl polyurethane foam PAS. PSRs of the cylindrical XAD-PAS therefore tend to vary much less between sampling sites exposed to different wind conditions.


Attenuated hepatitis A virus: genetic determinants of adaptation to growth in MRC-5 cells.
A live candidate hepatitis A virus vaccine, developed from the HM-175 strain and adapted to growth in primary African green monkey kidney (AGMK) cells, was adapted to growth in MRC-5 cells. The nucleotide sequence of the MRC-5 cell-adapted virus was determined and compared with the known sequence of the AGMK cell-adapted virus. Thirteen unique mutations, which occurred during passage in MRC-5 cells, were identified. Four of the unique mutations were located in a cluster in the 5' noncoding region (NC), and three of the remaining nine mutations encoded amino acid changes. Infectious chimeric cDNAs were constructed from infectious cDNA clones of the AGMK cell-adapted and wild-type HM-175 viruses and PCR-amplified cDNA segments of the MRC-5 cell-adapted virus. The viruses encoded by these plasmids were recovered after transfection of cultured cells with in vitro transcripts, and their growth phenotypes in fetal rhesus kidney 4 (FRhK-4) and MRC-5 cells were determined. The important growth-enhancing mutations could be divided into three sets. Two of these were located in the 5' NC region, and the third was located in the 2C nonstructural gene. The mutations in the 5' NC region that developed during passage in MRC-5 cells were indispensable for efficient growth in MRC-5 cells, but a combination of the two groups in the 5' NC region and one in the 2C gene were required to increase growth dramatically in MRC-5 cells.


Neurocognitive Impairment in a Chronically Well-Suppressed HIV-Infected Population: The Dutch TREVI Cohort Study.
We assessed the value of screening for cognitive abnormalities in a chronically infected HIV population (N = 388) and investigated the association with clinical correlates. The mean age was 48 years (±11), the majority of the patients were male (89%), the median duration of infection was 6 years [interquartile range (IQR) = 2-12], the median CD count was 600 (IQR = 450-780), and 326 (84%) had a viral load below 200 copies/mL. Screening for cognitive complaints was applied using the three Simioni questions and the international HIV dementia scale (iHDS). Neuropsychological assessment (NPA) included 13 well-validated tests assessing motor speed, concentration, and memory. A total of 69 patients completed the NPA. CD4 (nadir), viral load, combination antiretroviral therapy (cART) duration, and the presence of comorbidities were evaluated for associations with NPA result. A total of 127 (33%) reported cognitive complaints during screening. The sensitivity and specificity of the Simioni questions were 82% and 24%, respectively. Adding the iHDS resulted in a sensitivity of 50% and a specificity of 73%. A CD4 nadir count <50 cells/m3 was associated with an abnormal NPA (p = 0.01). Comorbidities were more prevalent in patients with an abnormal NPA, although not statistically significant (p = 0.276). Age, current CD4, viral load, and cART duration were not associated with abnormal NPA. The authors conclude that current screening strategies are insufficient in detecting HIV-associated neurocognitive disorder. A low CD4 nadir is associated with poor neurocognitive outcome in HIV.


Ursodeoxycholic acid as adjunctive therapy for problematic type 1 autoimmune hepatitis: a randomized placebo-controlled treatment trial.
To evaluate the efficacy of ursodeoxycholic acid as adjunctive therapy in type 1 autoimmune hepatitis, 37 patients who had experienced treatment failure, repeated relapse, or incomplete response were randomized to ursodeoxycholic acid (13-15 mg/kg daily) or placebo for 6 months in addition to their usual corticosteroid schedule. Serum aspartate transaminase (70% vs. 31%, P =.04) and alkaline phosphatase (47% vs. 7%, P =.02) levels improved more commonly in the 21 patients randomized to ursodeoxycholic acid. Mean serum levels, however, were similar before and after the treatment period. The frequency of dose reduction or corticosteroid withdrawal was comparable in both groups (29% versus 31%, P >.9), and clinical improvement (48% vs. 44%, P >.9) or its absence (52% vs. 56%, P >.9) occurred as commonly in patients receiving ursodeoxycholic acid or placebo. The modified histological activity score (3.5 +/- 0.8 vs. 3. 5 +/- 0.9) and the modified fibrosis score (2.4 +/- 0.4 vs. 2.4 +/- 0.4) were similar before and after treatment with ursodeoxycholic acid and no different than after placebo therapy. We conclude that ursodeoxycholic acid can improve certain laboratory tests in problematic patients with type 1 autoimmune hepatitis when administered adjunctively for 6 months. Short-term therapy, however, does not facilitate reduction in the dose of corticosteroids or its withdrawal, affect clinical outcome, or reduce histological activity.


[Circadian rhythm of parathyrin and calcitonin concentrations in the serum].
The circadian variations of serum intact parathyrin, C-terminal parathyrin (65-84), mid-region parathyrin (44-68) fragments, calcitonin, total calcium, ionized calcium, albumin and phosphate were measured in five healthy subjects. Intact parathyrin, parathyrin (44-68) and calcitonin show a synchronous diurnal fluctuation with a nocturnal increase to a maximum between 24.00 and 2.00 hours. Whereas phosphate has a marked circadian rhythmicity with a zenith between 1.00 and 8.00 hours, total calcium and albumin show a tendency to decrease between 20.00 and 6.00 hours. Ionized calcium concentration remains constant over the whole day.


Academic retainer medicine: an innovative business model for cross-subsidizing primary care.
Retainer-medicine primary care practices, commonly referred to as "luxury" or "concierge" practices, provide enhanced services to patients beyond those available in traditional practices for a yearly retainer fee. Adoption of retainer practices has been largely absent in academic health centers (AHCs). Reasons for this trend stem primarily from ethical concerns, such as the potential for patient abandonment when physicians downsize from larger, traditional practices to smaller, retainer-medicine practices.In 2004, the Department of Medicine at Tufts Medical Center developed an academic retainer-medicine primary care practice within the Division of General Medicine that not only generates financial support for the division but also incorporates a clinical and business model that is aligned with the mission and ethics of an academic institution.In contrast to private retainer-medicine practices, this unique business model addresses several of the ethical issues associated with traditional retainer practices-it does not restrict net access to care and it neutralizes concerns about patient abandonment. Addressing the growing primary care shortage, the model also presents the opportunity for a retainer practice to cross-subsidize the expansion of general medicine in an academic medical setting. The authors elucidate the benefits, as well as the inherent challenges, of embedding an academic retainer-medicine practice within an AHC.


[Characteristics of the changes in the natural killer activity of peripheral blood lymphocytes during pregnancy].
The influence of blood plasma of women of the 1st and 3rd pregnancy trimester and of cord blood on natural killer (NK) activity of peripheral blood lymphocytes of non-pregnant women was investigated. The maximum suppressive activity on NK was observed with plasma of pregnant women of the 3rd trimester with low NK-activity. Data suggesting the involvement of estradiol in this suppressive activity was obtained. No suppressive activity of cord blood plasma was found despite the low NK-activity of cord blood.


Catechol-O-methyltransferase: substrate-specificity and stereoselectivity for beta-adrenoceptor agents.
Isoprenaline, isoetharine, rimiterol, dobutamine and nadolol were investigated as substrates for purified pig-liver catechol-O-methyltransferase using a sensitive spectrophotometric assay. Kinetic parameters, Km and Vmax, were defined and the apparent first-order rate constant (Vmax/Km) was derived. On the basis of the apparent first-order rate constant, rimiterol was found to be a 1.5-fold and dobutamine a 5-fold better substrate for catechol-O-methyltransferase than isoprenaline; isoetharine shows no improvement over isoprenaline. Nadolol is not a substrate for catechol-O-methyltransferase. O-Methylation of isoprenaline- and noradrenaline-enantiomers was found to be stereoselective: catechol-O-methyltransferase shows selectivity towards the laevo (-) isomer with respect to the (+) form or racemic mixture. The investigation indicated stereochemical and steric determinants important in the interaction of catechol-O-methyltransferase with physiologically and clinically important beta-adrenoceptor agents.


Measurement of supramolecular effective molarities for intramolecular H-bonds in zinc porphyrin-imidazole complexes.
The association constants for formation of 1 : 1 complexes between five different imidazole ligands and eight different porphyrins have been measured by UV/vis titration experiments in two different solvents, toluene and 1,1,2,2-tetrachloroethane (TCE). Ligands equipped with H-bond acceptors (ester or amide) and porphyrins equipped with H-bond donors (phenol) can make H-bonds in addition to the zinc-nitrogen coordination interaction. The free energy contributions of these H-bonds to the overall stabilities of the complexes were determined using chemical double mutant cycles. Amide-phenol H-bonds contribute up to 5 kJ mol(-1) to the free energy change on complexation, and ester-phenol H-bonds contribute up to 3 kJ mol(-1). Porphyrin-ligand combinations with poor geometric complementarity do not make detectable H-bonding interactions. Effective molarities (EM) for the formation of H-bonds in the complexes were estimated by comparing the equilibrium constants for formation of the intramolecular interaction with the corresponding intermolecular interaction: the values are between 3 mM and 200 mM, which is comparable to previous results obtained for porphyrin-pyridine complexes. The values of EM measured for flexible and rigid ligand systems are comparable. This suggests that there is a trade off between restriction of conformational mobility in the flexible ligands and geometric strain in the rigid ligands, which results in similar binding affinities.


Chikungunya Virus Infection Impairs the Function of Osteogenic Cells.
Chikungunya virus (CHIKV) is a positive-sense, single-stranded RNA virus spread by the Aedes species of mosquito. Chikungunya virus causes a condition characterized by high fever, headache, rash, and joint pain. Recent investigations reveal the presence of bone lesions and erosive arthritis in the joints of CHIKV-infected patients, indicating an association of bone pathology with CHIKV infection. However, the molecular mechanism underlying CHIKV-induced bone pathology remains poorly defined. Bone marrow-derived mesenchymal stem cells (BMSCs) contribute to bone homeostasis by differentiating into osteogenic cells which later mature to form the bone. Disruption of osteogenic differentiation and function of BMSCs leads to bone pathologies. Studies show that virus infections can alter the properties and function of BMSCs. However, to date, pathogenesis of CHIKV infection in this context has not been studied. In the current study, we investigated the susceptibility of BMSCs and osteogenic cells to CHIKV and studied the effect of infection on these cells. For the first time to our knowledge, we report that CHIKV can productively infect BMSCs and osteogenic cells. We also observed decreased gene expression of the major regulator of osteogenic differentiation, RUNX2, in CHIKV-infected osteogenic cells. Furthermore, impaired functional properties of osteogenic cells, i.e., decreased production and activity of alkaline phosphatase (ALP) and matrix mineralization, were observed in the presence of CHIKV infection. Thus, we conclude that CHIKV likely impairs osteogenic differentiation of BMSCs, indicating a possible role of BMSCs in altering bone homeostasis during CHIKV infection.IMPORTANCE Presently, no vaccines or treatment options are available for CHIKV infection. Joint pain is one of the major concerns. Although studies have shown an association between bone pathology and infection, the molecular pathogenesis in the context of bone pathology is poorly defined. Here, we demonstrate for the first time that BMSCs and BMSC-derived osteogenic cells are susceptible to CHIKV infection, and that infection likely alters the function of osteogenic cells. This study highlights altered osteogenic differentiation as a possible mechanism for causing the bone pathology observed in CHIKV pathogenesis.


Expression of the non-gastric H+/K+ ATPase ATP12A in normal and pathological human prostate tissue.
Altered cellular proton handling and cell volume regulation are hallmarks of tumorigenesis. To investigate a possible involvement of the non-gastric H(+)/K(+) ATPase ATP12A (ATP1AL1) in prostate cancer, we performed immunohistochemistry in formalin-fixed, paraffin-embedded histological sections from benign and malignant human prostate lesions. Normal prostate tissue displayed a membrane-bound ATP12A staining with focal accumulated pattern, whereas in the benign prostate hyperplasia (BPH) and cancerous prostate tissue (tumor grade I-III) the protein appears to be displaced in the luminal cells of the glandular epithelium. Hence, the expression pattern of ATP12A is markedly altered in BPH and prostate cancer. To test for altered gene expression of ATP12A we performed quantitative reverse transcriptase PCR (QRT-PCR) in normal (tumor-free) prostate tissue, BPH and tumor stages I-III using a prostate cancer cDNA array. However, no significantly different expression levels could be detected in the various disease states compared to normal tissue, which contrasts the findings from immunohistochemistry and points to the possibility of altered post-translational processing and/or sorting of the protein. We further show that ATP12A mRNA is expressed at different levels in PC-3 and LNCaP prostate cancer cells, with a significant ~26-fold higher expression in the latter cell type. Protein expression in these tumor cell lines was verified by Western blot.


Neomycin inhibits catecholamine secretion by blocking nicotinic acetylcholine receptors in bovine adrenal chromaffin cells.
We investigated the effects of neomycin on nicotinic acetylcholine receptor-induced responses in bovine adrenal chromaffin cells. Neomycin inhibited the nicotinic agonist dimethylphenylpiperazinium iodide (DMPP)-induced norepinephrine secretion in a concentration-dependent manner. Neomycin had also an inhibitory effect on the DMPP-induced increase in cytosolic Ca++ concentration ([Ca++]i). This effect was further confirmed by inhibition of the DMPP-induced fluorescence quenching of fura-2 upon Mn++ entry. Under the same conditions, however, neomycin did not change the bradykinin-induced [Ca++]i increase, which follows the downstream signal of phospholipase C phospholipase C activation in this cell. The inhibitory effect of neomycin on the DMPP-induced [Ca++]i increase was apparent when the neomycin treatment was performed simultaneously with DMPP, suggesting a direct action on the nicotinic receptor. The direct inhibitory action of neomycin on the nicotinic receptor was also evident when neomycin inhibited the DMPP-induced cytosolic Ca++ increase, which is not affected by nifedipine nor omega-conotoxin MVIIC, and the cytosolic Na+ increase, which is not affected by tetrodotoxin. In addition, we observed that neomycin inhibited the binding of nicotine to the acetylcholine receptor in a noncompetitive manner. The data suggest that neomycin inhibits the nicotinic acetylcholine receptor directly, which results in blockage of the nicotinic receptor-mediated signaling without involvement of phospholipase C.


Flagella and pili-mediated near-surface single-cell motility mechanisms in P. aeruginosa.
Bacterial biofilms are structured multicellular communities that are responsible for a broad range of infections. Knowing how free-swimming bacteria adapt their motility mechanisms near a surface is crucial for understanding the transition from the planktonic to the biofilm phenotype. By translating microscopy movies into searchable databases of bacterial behavior and developing image-based search engines, we were able to identify fundamental appendage-specific mechanisms for the surface motility of Pseudomonas aeruginosa. Type IV pili mediate two surface motility mechanisms: horizontally oriented crawling, by which the bacterium moves lengthwise with high directional persistence, and vertically oriented walking, by which the bacterium moves with low directional persistence and high instantaneous velocity, allowing it to rapidly explore microenvironments. The flagellum mediates two additional motility mechanisms: near-surface swimming and surface-anchored spinning, which often precedes detachment from a surface. Flagella and pili interact cooperatively in a launch sequence whereby bacteria change orientation from horizontal to vertical and then detach. Vertical orientation facilitates detachment from surfaces and thereby influences biofilm morphology.


Sexual selection on wing interference patterns in Drosophila melanogaster.
Animals with color vision use color information in intra- and interspecific communication, which in turn may drive the evolution of conspicuous colored body traits via natural and sexual selection. A recent study found that the transparent wings of small flies and wasps in lower-reflectance light environments display vivid and stable structural color patterns, called "wing interference patterns" (WIPs). Such WIPs were hypothesized to function in sexual selection among small insects with wing displays, but this has not been experimentally verified. Here, to our knowledge we present the first experimental evidence that WIPs in males of Drosophila melanogaster are targets of mate choice from females, and that two different color traits--saturation and hue--experience directional and stabilizing sexual selection, respectively. Using isogenic lines from the D. melanogaster Genetic Reference Panel, we compare attractiveness of different male WIPs against black and white visual backgrounds. We show that males with more vivid wings are more attractive to females than are males with dull wings. Wings with a large magenta area (i.e., intermediate trait values) were also preferred over those with a large blue or yellow area. These experimental results add a visual element to the Drosophila mating array, integrating sexual selection with elements of genetics and evo-devo, potentially applicable to a wide array of small insects with hyaline wings. Our results further underscore that the mode of sexual selection on such visual signals can differ profoundly between different color components, in this case hue and saturation.


Application of a probabilistic microstructural model to determine reference length and toe-to-linear region transition in fibrous connective tissue.
This study shows how a probabilistic microstructural model for fibrous connective tissue behavior can be used to objectively describe soft tissue low-load behavior. More specifically, methods to determine tissue reference length and the transition from the strain-stiffening "toe-region" to the more linear region of the stress-strain curve of fibrous connective tissues are presented. According to a microstructural model for uniaxially loaded collagenous tissues, increasingly more fibers are recruited and bear load with increased tissue elongation. Fiber recruitment is represented statistically according to a Weibull probability density function (PDF). The Weibull PDF location parameter in this formulation corresponds to the stretch at which the first fibers begin to bear load and provides a convenient method of determining reference length. The toe-to-linear region transition is defined by utilizing the Weibull cumulative distribution function (CDF) which relates the fraction of loaded fibers to the tissue elongation. These techniques are illustrated using representative tendon and ligament data from the literature, and are shown to be applicable retrospectively to data from specimens that are not heavily preloaded. The reference length resulting from this technique provides an objective datum from which to calculate stretch, strain, and tangent modulus, while the Weibull CDF provides an objective parameter with which to characterize the limits of low-load behavior.


Cell proliferation in 3,800 node-negative breast cancers: consistency over time of biological and clinical information provided by 3H-thymidine labelling index.
For breast cancer, many prognostic markers that initially appeared promising have failed to maintain their clinical predictive value. Few reports have analyzed the consistency over time of biological and clinical information provided by biomarkers. Tumour cell proliferation has acquired relevance as an indicator of prognosis and of response to treatment. Since its clinical role has been investigated for some decades, cell proliferation represents an ideal marker for an over-time validation. In 3,800 node-negative breast cancers recruited between 1972 and 1991, the consistency of information provided by the 3H-thymidine labelling index (TLI), in terms of basic relations with other clinico-pathological and biological variables and clinical predictivity, was evaluated using a combined analysis of results previously published by our group for distinct series of patients. Clinical predictivity was analyzed on a subset of 2,067 patients given local-regional therapy until relapse and followed for a median time from 6 to 10 years. Over the entire period TLI maintained a weak direct relation with tumour size and an inverse strong relation with steroid receptors. An increase in TLI was observed for tumours in post-menopausal patients up to the mid 1980s. During 3 different accrual periods (1972-1983; 1984-1987; 1988-1991), TLI was a consistent and independent predictor of relapse-free time, distant metastasis and overall survival, regardless of its consideration as a continuous variable or with a cutoff value of 3%. The reproducibility of our results over time provides support to the consistency of the methodology used and of the biological and clinical information obtained when using TLI as an indicator of breast cancer cell proliferation.