Miracle-dz/UniFAR-FTUs huggingface dataset:
Intracellular survival and replication of Erysipelothrix rhusiopathiae within murine macrophages: failure of induction of the oxidative burst of macrophages

We investigated the ability of a virulent wild-type parent strain and acapsular avirulent transposon mutants to enter and survive intracellularly within murine peritoneal macrophages. In the presence of normal or immune serum, the parent and mutant strains were both ingested; however, the number of ingested bacteria was three- to fourfold greater in the case of mutant strains than in the case of the parent strain. The parent strain, but not the mutant strains, survived and replicated intracellularly when ingested in the presence of normal serum, whereas both the parent and the mutant strains were readily killed when ingested in the presence of immune serum. To further investigate the mechanism by which the parent strain can survive and replicate within macrophages, we studied the oxidative burst response of macrophages to these strains by measuring chemiluminescence and intracellular reduction of Nitro Blue Tetrazolium dye. Challenge exposure of macrophages with either the parent strain preopsonized with immune serum or the mutant strains preopsonized with normal or immune serum induced a strong oxidative burst, whereas the level was very low when the parent strain was preopsonized with normal serum. Phagocytosis of either the parent strain, in the presence of immune serum, or the mutant strains, in the presence of normal or immune serum, by macrophages reduced large amounts of intracellular Nitro Blue Tetrazolium, whereas minimal amounts were reduced by the parent strain in the presence of normal serum. These results suggest that virulent E. rhusiopathiae can survive and subsequently replicate within murine macrophages when ingested in the presence of normal serum and that the reduced production of reactive oxidative metabolites by macrophages may, in part, be responsible for this occurrence.
Functional Prokaryotic RubisCO from an Oceanic Metagenomic Library

ABSTRACT Culture-independent studies have indicated that there is significant diversity in the ribulose 1,5-bisphosphate carboxylase/oxygenase (RubisCO) enzymes used by marine, freshwater, and terrestrial autotrophic bacteria. Surprisingly, little is known about the catalytic properties of many environmentally significant RubisCO enzymes. Because one of the goals of RubisCO research is to somehow modify or select for RubisCO molecules with improved kinetic properties, a facile means to isolate functional and novel RubisCO molecules directly from the environment was developed. In this report, we describe the first example of functional RubisCO proteins obtained from genes cloned and characterized from metagenomic libraries derived from DNA isolated from environmental samples. Two form IA marine RubisCO genes were cloned, and each gene supported both photoheterotrophic and photoautotrophic growth of a RubisCO deletion strain of Rhodobacter capsulatus, strain SBI/II−, indicating that catalytically active recombinant RubisCO was synthesized. The catalytic properties of the metagenomic RubisCO molecules were further characterized. These experiments demonstrated the feasibility of studying the functional diversity and enzymatic properties of RubisCO enzymes without first cultivating the host organisms. Further, this “proof of concept” experiment opens the way for development of a simple functional screen to examine the properties of diverse RubisCO genes isolated from any environment, and subsequent further bioselection may be possible if the growth conditions of complemented R. capsulatus strain SBI/II− are varied.
The Effect of Temperature on Soft Contact Lens Diameter

Purpose: The diameters of soft contact lenses are conventionally measured at room temperature (RT), however, all soft lenses shrink when raised to eye temperature (ET). The purpose of this study was to evaluate RT to ET shrinkage in a representative sample of contemporary spherical soft contact lenses. Methods: Twenty-four reusable and daily disposable lens types (10 hydrogel and 14 silicone hydrogel) in three powers were measured for total diameter at RT (20°C±1°C) and ET (34°C±1°C). Lens diameter measurements were undertaken after equilibration in ISO saline in a temperature controlled lens analyzer (Optimec Ltd, Malvern, UK). Theoretical changes in base curve radii were also calculated. Results: All lens types reduced in diameter when raised to ET. The largest mean changes with silicone hydrogel and hydrogel lenses were with Avaria (&Dgr;0.33 mm) and SofLens Daily Disposable (&Dgr;0.69 mm), respectively. The smallest mean changes for silicone hydrogel and hydrogel lenses were with 1-DAY ACUVUE TruEye (&Dgr;0.04 mm) and SofLens 38 (&Dgr;0.11 mm), respectively. There was, however, a wide range in change between the different lenses tested. Conclusions: This study has identified the levels of shrinkage in current soft contact lenses. In many cases, these shrinkage levels can be expected to have significant effects on clinical performance.
Effect of acute immunoneutralization of endogenous leptin on prolactin and LH secretion during the afternoon of pro-oestrus or in steroid-treated ovariectomized female rats.

Recent data indicate that leptin is involved in the control of reproductive function. Experiments were carried out to analyse the role of endogenous leptin in the regulation of LH and prolactin secretion during the afternoon of pro-oestrus and that induced by ovarian steroids in ovariectomized rats. In the first experiment, cyclic female rats were implanted with intra-auricular and intracerebroventricular (i.c.v.) cannulae and, at pro-oestrus, were injected (i.c.v.) with 10 microliters normal rabbit serum or leptin antiserum (at 13:00 and 14:00 h). Blood samples were obtained at 10:00 h and at intervals of 1 h between 13:00 and 20:00 h. In the second experiment, female rats in pro-oestrus were injected with normal rabbit serum or leptin antiserum at 16:00 and 18:00 h and blood samples were taken every 10 min between 18:00 and 20:00 h. In the third experiment, adult female rats that had been ovariectomized 2 weeks before were implanted with intra-auricular and i.c. v. cannulae and treated with oestradiol benzoate (30 micrograms s.c.) at 10:00 h and progesterone (2 mg s.c.) 48 h later. Normal rabbit serum (10 microliters) or leptin antiserum (10 microliters) were injected (i.c.v.) at 13:00 and 14:00 h, and blood samples were obtained at 10:00 h and at intervals of 1 h between 13:00 and 20:00 h. In the fourth experiment, hemipituitaries from ovariectomized steroid-treated female rats were incubated in the presence of leptin116-130 (an active fragment of the native molecule), GnRH or leptin + GnRH. Prolactin and LH secretion during the afternoon of pro-oestrus in females treated with leptin antiserum was similar to that observed in animals injected with normal rabbit serum. In ovariectomized female rats, the steroid-induced LH surge increased slightly after administration of leptin antiserum, whereas the prolactin surge remained unchanged. In vitro, leptin116-130 (10(-5) to 10(-8) mol l-1) inhibited LH secretion and modulated the effect of GnRH on LH release, depending on the concentration of GnRH: leptin116-130 (10(-6) mol l-1) reduced the effectiveness of 10(-7) mol GnRH l-1 and increased that of 10(-9) mol GnRH l-1. In conclusion, these experiments indicate that acute immunoneutralization of endogenous leptin does not interfere with spontaneous or steroid-induced LH and prolactin surges. In addition, the finding that leptin116-130 inhibited LH release and modulated the effectiveness of GnRH in vitro provides evidence of the direct modulatory role of leptin on LH secretion acting at the pituitary.
Educating tomorrow’s doctors: A cross sectional survey of emotional intelligence and empathy in medical students of Lahore

Objective: Medical education in Pakistan traditionally emphasizes physician’s biomedical knowledge with less emphasis on interpersonal skills and ability to relate to the patients. This study explored the emotional intelligence & empathy of undergraduate medical students and investigated its relationship with various factors to act as baseline for future work in this area. Methodology: The Schutte Emotional Intelligence scale and Davis' Interpersonal Reactivity Index in addition to socio demographic questionnaire were administered to first year and final year medical undergraduates of two medical Institutions in Lahore, Pakistan. Data was analyzed by using SPSS 17 version. Results: The overall mean scores for medical students both on emotional intelligence and empathy is significantly lower than that found in previous literature, highlighting that medical students do not appear to fare better than average people in EQ. Women showed statistically significant higher scores on Appraisal of emotions , Regulation of emotions, Empathic concern Scale & Personal distress scale. Comparison of EI & IRI of students according to medical college year did not show any statistical significance. Conclusion: Current medical curriculum and training in Pakistan does not appear to increase EI abilities which are building blocks that may allow students and residents to develop competence. Medical educators in Pakistan should look for ways to incorporate emotional intelligence in medical curriculum which will ultimately contribute towards patient centered practice, patient satisfaction as well as effective communication skills.
Role of Cholesterol in Human Immunodeficiency Virus Type 1 Envelope Protein-Mediated Fusion with Host Cells

ABSTRACT In this study we examined the effects of target membrane cholesterol depletion and cytoskeletal changes on human immunodeficiency virus type 1 (HIV-1) Env-mediated membrane fusion by dye redistribution assays. We found that treatment of peripheral blood lymphocytes (PBL) with methyl-β-cyclodextrin (MβCD) or cytochalasin reduced their susceptibility to membrane fusion with cells expressing HIV-1 Env that utilize CXCR4 or CCR5. However, treatment of human osteosarcoma (HOS) cells expressing high levels of CD4 and coreceptors with these agents did not affect their susceptibility to HIV-1 Env-mediated membrane fusion. Removal of cholesterol inhibited stromal cell-derived factor-1α- and macrophage inflammatory protein 1β-induced chemotaxis of both PBL and HOS cells expressing CD4 and coreceptors. The fusion activity as well as the chemotactic activity of PBL was recovered by adding back cholesterol to these cells. Confocal laser scanning microscopy analysis indicated that treatment of lymphocytes with MβCD reduced the colocalization of CD4 or of CXCR4 with actin presumably in microvilli. These findings indicate that, although cholesterol is not required for HIV-1 Env-mediated membrane fusion per se, its depletion from cells with relatively low coreceptor densities reduces the capacity of HIV-1 Env to engage coreceptor clusters required to trigger fusion. Furthermore, our results suggest that coreceptor clustering may occur in microvilli that are supported by actin polymerization.
A Novel Approach for Repairing of Intestinal Fistula Using chitosan hydrogel

Intestinal fistula is associated with high morbidity and difficult to manage. Many fistulas require surgical treatment, which usually consists of segmental resection. In this study, using a rat model, the effectiveness of chitosan hydrogel as an intestinal fistula repair agent was investigated. Twenty rats underwent laparotomy under general anesthesia. The antimesentric portion of the cecum was incised (1 cm) and sutured to the abdominal wall. Chitosan hydrogel was applied daily to the fistula until it was completely closed. Blood samples taken from all animals were analyzed. After sacrifice, the cecum was removed and histopathologic investigation was performed. Spontaneous closure of the intestinal fistula was observed in all animals for both the control and chitosan hydrogel groups. Healing in the chitosan hydrogel group healing was faster than that in the control group. Blood analysis revealed significant differences between the chitosan hydrogel and control groups with regard to the total protein, albumin, total cholesterol and HDL before the surgery versus that on the day of sacrifice. Pathologic investigation also showed greater healing in the chitosan hydrogel group than the control group. This preliminary study showed the potential of chitosan hydrogel for repair of intestinal fistula. However further studies must be performed before we can approve testing chitosan hydrogel for intestinal fistula repair in humans.
Platinum agents in the treatment of osteosarcoma: efficacy of cisplatin vs. carboplatin in human osteosarcoma cell lines.

BACKGROUND Cisplatin (cDDP), when used either alone or, more often, in combination with other agents, especially adriamycin, achieves a high response rate in osteosarcoma. Its use, however, is limited by severe nephro- and neuro-toxicity. Second generation platinum compounds, most notably carboplatin (CBDCA), have been developed in order to attempt to reduce these dose-limiting toxicities, and thus improve the therapeutic ratio. Studies evaluating the role of combination CT containing CBDCA vs. cDDP have demonstrated differing results depending on the tumor type tested and its role in the treatment of osteosarcoma has yet to be clarified. PROCEDURE In this study, we compared the in vitro anti-tumor activity of cDDP and CBDCA in a panel of three human osteosarcoma cell lines (HOS, MG63, and U2OS). RESULTS cDDP and CBDCA (0-20 micromol) showed marked variation in cytotoxicity among the three cell lines. EC(50) values for CBDCA in HOS and MG63 cells were approximately two-fold higher than for cDDP and the ratio of AUC(CBDCA) to AUC(cDDP) varied from 1.8 in the HOS cell line to 2.3 in the MG63 cell line. Exposure of MG63 and HOS cells to either cDDP or CBDCA (1.67 and 13.5 micromol) caused a G2/M cell cycle arrest by 24 hr. Also evident was a sub G1 peak indicative of cell death by apoptosis. U2OS cells were relatively resistant to the cytotoxic effects of both drugs, although a cell cycle arrest in response to DNA damage was observed. This suggests that unlike MG63 and HOS cells, U2OS cells have either a more efficient repair pathway for platinum-induced DNA damage or are able to evade apoptosis. Examination of apoptotic events and cellular recovery demonstrated that both an 8-16-fold higher concentration and longer treatment period for CBDCA compared with cDDP was required to produce equivalent cell death and a loss of the ability of single cell clones to form colonies in both the HOS and MG63, but not the U2OS cell line. CONCLUSIONS Our findings suggest that CBDCA at a two- to four-fold higher concentration than cDDP has potential therapeutic activity in platinum sensitive osteosarcomas, particularly when cDDP cytotoxicity compromises therapeutic efficacy.
Free and supported stance in Parkinson's disease. The effect of posture and 'postural set' on leg muscle responses to perturbation, and its relation to the severity of the disease.

Upright stance and its reflex control were studied in parkinsonian patients and in age-matched normal subjects. They stood unperturbed on a force-measuring surface (static conditions), or were displaced by movement of a supporting platform (dynamic conditions). During quiet stance the following variables were analysed, with eyes open or closed: position of the centre of foot pressure (CFP), average sway area, length of sway path, amplitude and distribution of tonic leg muscle EMG activity. Perturbations of stance were induced by toe-up or toe-down rotations, and by backward or forward translations of the platform. Amplitude of short, medium and long-latency EMG responses to displacement were measured in the tibialis anterior (TA) and in the three heads of the triceps surae (TS) muscle. The perturbations were produced during both free and supported stance (holding onto a stable structure), under which condition normal subjects suppress medium and long-latency responses. Under static conditions, the only significant finding in parkinsonians was represented by a shift in the position of the CFP. This was correlated with the severity of the disease (Webster scale), the less affected patients being shifted backwards and the more affected patients forwards, with respect to normals. Under dynamic conditions, the reflex responses to perturbations of free stance were similar in both groups. Only the medium-latency burst of gastrocnemius lateralis and the long-latency burst TA evoked by TS stretch were larger in parkinsonians. The amplitude of these responses, as well as of all the others, was not related to the Webster score. Within the patients' group, a relationship between position of CFP and area of EMG burst was found for both medium and long-latency TA responses evoked by forward translation and toe-up rotation, respectively. Under supported conditions, the capability to suppress all medium and long-latency muscle responses to any perturbation was lost or impaired in the parkinsonians. The degree of impairment was unrelated to the position of the CFP, but was significantly related to the severity of the disease. The suppression to 40% (supported/nonsupported), of TA response to toe-down rotation is proposed as the point of separation between normals and parkinsonians. The forward projection of the CFP, occurring in the severe stages of the disease, and the increase in amplitude of some responses to perturbations of free stance might be a compensatory adaptation to the anomalous upright posture. (ABSTRACT TRUNCATED AT 400 WORDS)
Evidence for ship noise impacts on humpback whale foraging behaviour

Noise from shipping activity in North Atlantic coastal waters has been steadily increasing and is an area of growing conservation concern, as it has the potential to disrupt the behaviour of marine organisms. This study examines the impacts of ship noise on bottom foraging humpback whales (Megaptera novaeangliae) in the western North Atlantic. Data were collected from 10 foraging whales using non-invasive archival tags that simultaneously recorded underwater movements and the acoustic environment at the whale. Using mixed models, we assess the effects of ship noise on seven parameters of their feeding behaviours. Independent variables included the presence or absence of ship noise and the received level of ship noise at the whale. We found significant effects on foraging, including slower descent rates and fewer side-roll feeding events per dive with increasing ship noise. During 5 of 18 ship passages, dives without side-rolls were observed. These findings indicate that humpback whales on Stellwagen Bank, an area with chronically elevated levels of shipping traffic, significantly change foraging activity when exposed to high levels of ship noise. This measureable reduction in within-dive foraging effort of individual whales could potentially lead to population-level impacts of shipping noise on baleen whale foraging success.
Clinical Significance of Human Erythrocyte Glucose Transporter 1 Expression at the Deepest Invasive Site of Advanced Colorectal Carcinoma

Objective: Malignant cells exhibit increased glucose uptake and utilization in vitro and in vivo. This process is thought to be mediated by the glucose transporter (Glut) family. The aim of this study was to elucidate the clinical significance of Glut1 expression at the site of deepest invasion as a predictor of the invasive/metastatic potential and prognosis of advanced colorectal carcinoma (CRC). Methods: One hundred and fifty-two patients who had undergone surgical resection for advanced CRC were entered in this study. Histologic subclassifications at the deepest invasive site included well-differentiated (W), moderately to well-differentiated (Mw), moderately to poorly differentiated (Mp), poorly differentiated (Por) and mucinous (Muc) adenocarcinomas. Glut1 expression was examined immunohistochemically with a labeled streptavidin-biotin kit using anti-Glut1 polyclonal antibody MYM. As a marker of cell proliferation, Ki-67 expression was also examined. All immunoreactivity was analyzed at the deepest invasive site, central portion and superficial part. The immunohistochemical expression of Glut1 was defined as positive if distinct staining of the membrane or cytoplasm was observed in at least 30% of tumor cells. Results: Glut1 expression was detected in 56 of 152 lesions (36.8%) at the deepest invasive site. The incidence of Glut1 expression at the deepest invasive site correlated significantly with histologic grade (W/Mw grade, 28% vs. Mp/Por/Muc grade, 48%), depth of invasion (invasion of muscularis propria/invasion of subserosa or subadventitia, 29% vs. invasion of serosa or adventitia/invasion of adjacent structures, 52%), lymphatic invasion (absence of lymphatic invasion, 19% vs. presence of lymphatic invasion, 40%), lymph node metastasis (absence of lymph node metastasis, 25% vs. presence of lymph node metastasis, 41%) and Duke’s stage (Duke’s <D, 32% vs. Duke’s D, 51%). In the central portion and superficial part, there were no significant differences between Glut1, Ki-67 expression and clinicopathological findings. Glut1 expression at the deepest invasive site correlated significantly with the Ki-67 labeling index. In cases of curative surgery, patients with Glut1-positive lesions at the deepest invasive site showed a significantly poorer prognosis than those with Glut1-negative lesions. Multivariate analysis with logistic regression for 5-year survival in patients who had undergone curative surgery showed that lymph node metastasis and Glut1 expression were significant risk factors. Conclusion: These results indicate that Glut1 expression at the deepest site of tumor invasion can be a useful predictor of a high malignant potential and poor prognosis in advanced CRC.
Contextually Intelligent Leadership for Improving Schools Across Different Contexts and Regions

One of the main challenges facing school principals in many parts of the world today is how to create a meaningful balance between accountability and innovation. This challenge is rooted in the existing tension between managerialism and leadership theory prevailing across contexts and regions. It is manifested by two competing contextual demands put on school principals, namely, the demand for accountability for outcomes (expressed, among others, through standards-based accountability policies) on the one hand, and the demand for innovation and creativity current leadership theories postulate in view of the context of change schools are operating in. Based on the review of the literature, this chapter presents the tension between school leadership and management and how this manifests itself in both theory and practice. The chapter examines the potential for reducing the tension through contextual intelligence and concludes by outlining possible strategies to narrow the gap between the two and resolve the challenge.