Murine models to evaluate novel and conventional therapeutic strategies for cancer.
Animal models, by definition, are an approximation of reality, and their use in developing anti-cancer drugs is controversial. Positive retrospective clinical correlations have been identified with several animal models, in addition to limitations and a need for improvement. Model inadequacies include experimental designs that do not incorporate biological concepts, drug pharmacology, or toxicity. Ascites models have been found to identify drugs active against rapidly dividing tumors; however, neither ascitic nor transplantable subcutaneous tumors are predictive of activity for solid tumors. In contrast, primary human tumor xenografts have identified responsive tumor histiotypes if relevant pharmacodynamic and toxicological parameters were considered. Murine toxicology studies are also fundamental because they identify safe starting doses for phase I protocols. We recommend that future studies incorporate orthotopic and spontaneous metastasis models (syngeneic and xenogenic) because they incorporate microenvironmental interactions, in addition to confirmatory autochthonous models and/or genetically engineered models, for molecular therapeutics. Collectively, murine models are critical in drug development, but require a rational and hierarchical approach beginning with toxicology and pharmacology studies, progressing to human primary tumors to identify therapeutic targets and models of metastatic disease from resected orthotopic, primary tumors to compare drugs using rigorous, clinically relevant outcome parameters.


Increase in isolation of extended spectrum beta lactamase producing multidrug resistant non typhoidal Salmonellae in Pakistan.
Increasing resistance to quinolones and ceftriaxone in non typhoidal Salmonellae is a global concern. Resistance to quinolone and 3rd generation cephalosporin amongst non typhoidal Salmonellae (NTS) from Pakistan has been reported in this study. Retrospective analysis of laboratory data was conducted (1990-2006). NTS were isolated and identified from clinical samples using standard microbiological techniques. Antimicrobial susceptibility testing was performed by Kirby Bauer. Extended spectrum beta lactamase production (ESBL) was detected using combined disc method. Ciprofloxacin sensitivity was detected by nalidixic acid screening method. Minimum inhibitory concentration (MIC) of ciprofloxacin was determined by agar dilution method. Statistical analysis was performed using SPSS version 13. Analysis of 1967 NTS isolates showed a significant increase in ciprofloxacin resistance from 23% in 2002 to 50.5% in 2006, with increased mean MIC values from 0.6 to 1.3 ug/mL. Ceftriaxone resistant NTS also increased and ESBL production was seen in 98.7% isolates. These isolates exhibited high resistance against amoxicillin clavulanic acid (57%), gentamicin (69%), amikacin (44%) and piperacillin tazobactam (30%). No resistance to carbapenem was seen. Ceftriaxone resistance was significantly higher in children <1 year, in invasive isolates and in Salmonella Typhimurium. Increase in quinolone and ceftriaxone NTS is a serious threat to public health requiring continuous surveillance and use of appropriate screening tests for laboratory detection.


3D visualization of perianal fistulas using parametric models.
Magnetic resonance imaging (MRI) is used as a standard for assessment of complex perianal fistulas. Apart from textual description of the case, 3D reconstructed models from MRI further aid in understanding the entire anatomy of the fistula tract and its relation to the pelvic floor. This information is crucial as it helps surgeons to understand the extent and complexity of the disease before surgical treatment. However, 3D model generation from MRI is a time-consuming step for a radiologist as it requires tedious manual delineations to be performed on every slice of the images. The aim of this study was to develop a method that could enable radiologists to present enhanced information to surgeons for treatment of complex perianal fistulas while simultaneously reducing the manual efforts and time required to generate the information. A method was proposed to depict relevant anatomies of complex perianal fistula as parametric models in three-dimensional (3D) space. A plugin inside 3D Slicer software was developed for the generation of the parametric models from MRI. The levator ani muscle, internal sphincter, and external sphincter are represented as tubular structures, whereas fistula tracks and abscess are presented as splines. Parametric models were generated to depict three cases of complex perianal fistulas and similarity measures were computed for ten cases. Visual comparison of the parametric models was made with the 3D models generated by the standard approach. The parametric models took less time to create and were able to visually present enriched information as compared to the 3D models generated by the standard approach. The proposed method, using parametric models, shows potential for faster generation and better visualization of the 3D information required for the treatment of complex perianal fistula cases.


Morbidity risk for obsessive-compulsive spectrum disorders in first-degree relatives of patients with eating disorders.
A hypothesis that eating disorders are a phenomenological variant of obsessive-compulsive disorder (OCD) has been proposed. This study was conducted to determine whether anorexia nervosa and bulimia, the two main eating disorders, are familial and whether the risk for obsessive-compulsive spectrum disorders (OCD and tic disorders) is higher in families of patients with eating disorders. The morbidity risk for obsessive-compulsive spectrum disorders in first-degree relatives of 136 female probands with eating disorders (84 with anorexia nervosa, 52 with bulimia) was compared to that for first-degree relatives of 72 female comparison subjects. The morbidity risk for obsessive-compulsive spectrum disorders was significantly higher among the 436 relatives of the eating disorder probands than among the 358 relatives of the comparison subjects (9.69% versus 0%). This finding was independent of any comorbid diagnosis of an obsessive-compulsive spectrum disorder in the eating disorder probands. The eating disorder group and the comparison group did not differ in familial risk for eating disorders and tic disorders. To better understand the genetic components of eating disorders, these disorders should be considered as part of the obsessive-compulsive spectrum of disorders.


Seasonal body size reductions with warming covary with major body size gradients in arthropod species.
Major biological and biogeographical rules link body size variation with latitude or environmental temperature, and these rules are often studied in isolation. Within multivoltine species, seasonal temperature variation can cause substantial changes in adult body size, as subsequent generations experience different developmental conditions. Yet, unlike other size patterns, these common seasonal temperature-size gradients have never been collectively analysed. We undertake the largest analysis to date of seasonal temperature-size gradients in multivoltine arthropods, including 102 aquatic and terrestrial species from 71 global locations. Adult size declines in warmer seasons in 86% of the species examined. Aquatic species show approximately 2.5-fold greater reduction in size per °C of warming than terrestrial species, supporting the hypothesis that greater oxygen limitation in water than in air forces aquatic species to exhibit greater plasticity in body size with temperature. Total percentage change in size over the annual cycle appears relatively constant with annual temperature range but varies between environments, such that the overall size reduction in aquatic-developing species (approx. 31%) is almost threefold greater than in terrestrial species (approx. 11%). For the first time, we show that strong correlations exist between seasonal temperature-size gradients, laboratory responses and latitudinal-size clines, suggesting that these patterns share common drivers.


Analysis of endodontic therapy in patients irradiated for head and neck cancer.
The outcome of endodontic therapy in 16 patients irradiated for head and neck cancer was studied. Thirty-five postradiation endodontically treated teeth (54 roots) were included in the study. The follow-up period ranged from 6 months to 54 months. At the time of last follow-up, 46 of 54 roots were being maintained. No osteoradionecroses were seen in association with teeth that had been endodontically treated. From this review, it is clear that endodontic therapy is a viable method of treating diseased teeth in patients irradiated for oral neoplasms.


Mitotic Acetylation of Microtubules Promotes Centrosomal PLK1 Recruitment and Is Required to Maintain Bipolar Spindle Homeostasis.
Tubulin post-translational modifications regulate microtubule properties and functions. Mitotic spindle microtubules are highly modified. While tubulin detyrosination promotes proper mitotic progression by recruiting specific microtubule-associated proteins motors, tubulin acetylation that occurs on specific microtubule subsets during mitosis is less well understood. Here, we show that siRNA-mediated depletion of the tubulin acetyltransferase ATAT1 in epithelial cells leads to a prolonged prometaphase arrest and the formation of monopolar spindles. This results from collapse of bipolar spindles, as previously described in cells deficient for the mitotic kinase PLK1. ATAT1-depleted mitotic cells have defective recruitment of PLK1 to centrosomes, defects in centrosome maturation and thus microtubule nucleation, as well as labile microtubule-kinetochore attachments. Spindle bipolarity could be restored, in the absence of ATAT1, by stabilizing microtubule plus-ends or by increasing PLK1 activity at centrosomes, demonstrating that the phenotype is not just a consequence of lack of K-fiber stability. We propose that microtubule acetylation of K-fibers is required for a recently evidenced cross talk between centrosomes and kinetochores.


Anti-Inflammatory Diets in Fertility: An Evidence Review.
Infertility is a global health concern affecting 48 million couples and 186 million individuals worldwide. Infertility creates a significant economic and social burden for couples who wish to conceive and has been associated with suboptimal lifestyle factors, including poor diet and physical inactivity. Modifying preconception nutrition to better adhere with Food-Based Dietary Guidelines (FBDGs) is a non-invasive and potentially effective means for improving fertility outcomes. While several dietary patterns have been associated with fertility outcomes, the mechanistic links between diet and infertility remain unclear. A key mechanism outlined in the literature relates to the adverse effects of inflammation on fertility, potentially contributing to irregular menstrual cyclicity, implantation failure, and other negative reproductive sequelae. Therefore, dietary interventions which act to reduce inflammation may improve fertility outcomes. This review consistently shows that adherence to anti-inflammatory diets such as the Mediterranean diet (specifically, increased intake of monounsaturated and n-3 polyunsaturated fatty acids, flavonoids, and reduced intake of red and processed meat) improves fertility, assisted reproductive technology (ART) success, and sperm quality in men. Therefore, integration of anti-inflammatory dietary patterns as low-risk adjunctive fertility treatments may improve fertility partially or fully and reduce the need for prolonged or intensive pharmacological or surgical interventions.


Prophylactic alpha blockers fail to prevent postoperative urinary retention following orthopaedic procedures: evidence from a meta-analysis and trial sequential analysis of comparative studies.
Objective: The present systematic review and meta-analysis aimed to estimate the prophylactic effect of alpha blockers against postoperative urinary retention (POUR) in orthopaedic patients. Methods: PubMed, Embase, Web of Science and Cochrane Library databases were searched between 1 January 1990 and 1 March 2023. The studies reporting the preventive efficacy of alpha blockers on POUR after orthopaedic procedures were identified. The pooled rates of POUR in the Intervention group (patients receiving alpha blockers) and the Control group (patients not receiving alpha blockers) were estimated and compared. The risk ratios (RRs) were calculated using the random-effects model. Subgroup analysis was performed based on surgical type. Trial sequential analysis (TSA) was conducted to confirm the robustness of pooled results. Results: Seven studies containing 1,607 patients were identified. The rates of POUR were similar between the two groups (Intervention group: 126/748 [16.8%] VS. Control group: 168/859 [19.6%]; RR = 0.75; 95% confidence interval [CI] 0.51 to 1.09; p = 0.130; Heterogeneity: I2 = 67.1%; p = 0.006). No significant difference in the incidence of POUR was observed in either the Arthroplasty subgroup or Spine surgery subgroup. The result of TSA suggested that the total sample size of the existing evidence might be insufficient to draw conclusive results. Administrating alpha blockers was associated with a higher risk of complications (88/651 [13.5%] VS. 56/766 [7.3%]; RR = 1.73; 95% CI 1.27 to 2.37; p = 0.0005; Heterogeneity: I2 = 0%; p = 0.69). Conclusion: Prophylactic alpha blockers do not reduce the risk of POUR in orthopaedic procedures, and administrating these drugs was associated with a higher risk of complications. Systematic Review Registration: https://www.crd.york.ac.uk/PROSPERO/display_record.php?RecordID=409388.


Mast cells are novel independent prognostic markers in prostate cancer and represent a target for therapy.
Mast cells affect growth in various human tumors, but their role in prostate cancer (PC) is unclear. Here, we identify mast cells as independent prognostic markers in PC using a large cohort of untreated PC patients with a long follow-up. By analyzing mast cells in different tissue compartments, our data indicate that intratumoral and peritumoral mast cells have anti- opposed to protumor properties. Intratumoral mast cells negatively regulate angiogenesis and tumor growth, whereas peritumoral mast cells stimulate the expansion of human prostate tumors. We also observed mast cell recruitment particularly to the peritumoral compartment in men during the formation of castrate-resistant prostate tumors. In our ortothopic rat model, mast cells accumulated in the peritumoral tissue where they enhanced angiogenesis and tumor growth. In line with this, prostate mast cells expressed high levels of the angiogenic factor FGF-2. Similar to the situation in men, mast cells infiltrated rat prostate tumors that relapsed after initially effective castration treatment, concurrent with a second wave of angiogenesis and an up-regulation of FGF-2. We conclude that mast cells are novel independent prognostic markers in PC and affect tumor progression in animals and patients. In addition, peritumoral mast cells provide FGF-2 to the tumor micro environment, which may contribute to their stimulating effect on angiogenesis.


Axillary-femoral graft compression by prosthetic belt.
In severe peripheral vascular disease, a patient may need not only an amputation but an axillary-femoral bypass. In order to prevent the potential problem of axillary-femoral graft compression by a prosthetic belt, the importance of amputation length is stressed in this case report discussion.


Histochemical observations on the female reproductive tract of bats. III. Mucosubstances in the Fallopian tube of three species of bats.
The epithelium of the three species of bats studied shows the presence of ciliated and nonciliated cells both in the ovarian and uterine segments of the Fallopian tube of the functional uterine horn. The cytoplasm of these two cells types shows small amount of glycogen during anestrus, the amount increasing during proestrus and estrus. At estrus, the nonciliated cells show the presence of neutral and sialomucins in addition to glycogen in Pteropus giganteus giganteus and Megaderma lyra lyra. The concentration of these mucins was found to decline during pregnancy and parturition. In Taphozous longimanus, the mucins which are located in cytoplasmic projections or domes of the nonciliated cells show the presence of neutral, sialomucins, sulfomucins and glycogen. These mucins which are first seen in the proestrus period increase during estrus. However, during pregnancy, these domes regress concomitant with decline in the mucin content. Glycogen content of the ciliated cells increases during estrus and decreases during pregnancy. Since in all the three species of bats the mucin content is at its maximum at oestrus and shows a decline during pregnancy, it appears that its elaboration and increase is under the influence of estrogen, instead progesterone blocks its generation and brings about its release.


[Relationship of delusions to attempted suicide and suicide].
This article is a contribution to the psychopathology of communication. Using the communication model of orienting behavior the pathological phenomena of delusion, suicide attempt and suicide are dealt with in a communication-theoretical framework. On this basis, the relationship of delusion to suicide attempts and to suicides is examined. The results are that delusion and suicide attempt are subject to a communicative principle, whereas suicide is subject to an anticommunicative principle. Our numerous observations that the act of suicide leading to death is never carried out in a state of delusion are interpreted within the framework of the communication model presented here.


Chemotherapy and tumor immunity: an unexpected collaboration.
Chemotherapy directly targets the transformed tumor cell, and has long been a key component of therapy for most early and advanced cancers. However, its utility is ultimately limited by unavoidable toxicity to normal tissues, and by drug resistance pathways deeply embedded within the biology of the tumor cell itself. These limitations strongly argue for innovative strategies to treat and manage cancer. Engaging the power of the patient's own immune system is a highly attractive way to complement the activity of standard cancer treatment. Tumor vaccines offer the potential for preventing cancer in those at high risk for disease development, preventing relapse in those diagnosed with early cancer, and treating advanced disease. Notably, the barriers to tumor vaccine efficacy are distinct from the limitations of combination chemotherapy. The ability of vaccines to induce a response robust enough to mediate tumor rejection is limited by the extent of disease burden, the suppressive effect of the local tumor micronenvironment, and multiple layers of systemic immune tolerance established to keep the immune response turned off. Chemotherapy can be used with tumor vaccines in unexpected ways, breaking down these barriers and unleashing the full potential of the antitumor immune response.


Oral Health Status, Oral Health Behaviors, and Oral Health Care Utilization among Persons with Disabilities in Saudi Arabia.
Various studies have indicated that persons with special needs may face several barriers to dental treatment, which increases the prevalence of oral diseases in this population. Moreover, these studies suggested that Saudis with special needs have a higher prevalence of oral diseases, such as dental caries and periodontal disease. The aim of this review is to synthesize evidence regarding the current status, trends in oral health behaviors, and oral health care utilization among these individuals, as well as to assess the quality of the literature. Furthermore, this review seeks to recommend directions for future research and oral health care policymaking. An electronic search was conducted using the following databases and registers: PubMed/Medline, Embase, ISI Web of Science, Scopus, ClinicalTrials.gov, and CENTRAL. Gray literature, which included conference proceedings and unpublished literature, was searched via the library services and Google/Google Scholar, and the quality of studies was assessed using the AXIS scale for cross-sectional studies. A total of 38 studies were included in this review, with the majority of the studies graded as 'low'. Within the limitations of this study, it can be concluded that individuals with special needs have limited access to oral health care, poor oral health status, and a general lack of awareness in regard to oral health. Therefore, nationwide surveys should be carried out to ascertain the actual extent of the oral inequities among individuals with special needs.


Feedforward compensation for novel dynamics depends on force field orientation but is similar for the left and right arms.
There are well-documented differences in the way that people typically perform identical motor tasks with their dominant and the nondominant arms. According to Yadav and Sainburg's (Neuroscience 196: 153-167, 2011) hybrid-control model, this is because the two arms rely to different degrees on impedance control versus predictive control processes. Here, we assessed whether differences in limb control mechanisms influence the rate of feedforward compensation to a novel dynamic environment. Seventy-five healthy, right-handed participants, divided into four subsamples depending on the arm (left, right) and direction of the force field (ipsilateral, contralateral), reached to central targets in velocity-dependent curl force fields. We assessed the rate at which participants developed predictive compensation for the force field using intermittent error-clamp trials and assessed both kinematic errors and initial aiming angles in the field trials. Participants who were exposed to fields that pushed the limb toward ipsilateral space reduced kinematic errors more slowly, built up less predictive field compensation, and relied more on strategic reaiming than those exposed to contralateral fields. However, there were no significant differences in predictive field compensation or kinematic errors between limbs, suggesting that participants using either the left or the right arm could adapt equally well to novel dynamics. It therefore appears that the distinct preferences in control mechanisms typically observed for the dominant and nondominant arms reflect a default mode that is based on habitual functional requirements rather than an absolute limit in capacity to access the controller specialized for the opposite limb.


The impact of social media influencers on health outcomes: Systematic review.
A fast emerging area of public health interest is the potential role of social media influencers in spreading health information and affecting health behaviour. The aim of this research was to systematically synthesise evidence on the impact of social media influencers on health outcomes. A systematic search of six databases (Medline, Embase, PsycINFO, CINAHL, Science Citation Index and Sociology Collection) was conducted in October 2021, with an update search in January 2023. Two reviewers independently screened titles and abstracts. We included interventional study designs which examined the effect of social media influencer as the primary or only exposure on a quantified health outcome. We used narrative synthesis to summarise the characteristics and main findings of studies within each of four main topic areas. We assessed quality using a critical appraisal checklist for quasi-experimental studies. We identified twelve studies across four areas that met the inclusion criteria: children's dietary behaviour (n = 5), body image dissatisfaction (n = 4), influenza and COVID-19 (n = 2), and social comparison and anxiety (n = 1). Influencer marketing of unhealthy foods had a statistically significant effect on increasing children's immediate energy intake, but no effect was found for influencer promotion of vegetables. When an unhealthy food is marketed by an influencer of unhealthy appearance, this led to an increased preference for healthy snacks. Exposure to idealised influencer body imagery had statistically significant negative impacts on body image and mood. Anxiety caused by viewing idealised portrayals of motherhood was the same whether or not the portrayal was by an influencer. Public health campaigns harnessing influencers to promote hygiene habits in the context of COVID-19 and influenza showed some positive changes in health behaviours. Most of the published studies are of poor methodological quality with poor generalisability and statistical weaknesses. We conclude that social media influencers have both negative and positive impacts on health outcomes, with negative impacts seen consistently in studies of body image dissatisfaction. Further research is needed to harness the potential positive health impact of social media influencers, while mitigating against harmful effects.


Translational regulation of platelet-derived growth factor B chain (c-sis) mRNA by short open reading frames in the 5'-untranslated region.
Platelet-derived growth factor B chain (PDGF-B/c-SIS), the product of c-sis proto-oncogene, is a potent mitogen and chemoattractant for cells of mesenchymal origin. Expression of PDGF-B/c-SIS is regulated at the translational level, in addition to at the transcriptional level. The 5'-untranslated region (5'-UTR) of PDGF-B/c-sis mRNA is known to inhibit translation of the downstream coding sequences. The 5'-UTR contains putative influential elements, such as GC-rich elements, stem-loop structures and short open reading frames (SORFs). To clarify the inhibition mechanism of PDGF-B/c-sis mRNA translation, effects of three SORFs in the 5'-UTR on the translational regulation were investigated in transient expression assays. Introducing point mutation(s) in the initiation codons of SORFs affected the reporter gene expression in several cell lines (COS-1, U-2, JEG-3). Abrogation of three SORFs resulted in an increase of the reporter gene expression both in beta-galactosidase assay and Western blot analysis. These results suggest that SORFs in the 5'-UTR sequences have inhibitory effects on the translation of the downstream coding sequences.


Extracts from Flammulina velutipes Inhibit the Adhesion of Pathogenic Fungi to Epithelial Cells.
Recently, extracts from natural sources have been tested for their antifungal properties. In this aspect, Flammulina velutipes extracts possess a significant amount of branch-chained carbohydrates with mannose moieties that, hypothetically, can reduce the adhesion. In this study, we assessed the capacity of extracts from F. velutipes (wild-type AQF-1 and ATCC 34574 as the reference strain) to inhibit the adhesion of S. schenkii and C. albicans to epithelial cells. The aqueous extracts from F. velutipes strains were obtained by sonication, total carbohydrate and protein was analyzed by Dubois and Lowry methods respectively. Effect of the extracts (50, 100 and 150 μg/mL) on the fungi adhesion to host cells was evaluated after 1 h interaction, and the percentage of inhibition of adhesion was measured. After of interaction the cytoskeleton from cell was analyzed with phalloidin-FITC. The extract from strain AQF-1 (50, 100 and 150 μg/mL) inhibited the adhesion of: S. schenkii in a dose-dependent manner (4.9, 7.5 and 12.7%, respectively) and C. albicans in a dose-independent manner (5.2%). The percentage of inhibition by extracts from the strain ATCC34574 at the same concentrations, shown that are dose independent for both fungi: 3.9% for S. schenkii and 2.6% for C. albicans. The extracts from F. velutipes inhibit the adhesion of pathogenic fungi to host cells. The mechanism molecular is unknown; however, is probably an interaction between the polysaccharides from extracts with the fungi receptors. This aspect is currently analyzed. The yields of mycelium from two strains of F. velutipes and the extract from it were similar.Extracts from both strains have inhibited adhesion of S. schenkii and C. albicans to epithelial cells in vitro, but the extract from strain AQF-1 was more effective.The extracts have not prevented damage to epithelial cells caused by pathogenic fungi. Abbreviation Used: YPG: Yeast peptone glucose, DMEM: Dulbecco's Modified Eagle's medium, FITC: Fluorescein isothiocyanate.


Prevalence of gallstones in liver cirrhosis: a sonographic survey.
A sonographic prospective study of the prevalence of gallstones was performed in 140 patients with liver cirrhosis and in 140 controls. Gallstones were found more often in cirrhotic patients (29.2%) than in controls (13.6%) (p less than 0.01). Their prevalence increased with age. The ratio of women to men in cirrhotics was the same as in the general population, with a higher prevalence in women. The prevalence of gallstones increased in decompensated liver disease. There was a significantly higher prevalence of both hypersplenism and hemolysis in cirrhosis. No difference was found in gallstone prevalence in relation to cirrhosis etiology. This prospective study confirms, by means of sonography, the high prevalence of cholelithiasis in liver cirrhosis, and extends the previous data about the lithogenic risk factors in this disease.


Prevalence of myocardial bridging associated with coronary endothelial dysfunction in patients with chest pain and non-obstructive coronary artery disease.
Myocardial bridging (MB), characterised by the epicardial coronary vessel diving into the myocardium, is present in up to one third of adults and is associated with angina and acute coronary syndromes. MB is accompanied by altered blood flow mechanics and regional changes in wall sheer stress. The purpose of this study was to determine the association between myocardial bridging and coronary endothelial dysfunction. Patients presenting with chest pain and found to have non-obstructive CAD (stenosis <40%) on angiography underwent an invasive assessment of epicardial and microvascular endothelial function. Epicardial endothelial function was assessed by measuring the percent change in coronary artery diameter in response to intracoronary infusions of acetylcholine (%ΔCADAch). Epicardial endothelial dysfunction was defined as a %ΔCADAch of <-20%. Microvascular endothelial function was assessed by the percent change in coronary blood flow in response to intracoronary infusions of acetylcholine (%ΔCBFAch), and microvascular endothelial dysfunction was defined as a %ΔCBFAch of <50%. MB was diagnosed angiographically by identifying the characteristic reduction in minimal luminal diameter during systole. Patients were divided into those with and those without MB, and the frequency of epicardial endothelial dysfunction and microvascular endothelial dysfunction was compared between patients with versus those without MB. Between 1993 and 2012, 1,469 patients (mean age 50.4 years, 35% male) underwent coronary angiography and invasive testing of endothelial function. Two hundred and eight (14.2%) patients were found to have MB in the LAD. Patients with any MB had a significantly higher frequency of endothelial dysfunction within the mid and/or distal vessel segment compared to patients without MB (60.1% vs 50.4%, p=0.012). In multivariate analyses, mid and/or distal vessel MB was a significant predictor of mid and/or distal vessel epicardial endothelial dysfunction (OR 1.44, 95% CI: 1.04-2.00, p=0.029) and of microvascular endothelial dysfunction (OR 1.34, 95% CI: 1.00-1.82, p=0.050). MB co-localises with epicardial endothelial dysfunction and is significantly associated with microvascular endothelial dysfunction in symptomatic patients with non-obstructive CAD, supporting its potential role as a mechanism for angina in symptomatic patients with MB.


Reciprocal interferences of TNF-α and Wnt1/β-catenin signaling axes shift bone marrow-derived stem cells towards osteoblast lineage after ethanol exposure.
We have reported in a separate study that alcohol exposure triggers activation of the TNF-α signaling pathway leading to an adverse shift of multipotential mesenchymal stem cells in bone marrow (BMSCs) away from osteogenesis towards adipogenesis. However, inhibition of TNF-α signaling only yielded moderate inhibition of adipogenesis. Here we showed that in addition to promoting the TNF-α signaling, alcohol also suppressed the Wnt1/β-catenin signaling pathway. We treated primary BMSCs from human subjects with alcohol for 24 days. We measured changes of genes related to endoplasmic reticulum (ER) stress, adipogenic markers and osteogenic markers using quantitative real-time RT-PCR and Western blot analysis. We performed Alizarin red staining for osteogenesis. We also conducted assays for osteogenic biomarkers alkaline phosphatase, collagen-I and osteocalcin. Wnt/β-catenin signaling was markedly activated in BMSCs treated with osteogenic inducers relative to the control cells, as indicated by the increased levels of nuclear β-catenin along with reduced levels of cytosolic β-catenin, as well as increased protein levels of Wnt1. Activation of Wnt/β-catenin signaling was significantly suppressed in BMSCs exposed to alcohol, which was reflected by downregulated expression of osteogenic marker genes Osf2/Cbfa1, osteopontin and osteocalcin, upregulated adipogenic marker PPARγ2 and aP2, and reduced number of calcifcation nodules. In contrast, activation of Wnt/β-catenin signaling by BIO favored osteogenesis even in the presence of alcohol. Simultaneous activation of Wnt1 by BIO and inhibition of TNF-α by 3,6'-dithiothalidomide produced synergetic suppression of ethanol-induced adipogenic lineage compared to interference with either of them alone. This remarkable shift of BMSCs towards osteoblast lineage suggests the superiority of concordant and reciprocal interferences of the TNF-α and Wnt/β-catenin pathways for promoting osteogenesis.


Vancomycin and Gentamicin Removal with the HA380 Cartridge during Experimental Hemoadsorption.
Hemoadsorption has emerged as an adjunctive therapy for sepsis, but its impact on antibiotic levels remains poorly defined. We conducted an in vivo experimental study to investigate the removal of vancomycin and gentamicin during hemoadsorption using the HA380 cartridge, a novel styrene-divinylbenzene copolymer cartridge. Six surgically prepared sheep were administered 2 g of vancomycin and 400 mg of gentamicin over 30 min, followed by a continuous infusion of vancomycin (20 mg/h). Hemoadsorption was implemented with a styrene-divinylbenzene copolymer HA380 cartridge at a blood flow of 120 mL/min. The removal ratio, sorbent-based clearance, and the mass removal rate were calculated for each time point. The mean 10-min vancomycin removal ratio exceeded 90% and declined to 68.0% at 30 min; 52.8% at 60 min, and 28.0% by 4 h. Due to constant plasma flow, clearance varied proportionally with the removal ratio. Over 4 hours, the total mass removal was 556 mg (SD 106.3). For gentamicin, the mean 10-min removal ratio was 96.9% and the final ratio at 4 h remained 53.0%, with clearances changing proportionately. The total mass removal of gentamicin was 138 mg (SD 26.6) over 4 h. The sorbent-based clearance of vancomycin was significantly lower than that of gentamicin (Pgroup < 0.0001). The novel HA380 sorbent cartridge appears safe and achieves significant vancomycin and gentamicin removal over a four-hour period. This information can be used by clinicians to guide their prescription and consider the additional dosing of at least an extra 25-35% amount in patients receiving HA380 hemoadsorption therapy during sepsis.


Orthosiphon stamineus: traditional uses, phytochemistry, pharmacology, and toxicology.
Orthosiphon stamineus Benth. (Lambiaceae) is an important plant in traditional folk medicine. This review is a comprehensive summary of the currently available chemical, pharmacological, and toxicological investigations as well as the traditional and therapeutic uses of this plant. Different in vitro and in vivo models have been addressed along with a survey of all phytochemicals identified in this plant, including flavonoids, terpenoids, and essential oils. Previous studies revealed that O. stamineus possesses several pharmacological activities, which are attributed to its phytochemical content. It was found that O. stamineus exhibits diuretic, hypouricemic, renal protective, antioxidant, anti-inflammatory, hepatoprotective, gastroprotective, antihypertensive, antidiabetic, antihyperlipidemic, antimicrobial, and anorexic activities. In conclusion, O. stamineus has wide traditional and pharmacological uses in various pathophysiological conditions. Therefore, it is an attractive subject for further experimental and clinical investigations.


A first order system model of fracture healing.
A first order system model is proposed for simulating the influence of stress stimulation on fracture strength during fracture healing. To validate the model, the diaphyses of bilateral tibiae in 70 New Zealand rabbits were osteotomized and fixed with rigid plates and stress-relaxation plates, respectively. Stress shielding rate and ultimate bending strength of the healing bone were measured at 2 to 48 weeks postoperatively. Ratios of stress stimulation and fracture strength of the healing bone to those of intact bone were taken as the system input and output. The assumed first order system model can approximate the experimental data on fracture strength from the input of stress stimulation over time, both for the rigid plate group and the stress-relaxation plate group, with different system parameters of time constant and gain. The fitting curve indicates that the effect of mechanical stimulus occurs mainly in late stages of healing. First order system can model the stress adaptation process of fracture healing. This approach presents a simple bio-mathematical model of the relationship between stress stimulation and fracture strength, and has the potential to optimize planning of functional exercises and conduct parametric studies.


Epigenetic reprogramming of the germ cell nuclear factor gene is required for proper differentiation of induced pluripotent cells.
Somatic cells have been reprogrammed into induced pluripotent stem (iPS) cells that recapitulate the pluripotent nature of embryonic stem (ES) cells. Reduced pluripotency and variable differentiation capacities have hampered progress with this technology for applications in regeneration medicine. We have previously shown that germ cell nuclear factor (Gcnf) is required for the repression of pluripotency genes during ES cell differentiation and embryonic development. Here we report that iPS cell lines, in which the Gcnf gene was properly reprogrammed, allowing expression of Gcnf, repress pluripotency genes during subsequent differentiation. In contrast, iPS clones in which the Gcnf gene was not reprogrammed maintained pluripotency gene expression during differentiation and did not differentiate properly either in vivo or in vitro. These mal-reprogrammed cells recapitulated the phenotype of Gcnf knockout (Gcnf(-/-)) ES cells. Reintroduction of Gcnf into either the Gcnf negative iPS cells or the Gcnf(-/-) ES cells rescued repression of Oct4 during differentiation. Our findings establish a key role for Gcnf as a regulator of iPS cell pluripotency gene expression. It also demonstrates that reactivation of the Gcnf gene may serve as a marker to distinguish completely reprogrammed iPS cells from incompletely pluripotent cells, which would make therapeutic use of iPS cells safer and more practical as it would reduce the oncogenic potential of iPS cells.


Phencyclidine-induced head-twitch response in rats treated chronically with methysergide.
This study was designed to assess whether phencyclidine (PCP)-induced behaviors in rats were potentiated after two days' withdrawal from chronic methysergide (a 5-HT2 receptor blocker) treatment (10 mg/kg per day i.p. for 12 days), in order to confirm the involvement of 5-hydroxytryptamine (5-HT) neurons in PCP actions. The PCP (10 mg/kg)-induced behaviors (head-twitch, head-weaving, turning and backpedalling) were attenuated by successive pretreatment with PCP (10 mg/kg per day i.p. for 12 days), while PCP- and 5-methoxy-N,N-dimethyltryptamine (2 mg/kg)-induced head-twitch increased significantly after the repeated methysergide treatment was stopped. The development of tolerance to PCP-induced head-twitch was antagonized by pretreatment with methysergide. Furthermore, Scatchard plots of specific [3H]ketanserin binding at the 5-HT2 receptors and [3H]PCP binding at the PCP receptors in the methysergide group revealed significant increases in binding capacity (Bmax) with no change in affinity (Kd). On the contrary, after development of tolerance to PCP, there were significant decreases in Bmax of [3H]ketanserin binding with no change in affinity. PCP can thus displace [3H]ketanserin at the 5-HT2 receptor site, but not [3H]5-HT at the 5-HT1 receptor site. These facts indicate that PCP may produce head-twitch via an agonistic interaction with 5-HT2 receptor sites.


In-vitro evaluation of antimicrobial and insect repellent potential of supercritical-carbon dioxide (SCF-CO2) extracts of selected botanicals against stored product pests and foodborne pathogens.
In the present study, the antimicrobial and the insect repellent activity of 16 botanical extracts obtained by supercritical CO2 (SCF) extraction were evaluated. The present investigation was conducted as there is a necessity for exploration of natural botanical extracts that target both stored product insects and microbes. The antimicrobial activity was studied by disc diffusion and broth microdilution methods against ten microbial species, including Gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis and Listeria monocytogenes), Gram-negative bacteria (Escherichia coli and Salmonella enterica), and fungi (Aspergillus flavus, Aspergillus paraciticus, Aspergillus ochraceous, Aspergillus niger and Penicillium verrucosum). Repellency assay was carried out by area preference method against three coleopteran insects (Tribolium castaneum, Rhyzopertha dominica and Sitophilus oryzae). Among all the extracts, thyme and ajwain were effective against all the tested bacteria with a minimum inhibition concentration (MIC) of 256-1024 µg/mL. Hop extract resulted in better antibacterial activity against all the tested Gram-positive bacteria with a MIC of 32-64 µg/mL. Oregano, thyme and ajwain extracts showed broad-spectrum antifungal activity against all the tested fungi with MIC of 128-1024 µg/mL. Most of the extracts exhibited class V (80.1-100%) repellency against T. castaneum. Extracts of hop, ajwain and thyme were found to have strong repellency against T. castaneum and R. dominica. Therefore, SCF extracts of ajwain and thyme can be explored further for the application of bio-extracts as a growth limiting factors in a microcosm where such consortia thrive.


Thermotolerant hydrogenases: biological diversity, properties, and biotechnological applications.
Hydrogenases are metalloproteins that catalyze the oxidation and reduction of molecular hydrogen and play a crucial role in many microbial metabolic processes. A subset of hydrogenases capable of functioning at temperatures from 50 to 125 degrees C is found in thermophilic microorganisms. Most known thermotolerant hydrogenases contain a [NiFe] active site and are either bidirectional or uptake type. Although no exhaustive survey has been done of the ecological diversity of thermophilic hydrogen-reducing or oxidizing bacteria, they appear to exist in virtually every thermophilic environment examined to date. Thermotolerant hydrogenases share many similarities with their mesophilic counterparts, but they have several features in addition to thermotolerance that make them especially well suited for biotechnological applications. Ongoing research is focused on potential applications of thermotolerant H2 ases in biosynthesis, H2 production, bioremediation, and biosensors.


Evaluation of Tiletamine-Zolazepam as an Anesthetic in Quail (Coturnix coturnix japonica).
The objective of the present study was to evaluate the effects of tiletamine-zolazepam (TZ) administered alone or in combination with atropine, xylazine, and levomepromazine to quail (Coturnix coturnix japonica). The induction time, duration of hypnosis and anesthesia, and time to recovery were determined. The presence or absence of tremor, upper respiratory tract secretions, and excitability and the degree of muscular tone were also observed. The results showed that doses from 10 to 100 mg/kg TZ administered alone or in combination with xylazine or levomepromazine failed to produce anesthesia; only hypnosis was obtained in a dose-dependent manner. Immediately after injection of the drug, histopathologic examination of the site of drug injection indicated the presence of discrete acute focal myositis. After 21 days, a discrete fibrosis between muscle fibers was detected in the pectoral muscle as a sign of scarring. We conclude that the administration of TZ to a dose of 100 mg/kg does not produce anesthesia in quail. For noninvasive and minimally painful procedures requiring chemical restraint and recumbency, the recommended dose is 30 mg/kg.


Rubus idaeus Inhibits Migration and Invasion of Human Nasopharyngeal Carcinoma Cells by Suppression of MMP-2 through Modulation of the ERK1/2 Pathway.
Nasopharyngeal carcinoma (NPC) is characterized by a high incidence of metastasis in the neck lymph nodes, resulting in a poor prognosis and posing challenges for treatment. In this study, we investigated the in vitro antimetastatic properties of Rubus idaeus extract (RIE) on human nasopharyngeal carcinoma cells. HONE-1, NPC-39 and NPC-BM cells were subjected to RIE treatment, and effects on the migration and invasion of tumor cells were analyzed. The results showed that RIE suppressed the migration and invasion of NPC cells. Gelatin zymography assay, Western blotting and real-time PCR showed that matrix metalloproteinases-2 (MMP-2) enzyme activity, protein expression and mRNA levels were down-regulated by RIE treatment. To identify the signaling pathway, mitogen-activated protein kinase proteins were examined, which showed that phosphorylation of ERK1/2 was inhibited after the treatment of RIE. In summary, our data showed that RIE inhibited the migration and invasion of NPC cells by suppressing the expression of MMP-2 by down-regulating the ERK1/2 signaling pathway, suggesting that Rubus idaeus may serve as chemotherapeutic and chemopreventive agent for NPC.


Understanding One Half of the Sex Difference Equation: The Modulatory Effects of Testosterone on Diabetic Cardiomyopathy.
Diabetes is a prevalent disease, primarily characterized by high blood sugar (hyperglycemia). Significantly higher rates of myocardial dysfunction have been noted in individuals with diabetes, even in those without coronary artery disease or high blood pressure (hypertension). Numerous molecular mechanisms have been identified, through which diabetes contributes to the pathology of diabetic cardiomyopathy, which presents as cardiac hypertrophy and fibrosis. At the cellular level, oxidative stress and inflammation in cardiomyocytes are triggered by hyperglycemia. While males are generally more likely to develop cardiovascular disease than females, diabetic males are less likely to develop diabetic cardiomyopathy than are diabetic females. One reason for these differences may be the higher levels of serum testosterone in males compared to females. While testosterone appears to protect against cardiomyocyte oxidative stress and exacerbate hypertrophy, its role in inflammation and fibrosis are much less clear. Additional pre-clinical and clinical studies will be required to delineate testosterone's effect on the diabetic heart.


Challenge Test as Special Tool to Estimate the Dynamic of Listeria monocytogenes and Other Foodborne Pathogens.
Over 23 million cases of foodborne disease (FBD) occur in Europe each year, with over 4700 deaths. Outbreaks of FBD have a significant impact on our society due to the high economic losses they cause (hospital treatment of affected patients and destruction of contaminated food). Among its health objectives, the European Union has set itself the goal of reducing the incidence of the main FBDs, approving various regulations that codify requirements in order to produce food that is "safe" for human consumption. Among these rules, Regulation 2005/2073 establishes precise food safety criteria for foods that are judged to be most at risk of causing episodes of FBD. The food business operator (FBO) must know their food better and know how to estimate whether a food can support the growth of food pathogens or if they are able to hinder it during the food's shelf life. It is becoming crucial for each FBO to schedule specific laboratory tests (challenge tests) to establish the growth potential of individual pathogens and their maximum growth rate. In 2008 the European Union published the guidelines for programming the challenge tests for Listeria monocytogenes in RTE foods. These guidelines were further implemented in 2014 and again in 2019. In June 2019 the UNI EN ISO 20976-1 was published, which contains indications for setting up and carrying out challenge tests for all foodborne pathogens in all foods. In this article, we compare the three official documents to highlight their common aspects and differences, highlighting the advantages and disadvantages that each of them offers for those who have to set up a challenge test for the various foodborne pathogens. Our conclusion is that the challenge test is today the most effective tool to estimate the dynamics and growth potential of pathogenic microorganisms in food, if it is designed and implemented in a scrupulous way. It is important to develop a rational experimental design for each challenge test, and for each food, and this requires professionals who are experts in this specific field of study and who must be properly trained.


A Clinicopathological Investigation of White Matter Hyperintensities and Alzheimer's Disease Neuropathology.
White matter hyperintensities (WMH) on magnetic resonance imaging (MRI) have been postulated to be a core feature of Alzheimer's disease. Clinicopathological studies are needed to elucidate and confirm this possibility. This study examined: 1) the association between antemortem WMH and autopsy-confirmed Alzheimer's disease neuropathology (ADNP), 2) the relationship between WMH and dementia in participants with ADNP, and 3) the relationships among cerebrovascular disease, WMH, and ADNP. The sample included 82 participants from the National Alzheimer's Coordinating Center's Data Sets who had quantitated volume of WMH from antemortem FLAIR MRI and available neuropathological data. The Clinical Dementia Rating (CDR) scale (from MRI visit) operationalized dementia status. ADNP+ was defined by moderate to frequent neuritic plaques and Braak stage III-VI at autopsy. Cerebrovascular disease neuropathology included infarcts or lacunes, microinfarcts, arteriolosclerosis, atherosclerosis, and cerebral amyloid angiopathy. 60/82 participants were ADNP+. Greater volume of WMH predicted increased odds for ADNP (p = 0.037). In ADNP+ participants, greater WMH corresponded with increased odds for dementia (CDR≥1; p = 0.038). WMH predicted cerebral amyloid angiopathy, microinfarcts, infarcts, and lacunes (ps < 0.04). ADNP+ participants were more likely to have moderate-severe arteriolosclerosis and cerebral amyloid angiopathy compared to ADNP-participants (ps < 0.04). This study found a direct association between total volume of WMH and increased odds for having ADNP. In patients with Alzheimer's disease, FLAIR MRI WMH may be able to provide key insight into disease severity and progression. The association between WMH and ADNP may be explained by underlying cerebrovascular disease.


Expression of DinJ-YafQ System of Lactobacillus casei Group Strains in Response to Food Processing Stresses.
Toxin-antitoxin (TA) systems are widely distributed in bacterial genomes and are involved in the adaptive response of microorganisms to stress conditions. Few studies have addressed TA systems in Lactobacillus and their role in the adaptation to food environments and processes. In this work, for six strains belonging to L. casei group isolated from dairy products, the expression of DinJ-YafQ TA system was investigated after exposure to various food-related stresses (nutrient starvation, low pH, high salt concentration, oxidative stress, and high temperature), as well as to the presence of antibiotics. In particular, culturability and DinJ-YafQ expression were evaluated for all strains and conditions by plate counts and RT qPCR. Among all the food-related stress conditions, only thermal stress was capable to significantly affect culturability. Furthermore, exposure to ampicillin significantly decreased the culturability of two L. rhamnosus strains. The regulation of DinJ-YafQ TA system resulted strain-specific; however, high temperature was the most significant stress condition able to modulate DinJ-YafQ expression. The increasing knowledge about TA systems activity and regulation might offer new perspectives to understand the mechanisms that L. casei group strains exploit to adapt to different niches or production processes.


[Electron microscopy in the differential diagnosis of uterine sarcoma].
Three uterine sarcomas histologically diagnosed as differentiated leiomyosarcomas were studied electron microscopically for differential diagnosis. The study showed not all highly differentiated uterine sarcomas to be of myogenic origin. The histological diagnosis of differentiated leiomyosarcoma which consists mostly of spindle-shaped and elongated cells may often cover tumours of different histogenetic composition. In our material, electron microscopy established leiomyosarcoma in 2 cases, and sarcoma of the fibroblastic type in 1 case.


Maternal serum lipidomics identifies lysophosphatidic acid as a predictor of small for gestational age neonates.
To discover lipidomic alterations during pregnancy in mothers who subsequently delivered small for gestational age (SGA) neonates and identify predictive lipid markers that can help recognize and manage these mothers, we carried out untargeted lipidomics on maternal serum samples collected between 24-28 weeks of gestation. We used a nested case-control study design and serum from mothers who delivered SGA and appropriate for gestational age babies. We applied untargeted lipidomics using mass spectrometry to characterize lipids and discover changes associated with SGA births during pregnancy. Multivariate pattern recognition software Collaborative Laboratory Integrated Reports (CLIR) was used for the post-analytical recognition of range differences in lipid ratios that could differentiate between SGA and control mothers and their integration for complete separation between the two groups. Here, we report changes in lipids from serum collected during pregnancy in mothers who delivered SGA neonates. In contrast to normal pregnancies where lysophosphatidic acid increased over the course of the pregnancy owing to increased activity of lysophospholipase D, we observed a decrease (32%; P = 0.05) of 20:4-lysophosphatidic acid in SGA mothers, which could potentially compromise fetal growth and development. Integration of lipid ratios in an interpretive tool (CLIR) could completely separate SGA mothers from controls demonstrating the power of untargeted lipidomic analyses for identifying novel predictive biomarkers. Additional studies are required for further assessment of the lipid biomarkers identified in this report.


Graph-based homogenisation for modelling cardiac fibrosis.
Fibrosis, the excess of extracellular matrix, can affect, and even block, propagation of action potential in cardiac tissue. This can result in deleterious effects on heart function, but the nature and severity of these effects depend strongly on the localisation of fibrosis and its by-products in cardiac tissue, such as collagen scar formation. Computer simulation is an important means of understanding the complex effects of fibrosis on activation patterns in the heart, but concerns of computational cost place restrictions on the spatial resolution of these simulations. In this work, we present a novel numerical homogenisation technique that uses both Eikonal and graph approaches to allow fine-scale heterogeneities in conductivity to be incorporated into a coarser mesh. Homogenisation achieves this by deriving effective conductivity tensors so that a coarser mesh can then be used for numerical simulation. By taking a graph-based approach, our homogenisation technique functions naturally on irregular grids and does not rely upon any assumptions of periodicity, even implicitly. We present results of action potential propagation through fibrotic tissue in two dimensions that show the graph-based homogenisation technique is an accurate and effective way to capture fine-scale domain information on coarser meshes in the context of sharp-fronted travelling waves of activation. As test problems, we consider excitation propagation in tissue with diffuse fibrosis and through a tunnel-like structure designed to test homogenisation, interaction of an excitation wave with a scar region, and functional re-entry.


Control of Arabidopsis meristem development by thioredoxin-dependent regulation of intercellular transport.
Cell-to-cell transport in plants occurs through cytoplasmic channels called "plasmodesmata" and is regulated by developmental and environmental factors. Callose deposition modulates plasmodesmal transport in vivo, but little is known about the mechanisms that regulate this process. Here we report a genetic approach to identify mutants affecting plasmodesmal transport. We isolated 5 mutants, named gfp arrested trafficking (gat), affected in GFP unloading from the phloem into the meristem. gat1 mutants were seedling lethal and carried lesions in an m-type thioredoxin that is expressed in non-green plastids of meristems and organ primordia. Callose and hydrogen peroxide accumulated in gat1 mutants, and WT plants subjected to oxidative conditions phenocopied the gat1 trafficking defects. Ectopic expression of GAT1 in mature leaves increased plasmodesmal permeability and led to a delay in senescence and flowering time. We propose a role for the GAT1 thioredoxin in the redox regulation of callose deposition and symplastic permeability that is essential for meristem maintenance in Arabidopsis.


Imipramine and pregabalin combination for painful polyneuropathy: a randomized controlled trial.
Monotherapy with first-line drugs for neuropathic pain often fails to provide sufficient pain relief or has unacceptable side effects because of the need for high doses. The aim of this trial was to test whether the combination of imipramine and pregabalin in moderate doses would relieve pain more effectively than monotherapy with either of the drugs. This was a randomized, double-blind, placebo-controlled, crossover, multicenter trial consisting of four 5-week treatment periods in patients with painful polyneuropathy. Treatment arms were imipramine 75 mg/d vs pregabalin 300 mg/d vs combination therapy vs placebo. Patients with polyneuropathy and symptoms for more than 6 months, age 20 to 85 years, pain intensity ≥4 on a 0- to 10-point numeric rating scale (NRS) and pain at least 4 days a week were included in the trial. A total of 262 patients were screened for participation, 73 patients were randomized, and 69 patients were included in the data analysis. The effect on average pain in comparison with placebo was: combination (-1.67 NRS points, P < 0.001), imipramine (-1.08 NRS points, P < 0.001), and pregabalin (-0.48 NRS points, P = 0.03). The combination therapy had significantly lower pain scores than both monotherapies: combination vs imipramine (P = 0.009), combination vs pregabalin (P < 0.001). During combination therapy, the dropout rate was higher and the patients reported a higher rate and severity of side effects. Combination of moderate doses of the tricyclic antidepressant imipramine and pregabalin could be considered as an alternative to high-dosage monotherapy. However, the trial also emphasized that balance between efficacy and safety is an issue.


A simple method for the preparation of single cells and regeneration of colonies of Botryococcus braunii NIES836.
Botryococcus braunii, a colonial alga, is known for notably slow growth; however, the growth rate and hydrocarbon productivity are expected to be improved using genetic modification techniques. Nevertheless, B. braunii has a hydrocarbon-rich extracellular matrix (ECM), and the ECM is a major barrier to DNA transformation. To analyse and utilize genetically modified B. braunii, it is essential to regenerate genetically homogeneous colonies derived from single cells. In this study, we developed a novel, simple method for harvesting viable single cells of B. braunii by centrifugation of the culture and subsequent filtration alone. The harvest of single cells was made possible by culturing B. braunii colonies in AF6 medium until the depletion of nitrogen and phosphorus sources and then releasing the single cells in colonies into the medium. Twenty-day culture of single cells in a 96-well plate resulted in 96% regeneration of colonies, and the regeneration of colonies was also confirmed on agar medium. This is the first report of colony regeneration from single cells of B. braunii. We believe that our method developed in this study will contribute greatly to the advancement of genetic modification techniques for B. braunii.


Recapitulating human cardio-pulmonary co-development using simultaneous multilineage differentiation of pluripotent stem cells.
The extensive crosstalk between the developing heart and lung is critical to their proper morphogenesis and maturation. However, there remains a lack of models that investigate the critical cardio-pulmonary mutual interaction during human embryogenesis. Here, we reported a novel stepwise strategy for directing the simultaneous induction of both mesoderm-derived cardiac and endoderm-derived lung epithelial lineages within a single differentiation of human-induced pluripotent stem cells (hiPSCs) via temporal specific tuning of WNT and nodal signaling in the absence of exogenous growth factors. Using 3D suspension culture, we established concentric cardio-pulmonary micro-Tissues (μTs), and expedited alveolar maturation in the presence of cardiac accompaniment. Upon withdrawal of WNT agonist, the cardiac and pulmonary components within each dual-lineage μT effectively segregated from each other with concurrent initiation of cardiac contraction. We expect that our multilineage differentiation model will offer an experimentally tractable system for investigating human cardio-pulmonary interaction and tissue boundary formation during embryogenesis.


Metabolic Heterogeneity in Patient Tumor-Derived Organoids by Primary Site and Drug Treatment.
New tools are needed to match cancer patients with effective treatments. Patient-derived organoids offer a high-throughput platform to personalize treatments and discover novel therapies. Currently, methods to evaluate drug response in organoids are limited because they overlook cellular heterogeneity. In this study, non-invasive optical metabolic imaging (OMI) of cellular heterogeneity was characterized in breast cancer (BC) and pancreatic cancer (PC) patient-derived organoids. Baseline heterogeneity was analyzed for each patient, demonstrating that single-cell techniques, such as OMI, are required to capture the complete picture of heterogeneity present in a sample. Treatment-induced changes in heterogeneity were also analyzed, further demonstrating that these measurements greatly complement current techniques that only gauge average cellular response. Finally, OMI of cellular heterogeneity in organoids was evaluated as a predictor of clinical treatment response for the first time. Organoids were treated with the same drugs as the patient's prescribed regimen, and OMI measurements of heterogeneity were compared to patient outcome. OMI distinguished subpopulations of cells with divergent and dynamic responses to treatment in living organoids without the use of labels or dyes. OMI of organoids agreed with long-term therapeutic response in patients. With these capabilities, OMI could serve as a sensitive high-throughput tool to identify optimal therapies for individual patients, and to develop new effective therapies that address cellular heterogeneity in cancer.


Doctors and the assessment of blood glucose testing sticks: does colour blindness matter?
A group of doctors with congenital colour vision deficiency (CCVD) were compared with a group of controls in their assessment of colour blocks in the colour range of a widely available blood glucose testing stick. The majority of doctors with CCVD agreed with controls on colour matching. However, subjects with severe CCVD tended to match test blocks to a wider range of options than either those with a less severe defect or controls. This paper discusses the implications of these findings.


Foot-and-mouth disease vaccination in South Sudan: benefit-cost analysis and livelihoods impact.
A benefit-cost analysis of vaccination for foot-and-mouth disease (FMD) was conducted in an area of South Sudan, which due to chronic conflict, had been subject to large-scale humanitarian assistance for many years. The study used participatory epidemiology (PE) methods to estimate the prevalence and mortality of acute and chronic FMD in different age groups of cattle, and the reduction in milk off-take in cows affected by FMD. The benefit-cost of FMD vaccination was 11.5. Losses due to the chronic form of FMD accounted for 28.2% of total FMD losses, indicating that future benefit-cost analyses for FMD control in pastoral and agropastoral areas of Africa need to consider losses caused by chronic disease. Participatory epidemiological methods were also used to assess the importance of milk in the diet of Nuer agropastoralists, and seasonal variations in diet in relation to cattle movements and FMD outbreaks. Marked seasonal variation in diet included a 'hunger gap' period during which households were highly dependent on milk as their main source of food. Outbreaks of FMD occurred immediately before this period of milk dependency, with chronic losses extending through this period and affecting human food security. The paper discusses the need and feasibility of mass vaccination and strategic vaccination for FMD in South Sudan. The paper also discusses the value of combining conventional benefit-cost analysis with livelihoods analysis to inform disease control efforts and funding commitments in humanitarian contexts.


Predicting oral druglikeness by iterative stochastic elimination.
Integration of computational methods in the early stages of drug discovery has been one of the key trends in the pharmaceutical industry. Starting with high quality drug candidates should ultimately minimize clinical attrition rates and give rise to higher success rates. In this paper, we present a novel approach for indexing oral druglikeness of compounds. With the Iterative Stochastic Elimination (ISE) Algorithm, we distinguish between orally available drugs and nondrugs by generating sets of optimized descriptors' ranges, each set constituting a "filter". We delineate in this paper how to produce an ensemble of best k-descriptor sets out of the huge number of possibilities, and how to construct a "filter bank" that retains diverse filters by clustering. Finally, we define the "orally bioavailable drug-like" character of individual molecules by combining the filters into an "Orally Bioavailable Druglike Index" (OB-DLI) which may be used to prioritize molecules in databases and discuss its uses in several potential applications. The predictive power with sets of 4-6 descriptors is high (i.e., one filter of 5 descriptors retrieved 81% true positives and >77% true negatives). Thus, OB-DLI has advantages over binary decisions (that use only one filter) not only in raising discriminative power but also in ranking drug candidates according to their chance to be successful oral drugs. We demonstrate the ability of our approach to discover molecular entities with the required property, orally bioavailable drug likeness, that are structurally dissimilar to those of the training set. Comparison of this ISE application to some of the current main methods for classification reveals that our approach has >13% improvement in the Matthews Correlation Coefficient, which measures the success of identifying true and false positives and negatives.


Use of dithiothreitol to improve the diagnosis of prosthetic joint infections.
Diagnosis of prosthetic joint infections (PJIs) remains a challenge for microbiologists, despite new techniques for bacteria isolation have been developed in recent years. A widely recognized standard method has not yet been indicated mainly because of limitations due difficult procedures and need of dedicated instrumentation. We evaluated the ability of a sulfhydryl compound routinely used in microbiology laboratories, dithiothreitol (DTT), to dislodge bacteria from biofilm, keeping them alive and cultivable for identification and antibiotic susceptibility testing. We compared DTT treatment against sonication of prosthesis and culture of periprosthetic tissues, in order to establish if it could be introduced in routine microbiological diagnosis of PJIs. The study was conducted on 76 patients, 34 with aseptic loosening of their prosthesis and 42 who were diagnosed for PJI. DTT treatment gave results similar to sonication in terms of bacterial yielding. Sonication provided higher sensitivity (71.4%) and specificity (94.1%) respect to periprosthetic tissue culture, while DTT showed the same specificity of sonication but a better sensitivity (85.7%), especially when the causative microorganism was Staphylococcus epidermidis. In conclusion, we demonstrated that DTT could be used for PJIs diagnosis, thanks to its ease of use and its high sensitivity and specificity.


Return Migrants' Experience of Access to Care in Corrupt Healthcare Systems: The Bosnian Example.
Equal and universal access to healthcare services is a core priority for a just health system. A key societal determinant seen to create inequality in access to healthcare is corruption in the healthcare system. How return migrants' access to healthcare is affected by corruption is largely unstudied, even though return migrants may be particularly vulnerable to problems related to corruption due to their period of absence from their country of origin. This article investigates how corruption in the healthcare sector affects access to healthcare for refugees who repatriated to Bosnia, a country with a high level of corruption, from Denmark, a country with a low level of corruption. The study is based on 18 semi-structured interviews with 33 refugees who returned after long-term residence in Denmark. We found that the returned refugees faced greater problems with corruption than was the case for those who had not left the country, as doctors considered them to be better endowed financially and therefore demanded larger bribes from them than they did from those who had remained in Bosnia. Moreover, during their stay abroad the returnees had lost the connections that could have helped them sidestep the corruption. Returned refugees are thus particularly vulnerable to the effects of corruption.


Antigen-specific murine T cell clones produce soluble interleukin 2 receptor on stimulation with specific antigens.
In this study, we used monoclonal antibodies to the murine IL 2 receptor (IL 2R) termed 3C7 and 7D4, which bind to different epitopes on the murine IL 2R, to develop an ELISA to measure soluble murine IL 2R. Surprisingly, stimulated murine spleen cells not only expressed cell-associated IL 2R, but also produced a considerable level of cell free IL 2R in the culture supernatant fluid. To assess the fine specificity of this response, myoglobin-immune murine T cell clones were stimulated with appropriate or inappropriate antigen and syngeneic or allogeneic presenting cells. Proliferation, measured by [3H] thymidine incorporation, and levels of soluble IL 2R were determined at day 4. The production of soluble IL 2R displayed the same epitope fine specificity, genetic restriction, and antigen dose-response as the proliferative response. Indeed, in some cases there was sharper discrimination of epitope specificity and genetic restriction with the soluble IL 2R levels. There was also reproducible clone-to-clone variation in the amount of soluble receptor produced in response to antigen among 12 T cell clones and lines tested. In time course experiments, proliferation was greatest at day 3, whereas soluble IL 2R levels continued to rise in subsequent days. To our knowledge, this is the first demonstration of release or secretion of soluble IL 2R by murine T cells, and the first demonstration of the fine specificity and genetic restriction of the induction of soluble IL 2R by specific antigen.


An assessment of the intestinal lumen as a site for intervention in reducing body burdens of organochlorine compounds.
Many individuals maintain a persistent body burden of organochlorine compounds (OCs) as well as other lipophilic compounds, largely as a result of airborne and dietary exposures. Ingested OCs are typically absorbed from the small intestine along with dietary lipids. Once in the body, stored OCs can mobilize from adipose tissue storage sites and, along with circulating OCs, are delivered into the small intestine via hepatic processing and biliary transport. Retained OCs are also transported into both the large and small intestinal lumen via non-biliary mechanisms involving both secretion and desquamation from enterocytes. OCs and some other toxicants can be reabsorbed from the intestine, however, they take part in enterohepatic circulation(EHC). While dietary fat facilitates the absorption of OCs from the small intestine, it has little effect on OCs within the large intestine. Non-absorbable dietary fats and fat absorption inhibitors, however, can reduce the re-absorption of OCs and other lipophiles involved in EHC and may enhance the secretion of these compounds into the large intestine--thereby hastening their elimination. Clinical studies are currently underway to determine the efficacy of using non-absorbable fats and inhibitors of fat absorption in facilitating the elimination of persistent body burdens of OCs and other lipophilic human contaminants.


The psycho-social malaise of migrant private carers in Italy: a rampant, but hidden health demand.
Despite having become an essential part of the national welfare system, the Italian private care market has developed in a situation of institutional disengagement. As a consequence, this sector, which has a high presence of female migrant workers, is currently characterised by serious flaws. What are the consequences for the psycho-social well-being of migrant private carers (MPCs)? This article highlights and analyses the high correlation between migrant women's involvement in the Italian private care market and their manifestations of psycho-social malaise, the latter being one of the main factors motivating the access to health services for MPCs employed in Italy. Based on qualitative data collected in Italy in 2009-2010: 32 in-depth interviews to MPCs, a focus group discussion involving seven MPCs, and 23 semi-structured interviews to providers working in close contact with MPCs, mostly in health services, it describes how the malaise of MPCs is generated, and how it is interpreted and expressed (or not expressed) by them. In particular, it investigates the social dynamics determining the latent nature of this phenomenon, and its tendency to work silently on MPCs' subjectivities without coming to the attention of health services, which are therefore not able to prevent or limit its negative consequences. Nevertheless, this article also shows that this malaise sometimes surfaces in the health service (albeit too rarely, or too late) and how it is generally managed by health providers. In particular, a timely encounter with the health system involving the simple externalisation and acknowledgement of this psycho-social malaise can have a crucial therapeutic value, meaningfully helping to prevent it from becoming a serious condition. On this basis, the importance is highlighted of creating channels for facilitating the meeting between this pressing but unexpressed health need and the institutions in charge of the health of society.


Overcoming Barriers Associated with Oral Delivery of Differently Sized Fluorescent Core-Shell Silica Nanoparticles.
Oral delivery, while a highly desirable form of nanoparticle-drug administration, is limited by challenges associated with overcoming several biological barriers. Here, the authors study how fluorescent and poly(ethylene glycol)-coated (PEGylated) core-shell silica nanoparticles sized 5 to 50 nm interact with major barriers including intestinal mucus, intestinal epithelium, and stomach acid. From imaging fluorescence correlation spectroscopy studies using quasi-total internal reflection fluorescence microscopy, diffusion of nanoparticles through highly scattering mucus is progressively hindered above a critical hydrodynamic size around 20 nm. By studying Caco-2 cell monolayers mimicking the intestinal epithelia, it is observed that ultrasmall nanoparticles below 10 nm diameter (Cornell prime dots, [C' dots]) show permeabilities correlated with high absorption in humans from primarily enhanced passive passage through tight junctions. Particles above 20 nm diameter exclusively show active transport through cells. After establishing C' dot stability in artificial gastric juice, in vivo oral gavage experiments in mice demonstrate successful passage through the body followed by renal clearance without protein corona formation. Results suggest C' dots as viable candidates for oral administration to patients with a proven pathway towards clinical translation and may generate renewed interest in examining silica as a food additive and its effects on nutrition and health.


A Case Of Tubal Ectopic Pregnancy.
A healthy 27 year old Para 3 presenting with abnormal menstruation without a period of amenorrhoea was diagnosed to have left tubal ectopic pregnancy after vaginal examination and abdominal ultrasonography. The case illustrates the need for careful history taking and the need for considering ectopic pregnancy in women in the reproductive age group, who have abnormal menstruation even if they are on contraception.


Local Riemannian geometry of model manifolds and its implications for practical parameter identifiability.
When non-linear models are fitted to experimental data, parameter estimates can be poorly constrained albeit being identifiable in principle. This means that along certain paths in parameter space, the log-likelihood does not exceed a given statistical threshold but remains bounded. This situation, denoted as practical non-identifiability, can be detected by Monte Carlo sampling or by systematic scanning using the profile likelihood method. In contrast, any method based on a Taylor expansion of the log-likelihood around the optimum, e.g., parameter uncertainty estimation by the Fisher Information Matrix, reveals no information about the boundedness at all. In this work, we present a geometric approach, approximating the original log-likelihood by geodesic coordinates of the model manifold. The Christoffel Symbols in the geodesic equation are fixed to those obtained from second order model sensitivities at the optimum. Based on three exemplary non-linear models we show that the information about the log-likelihood bounds and flat parameter directions can already be contained in this local information. Whereas the unbounded case represented by the Fisher Information Matrix is embedded in the geometric framework as vanishing Christoffel Symbols, non-vanishing constant Christoffel Symbols prove to define prototype non-linear models featuring boundedness and flat parameter directions of the log-likelihood. Finally, we investigate if those models could allow to approximate and replace computationally expensive objective functions originating from non-linear models by a surrogate objective function in parameter estimation problems.


The effect of beta 2-adrenoceptor stimulation and blockade of L-type calcium channels on in vivo Na+/H+ antiporter activity in rat skeletal muscle.
We have studied the in vivo response of the Na+/H+ antiporter in skeletal muscle to beta 2-adrenoceptor stimulation with isoprenaline and the effect of blocking L-type calcium channels with nifedipine. Na+/H+ antiporter activity in skeletal muscle in vivo increased after beta 2-adrenoceptor stimulation with isoprenaline; nifedipine attenuated that effect. This suggests that opening of L-type calcium channels is necessary for full activation of the Na+/H+ antiporter in skeletal muscle. Bleeding also increased Na/H+ antiporter activity, which we believe could be explained by an increase in sympathetic nervous system activity as a result of hypotension. This may be one of the mechanisms by which animals under stress prepare their skeletal muscle for exercise as part of the 'fright and flight' reaction.


[Roaming through methodology. VII. Reproducibility of measurements].
Reproducibility measurements are important for a proper interpretation of medical data. Kappa is the most adequate measure for categorical variables. Kappa adjusts the observed agreement for chance agreement. The interpretation of kappa is rather difficult. The kappa value is influenced by the number of categories used for classification and the prevalence of scores of the observers. For continuous variables the Pearson correlation coefficient can be used, keeping in mind its ignoring systematic errors and its dependence on the heterogeneity of the data. Another method to assess reproducibility for continuous variables is the method of limits of agreement. This method distinguishes systematic and random errors, and quantifies the differences in the dimension of the measurements. In general, the interpretation of the various measures of agreement is helped by a visual presentation of the data in a table or figure.


Highlights of glycosylation and adhesion related genes involved in myogenesis.
Myogenesis is initiated by myoblast differentiation and fusion to form myotubes and muscle fibres. A population of myoblasts, known as satellite cells, is responsible for post-natal growth of muscle and for its regeneration. This differentiation requires many changes in cell behaviour and its surrounding environment. These modifications are tightly regulated over time and can be characterized through the study of changes in gene expression associated with this process. During the initial myogenesis steps, using the myoblast cell line C2C12 as a model, Janot et al. (2009) showed significant variations in expression for genes involved in pathways of glycolipid synthesis. In this study we used murine satellite cells (MSC) and their ability to differentiate into myotubes or early fat storage cells to select glycosylation related genes whose variation of expression is myogenesis specific. The comparison of variant genes in both MSC differentiation pathways identified 67 genes associated with myogenesis. Comparison with data obtained for C2C12 revealed that only 14 genes had similar expression profiles in both cell types and that 17 genes were specifically regulated in MSC. Results were validated statistically by without a priori clustering. Classification according to protein function encoded by these 31 genes showed that the main regulated cellular processes during this differentiation were (i) remodeling of the extracellular matrix, particularly, sulfated structures, (ii) down-regulation of O-mannosyl glycan biosynthesis, and (iii) an increase in adhesion protein expression. A functional study was performed on Itga11 and Chst5 encoding two highly up-regulated proteins. The inactivation of Chst5 by specific shRNA delayed the fusion of MSC. By contrast, the inactivation of Itga11 by specific shRNA dramatically decreased the fusion ability of MSC. This result was confirmed by neutralization of Itga11 product by specific antibodies. Our screening method detected 31 genes specific for myogenic differentiation out of the 383 genes studied. According to their function, interaction networks of the products of these selected genes converged to cell fusion. Functional studies on Itga11 and Chst5 demonstrated the robustness of this screening.


Genome composition analysis of multipartite BNYVV reveals the occurrence of genetic re-assortment in the isolates of Asia Minor and Thrace.
Beet necrotic yellow vein virus (BNYVV) is the cause of rhizomania, an important disease of sugar beet around the world. The multipartite genome of the BNYVV contains four or five single-stranded RNA that has been used to characterize the virus. Understanding genome composition of the virus not only determines the degree of pathogenicity but also is required to development of resistant varieties of sugar beet. Resistance to rhizomania has been conferred to sugar beet varieties by conventional breeding methods or modern genome engineering tools. However, over time, viruses undergo genetic alterations and develop new variants to break crop resistance. Here, we report the occurrence of genetic reassortment and emergence of new variants of BNYVV among the isolates of Thrace and Asia Minor (modern-day Turkey). Our findings indicate that the isolates harbor European A-type RNA-2 and RNA-3, nevertheless, RNA-5 is closely related to East Asian J-type. Furthermore, RNA-1 and RNA-4 are either derived from A, B, and P-types or a mixture of them. The RNA-5 factor which enhance the pathogenicity, is rarely found in the isolates studied (20%). The creation of new variants of the virus emphasizes the necessity to develop new generation of resistant crops. We anticipate that these findings will be useful for future genetic characterization and evolutionary studies of BNYVV, as well as for developing sustainable strategies for the control of this destructive disease.


High-dose intravenous gamma globulin improves responses to single-donor platelets in patients refractory to platelet transfusion.
Ten patients, with bone marrow failure or malignant disorders, became refractory to platelet transfusions using random, as well as partial or fully HLA-matched, single-donor platelets. To determine its effect on platelet refractoriness, intravenous gamma globulin (IV IgG) was administered at 400 or 800 mg/kg/d for five days, and postinfusion platelet responses were monitored. Platelet transfusion responses following intravenous gamma globulin (IV IgG) were graded as follows: Excellent, 48-hour posttransfusion count greater than 50,000/microL; good, 48-hour count greater than 20,000 but less than 50,000/microL; Fair, increased increment, 48-hour count less than 20,000; and failed, no increased increment. Six of ten patients (60%) had improved responses to selected single-donor platelets (two were excellent, three were good, and one was fair). The time to achieve a platelet transfusion count greater than 25,000/microL ranged from one to nine days of IgG therapy. One individual had sustained benefit (greater than 1 year); the remaining responses persisted for 6 to 8 weeks. These results suggest that IV IgG may be useful in the management of platelet refractoriness, especially in patients receiving single-donor platelets.


A genetic instrumental variables analysis of the effects of prenatal smoking on birth weight: evidence from two samples.
There is a large literature showing the detrimental effects of prenatal smoking on birth and childhood health outcomes. It is somewhat unclear though, whether these effects are causal or reflect other characteristics and choices by mothers who choose to smoke that may also affect child health outcomes or biased reporting of smoking. In this paper we use genetic markers that predict smoking behaviors as instruments to address the endogeneity of smoking choices in the production of birth and childhood health outcomes. Our results indicate that prenatal smoking produces more dramatic declines in birth weight than estimates that ignore the endogeneity of prenatal smoking, which is consistent with previous studies with non-genetic instruments. We use data from two distinct samples from Norway and the United States with different measured instruments and find nearly identical results. The study provides a novel application that can be extended to study several behavioral impacts on health and social and economic outcomes.


Quantitative analysis of the cooperative effect between inducers of Epstein-Barr virus antigen synthesis.
Induction of Epstein-Barr virus antigens in Raji cells by 12-O-tetradecanoylphorbol 13-acetate (TPA), 5'-iodo-2'-deoxyuridine (IUdR), n-butyric acid (n-BA) antibody to IgM (anti-IgM) and a recently described serum factor was studied after single applications of individual inducers or of their combinations. Whereas individual inducers led to activation only in a limited number of cells, certain combinations of inducers greatly enlarged the reactive cell population. In synergistically acting combinations of drugs, the dose necessary for half-maximal induction of the first inducer was shifted to a much lower value by the second inducer. The serum factor cooperated with any of the four inducers TPA, IUdR, anti-IgM and n-butyric acid, whereas TPA only interacted with IUdR and n-butyric acid. The effects of the serum factor and of TPA showed some similarities but were not identical, whereas IUdR and n-BA showed identical effects.


Evaluation of polyphenol contents in differently processed apricots using accelerated solvent extraction followed by high-performance liquid chromatography-diode array detector.
Concentrations of 17 polyphenols in ripe-fresh, sun-dried and sulfited-dried apricots either harvested from organic or pesticide-treated trees before harvest were determined using reverse-phase high-performance liquid chromatography with UV-Vis diode array detector, and the change of polyphenol profile with origin of the apricot product was established. Extraction of polyphenols was achieved using the accelerated solvent extraction technique (ASE). A mixture of methanol, water (70:30 v/v) including 0.1 g of tert-butylhydroquinone as solvent, 60 min extraction time, 60°C temperature and 1,500 psi pressure were found the most productive operating conditions for ASE. Concentrations of polyphenols in organic apricots were higher than pesticide-treated samples, and roughly the same in ripe-fresh and sun-dried apricots. Polyphenol concentrations of either organic or pesticide-treated sulfited-dried apricots were lower than the ripe-fresh apricots. Considering the organic ripe-fresh apricots, there was a decrease in some polyphenol concentration in the sulfited-dried apricots compared with the ripe-fresh apricot, and the range of decrease is between 1.4 and 53%.


[Recent methods for the detection of airborne microorganisms and source identification].
The detection of airborne microorganisms including selected cell constituents (e.g. allergens or endotoxins) depends on suitable methods and instruments for their collection. Furthermore, microbiological methods are necessary for their quantification and qualification. In the past these methods were largely based on the classical cultivation dependent approach. Modern molecular methods, e.g. direct staining procedures, hybridization assays with nucleic acids including the PCR-technology or immunological assays are promising new tools for a more sophisticated detection of bioaerosols. They allow a better detection rate, a more precise identification of certain members of the aerosol including cell constituents. With respect to speed and lower costs they are an important alternative to established detection methods.


Harriet Tubman's Hypersomnia: Insights from Historical and Medical Perspectives.
Harriet Tubman, a hero of the abolitionist movement and early civil rights advocate, suffered a head injury in childhood and subsequently developed sleep attacks associated with visions that were extensively documented in historical accounts. Her contemporaries perceived these visions together with unpredictable and unavoidable urges to sleep as manifestations of her deep faith, rather than as symptoms of an illness. While religious perspectives remain crucial to understanding Tubman's sleep-related experiences, some may consider them insufficient in view of modern medical advances. We propose the parallel explanation that her sleep attacks, usually attributed to temporal lobe epilepsy, actually represent a hypersomnia that is most consistent with the modern diagnosis of post-traumatic narcolepsy. Using historical analysis as well as current understandings of sleep medicine, we aim to shed light on this under-recognized aspect of Tubman's life. In addition, this case study allows us to review the potential long-term effects of severe traumatic brain injuries; consider a differential for excessive daytime sleepiness and hypnagogic hallucinations; and familiarize readers with the pathophysiology, diagnosis, and treatment of narcolepsy. Whether her symptoms are viewed through the lens of the past or measured against current biomedical standards, Tubman demonstrated an inspiring ability to persevere despite intrusive sleep episodes and to realize her dreams for the betterment of others.


The effects of frequent ejaculation on seminal spermatozoal number and calculation of daily spermatozoal output.
Spermatozoal depletion of the extra-gonadal reserve (EGR) has been monitored in 14 men by analysing ejaculates collected daily for 5 days. It has been concluded that EGR is stabilized within 2 days and that an estimate of daily spermatozoal output can be obtained either by averaging the values from days 3 to 5 or by using the day 5 value alone. This latter parameter may prove useful clinically.


The children of preterm survivors: shyness, parenting, and parental stress.
Extremely low birth weight (ELBW) survivors have higher rates of shyness, a risk factor for poorer outcomes across the life span. Due to advances in fetal and neonatal medicine, the first generation of ELBW survivors have survived to adulthood and become parents. However, no studies have investigated the transmission of their stress vulnerability to their offspring. We explored this phenomenon using a population-based cohort of ELBW survivors and normal birth weight (NBW) controls. Using data from three generations, we examined whether the shyness and parenting stress of ELBW and NBW participants (Generation 2) mediated the relation between the parenting style of their parents (Generation 1) and shyness in their offspring (Generation 3), and the extent to which exposure to perinatal adversity (Generation 2) moderated this mediating effect. We found that among ELBW survivors, parenting stress (in Generation 2) mediated the relation between overprotective parenting style in Generation 1 (grandparents) and child shyness in Generation 3. These findings suggest that perinatal adversity and stress may be transmitted to the next generation in humans, as reflected in their perceptions of their children as shy and socially anxious, a personality phenotype that may subsequently place their children at risk of later mental and physical health problems.


Embryo transfers from mares in athletic competition.
The objective of this project was to produce multiple pregnant recipient mares during one year from each of four mares which were in athletic training and competition. Twenty-six embryos were recovered from the four mares in 38 collection attempts. Each embryo was surgically transferred to one of 20 recipient mares. Nine of 10 recipient mares which were transferred to on one occasion were detected pregnant at the one week post transfer pregnancy examination and six of eight mares which were transferred to on two occasions were detected pregnant at one of the one-week post transfer pregnancy examinations. Three of the 15 pregnant recipient mares lost their pregnancies by day 150. The 12 resulting pregnant recipient mares from the four competing donor mares demonstrated that donor mares can remain in athletic competition and at the same time provide viable embryos for embryo transfer.


Trypan blue/giemsa staining to assess sperm membrane integrity in salernitano stallions and its relationship to pregnancy rates.
Aim of this study was to test the reliability of Trypan blue/Giemsa staining to evaluate sperm membrane integrity, acrosomal intactness and morphology in stallion to verify whether it could be applied in vitro as useful tool for sperm fertilizing ability. Fertility data on inseminated mares were collected to evaluate the relationship of sperm quality to pregnancy rates. Forty-one ejaculates were collected from 3 stallions of Salernitano Horse Breed and evaluated for gross appearance, volume, visual motility and membrane integrity with Trypan blue/Giemsa staining and thirty-five mares were inseminated during the breeding season from April to July. Differences among stallions were found in volume, sperm concentration (p < 0.05) and visual motility (p < 0.01). A decrease in sperm motility, concentration (p < 0.05) and total sperm number was found in June-July (p < 0.01). Live sperm with intact acrosome (LSIA) and proximal droplets (PD) were lower (p < 0.01) in June-July, while acrosome reacted sperm (ARS) percentage increased (p < 0.05). No fertility differences were found among stallions with an average fertility per cycle of 44.6% and a pregnancy rate of 68.6%. Higher percentages of LSIA were found in the ejaculates used to inseminate mares that became pregnant vs those used in mares not pregnant (p < 0.05). The significance of LSIA as test variable to verify the reliability of Trypan blue/Giemsa staining was confirmed by Receiver operating characteristic ROC analysis and the sensitivity of the test was 85% at a cut-off value of 48% LSIA. Trypan blue-Giemsa showed to be an accurate method that can be applied on field to evaluate sperm membrane integrity and to identify poor-quality ejaculates.