Bacteria from diverse habitats colonize and compete in the mouse gut.
To study how microbes establish themselves in a mammalian gut environment, we colonized germ-free mice with microbial communities from human, zebrafish, and termite guts, human skin and tongue, soil, and estuarine microbial mats. Bacteria from these foreign environments colonized and persisted in the mouse gut; their capacity to metabolize dietary and host carbohydrates and bile acids correlated with colonization success. Cohousing mice harboring these xenomicrobiota or a mouse cecal microbiota, along with germ-free "bystanders," revealed the success of particular bacterial taxa in invading guts with established communities and empty gut habitats. Unanticipated patterns of ecological succession were observed; for example, a soil-derived bacterium dominated even in the presence of bacteria from other gut communities (zebrafish and termite), and human-derived bacteria colonized germ-free bystander mice before mouse-derived organisms. This approach can be generalized to address a variety of mechanistic questions about succession, including succession in the context of microbiota-directed therapeutics.


WHO multicenter evaluation of FACSCount CD4 and Pima CD4 T-cell count systems: instrument performance and misclassification of HIV-infected patients.
CD4⁺ T-cell counts are used to screen and follow-up HIV-infected patients during treatment. As part of the World Health Organization prequalification program of diagnostics, we conducted an independent multicenter evaluation of the FACSCount CD4 and the Pima CD4, using the FACSCalibur as reference method. A total of 440 paired capillary and venous blood samples were collected from HIV-infected patients attending the HIV outpatient clinic in Antwerp, Belgium, and the HIV care and treatment center in Dar es Salam, Tanzania. Capillary blood was run on Pima analyzer, whereas venous blood was analyzed on FACSCount, Pima, and FACSCalibur instruments. Precision and agreement between methods were assessed. The FACSCount CD4 results were in agreement with the FACSCalibur results with relative bias of 0.4% and 3.1% on absolute CD4 counts and an absolute bias of -0.6% and -1.1% on CD4% in Antwerp and Dar es Salam, respectively. The Pima CD4 results were in agreement with the FACSCalibur results with relative bias of -4.1% and -9.4% using venous blood and of -9.5% and -0.9% using capillary blood in Antwerp and Dar es Salam, respectively. At the threshold of 350 cells per microliter, the FACSCount CD4 and Pima CD4 using venous and capillary blood misclassified 7%, 9%, and 13% of patients, respectively. The FACSCount CD4 provides reliable CD4 counts and CD4% and is suitable for monitoring adult and pediatric HIV patients in moderate-volume settings. The Pima CD4 is more suitable for screening eligible adult HIV patients for antiretroviral treatment initiation in low-volume laboratories.


Clinical effects and plasma levels of delta 9-tetrahydrocannabinol (delta 9-THC) in heavy and light users of cannabis.
delta 9-Tetrahydrocannabinol (delta 9-THC) was administered in a crossover design by smoking and IV injection to groups of heavy and light users of marihuana. Plasma concentrations of delta 9-THC were similar for the groups after IV injection of 5.0 mg delta 9-THC, but the AUC0-240 min showed a trend towards lower values for the heavy user group. To achieve a maximum desired "high", both groups smoked similar amounts (about 13 mg) of delta 9-THC. Heavy users tended to have higher plasma levels than light users. The systemic availability of smoked delta 9-THC was significantly higher for the heavy users (heavy users 23 +/- 16% vs 10 +/- 7% for light users). These results also indicate that heavy cannabis users smoke more efficiently than casual smokers. Both light and heavy users showed more clinical effect following IV administration than after smoking. The response of the heavy users, both with respect to effect on heart and "high", was quite comparable to that of light users. The present study does not suggest that tolerance readily develops in heavy users.


[A researcher's itinary on the history of the body].
As a pioneer of a new field of research, history of the body, Georges Vigarello goes back over his career as a historian, not so much in search of a hypothetical or ultimate unity of the body, but beyond the original dipersion is rather aiming at relating the existence of coherences and unified logics passing through different fields: the body as an object of thought, of representations and practices. In such a perspective, the history of the body sheds light on a specific era, underlines its culture, while indicating its possible homogeneity.


Review of clinical studies on cholesterol-lowering response to soy protein.
Experiments on animals have shown that soybean protein has hypocholesterolemic and antiatherogenic properties. In human beings, substitution of soy protein for dietary animal protein or addition of soy protein to the diet lowers total and low-density-lipoprotein cholesterol levels in individuals with hypercholesterolemia. Reductions of 20% or more have been obtained with diets high in protein (about 20% of total energy) and relatively low in fat. Triglycerides are also decreased, particularly in subjects with hypertriglyceridemia, but soy-protein diets appear to have little effect on high-density-lipoprotein cholesterol levels. Responses are similar in men and women, but may be greater in younger than in older subjects. The hypocholesterolemic effect is thought to be mainly attributable to the protein itself rather than to nonprotein components of soy-protein preparations. The mechanism of action is not known, and it may not be possible to explain the observed effects in human beings and in experimental animal models by the same mechanism. Although the hypocholesterolemic response to dietary soy protein has been observed by a number of European research groups, substitution of soy protein for animal protein in North American diets has generally had little effect, for reasons that are still not clear.


Spatial arrangement in texture discrimination and texture segregation.
We investigated the role of spatial arrangement of texture elements in three psychophysical experiments on texture discrimination and texture segregation. In our stimuli, oriented Gabor elements formed an iso-oriented and a randomly oriented texture region. We manipulated (1) the orientation similarity in the iso-oriented region by adding orientation jitter to the orientation of each Gabor; (2) the spatial arrangement of the Gabors: quasi-random or regular; and (3) the shape of the edge between the two texture regions: straight or curved. In Experiment 1, participants discriminated an iso-oriented stimulus from a stimulus with only randomly oriented elements. Experiment 2 required texture segregation to judge the shape of the texture edge. Experiment 3 replicated Experiment 2 with Gabors of a smaller spatial extent in a denser arrangement. We found comparable performance levels with regular and quasi-random Gabor positions in the discrimination task but not in the segregation tasks. We conclude that spatial arrangement plays a role in a texture segregation task requiring shape discrimination of the texture edge but not in a texture discrimination task in which it is sufficient to discriminate an iso-oriented region from a completely random region.


Activity, Abundance, and Localization of Quorum Sensing Receptors in Vibrio harveyi.
Quorum sensing (QS) is a process enabling a bacterial population to communicate via small molecules called autoinducers (AIs). This intercellular communication process allows single cells to synchronize their behavior within a population. The marine bacterium Vibrio harveyi ATCC BAA-1116 channels the information of three AI signals into one QS cascade. Three receptors perceive these AIs, the hybrid histidine kinases LuxN, Lux(P)Q and CqsS, to transduce the information to the histidine phosphotransfer (HPt) protein LuxU via phosphorelay, and finally to the response regulator LuxO. Hence, the level of phosphorylated LuxO depends on the AI concentrations. The phosphorylated LuxO (P-LuxO) controls the expression of small regulatory RNAs (sRNAs), which together with the RNA chaperon Hfq, destabilize the transcript of the master regulator luxR. LuxR is responsible for the induction and repression of several genes (e.g., for bioluminescence, exoprotease and siderophore production). In vivo studies with various mutants have demonstrated that the ratio between kinase and phosphatase activities of the individual QS receptors and therefore the P-LuxO/LuxO ratio is crucial not only for the output strength but also for the degree of noise. This study was undertaken to better understand the inherent design principles of this complex signaling cascade, which allows sensing and integration of different signals, but also the differentiated output in individual cells. Therefore, we quantitatively analyzed not only the enzymatic activities, but also the abundance and localization of the three QS receptors. We found that LuxN presents the highest capacity to phosphorylate LuxU, while the phosphatase activity was comparable to LuxQ and CqsS in vitro. In whole cells the copy number of LuxN was higher than that of LuxQ and CqsS, and further increased in the late exponential growth phase. Microscopy experiments indicate that LuxN and LuxQ form independent clusters. Altogether, these results suggest, that the three QS receptors act in parallel, and V. harveyi has developed with LuxN the most dynamic sensing range for HAI-1, the species-specific AI.


Lung surfactant gelation induced by epithelial cells exposed to air pollution or oxidative stress.
Lung surfactant lowers surface tension and adjusts interfacial rheology to facilitate breathing. A novel instrument, the interfacial stress rheometer (ISR), uses an oscillating magnetic needle to measure the shear viscosity and elasticity of a surfactant monolayer at the air-water interface. The ISR reveals that calf lung surfactant, Infasurf, exhibits remarkable fluidity, even when exposed to air pollution residual oil fly ash (ROFA), hydrogen peroxide (H2O2), or conditioned media from resting A549 alveolar epithelial cells (AEC). However, when Infasurf is exposed to a subphase of the soluble fraction of ROFA- or H2O2-treated AEC conditioned media, there is a prominent increase in surfactant elasticity and viscosity, representing two-dimensional gelation. Surfactant gelation is decreased when ROFA-AEC are pretreated with inhibitors of cellular reactive oxygen species (ROS), or with a mitochondrial anion channel inhibitor, as well as when A549-rho0 cells that lack mitochondrial DNA and functional electron transport are investigated. These results implicate both mitochondrial and nonmitochondrial ROS generation in ROFA-AEC-induced surfactant gelation. A549 cells treated with H2O2 demonstrate a dose-dependent increase in lung surfactant gelation. The ISR is a unique and sensitive instrument to characterize surfactant gelation induced by oxidatively stressed AEC.


High-order harmonic generation from a dual-gas, multi-jet array with individual gas jet control.
We present a gas jet array for use in high-order harmonic generation experiments. Precise control of the pressure in each individual gas jet has allowed a thorough investigation into mechanisms contributing to the selective enhancement observed in the harmonic spectra produced by dual-gas, multi-jet arrays. Our results reveal that in our case, the dominant enhancement mechanism is the result of a compression of the harmonic-producing gas jet due to the presence of other gas jets in the array. The individual control of the gas jets in the array also provides a promising method for enhancing the harmonic yield by precise tailoring of the length and pressure gradient of the interaction region.


Autonomy, religious values, and refusal of lifesaving medical treatment.
The principal question of this paper is: Why are religious values special in refusal of lifesaving medical treatment? This question is approached through a critical examination of a common kind of refusal of treatment case, one involving a rational adult. The central value cited in defence of honouring such a patient's refusal is autonomy. Once autonomy is isolated from other justificatory factors, however, possible cases can be imagined which cast doubt on the great valuational weight assigned it by strong anti-paternalists. This weight is sufficient, in their estimation, to justify honouring the patient's refusal. There is thus a tension between the strong anti-paternalist's commitment to the sufficiency of autonomy and our intuitions respecting such cases. Attempts can be made to relieve this tension, such as arguing that patients aren't really rational in the circumstances envisaged, or that other values, such as privacy or bodily integrity, if added to autonomy, are sufficient to justify an anti-paternalistic stance. All such attempts fail, however. But what does not fail is the addition of religious freedom, freedom respecting a patient's religious beliefs and values. Why religious freedom reduces the tension is then explained, and the specialness of religious beliefs and values examined.


A practical approach to prescribing antidepressants.
Although antidepressant drugs do not differ much in their efficacy rates, the particular characteristics of one drug may make it a better choice in a given patient. This article provides insight into the art of prescribing antidepressants in primary care, with recommendations for prescribing for patients with chronic pain, sexual dysfunction, anxiety, chronic fatigue syndrome, fibromyalgia, severe insomnia, old age, diabetes, and heart problems.


Microbial volatile organic compounds.
Microbial volatile organic compounds (MVOCs) are a variety of compounds formed in the metabolism of fungi and bacteria. Of more than 200 compounds identified as MVOCs in laboratory experiments, none can be regarded as exclusively of microbial origin or as specific for certain microbial species. Thus, the recognition of microbially contaminated areas by MVOC measurements is not successful with current methods. In this review, the basic physical and chemical properties of 96 typical MVOCs have been summarised. Of these, toxicological and exposure data were gathered for the 15 MVOCs most often analysed and reported in buildings with moisture and microbial damage. The most obvious health effect of MVOC exposure is eye and upper-airway irritation. However, in human experimental exposure studies, symptoms of irritation have appeared at MVOC concentrations several orders of magnitude higher than those measured indoors (single MVOC levels in indoor environments have ranged from a few ng/m(3) up to 1 mg/m(3)). This is also supported by dose-dependent sensory-irritation response, as determined by the American Society for Testing and Materials mouse bioassay. On the other hand, the toxicological database is poor even for the 15 examined MVOCs. There may be more potent compounds and other endpoints not yet evaluated.


Celiac Disease is Misdiagnosed Based on Serology Only in a Substantial Proportion of Patients.
Although the diagnostic process in celiac disease (CeD) has been addressed in several international guidelines, little is known about the actual proceeding in current clinical practice. This study investigated the initial presentation, the diagnostic process, follow-up evaluations, and adherence to a gluten-free diet in CeD patients in a real-life setting in Switzerland from a patient's perspective. We performed a large patient survey among unselected CeD patients in Switzerland. A total of 1689 patients were analyzed. The vast majority complained of both gastrointestinal and nonspecific symptoms (71.5%), whereas 1.8% reported an asymptomatic disease course. A total of 35.8% CeD patients were diagnosed by a nongastroenterologist. The diagnostic process differed between nongastroenterologists and gastroenterologists, with the latter more often using duodenal biopsy alone or in combination with serology (94.7% vs. 63.0%) and nongastroenterologists more frequently establishing the diagnosis without endoscopy (37.0% vs. 5.3%, P<0.001). Follow-up serology after 6 months was performed only in half of all patients (49.4%), whereas 69.9% had at least 1 follow-up serology within the first year after diet initiation. About 39.7% had a follow-up endoscopy with duodenal biopsies (after a median of 12 mo; range, 1 to 600 mo). The likelihood of receiving any follow-up examination was higher in patients initially diagnosed by a gastroenterologist. A significant proportion of CeD patients are diagnosed by nongastroenterologists. Under the diagnostic lead of the latter, more than a third of the patients receive their diagnosis on the basis of a positive serology and/or genetics only, in evident violation of current diagnostic guidelines, which may lead to an overdiagnosis of this entity.


[Best knee flexion angle through anteromedial portal during anterior cruciate ligament reconstruction].
To investigate the best knee flexion angle by analyzing the length and orientation of the femoral tunnel through anteromedial portal (AM) at different flexion angles during anterior cruciate ligament (ACL) reconstruction. Twelve fresh cadaveric knees were selected to locate the center of ACL femoral footprint through AM using the improved hook slot vernier caliper, and to locate the posterior bone cortex using a diameter 3 mm ball at flexion of 90, 100, 110, 120, and 130 degrees. The femoral tunnel length, standard coronal and sagittal plane angles, and the position relation between exit point and the lateral epicondyle were measured; the tunnel orientation on the anteroposterior and lateral X-ray films was also measured. With increasing flexion of the knee, the femoral tunnel length showed a first increasing and then stable tendency; significant difference was found between at flexion of 90 degrees and at flexions of 100, 110, 120, and 130 degrees, and between flexions of 100 degrees and 120 degrees (P < 0.05). The femoral tunnel showed a trend of decreasing with coronal angle, whereas gradually increasing with sagittal angle. The knee flexion angle had significant difference either among flexions of 90, 110, and 130 degrees or between flexions of 100 degrees and 120 degrees (P < 0.05). The exit point of the femoral tunnel located at the lateral epicondyle of the femur proximal to posterior region at flexion of 90 degrees in all knees, and at flexion of 100 degrees in 7 knees, but it located at the lateral epicondyle of the femur proximal to anterior region at flexion of 110, 120, and 130 degrees in all knees. As the knee flexion angle increasing, the angle between femoral tunnel with the tangent of internal-external femoral condyle on anteroposterior X-ray films showed a trend of decreasing gradually, but a trend of increasing gradually on lateral X-ray films. On the anteroposterior X-ray films, significant differences were found in the angle either among flexions of 90, 110, and 130 degrees or between flexions of 100 degrees and 120 degrees (P < 0.05). On the lateral X-ray films, there were significant differences in the angle among flexions of 90, 100, 110, 120, and 130 degrees (P < 0.05). During ACL reconstruction by AM, 110 degrees is the best flexion angle, which can get the ideal femoral tunnel.


Fos expression following isotonic volume expansion of the unanesthetized male rat.
Cardiopulmonary afferents, baroreceptor afferents, or atrial natriuretic peptide binding to circumventricular organs may mediate the central response to volume expansion, a condition common to pregnancy, exercise training, and congestive heart failure. This study used Fos immunocytochemistry to examine brain regions activated by volume expansion. Male Sprague-Dawley rats were infused with isotonic saline equal to 10% of their body weight in 10 min followed by a maintenance infusion of 0.5 ml/min for 110 min. Control animals received 2-h infusions at 0.01 ml/min. Five minutes after the start of volume expansion, central venous pressure of expanded animals was significantly greater than control animals. The volume-expanded group exhibited significantly greater Fos activation (P < 0.05) in the area postrema, nucleus of the solitary tract, caudal ventrolateral medulla, paraventricular nucleus, supraoptic nucleus, and perinuclear zone of the supraoptic nucleus. Double labeling indicates that oxytocinergic neurons in the supraoptic nucleus are activated. Neurons in brain regions known to inhibit both sympathetic activity and vasopressin release show increased Fos expression following isotonic volume expansion.


Dissociation and reconstitution of a DNA polymerase alpha-primase complex.
The conditions for dissociation of the DNA polymerase alpha-primase complex (DNA polymerase alpha 1) have been examined. It was revealed that 50% ethylene glycol effectively dissociated the complex. The dissociated DNA polymerase and primase were purified to eliminate cross-contaminating activities by column chromatography using buffers containing 50% ethylene glycol. The sedimentation coefficients of the purified DNA polymerase and primase were 7.1S and 5.7S, respectively. These two enzymes were mixed in the presence of 20% ethylene glycol and the mixture was sedimented through a glycerol gradient containing no ethylene glycol. The DNA polymerase and primase activities co-sedimented at 9.1S which corresponds to the S value of intact alpha 1, indicating the reconstitution of the DNA polymerase alpha-primase complex.


Sensory neurons expressing calcitonin gene-related peptide α regulate adaptive thermogenesis and diet-induced obesity.
Heat-sensory neurons from the dorsal root ganglia (DRG) play a pivotal role in detecting the cutaneous temperature and transmission of external signals to the brain, ensuring the maintenance of thermoregulation. However, whether these thermoreceptor neurons contribute to adaptive thermogenesis remains elusive. It is also unknown whether these neurons play a role in obesity and energy metabolism. We used genetic ablation of heat-sensing neurons expressing calcitonin gene-related peptide α (CGRPα) to assess whole-body energy expenditure, weight gain, glucose tolerance, and insulin sensitivity in normal chow and high-fat diet-fed mice. Exvivo lipolysis and transcriptional characterization were combined with adipose tissue-clearing methods to visualize and probe the role of sensory nerves in adipose tissue. Adaptive thermogenesis was explored using infrared imaging of intrascapular brown adipose tissue (iBAT), tail, and core temperature upon various stimuli including diet, external temperature, and the cooling agent icilin. In this report, we show that genetic ablation of heat-sensing CGRPα neurons promotes resistance to weight gain upon high-fat diet (HFD) feeding and increases energy expenditure in mice. Mechanistically, we found that loss of CGRPα-expressing sensory neurons was associated with reduced lipid deposition in adipose tissue, enhanced expression of fatty acid oxidation genes, higher exvivo lipolysis in primary white adipocytes, and increased mitochondrial respiration from iBAT. Remarkably, mice lacking CGRPα sensory neurons manifested increased tail cutaneous vasoconstriction at room temperature. This exacerbated cold perception was not associated with reduced core temperature, suggesting that heat production and heat conservation mechanisms were engaged. Specific denervation of CGRPα neurons in intrascapular BAT did not contribute to the increased metabolic rate observed upon global sensory denervation. Taken together, these findings highlight an important role of cutaneous thermoreceptors in regulating energy metabolism by triggering counter-regulatory responses involving energy dissipation processes including lipid fuel utilization and cutaneous vasodilation.


Dual role for microtubules in regulating cortical contractility during cytokinesis.
Microtubules stimulate contractile-ring formation in the equatorial cortex and simultaneously suppress contractility in the polar cortex; how they accomplish these differing activities is incompletely understood. We measured the behavior of GFP-actin in mammalian cells treated with nocodazole under conditions that either completely eliminate microtubules or selectively disassemble astral microtubules. Selective disassembly of astral microtubules resulted in functional contractile rings that were wider than controls and had altered dynamic activity, as measured by FRAP. Complete microtubule disassembly or selective loss of astral microtubules resulted in wave-like contractile behavior of actin in the non-equatorial cortex, and mislocalization of myosin II and Rho. FRAP experiments showed that both contractility and actin polymerization contributed to the wave-like behavior of actin. Wave-like contractile behavior in anaphase cells was Rho-dependent. We conclude that dynamic astral microtubules function to suppress Rho activation in the non-equatorial cortex, limiting the contractile activity of the polar cortex.


Natural indoles, indole-3-carbinol (I3C) and 3,3'-diindolylmethane (DIM), attenuate staphylococcal enterotoxin B-mediated liver injury by downregulating miR-31 expression and promoting caspase-2-mediated apoptosis.
Staphylococcal enterotoxin B (SEB) is a potent superantigen capable of inducing inflammation characterized by robust immune cell activation and proinflammatory cytokine release. Exposure to SEB can result in food poisoning as well as fatal conditions such as toxic shock syndrome. In the current study, we investigated the effect of natural indoles including indole-3-carbinol (I3C) and 3,3'-diindolylmethane (DIM) on SEB-mediated liver injury. Injection of SEB into D-galactosamine-sensitized female C57BL/6 mice resulted in liver injury as indicated by an increase in enzyme aspartate transaminase (AST) levels, induction of inflammatory cytokines, and massive infiltration of immune cells into the liver. Administration of I3C and DIM (40 mg/kg), by intraperitonal injection, attenuated SEB-induced acute liver injury, as evidenced by decrease in AST levels, inflammatory cytokines and cellular infiltration in the liver. I3C and DIM triggered apoptosis in SEB-activated T cells primarily through activation of the intrinsic mitochondrial pathway. In addition, inhibitor studies involving caspases revealed that I3C and DIM-mediated apoptosis in these activated cells was dependent on caspase-2 but independent of caspase-8, 9 and 3. In addition, I3C and DIM caused a decrease in Bcl-2 expression. Both compounds also down-regulated miR-31, which directly targets caspase-2 and influences apoptosis in SEB-activated cells. Our data demonstrate for the first time that indoles can effectively suppress acute hepatic inflammation caused by SEB and that this may be mediated by decreased expression of miR-31 and consequent caspase-2-dependent apoptosis in T cells.


Molecular mechanism of EAG1 channel inhibition by imipramine binding to the PAS domain.
Ether-a-go-go (EAG) channels are key regulators of neuronal excitability and tumorigenesis. EAG channels contain an N-terminal Per-Arnt-Sim (PAS) domain that can regulate currents from EAG channels by binding small molecules. The molecular mechanism of this regulation is not clear. Using surface plasmon resonance and electrophysiology we show that a small molecule ligand imipramine can bind to the PAS domain of EAG1 channels and inhibit EAG1 currents via this binding. We further used a combination of molecular dynamics (MD) simulations, electrophysiology, and mutagenesis to investigate the molecular mechanism of EAG1 current inhibition by imipramine binding to the PAS domain. We found that Tyr71, located at the entrance to the PAS domain cavity, serves as a "gatekeeper" limiting access of imipramine to the cavity. MD simulations indicate that the hydrophobic electrostatic profile of the cavity facilitates imipramine binding and in silico mutations of hydrophobic cavity-lining residues to negatively charged glutamates decreased imipramine binding. Probing the PAS domain cavity-lining residues with site-directed mutagenesis, guided by MD simulations, identified D39 and R84 as residues essential for the EAG1 channel inhibition by imipramine binding to the PAS domain. Taken together, our study identified specific residues in the PAS domain that could increase or decrease EAG1 current inhibition by imipramine binding to the PAS domain. These findings should further the understanding of molecular mechanisms of EAG1 channel regulation by ligands and facilitate the development of therapeutic agents targeting these channels.


Isolation, Characterization, and Comparative Genomic Analysis of Bacteriophage Ec_MI-02 from Pigeon Feces Infecting Escherichia coli O157:H7.
The most significant serotype of Shiga-toxigenic Escherichia coli that causes foodborne illnesses is Escherichia coli O157:H7. Elimination of E. coli O157:H7 during food processing and storage is a possible solution. Bacteriophages have a significant impact on bacterial populations in nature due to their ability to lyse their bacterial host. In the current study, a virulent bacteriophage, Ec_MI-02, was isolated from the feces of a wild pigeon in the United Arab Emirates (UAE) for potential future use as a bio-preservative or in phage therapy. Using a spot test and an efficiency of plating analysis, Ec_MI-02 was found to infect in addition to the propagation host, E. coli O157:H7 NCTC 12900, five different serotypes of E. coli O157:H7 (three clinical samples from infected patients, one from contaminated green salad, and one from contaminated ground beef). Based on morphology and genome analysis, Ec_MI-02 belongs to the genus Tequatrovirus under the order Caudovirales. The adsorption rate constant (K) of Ec_MI-02 was found to be 1.55 × 10-8 mL/min. The latent period was 50 min with a burst size of almost 10 plaque forming units (pfu)/host cell in the one-step growth curve when the phage Ec_MI-02 was cultivated using the propagation host E. coli O157:H7 NCTC 12900. Ec_MI-02 was found to be stable at a wide range of pH, temperature, and commonly used laboratory disinfectants. Its genome is 165,454 bp long with a GC content of 35.5% and encodes 266 protein coding genes. Ec_MI-02 has genes encoding for rI, rII, and rIII lysis inhibition proteins, which supports the observation of delayed lysis in the one-step growth curve. The current study provides additional evidence that wild birds could also be a good natural reservoir for bacteriophages that do not carry antibiotic resistance genes and could be good candidates for phage therapy. In addition, studying the genetic makeup of bacteriophages that infect human pathogens is crucial for ensuring their safe usage in the food industry.


Validity of thermal ramping assays used to assess thermal tolerance in arthropods.
Proper assessment of environmental resistance of animals is critical for the ability of researchers to understand how variation in environmental conditions influence population and species abundance. This is also the case for studies of upper thermal limits in insects, where researchers studying animals under laboratory conditions must select appropriate methodology on which conclusions can be drawn. Ideally these methods should precisely estimate the trait of interest and also be biological meaningful. In an attempt to develop such tests it has been proposed that thermal ramping assays are useful assays for small insects because they incorporate an ecologically relevant gradual temperature change. However, recent model-based papers have suggested that estimates of thermal resistance may be strongly confounded by simultaneous starvation and dehydration stress. In the present study we empirically test these model predictions using two sets of independent experiments. We clearly demonstrate that results from ramping assays of small insects (Drosophila melanogaster) are not compromised by starvation- or dehydration-stress. Firstly we show that the mild disturbance of water and energy balance of D. melanogaster experienced during the ramping tests does not confound heat tolerance estimates. Secondly we show that flies pre-exposed to starvation and dehydration have "normal" heat tolerance and that resistance to heat stress is independent of the energetic and water status of the flies. On the basis of our results we discuss the assumptions used in recent model papers and present arguments as to why the ramping assay is both a valid and ecologically relevant way to measure thermal resistance in insects.


Neurosensory impairment after surgical closure of patent ductus arteriosus in extremely low birth weight infants: results from the Trial of Indomethacin Prophylaxis in Preterms.
To determine whether surgical closure of a patent ductus arteriosus (PDA) is a risk factor for bronchopulmonary dysplasia (BPD), severe retinopathy of prematurity (ROP), and neurosensory impairment in extremely low birth weight (ELBW) infants. We studied 426 infants with a symptomatic PDA, 110 of whom underwent PDA ligation and 316 of whom received medical therapy only. All infants participated in the multicenter Trial of Indomethacin Prophylaxis in Preterms (TIPP) and were observed to a corrected age of 18 months. Of the 95 infants who survived after PDA ligation, 50 (53%) had neurosensory impairment, compared with 84 of the 245 infants (34%) who survived after receiving only medical therapy (adjusted odds ratio, 1.98; 95% CI, 1.18-3.30; P = .0093). BPD (adjusted odds ratio, 1.81; 95% CI, 1.09-3.03; P = .023) and severe ROP (adjusted odds ratio, 2.20; 95% CI, 1.19-4.07; P = .012) were also more common after surgical PDA closure. PDA ligation may be associated with increased risks of BPD, severe ROP, and neurosensory impairment in ELBW infants.


Variceal haemorrhage: An update on treatment.
Recent advances in the management of variceal haemorrhage include injection of cyanoacrylate and the use of banding ligation. New pharmacological agents, such as somatostatin and octreotide, have also proved to be effective in controlling variceal haemorrhage. The transjugular intrahepatic porto-systemic shunt (TIPS) represents an effective rescue when other treatment modalities fail. A multi-disciplinary approach should be emphasized in the management of this high-risk complication of portal hypertension.


A Zebrafish Model of Mycobacterium leprae Granulomatous Infection.
Understanding the pathogenesis of leprosy granulomas has been hindered by a paucity of tractable experimental animal models. Mycobacterium leprae, which causes leprosy, grows optimally at approximately 30°C, so we sought to model granulomatous disease in the ectothermic zebrafish. We found that noncaseating granulomas develop rapidly and eventually eradicate infection. rag1 mutant zebrafish, which lack lymphocytes, also form noncaseating granulomas with similar kinetics, but these control infection more slowly. Our findings establish the zebrafish as a facile, genetically tractable model for leprosy and reveal the interplay between innate and adaptive immune determinants mediating leprosy granuloma formation and function.


An indirect route to repetitive actions.
It is increasingly evident that there is a genetic contribution to autism spectrum disorders (ASDs) and other neural disorders involving excessive repetition of action sequences. Among the implicated genes in these disorders are those encoding postsynaptic scaffolding proteins with roles in synaptic transmission and plasticity. Several mouse models harboring synonymous mutations have shown alterations in synaptic transmission within the striatum, which has key roles in controlling actions and action sequences. In this issue of the JCI, Wang and coworkers show that glutamatergic synaptic transmission onto striatal projection neurons is weakened in mutant mice lacking the SH3 and multiple ankyrin repeat domains 3 (SHANK3B) scaffolding protein, defective expression of which has been implicated in ASDs. This synaptic alteration gives rise to stronger activity in the indirect pathway accompanied by decreased dendritic spines on the indirect pathway medium spiny projection neuron, indicative of decreased numbers of glutamatergic synapses. Selectively enhancing activity in this pathway reduced excessive repetitive grooming in the mutant mice. Changes in glutamatergic input to striatal projection neurons have been observed in several other murine ASD models and associated disorders. Thus, manipulation of the function of the striatal indirect pathway may be a useful therapeutic target for treating disorders characterized by excessive repetitive behaviors.


Assessing obstetric risk. A review of obstetric risk-scoring systems.
The primary purpose of formal risk assessment in obstetrics is the prevention and consequent reduction of perinatal morbidity and mortality through early identification and intervention. Obstetric risk scoring quantifies identified risk factors according to their relative contribution to adverse perinatal outcomes and aggregates individual factor scores. A review of existing scoring methods reveals consistently low positive predictive values and more accurate prediction when the assessment occurs closer to the time of actual delivery. While numerous scoring systems exist in the literature, few are convenient in practice, and none appear to assess effectively the dynamic character of pregnancy.


Rating communication skills in dental practice: the impact of different sociodemographic factors.
Communication abilities are essential for the successful operation of a dental business and significantly influence outcomes, compliance, and patient satisfaction. The aim of our study was to evaluate the knowledge and practice of doctor-patient communication among Jordanian dentists. This evaluation was conducted through a survey based on the key components of the Calgary Cambridge Observation Guides. Additionally, the impact of several sociodemographic characteristics on communication abilities was investigated. This cross-sectional study was conducted from January to June 2022. The data collection tool was an online questionnaire developed by the researchers, consisting of three sections: self-reported demographic and professional data, the practice of doctor-patient communication, and knowledge of doctor-patient communication. The study included 305 dentists, comprising 106 males and 199 females, with a mean age of 32.9 ± 9.0 years. The mean score for communication skills knowledge was 41.5, indicating a moderate level of communication skills knowledge. Female dentists demonstrated significantly higher communication scores compared to their male counterparts, and those working in the private sector scored significantly higher than those in the governmental sector or in both sectors (P ≤ 0.05). In general, older and more experienced dentists exhibited better communication skills. Educational level had a positive impact on certain communication skills items. 58.4% believed that communication skills can always be developed and improved through training sessions, while 48.9% reported never having attended such courses. 95.1% believed that training courses on communication skills are always necessary as part of the educational curriculum. The main obstacles that may deter dentists from considering communication skills courses were limited time (62.3%), course availability (37.7%), cost (28.2%), and perceived lack of importance (8.2%). Among a sample of Jordanian dentists, there appears to be a discrepancy between knowledge and self-reported practices regarding communication abilities. In certain crucial, evidence-based areas of doctor-patient communication, there are fundamental deficiencies. Considering the significant role dentists play in oral health and prevention, communication skills should be a top educational priority for them.


Extending the durability of cultivar resistance by limiting epidemic growth rates.
Cultivar resistance is an essential part of disease control programmes in many agricultural systems. The use of resistant cultivars applies a selection pressure on pathogen populations for the evolution of virulence, resulting in loss of disease control. Various techniques for the deployment of host resistance genes have been proposed to reduce the selection for virulence, but these are often difficult to apply in practice. We present a general technique to maintain the effectiveness of cultivar resistance. Derived from classical population genetics theory; any factor that reduces the population growth rates of both the virulent and avirulent strains will reduce selection. We model the specific example of fungicide application to reduce the growth rates of virulent and avirulent strains of a pathogen, demonstrating that appropriate use of fungicides reduces selection for virulence, prolonging cultivar resistance. This specific example of chemical control illustrates a general principle for the development of techniques to manage the evolution of virulence by slowing epidemic growth rates.


RNA Interference in the Egg Parasitoid, Trichogramma dendrolimi Matsumura.
The egg parasitoids, Trichogramma spp, are recognized as efficient biological control agents against various lepidopteran pests in agriculture and forests. The immature stages of Trichogramma offspring develop within the host egg, exhibiting remarkable diminutiveness (approximately 0.5 mm in adult length). RNA-interference (RNAi) methodology has emerged as a crucial tool for elucidating gene functions in numerous organisms. However, manipulating RNAi in certain small parasitoid species, such as Trichogramma, has generally posed significant challenges. In this study, we present an efficient RNAi method in Trichogramma denrolimi. The outlined procedure encompasses the acquisition and isolation of individual T. dendrolimi specimens from host eggs, the design and synthesis of double-stranded RNA (dsRNA), the in vitro transplantation and cultivation of T. dendrolimi pupae, the micro-injection of dsRNA, and the subsequent assessment of target gene knockdown through RT-qPCR analysis. This study furnishes a comprehensive, visually detailed procedure for conducting RNAi experiments in T. dendrolimi, thereby enabling researchers to investigate the gene regulation in this species. Furthermore, this methodology is adaptable for RNAi studies or micro-injections in other Trichogramma species with minor adjustments, rendering it a valuable reference for conducting RNAi experiments in other endoparasitic species.


Surface Functionalization with Polyethylene Glycol and Polyethyleneimine Improves the Performance of Graphene-Based Materials for Safe and Efficient Intracellular Delivery by Laser-Induced Photoporation.
Nanoparticle mediated laser-induced photoporation is a physical cell membrane disruption approach to directly deliver extrinsic molecules into living cells, which is particularly promising in applications for both adherent and suspension cells. In this work, we explored surface modifications of graphene quantum dots (GQD) and reduced graphene oxide (rGO) with polyethylene glycol (PEG) and polyethyleneimine (PEI) to enhance colloidal stability while retaining photoporation functionality. After photoporation with FITC-dextran 10 kDa (FD10), the percentage of positive HeLa cells (81% for GQD-PEG, 74% for rGO-PEG and 90% for rGO-PEI) increased approximately two-fold compared to the bare nanomaterials. While for Jurkat suspension cells, the photoporation efficiency with polymer-modified graphene-based nanomaterial reached as high as 80%. Cell viability was >80% in all these cases. In addition, polymer functionalization proved to be beneficial for the delivery of larger macromolecules (FD70 and FD500) as well. Finally, we show that rGO is suitable for photoporation using a near-infrared laser to reach 80% FD10 positive HeLa cells at 80% cell viability. We conclude that modification of graphene-based nanoparticles with PEG and especially PEI provide better colloidal stability in cell medium, resulting in more uniform transfection and overall increased efficiency.


First do no harm: A patient-reported survey of split skin graft donor site morbidities following thin and super-thin graft harvest.
The split-thickness skin graft (STSG) donor site is the commonest used during burn surgery which has its own complications and as such the focus should be on minimizing it. Modifications to practice in our unit which we believe aid this include limiting the amount of STSG taken and the harvest of super-thin STSGs, with 0.003-0.005 in. (0.08-0.13 mm) being the commonest dermatome settings used. A patient-reported survey via a mobile phone link to a questionnaire was sent to 250 patients who had a STSG for an acute burn between 1st August 2020 and 31st July 2021. Patient demographics were collected from electronic records including the thickness of the FTSG taken when recorded. Patient responses were statistically analyzed and logistic regression with backwards elimination was performed to explore which contributing factors led to an improved experience of the donor site. Questionnaire responses were obtained from 107 patients (43%). These were between one and two and a half years after the injury. Concerning early donor site issues, itch was a problem for 52% of patients, pain was a problem for 48% of patients. Less common problems (fewer than 25% of patients) were leaking donor sites, wound breakdown, and over-granulation. Regarding long-term outcomes, increased, decreased or mixed pigmentation at the donor site was reported by 32% patients at the time of the survey. Hyper-vascular donor sites were reported by 24% patients. Raised or uneven feeling donor sites were reported by 19% patients, firm or stiff donor sites by 13% patients, and altered sensation by 10% patients. At the time of the survey, 70% responders reported their donor site looked "the same or about the same as my normal skin". Of these, 62 reported how long it took for this to happen, and it equates to a third looking normal at 6 months and half looking normal at a year. For the 32 patients who reported their donor site looking abnormal, 72% were "not bothered" by it. Patients with super-thin grafts (0.003-0.005 in.) were significantly more likely to have normal sensation, normal stiffness, and be less raised at their donor sites than those who had thin grafts (0.006-0.008 in.). This survey gives important information on patients' experiences of donor site morbidity that may form part of an informed consent process and allow tailored advice. Furthermore, it suggests that super-thin grafts may provide a superior donor site experience for patients.


Bomb blast injuries to the ear: the London Bridge incident series.
Twelve patients who were treated for ear injuries at Guy's Hospital following the London Bridge bomb blast in February 1992 were reviewed. Among three there were four perforated eardrums, two of which closed spontaneously (50%). All three patients had a persistent mixed hearing loss. The remaining nine patients had acute sensorineural hearing loss and/or tinnitus only. Four of these had resolved completely by 4h, another one by 48h, and two by 4 weeks. Two patients had a residual high frequency hearing loss. In total, five patients (42%) have a persistent hearing loss. None of the patients suffered from balance problems. In summary, the ear is very susceptible to bomb blast injury, but there is a high rate of spontaneous closure of perforations and improvement of sensorineural hearing loss and tinnitus.


New Host-Directed Therapeutics for the Treatment of Clostridioides difficile Infection.
Frequent and excessive use of antibiotics primes patients to Clostridioides difficile infection (CDI), which leads to fatal pseudomembranous colitis, with limited treatment options. In earlier reports, we used a drug repurposing strategy and identified amoxapine (an antidepressant), doxapram (a breathing stimulant), and trifluoperazine (an antipsychotic), which provided significant protection to mice against lethal infections with several pathogens, including C. difficile However, the mechanisms of action of these drugs were not known. Here, we provide evidence that all three drugs offered protection against experimental CDI by reducing bacterial burden and toxin levels, although the drugs were neither bacteriostatic nor bactericidal in nature and had minimal impact on the composition of the microbiota. Drug-mediated protection was dependent on the presence of the microbiota, implicating its role in evoking host defenses that promoted protective immunity. By utilizing transcriptome sequencing (RNA-seq), we identified that each drug increased expression of several innate immune response-related genes, including those involved in the recruitment of neutrophils, the production of interleukin 33 (IL-33), and the IL-22 signaling pathway. The RNA-seq data on selected genes were confirmed by quantitative real-time PCR (qRT-PCR) and protein assays. Focusing on amoxapine, which had the best anti-CDI outcome, we demonstrated that neutralization of IL-33 or depletion of neutrophils resulted in loss of drug efficacy. Overall, our lead drugs promote disease alleviation and survival in the murine model through activation of IL-33 and by clearing the pathogen through host defense mechanisms that critically include an early influx of neutrophils.IMPORTANCEClostridioides difficile is a spore-forming anaerobic bacterium and the leading cause of antibiotic-associated colitis. With few therapeutic options and high rates of disease recurrence, the need to develop new treatment options is urgent. Prior studies utilizing a repurposing approach identified three nonantibiotic Food and Drug Administration-approved drugs, amoxapine, doxapram, and trifluoperazine, with efficacy against a broad range of human pathogens; however, the protective mechanisms remained unknown. Here, we identified mechanisms leading to drug efficacy in a murine model of lethal C. difficile infection (CDI), advancing our understanding of the role of these drugs in infectious disease pathogenesis that center on host immune responses to C. difficile Overall, these studies highlight the crucial involvement of innate immune responses, as well as the importance of immunomodulation as a potential therapeutic option to combat CDI.


Increased FADS2-derived n-6 PUFAs and reduced n-3 PUFAs in plasma of atopic dermatitis patients.
Fatty acid concentrations, in particular n-3 and n-6 polyunsaturated fatty acids (PUFAs), have been described to be dysregulated in atopic dermatitis (AD) patients. The role of genetic polymorphisms of fatty acid enzymes in AD is controversial. We determined in a Hungarian cohort of healthy volunteers (n = 20) and AD patients (n = 20) triglyceride-, sterol- and phospholipid-bound fatty acids in the plasma, mRNA expression of fatty acid desaturase 2 (FADS2) and stearoyl-coenzyme A desaturase 1 in peripheral blood mononuclear cells (PBMCs) and FADS2 concentrations in plasma. We observed higher levels of monounsaturated fatty acids, 16:1 versus 16:0 ratios in phospholipids, triglycerides and sterol esters in patients compared to healthy subjects. In addition higher levels of the FADS2-derived n-6 PUFAs γ-linolenic acid and dihomo-γ-linolenic acid were observed in PBMCs of patients as well as lower levels of n-3 PUFAs. We conclude that the increased expression of FADS2 in PBMCs, as a representative tissue accessible from human blood of AD patients, might be responsible for higher levels of FADS2-derived n-6 PUFAs and lower n-3 PUFA levels in patients.


Deep Brain Stimulation for Obesity.
Obesity is now the third leading cause of preventable death in the US, accounting for 216,000 deaths annually and nearly 100 billion dollars in health care costs. Despite advancements in bariatric surgery, substantial weight regain and recurrence of the associated metabolic syndrome still occurs in almost 20-35% of patients over the long-term, necessitating the development of novel therapies. Our continually expanding knowledge of the neuroanatomic and neuropsychiatric underpinnings of obesity has led to increased interest in neuromodulation as a new treatment for obesity refractory to current medical, behavioral, and surgical therapies. Recent clinical trials of deep brain stimulation (DBS) in chronic cluster headache, Alzheimer's disease, and depression and obsessive-compulsive disorder have demonstrated the safety and efficacy of targeting the hypothalamus and reward circuitry of the brain with electrical stimulation, and thus provide the basis for a neuromodulatory approach to treatment-refractory obesity. In this study, we review the literature implicating these targets for DBS in the neural circuitry of obesity. We will also briefly review ethical considerations for such an intervention, and discuss genetic secondary-obesity syndromes that may also benefit from DBS. In short, we hope to provide the scientific foundation to justify trials of DBS for the treatment of obesity targeting these specific regions of the brain.


Peri-implant mucositis.
This narrative review was prepared for the 2017 World Workshop of the American Academy of Periodontology and European Federation of Periodontology to address key questions related to the clinical condition of peri-implant mucositis, including: 1) the definition of peri-implant mucositis, 2) conversion of peri-implant health to the biofilm-induced peri-implant mucositis lesion, 3) reversibility of peri-implant mucositis, 4) the long-standing peri-implant mucositis lesion, 5) similarities and differences between peri-implant mucositis at implants and gingivitis at teeth, and 6) risk indicators/factors for peri-implant mucositis. A literature search of MEDLINE (PubMed) and The Cochrane Library up to and including July 31, 2016, was carried out using the search strategy (peri-implant[All Fields] AND ("mucositis"[MeSH Terms] OR "mucositis"[All Fields])) OR (periimplant[All Fields] AND mucosits[All Fields]). Prospective, retrospective, and cross-sectional studies and review papers that focused on risk factors/indicators for peri-implant mucositis as well as experimental peri-implant mucositis studies in animals and humans were included. Peri-implant mucositis is an inflammatory lesion of the soft tissues surrounding an endosseous implant in the absence of loss of supporting bone or continuing marginal bone loss. A cause-and-effect relationship between experimental accumulation of bacterial biofilms around titanium dental implants and the development of an inflammatory response has been demonstrated. The experimental peri-implant mucositis lesion is characterized by an inflammatory cell infiltrate present within the connective tissue lateral to the barrier epithelium. In long-standing peri-implant mucositis, the inflammatory cell infiltrate is larger in size than in the early (3-week) experimental peri-implant mucositis lesion. Biofilm-induced peri-implant mucositis is reversible at the host biomarker level once biofilm control is reinstituted. Reversal of the clinical signs of inflammation may take longer than 3 weeks. Factors identified as risk indicators for peri-implant mucositis include biofilm accumulation, smoking, and radiation. Further evidence is required for potential risk factors, including diabetes, lack of keratinized mucosa, and presence of excess luting cement. Peri-implant mucositis is caused by biofilm accumulation which disrupts the host-microbe homeostasis at the implant-mucosa interface, resulting in an inflammatory lesion. Peri-implant mucositis is a reversible condition at the host biomarker level. Therefore, the clinical implication is that optimal biofilm removal is a prerequisite for the prevention and management of peri-implant mucositis. An understanding of peri-implant mucositis is important because it is considered a precursor for peri-implantitis.


The biology of bacterial peptidoglycans and their impact on host immunity and physiology.
Peptidoglycans (PGN) are a constituent of the bacterial cell wall, and are shed as bacteria divide. The presence of PGN is therefore a marker of bacterial activity that has been exploited by both plants and animals to induce defence mechanisms. Pattern recognition receptors that recognize PGN are extremely well conserved throughout evolution and shown to play important and diverse role in the development, homeostasis and activation of the immune system. In addition, PGN can be detected beyond mucosal surfaces, and their receptor can be expressed in tissues and cells that are far from the niches where bacteria reside. Thus, PGN affects not only the host's immunity, but also more generally the host's physiology. In this review, we discuss the biochemistry and biology of PGN, and their intriguing effects on the development of the immune system and the host physiology.


Rapid generation of OPC-like cells from human pluripotent stem cells for treating spinal cord injury.
Remyelination via the transplantation of oligodendrocyte precursor cells (OPCs) has been considered as a strategy to improve the locomotor deficits caused by traumatic spinal cord injury (SCI). To date, enormous efforts have been made to derive OPCs from human pluripotent stem cells (hPSCs), and significant progress in the transplantation of such cells in SCI animal models has been reported. The current methods generally require a long period of time (>2 months) to obtain transplantable OPCs, which hampers their clinical utility for patients with SCI. Here we demonstrate a rapid and efficient method to differentiate hPSCs into neural progenitors that retain the features of OPCs (referred to as OPC-like cells). We used cell sorting to select A2B5-positive cells from hPSC-derived neural rosettes and cultured the selected cells in the presence of signaling cues, including sonic hedgehog, PDGF and insulin-like growth factor-1. This method robustly generated neural cells positive for platelet-derived growth factor receptor-α (PDGFRα) and NG2 (~90%) after 4 weeks of differentiation. Behavioral tests revealed that the transplantation of the OPC-like cells into the spinal cords of rats with contusive SCI at the thoracic level significantly improved hindlimb locomotor function. Electrophysiological assessment revealed enhanced neural conduction through the injury site. Histological examination showed increased numbers of axon with myelination at the injury site and graft-derived myelin formation with no evidence of tumor formation. Our method provides a cell source from hPSCs that has the potential to recover motor function following SCI.


Commonly Used Insect Repellents Hide Human Odors from Anopheles Mosquitoes.
The mode of action for most mosquito repellents is unknown. This is primarily due to the difficulty in monitoring how the mosquito olfactory system responds to repellent odors. Here, we used the Q-system of binary expression to enable activity-dependent Ca2+ imaging in olfactory neurons of the African malaria mosquito Anopheles coluzzii. This system allows neuronal responses to common insect repellents to be directly visualized in living mosquitoes from all olfactory organs, including the antenna. The synthetic repellents N,N-diethyl-meta-toluamide (DEET) and IR3535 did not activate Anopheles odorant receptor co-receptor (Orco)-expressing olfactory receptor neurons (ORNs) at any concentration, and picaridin weakly activated ORNs only at high concentrations. In contrast, natural repellents (i.e. lemongrass oil and eugenol) strongly activated small numbers of ORNs in the Anopheles mosquito antennae at low concentrations. We determined that DEET, IR3535, and picaridin decrease the response of Orco-expressing ORNs when these repellents are physically mixed with activating human-derived odorants. We present evidence that synthetic repellents may primarily exert their olfactory mode of action by decreasing the amount of volatile odorants reaching ORNs. These results suggest that synthetic repellents disruptively change the chemical profile of host scent signatures on the skin surface, rendering humans invisible to Anopheles mosquitoes.


Regulation of protein synthesis in isolated hepatocytes by calcium-mobilizing hormones.
The incorporation of leucine into protein was studied in Ca2+-depleted and Ca2+-restored preparations of normal liver cells isolated from fed, adult male rats. Ca2+-restored cells incorporated amino acid 5-10-fold more rapidly than did Ca2+-depleted cells for incubation periods up to 1 hr. Readdition of Ca2+ at supraphysiologic concentrations (3 mM) to depleted cells restored the rate of incorporation within 8-10 min, whereas lesser concentrations of the cation acted more slowly. Vasopressin and alpha-adrenergic agonists rapidly (in minutes) inhibited amino acid incorporation to variable degrees in liver cells, with pronounced inhibitions (40-75%) occurring at moderate (0.1-1 mM) extracellular Ca2+ concentrations and smaller inhibitions (10-30%) occurring at supraphysiologic concentrations of the cation. Hormonally produced inhibitions were more intense at acid pH than at alkaline pH. The effects of epinephrine were mediated through alpha 1-adrenergic receptors and were not additive with those of vasopressin at saturating concentrations. It is proposed that these hormones, which are known to mobilize sequestered Ca2+ within liver cells, inhibit amino acid incorporation by influencing a Ca2+ requirement associated with protein synthesis.


Association of smoking, alcohol drinking and dietary factors with esophageal cancer in high- and low-risk areas of Jiangsu Province, China.
To study the main environmental and lifestyle factors that account for the regional differences in esophageal cancer (EC) risk in low- and high-risk areas of Jiangsu Province, China. Since 2003, a population-based case-control study has been conducted simultaneously in low-risk (Ganyu County) and high-risk (Dafeng County) areas of Jiangsu Province, China. Using identical protocols and pre-tested standardized questionnaire, following written informed consent, eligible subjects were inquired about their detail information on potential determinants of EC, including demographic information, socio-economic status, living conditions, disease history, family cancer history, smoking, alcohol drinking, dietary habits, frequency, amount of food intake, etc. Conditional logistic regression with maximum likelihood estimation was used to obtain Odds ratio (OR) and 95 % confidence interval (95% CI), after adjustment for potential confounders. In the preliminary analysis of the ongoing study, we recruited 291 pairs of cases and controls in Dafeng and 240 pairs of cases and controls in Ganyu, respectively. In both low-risk and high-risk areas, EC was inversely associated with socio-economic status, such as level of education, past economic status and body mass index. However, this disease was more frequent among those who had a family history of cancer or encountered misfortune in the past 10 years. EC was also more frequent among smokers, alcohol drinkers and fast eaters. Furthermore, there was a geographic variation of the associations between smoking, alcohol drinking and EC risk despite the similar prevalence of these risk factors in both low-risk and high-risk areas. The dose-response relationship of smoking and smoking related variables, such as age of the first smoking, duration and amount were apparent only in high-risk areas. On the contrary, a dose-response relationship on the effect of alcohol drinking on EC was observed only in low-risk areas. The environmental risk factors, together with genetic factors and gene-environmental interactions might be the main reason for this high-risk gradient in Jiangsu Province, China.


A sex/age anomaly in thermal comfort observed in an office worker field study: A menopausal effect?
In a field study conducted in office settings in Sydney, Australia, background survey and right-here-right-now thermal comfort questionnaires were collected from a sample of office workers. Indoor environmental observations, including air temperature, mean radiant temperature, air velocity, and relative humidity, were also recorded and matched with each questionnaire according to the time and location. During exploratory data analyses, we observed that female subjects aged over 40 and 50 or younger registered significantly warmer sensations than other subjects, male and female, from other age ranges. To further explore this phenomenon, the sample of building occupants was classified into two groups-women of perimenopausal age (over 40 and 50 or younger) while the remaining respondents served as a reference group for comparison. Women in the perimenopausal age range demonstrated an increased perception of warmth (p < 0.01) and expressed thermal dissatisfaction more frequently (p < 0.01) than the reference group respondents who were exposed to the same indoor environmental conditions. Furthermore, women of perimenopausal age also expressed preference for cooler thermal environments, that is, lower air temperature (p < 0.01) and greater air movement (p<0.01) than the reference group, and their thermal neutrality (ie, the room temperature corresponding to a neutral thermal sensation) was approximately 2°C cooler than that of the reference group (20.7°C vs 22.4°C). A potential physiological explanation for the distinct thermal perception of women aged over 40 and 50 or younger observed in this study could stem from menopausal symptoms-the presence of hot flushes and dysregulation of the thermoregulatory system.


In vitro and in vivo evaluation of novel immediate release carbamazepine tablets: complexation with hydroxypropyl-β-cyclodextrin in the presence of HPMC.
Carbamazepine (CBZ)-hydroxypropyl-β-cyclodextrin (HP-β-CD) complex in the presence of HPMC was prepared and characterized by differential scanning calorimetry (DSC) and X-ray diffractometer intended for improving the dissolution rate of CBZ. The phase-solubility method was used to investigate the effect of HP-β-CD and HPMC on the solubility of CBZ. Tablets of the resulting complex were prepared using direct compression method and the bioavailability was evaluated in beagle dogs using a UPLC/MS/MS method. The results showed solubility of CBZ was increased up to 95 times by complexation with HP-β-CD in the presence of 0.1% HPMC. The results of DSC and X-ray diffraction proved a formation of complex between CBZ and HP-β-CD. Dissolution rate of CBZ was notably improved from complex tablets with more than 97.39% released within 10 min; whereas for the commercial tablets, around 60% was released within 30 min. Using commercial tablets as the reference formulation, the bioavailability of complex tablets was considerably increased by 1.5-fold (P<0.05) and T(max) was reduced to 0.88 h compared with 1.25 h for commercial tablets. Furthermore, a lower inter-subject variability (49.9%) was observed compared with that of the commercial tablets (39.7%). It is evident from the results herein that complexation with HP-β-CD in the presence of HPMC is a feasible way to prepare a rapidly acting and better absorbed CBZ oral product.


Spatio-temporal dynamics of gastrointestinal helminths infecting four lake whitefish (Coregonus clupeaformis) stocks in northern lakes Michigan and Huron, U.S.A.
This study was undertaken to identify the community composition, structure, and dynamics of helminths infecting the gastrointestinal tract (GIT) of lake whitefish (Coregonus clupeaformis) collected from 4 sites in northern lakes Huron (Cheboygan and De Tour Village) and Michigan (Big Bay de Noc and Naubinway) from fall 2003 through summer 2006. A total of 21,203 helminths was retrieved from the GITs of 1,284 lake whitefish. Approximately 42% (SE = 1.4%) of the examined lake whitefish were infected with at least 1 helminth species in their GIT, with a mean intensity of 39.4 worms/fish (SE = 0.3) and a mean abundance of 16.4 worms/fish (SE = 0.1). Collected helminths appeared to be generalists and consisted of 2 phyla (Acanthocephala and Cestoda) and 5 species (Acanthocephalus dirus, Neoechinorhynchus tumidus, Echinorhynchus salmonis, Cyathocephalus truncatus, and Bothriocephalus sp.). Lake whitefish from Lake Huron on average had greater infection prevalences, abundances, and intensities than did fish from Lake Michigan. Infection parameters for each of the helminth species generally followed the same pattern observed for the combined data. Acanthocephalus dirus was the most prevalent and abundant helminth in lake whitefish GITs, although intensity of infection was the greatest for C. truncatus. Helminth infection parameters often peaked in the spring while diversity was greatest in the winter samples. There was substantial temporal variability in helminth infections with prevalences, abundances, and intensities often fluctuating widely on consecutive sampling occasions. Analysis of the GIT helminth community composition suggested that 3 (Big Bay de Noc, De Tour Village, and Cheboygan) of the 4 primary spawning sites, overall, had similar community compositions. The reason for the observed spatial and temporal variability in the lake whitefish GIT helminth infections remains to be elucidated. The findings of this study represent the most comprehensive parasitological study ever conducted on lake whitefish in the Great Lakes and will provide valuable information for future comparisons.


A cyclic AMP-independent protein kinase from Candida albicans.
A cyclic AMP-independent protein kinase which phosphorylates casein was purified to homogeneity from Candida albicans by affinity and ion-exchange chromatography. This protein kinase exhibits maximal activity with casein as substrate and is not stimulated by cyclic AMP or cyclic GMP. The Mr of the purified enzyme is 115,000, as determined by h.p.l.c. It migrates as a single band on gel electrophoresis and has three non-identical subunits, of Mr 44,000, 28,500 and 26,000, as determined by SDS/polyacrylamide-gel electrophoresis. This enzyme is insensitive to heparin, but is inhibited by polyamines. Furthermore, it is sensitive to thermal denaturation and to thiol reagents.


Designing one-step reverse transcriptase loop-mediated isothermal amplification for serotype O foot-and-mouth disease virus detection during the 2022 outbreak in East Java, Indonesia.
Various methods can detect foot-and-mouth disease (FMD) in cows, but they necessitate resources, time, costs, laboratory facilities, and specific clinical specimen submission, often leading to FMD virus (FMDV) diagnosis delays. The 2022 FMD outbreak in East Java, Indonesia, highlighted the need for an easy, inexpensive, rapid, and accurate detection approach. This study aims to devise a one-step reverse transcriptase loop-mediated isothermal amplification (RT-LAMP) technique and phylogenetic analysis to detect the serotype O FMDV outbreak in East Java. Swab samples were collected from the foot vesicles, nasal secretions, and saliva of five suspected FMDV-infected cows in East Java between June and July 2022. The RT-LAMP design used hydroxy naphthol blue dye or SYBR Green I dye, with confirmatory analysis through reverse transcriptase polymerase chain reaction (RT-PCR) targeting 249 base pairs. PCR products underwent purification, sequencing, and nucleotide alignment, followed by phylogenetic analysis. The RT-LAMP method using hydroxy naphthol blue dye displayed a positive reaction through a color shift from purple to blue in the tube. Naked-eye observation in standard light or ultraviolet (UV) light at 365 nm, with SYBR Green I stain, also revealed color change. Specifically, using SYBR Green I dye, UV light at 365 nm revealed a color shift from yellow to green, signifying a positive reaction. Nucleotide alignment revealed mutations and deletion at the 15th sequence in the JT-INDO-K3 isolate from the East Java FMDV outbreak. Despite differing branches, the phylogenetic tree placed it in the same cluster as serotype O FMDV from Malaysia and Mongolia. JT-INDO-K3 exhibited distinctions from Indonesian serotype O FMDV isolates and those documented in GenBank. Then, the RT-LAMP method used in this study has a detection limit 10 times higher latter than the conventional RT-PCR limit, without any cross-reactivity among strains.


Entamoeba histolytica: electrophoretic isoenzyme patterns of strains and their virulence in the cecum of gerbils (Meriones unguiculatus).
Six strains of Entamoeba histolytica isolated from humans were characterized by their electrophoretic isoenzyme patterns and by virulence in the cecum of Mongolian gerbils (Meriones unguiculatus). Amebae from symptomatic cases of amebiasis were found to belong to zymodeme group II, whereas strains isolated from asymptomatic infections belonged to zymodeme groups I or V. Strains recently isolated from symptomatic cases of amebiasis were highly invasive in the cecum of gerbils. The most prominent pathologic changes present in these infections were the destruction of the interglandular epithelium, crypt hyperplasia and trophozoite invasion of the crypts and submucosa. Strains from asymptomatic carriers were of low or moderate invasiveness in the animal model. Two of the carrier strains expressing a zymodeme group I pattern caused diffuse or transient focal erosions of the interglandular epithelium in some animals. Trophozoites of carrier strains did not invade the crypts nor cause sloughing of the surface epithelium. Presence of colonic lesions was correlated with passage of motile trophozoites in the feces. The severity of histologically demonstrable lesions in the cecum of gerbils was positively correlated with the isoenzyme patterns characteristic of the more virulent amebic strains.


[Hormonal balance in women with lung cancer and its changes after combined treatment].
A relatively more favorable course of lung cancer and higher survival rates observed in female patients than in males make the case for further study of the problem. Endocrine system study group included female lung cancer patients. Blood-ACTH, FSH, LH, STH, T3 and T4 as well as F, 17-CS and estrogen excretion with urine were followed prior to, immediately and long after treatment in 25 menopausal and 11 premenopausal cases (at folliculin phase). Most patients revealed increased levels of free hydrocortisone in blood and urine matched by a lowered excretion of 17-CS. Hyperproduction of ACTH was registered in 40%, whereas 60% showed subnormal concentrations of said hormone. On the average, the levels of estrogens were increased, gonadotropin--decreased, T3--below normal concentrations and T4 and STH--within norm. TTH levels showed a wide-range variation: half that in healthy females-20%, and much higher than in healthy one-65%. Concentration of F was close to normal 15-20 days after surgery while ACTH and 17-CS--after one year. The levels of the other hormones remained virtually unchanged within 1-3 years.


Estradiol increases VEGF in human breast studied by whole-tissue culture.
Sex steroid exposure constitutes a risk factor for breast cancer, but little is known about the effects of sex steroids on the normal breast, largely because of the lack of convenient models. We have developed a method of culturing normal breast tissue ex vivo. We have applied this method to investigate the effects of estradiol and progesterone on the key angiogenic mediator, vascular endothelial growth factor (VEGF), in the breast. Whole breast tissue was obtained from routine reduction mammoplasty. Tissue biopsies were cultured in vitro for 1-3 weeks, and the expression of luminal cytokeratin 18 was determined by immunohistochemistry. As an application, tissue biopsies were treated in vitro for 1 week with or without estradiol or estradiol and progesterone. Estrogen receptor, progesterone receptor, and Ki-67 were analyzed, and VEGF levels were examined by quantitative immunoassay and immunohistochemistry. Whole breast tissue was cultured ex vivo for 1 week with preserved morphology. Increased detachment of the luminal epithelium was observed after 2 weeks. Estradiol increased extracellular levels of VEGF in normal breast tissue biopsy medium. The addition of progesterone had neither stimulatory nor inhibitory effects on secreted VEGF. The method of whole breast tissue culturing thus provide a means by which to explore the biology of normal breast tissue. Our results suggest that estradiol exerts pro-angiogenic effects in normal breast by increasing levels of biologically active VEGF.


Stable imprinting of long-period gratings in microstructured polymer optical fibre.
A simple heat imprinting method for producing stable long-period gratings (LPGs) in microstructured polymer optical fibre (mPOF) is presented as well as the examination of their lifetime and the modelling results of these gratings. Writing LPGs in mPOF presents opportunities for sensors in fibre that can withstand greater bending and strain and are adaptable to specific applications through modification of the cladding structure.


Dacarbazine in combination with bevacizumab for the treatment of unresectable/metastatic melanoma: a phase II study.
The combined treatment of dacarbazine with an antiangiogenic drug such as bevacizumab may potentiate the therapeutic effects of dacarbazine in metastatic melanoma (MM). Preliminary antitumour activity of dacarbazine plus bevacizumab is evaluated, together with the toxicity and safety profile, in MM patients. This prospective, open-label, phase II study included patients with previously untreated MM or unresectable melanoma. Patients received dacarbazine and bevacizumab until progressive disease or unacceptable toxicity. The primary efficacy variable was the overall response rate. The secondary efficacy parameters included duration of response, duration of stable disease, time to progression/progression-free survival, time to treatment failure and overall survival. The safety analysis included recordings of adverse events and exposure to study treatment. The intention-to-treat population included 37 patients (24 men and 13 women, mean age 54.2±13.1 years). Overall response rate was 18.9% (seven patients achieved a response) and clinical benefit was 48.6%. In patients who achieved a response, the median duration of response was 16.9 months and the median duration of stable disease was 12.5 months. The median time to progression/progression-free survival and time to treatment failure were 5.5 and 3.1 months, respectively. The median overall survival was 11.4 months. Almost all patients (94.6%) experienced at least one adverse event; however, no new area of toxicity of bevacizumab emerged. The dacarbazine/bevacizumab combination provides benefits compared with dacarbazine monotherapy in historical controls, with an acceptable safety profile. This combination appears to be a valid option in specific subgroups of patients, namely, those triple negative (BRAF, C-KIT and NRAS wild type) or with a BRAF mutation who have already received, or are not eligible for, immunomodulating or targeted agents.


Cannabidiol attenuates cardiac dysfunction, oxidative stress, fibrosis, and inflammatory and cell death signaling pathways in diabetic cardiomyopathy.
In this study, we have investigated the effects of cannabidiol (CBD) on myocardial dysfunction, inflammation, oxidative/nitrative stress, cell death, and interrelated signaling pathways, using a mouse model of type I diabetic cardiomyopathy and primary human cardiomyocytes exposed to high glucose. Cannabidiol, the most abundant nonpsychoactive constituent of Cannabis sativa (marijuana) plant, exerts anti-inflammatory effects in various disease models and alleviates pain and spasticity associated with multiple sclerosis in humans. Left ventricular function was measured by the pressure-volume system. Oxidative stress, cell death, and fibrosis markers were evaluated by molecular biology/biochemical techniques, electron spin resonance spectroscopy, and flow cytometry. Diabetic cardiomyopathy was characterized by declined diastolic and systolic myocardial performance associated with increased oxidative-nitrative stress, nuclear factor-κB and mitogen-activated protein kinase (c-Jun N-terminal kinase, p-38, p38α) activation, enhanced expression of adhesion molecules (intercellular adhesion molecule-1, vascular cell adhesion molecule-1), tumor necrosis factor-α, markers of fibrosis (transforming growth factor-β, connective tissue growth factor, fibronectin, collagen-1, matrix metalloproteinase-2 and -9), enhanced cell death (caspase 3/7 and poly[adenosine diphosphate-ribose] polymerase activity, chromatin fragmentation, and terminal deoxynucleotidyl transferase dUTP nick end labeling), and diminished Akt phosphorylation. Remarkably, CBD attenuated myocardial dysfunction, cardiac fibrosis, oxidative/nitrative stress, inflammation, cell death, and interrelated signaling pathways. Furthermore, CBD also attenuated the high glucose-induced increased reactive oxygen species generation, nuclear factor-κB activation, and cell death in primary human cardiomyocytes. Collectively, these results coupled with the excellent safety and tolerability profile of CBD in humans, strongly suggest that it may have great therapeutic potential in the treatment of diabetic complications, and perhaps other cardiovascular disorders, by attenuating oxidative/nitrative stress, inflammation, cell death and fibrosis.


Cardiac output measurement in children: comparison of the Ultrasound Cardiac Output Monitor with thermodilution cardiac output measurement.
To compare the assessment of cardiac output (CO) in children using the noninvasive Ultrasound Cardiac Output Monitor (USCOM) with the invasive pulmonary artery catheter (PAC) thermodilution cardiac output measurement. Prospective observational study in a tertiary center for pediatric cardiology of a university children's hospital. Twenty-four pediatric patients with congenital heart disease without shunt undergoing cardiac catheterization under general anesthesia. CO was measured by USCOM using a suprasternal CO Doppler probe in children undergoing cardiac catheterization. USCOM data were compared to CO simultaneously measured by PAC thermodilution technique. Measurements were repeated three times within 5 min in each patient. A mean percentage error not exceeding 30% was defined as indicating clinical useful reliability of the USCOM. CO values measured by PAC ranged from 1.3 to 5.3 l/min (median 3.6 l/min). Bias and precision were -0.13 and 1.34 l/min, respectively. The mean percentage error of CO measurement by the USCOM compared to PAC thermodilution technique was 36.4% for USCOM. Our preliminary data demonstrate that cardiac output measurement in children using the USCOM does not reliably represent absolute CO values as compared to PAC thermodilution. Further studies must evaluate the impact of incorporating effective aortic valve diameters on CO measurement using the USCOM.


Drug Reaction with Eosinophilia and Systemic Symptom Syndrome Due to Everolimus: A Case Report.
Drug reaction with eosinophilia and systemic symptom (DRESS) is a life-threatening drug hypersensitivity reaction that is characterized by skin rash, hematological abnormalities (eosinophilia, atypical lymphocytosis), lymphadenopathy, and internal organ involvement (liver, kidneys, and lung). Many drugs may cause DRESS syndrome, the most frequently reported of which are antiepileptics and allopurinol. In this case report, a patient who developed DRESS syndrome due to everolimus was presented herein.


[Detection of nitrite and nitrosocompounds in chemical systems and biological liquids by the calorimetric method].
The capacity of nitrite, S-nitrosothiols (RS-NO), dinitrosyl iron complexes (DNICs) with thiol-containing ligands, and nitrosoamines to inhibit catalase has been used for the selective determination of these compounds in purely chemical systems and biological liquids: cow milk and colostram. The limiting sensitivity of the method is 50 nM. A comparison of the results of the determinations of RS-NO, DNIC, and nitrite by the catalase method and the Greese method conventionally used for nitrite detection showed that, firstly, Greese reagents decompose DNIC and RS-NO to form nitrite. Therefore, the Greese method cannot be used for nitrite determination in solutions of these substances. Secondly, Greese reagents interact with complexes of mercury ions with RS-NO, inducing the release of nitrosonium ions from the complex followed by the hydrolysis of nitrosonium to nitrite. Thus, the proposition about the spontaneous decay of the complexes of mercury ions with RS-NO is incorrect. Keeping in mind a high sensitivity of the method, the use of catalase as an enzyme detector of nitrosocompounds allows one to detect these compounds in neutral medium without prior purification of the object, thereby preventing artificial effects due to noncontrolled modifications of the compounds under study.


Application of diagnostic network optimization in Kenya and Nepal to design integrated, sustainable and efficient bacteriology and antimicrobial resistance surveillance networks.
Antimicrobial resistance (AMR) is a major global public health concern, particularly in low- and middle-income countries, which experience the highest burden of AMR. Critical to combatting AMR is ensuring there are effective, accessible diagnostic networks in place to diagnose, monitor and prevent AMR, but many low- and middle-income countries lack such networks. Consequently, there is substantial need for approaches that can inform the design of efficient AMR laboratory networks and sample referral systems in lower-resource countries. Diagnostic network optimization (DNO) is a geospatial network analytics approach to plan diagnostic networks and ensure greatest access to and coverage of services, while maximizing the overall efficiency of the system. In this intervention, DNO was applied to strengthen bacteriology and AMR surveillance network design in Kenya and Nepal for human and animal health, by informing linkages between health facilities and bacteriology testing services and sample referral routes between farms, health facilities and laboratories. Data collected from the target settings in each country were entered into the open-access DNO tool OptiDx, to generate baseline scenarios, which depicted the current state of AMR laboratory networks and sample referral systems in the countries. Subsequently, baselines were adjusted to evaluate changing factors such as samples flows, transport frequency, transport costs, and service distances. Country stakeholders then compared resulting future scenarios to identify the most feasible solution for their context. The DNO analyses enabled a wealth of insights that will facilitate strengthening of AMR laboratory and surveillance networks in both countries. Overall, the project highlights the benefits of using a data-driven approach for designing efficient diagnostic networks, to ensure better health resource allocation while maximizing the impact and equity of health interventions. Given the critical need to strengthen AMR laboratory and surveillance capacity, DNO should be considered an integral part of diagnostic strategic planning in the future.


Curcumin enhances wound healing in streptozotocin induced diabetic rats and genetically diabetic mice.
Tissue repair and wound healing are complex processes that involve inflammation, granulation and tissue remodeling. Interactions of different cells, extracellular matrix proteins and their receptors are involved in wound healing, and are mediated by cytokines and growth factors. Previous studies from our laboratory have shown that curcumin (diferuloylmethane), a natural product obtained from the rhizomes of Curcuma longa, enhanced cutaneous wound healing in rats and guinea pigs. In this study, we have evaluated the efficacy of curcumin treatment by oral and topical applications on impaired wound healing in diabetic rats and genetically diabetic mice using a full thickness cutaneous punch wound model. Wounds of animals treated with curcumin showed earlier re-epithelialization, improved neovascularization, increased migration of various cells including dermal myofibroblasts, fibroblasts, and macrophages into the wound bed, and a higher collagen content. Immunohistochemical localization showed an increase in transforming growth factor-beta1 in curcumin-treated wounds compared to controls. Enhanced transforming growth factor-beta1 mRNA expression in treated wounds was confirmed by in situ hybridization, and laser scan cytometry. A delay in the apoptosis patterns was seen in diabetic wounds compared to curcumin treated wounds as shown by terminal deoxynucleotidyl transferase-mediated deoxyuridyl triphosphate nick end labeling analysis. Curcumin was effective both orally and topically. These results show that curcumin enhanced wound repair in diabetic impaired healing, and could be developed as a pharmacological agent in such clinical settings.


High-resolution 4-D acquisition of freely swimming human sperm cells without staining.
We present a new acquisition method that enables high-resolution, fine-detail full reconstruction of the three-dimensional movement and structure of individual human sperm cells swimming freely. We achieve both retrieval of the three-dimensional refractive-index profile of the sperm head, revealing its fine internal organelles and time-varying orientation, and the detailed four-dimensional localization of the thin, highly-dynamic flagellum of the sperm cell. Live human sperm cells were acquired during free swim using a high-speed off-axis holographic system that does not require any moving elements or cell staining. The reconstruction is based solely on the natural movement of the sperm cell and a novel set of algorithms, enabling the detailed four-dimensional recovery. Using this refractive-index imaging approach, we believe that we have detected an area in the cell that is attributed to the centriole. This method has great potential for both biological assays and clinical use of intact sperm cells.


[Study of the possible harmful action of meat containing ammonia on experimental animals].
The influence of meat containing ammonia in concentrations of 0.1 and 0.3% was studied in chronic experiments on 3 generations of rats (males and females). To investigate the function of the liver, kidneys and CNS, alanine-aminotransferase, lactate dehydrogenase activity, and the content of the total protein and its fractions were assayed in the blood serum; cholinesterase activity, sulfhydryl groups, urea and residual nitrogen were assayed in the blood, as well as the parameters of rheobase, chronaxie and summation of subliminal impulses. The results of the investigation have shown that meat containing ammonia in concentrations of 0.1 and 0.3% produces a detrimental effect on the experimental animals, the highest effect being recorded with an ammonia concentration of 0.3%.


The Case for Anthocyanin Consumption to Promote Human Health: A Review.
Anthocyanins belong to the flavonoid group of polyphenolic compounds, which are responsible for the red and blue colors of plant organs such as fruits, flowers, and leaves. Due to their frequent presence in plants, particularly berry fruits, vegetables, and grapes, they are key components of the human diet. Interest in anthocyanins has increased widely during the past decade. Numerous studies have suggested that anthocyanins have a wide range of health-promoting properties. These compounds are therefore considered to be a functional food factor, which may have important implications in the prevention of chronic diseases. The aim of this body of work is to investigate and review the current literature on anthocyanins, and particularly their pharmacokinetics and any health-promoting properties, in order to summarize existing knowledge and highlight any aspects that require further study and analysis.


Feasibility of using the ProtekDuo cannula in V-P ECMO and PROpella configurations during ground and air transport.
Use of the ProtekDuo cannula has been described for right ventricular assist devices (RVADs) and extracorporeal membrane oxygenation (ECMO) systems. We describe remote cannulation and transport of two patients with ProtekDuo cannula. One patient had isolated acute right ventricular failure (aRVF), was cannulated with ProtekDuo cannula in venopulmonary (V-P) configuration and transported by ambulance. Another patient had biventricular failure after myocardial infarction, was supported with ProtekDuo and Impella CP in PROpella configuration, and transported by helicopter. We appear to be the first group to report remote cannulation using the ProtekDuo cannula followed by ambulance and helicopter transport, which were performed without complication. We describe the pros and cons of these configurations in comparison to the gold standard of shock management with venoarterial ECMO, as well as important considerations for transport. Use of the ProtekDuo cannula for remote cannulations and transport is feasible and appears safe.


Bioactive decellularized adipose matrix prepared using a rapid, nonchemical/enzymatic method for adipogenesis.
Recent developments in the field of regenerative surgeries and medical applications have led to a renewed interest in adipose tissue-enriched mesenchymal stem cell scaffolds. Various advantages declared for the decellularized adipose matrix (DAM) have caused its extensive use in the transfer of stem cells or growth factors for soft tissue regeneration induction. Meanwhile, the long-term application of detergents toward DAM regeneration has been assumed as a risky obstacle in this era. Herein, a rapid, mechanical protocol was developed to prepare DAM (M-DAM) without chemicals/enzymes and was comprehensively compared with the ordinary DAM (traditional chemical method). Accordingly, this method could effectively hinder oils and cells, sustain the structural and biological elements, and contain a superior level of collagen content. In addition, more protein numbers, as well as higher basement membrane elements, glycoproteins, and extracellular matrix-related proteins were detected in the regenerated M-DAM. Also, superior adipogenesis and angiogenesis proteins were distinguished. The noncytotoxicity of the M-DAM was also approved, and a natural ecological niche was observed for the proliferation and differentiation of stem cells, confirming its great potential for vascularization and adipogenesis in vivo. The suggested technique could effectively prepare the modified DAM in variant constructions of tablets, powders, emulsions, hydrogels, and different three-dimensional-printed structures. Hence, this rapid, mechanical process can produce bioactive DAM, which has the potential to be widely used in various research fields of regenerative medicine.


[Cation-anion selectivity and conductivity of the channels formed by black widow spider whole venom in lipid bilayer].
It is established that BWSV (black widow spider venom) is capable of forming ion channels in lipid bilayer (BPhM). This effect is minimal upon pH value close to 5.0 and is increased either upon oxidation or alkalization of the medium. The amplitudes of the channels are monotonously decreased upon the reduction of pH with simultaneous narrowing of their dispersion. The degree of the change of BPhM conductivity by concentration of BWSV is close to 2 at different pH. Apparently the absorption of the active component of venom on bilayer surface is accompanied by doubling of the degree of its aggregation. Almost ideal cation selectivity of BWSV channels (pH 7.5) is significantly decreased upon the increase in ionic strength. Ionic channel is thought to be a zone of interaction of lipid domains. Possible structure of such a channel is discussed.


[Bovine herpes mammillitis: clinical symptoms and serologic course].
Bovine herpes mammillitis was diagnosed in a dairy herd with udder and teat skin lesions. Clinical symptoms seen in 6 cows consisted of round dry areas at the teats as well as large red and painful areas with crust formation at the teats, the teat basis and the udder. Diagnosis was verified by demonstrating numerous virus particles with the typical herpes structure and by BHV-2 serum neutralization test. Prevalence of BHV-2 in the herd was determined by using BHV-2 SNT at 7 occasions during a period of 15 months. The relatively low BHV-2 SNT-titres as well as the seasonal increase of BHV-2 titres and seroprevalence in the month of September were indicative of a chronic and latent BHV-2 infection in the herd.


Race, Social Determinants of Health, and Length of Stay Among Hospitalized Patients With Heart Failure: An Analysis From the Get With The Guidelines-Heart Failure Registry.
Racial disparities in heart failure hospitalization and mortality are well established; however, the association between different social determinants of health (SDOH) and length of stay (LOS) and the extent to which this association may differ across racial groups is not well established. We utilized data from the Get With The Guidelines-Heart Failure registry to evaluate the association between SDOH, as determined by patients' residential ZIP Code and LOS among patients hospitalized with heart failure. We also assessed the race-specific contribution of the ZIP Code-level SDOH to LOS in patients of Black and non-Black races. Finally, we evaluated SDOH predictors of racial differences in LOS at the hospital level. Among 301 500 patients (20.2% Black race), the median LOS was 4 days. In adjusted analysis accounting for patient-level and hospital-level factors, SDOH parameters of education, income, housing instability, and foreign-born were significantly associated with LOS after adjusting for clinical status and hospital-level factors. SDOH parameters accounted for 25.8% of the total attributable risk for prolonged LOS among Black patients compared with 10.1% in patients of non-Black race. Finally, hospitals with disproportionately longer LOS for Black versus non-Black patients were more likely to care for disadvantaged patients living in ZIP Codes with a higher percentage of foreign-born and non-English speaking areas. ZIP Code-level SDOH markers can identify patients at risk for prolonged LOS, and the effects of SDOH parameters are significantly greater among Black adults with heart failure as compared with non-Black adults.


The child with repeated injuries--a family problem.
Recurrent injuries in children should be regarded as a symptom complex rather than a diagnosis of ;accident proneness'. Investigation, which may include much listening will often reveal a family with emotional problems. The family doctor who can see the family as a whole is often best placed to offer or co-ordinate help.