The problem of prozone in serum antibody titration and its mathematical interpretation.
The question of the mechanism of "prozone" creation was considered from the point of view of the concentrations of free and semi-blocked bivalent antibodies in the mixture of these antibodies with monovalent antigen. Using the so called "coordinates of dilution", suggested by the author earlier, it was possible to calculate the relationships between the concentrations of either free or semi-blocked bivalent antibodies and the dilution of the antigen-antibody mixture. It was shown that dilution of antigen-antibody mixture leads to an increase of the concentration of free bivalent antibodies and simultaneous sharp decrease of the concentration of semi-blocked antibodies. It is suggested, that such a relationship is quite enough for the creation of prozone effect in reactions, when only bivalent antibodies are active and semi-blocked antibodies compete with free antibodies, providing inhibition of the reaction.


Forecasting asylum-related migration flows with machine learning and data at scale.
The sudden and unexpected migration flows that reached Europe during the so-called 'refugee crisis' of 2015-2016 left governments unprepared, exposing significant shortcomings in the field of migration forecasting. Forecasting asylum-related migration is indeed problematic. Migration is a complex system, drivers are composite, measurement incorporates uncertainty, and most migration theories are either under-specified or hardly actionable. As a result, approaches to forecasting generally focus on specific migration flows, and the results are often inconsistent and difficult to generalise. Here we present an adaptive machine learning algorithm that integrates administrative statistics and non-traditional data sources at scale to effectively forecast asylum-related migration flows. We focus on asylum applications lodged in countries of the European Union (EU) by nationals of all countries of origin worldwide, but the same approach can be applied in any context provided adequate migration or asylum data are available. Uniquely, our approach (a) monitors drivers in countries of origin and destination to detect early onset change; (b) models individual country-to-country migration flows separately and on moving time windows; (c) estimates the effects of individual drivers, including lagged effects; (d) delivers forecasts of asylum applications up to four weeks ahead; (e) assesses how patterns of drivers shift over time to describe the functioning and change of migration systems. Our approach draws on migration theory and modelling, international protection, and data science to deliver what is, to our knowledge, the first comprehensive system for forecasting asylum applications based on adaptive models and data at scale. Importantly, this approach can be extended to forecast other social processes.


Potential Cost Savings in Medicare Part D Across Gastroenterology; An Assessment of Alternative Drug Sourcing.
Until recently the Centers of Medicare and Medicaid were restricted from negotiating drug cost. We assessed the potential impact of alternative drug sourcing models on Medicare Part D spending. 27 drugs were extracted from 2021 Medicare Part D claims. Drug-specific/total spending was compared against cost at Mark Cuban Cost Plus Drugs Company, Costco Member Prescription Program, and Veterans Health Administration price point. Potential Part D savings were $798.99 million, $573.84 million, and $1.02 billion (Mark Cuban Cost Plus Drugs Company, Costco Member Prescription Program, and Veterans Health Administration, respectively). Disproportionate Part D spending likely reflects less competitive acquisition cost. Provider awareness of medications with advantaged price may promote targeted prescribing with potentially tremendous healthcare savings.


Experience-dependent plasticity of excitatory and inhibitory intertectal inputs in Xenopus tadpoles.
Communication between optic tecta/superior colliculi is thought to be required for sensorimotor behaviors by comparing inputs across the midline, however the development of and the role of visual experience in the function and plasticity of intertectal connections are unclear. We combined neuronal tracing, in vivo time-lapse imaging, and electrophysiology to characterize the structural and functional development of intertectal axons and synapses in Xenopus tadpole optic tectum. We find that intertectal connections are established early during optic tectal circuit development. We determined the neurotransmitter identity of intertectal neurons using both rabies virus-mediated tracing combined with post-hoc immunohistochemistry, and electrophysiology. Excitatory and inhibitory intertectal neuronal somata are similarly distributed throughout the tectum. Excitatory and inhibitory intertectal axons are structurally similar and elaborate broadly in the contralateral tectum. We demonstrate that intertectal and retinotectal axons converge onto tectal neurons by recording postsynaptic currents after stimulating intertectal and retinotectal inputs. Cutting the intertectal commissure removes synaptic responses to contralateral tectal stimulation. In vivo time-lapse imaging demonstrated that visual experience drives plasticity in intertectal bouton size and dynamics. Finally, visual experience coordinately drives the maturation of excitatory and inhibitory intertectal inputs by increasing AMPA- and GABA-receptor mediated currents, comparable to experience-dependent maturation of retinotectal inputs. These data indicate that visual experience regulates plasticity of excitatory and inhibitory intertectal inputs, maintaining the excitatory: inhibitory ratio of intertectal input. These studies place intertectal inputs as key players in tectal circuit development and suggest that they may play a role in sensory information processing critical to sensorimotor behaviors.


Mammographic Density Change With Estrogen and Progestin Therapy and Breast Cancer Risk.
Estrogen plus progestin therapy increases both mammographic density and breast cancer incidence. Whether mammographic density change associated with estrogen plus progestin initiation predicts breast cancer risk is unknown. We conducted an ancillary nested case-control study within the Women's Health Initiative trial that randomly assigned postmenopausal women to daily conjugated equine estrogen 0.625 mg plus medroxyprogesterone acetate 2.5 mg or placebo. Mammographic density was assessed from mammograms taken prior to and one year after random assignment for 174 women who later developed breast cancer (cases) and 733 healthy women (controls). Logistic regression analyses included adjustment for confounders and baseline mammographic density when appropriate. Among women in the estrogen plus progestin arm (97 cases/378 controls), each 1% positive change in percent mammographic density increased breast cancer risk 3% (odds ratio [OR] = 1.03, 95% confidence interval [CI] = 1.01 to 1.06). For women in the highest quintile of mammographic density change (>19.3% increase), breast cancer risk increased 3.6-fold (95% CI = 1.52 to 8.56). The effect of estrogen plus progestin use on breast cancer risk (OR = 1.28, 95% CI = 0.90 to 1.82) was eliminated in this study, after adjusting for change in mammographic density (OR = 1.00, 95% CI = 0.66 to 1.51). We found the one-year change in mammographic density after estrogen plus progestin initiation predicted subsequent increase in breast cancer risk. All of the increased risk from estrogen plus progestin use was mediated through mammographic density change. Doctors should evaluate changes in mammographic density with women who initiate estrogen plus progestin therapy and discuss the breast cancer risk implications.


Design Considerations for the Development and Implementation of a Medical Respite for Older Adults Experiencing Homelessness in Metro Vancouver.
Older persons with lived or living experience of homelessness (PWLEs) often live with complex physical and/or mental health conditions which are challenged by poor access to health services, especially primary care. To fill the gap in the continuum of care following hospital discharge for PWLEs, medical respite provides health and shelter support for PWLEs who do not have acute care needs that qualify for a stay in a hospital bed, yet are too sick or frail to recover on the streets or in a traditional shelter. This study examines how a medical respite could be designed for older PWLEs in Metro Vancouver, BC. Using a community-based participatory research approach, in-depth interviews and focus groups were conducted with PWLE (n=15) and service provider (n=11) participants. Participants offered rich suggestions about (a) the culture of the medical respite, (b) the physical design of a medical respite, (c) individuals who should be involved in medical respite delivery, (d) services a medical respite should provide, and (e) who the medical respite should serve. When designing a medical respite for older PWLEs, considerations include providing an environment where patients can rest, but also feel safe and be surrounded by persons who they trust and who care for them. Developing a medical respite that adheres to the tenets of trauma-informed and patient-centered care acknowledges the mistrust and traumatization that often accompanies homeless patients presenting to health care.


The occurrence of Toxoplasma gondii infection in people and animals from rural environment of Lublin region - estimate of potential role of water as a source of infection.
A total of 254 humans and 489 domestic animals living on farms in the Lublin province (eastern Poland) were examined for the presence of antibodies against Toxoplasma gondii using, respectively, the ELFA and direct agglutination tests. In parallel, 182 samples of potable water, mostly from shallow household wells, were taken on farms and examined for the presence of T. gondii by microscopy and PCR. The frequency of seropositive reactions in farm inhabitants (66.9 percent) was significantly greater (p < 0.01) compared to the reference group of 39 healthy urban dwellers (41.0 percent). A highly significant positive correlation was found between the age of examined farm inhabitants and the rate of positive reactions with Toxoplasma antigen (p < 0.0001). Among domestic animals, the greatest frequency of seropositive reactions to T. gondii occurred in cats (75.0 percent) and dogs (53.6 percent), less frequent in cattle (33.8 percent) and hens (33.5 percent) and the least frequent in pigs (17.9 percent) and ducks (21.2 percent). The presence of T. gondii was found in potable water samples taken from water intakes on farms: in 12.6 percent of samples by microscopy, and in 22.5 percent of samples by PCR. Among 19 water samples taken from bathing places on the territory of the Lublin province, 2 samples positive for T. gondii (10.5 percent) were found by microscopic examination and confirmed by PCR . The presence of live parasites in water samples was demonstrated by the isolation of Toxoplasma gondii strains in mice. By use of RFLP -PCR it was found that the majority of isolated Toxoplasma strains (78.0 percent) belonged to clonal type I which is most virulent for humans and animals. Although no statistically significant relationship between the presence of T. gondii in water and occurrence of seropositive reactions in farm inhabitants and/or domestic animals could be found, the above-mentioned data suggest a potential role of potable water in the spread of toxoplasmosis in the rural environment.


Covalent modification of hemoglobin by carbon disulfide: III. A potential biomarker of effect.
Although the neurotoxicity of CS2 has been recognized for over a century, presently there is no accepted biomarker of effect for CS2 exposure. Previous investigations have supported covalent cross-linking of erythrocyte spectrin as a potential preneurotoxic marker reflective of the biochemical changes occurring within the axon. In the present investigation, the potential of using CS2 promoted modification of hemoglobin as a dosimeter for quantifying exposure to CS2 was evaluated. Liquid chromatography was used to isolate and measure alpha and beta chains of globin in blood obtained from rats exposed to CS2 by inhalation as a function of exposure level and duration. The degree of globin modification was compared to light microscopic and ultrastructural changes in the central and peripheral nervous systems to determine the temporal relationship of globin modification to the structural changes in the axon. Samples obtained from rats exposed to CS2 contained a globin chain not present in control samples. Analysis of the peak corresponding to the new chain using electrospray mass spectrometry was consistent with the generation of a single dithiocarbamate ester or thiourea intramolecular cross-link in the alpha 1 major chain. This altered globin chain was detectable both at the subneurotoxic level of exposure and prior to axonal structural changes at the neurotoxic levels of exposure used. The extent of modification was positively correlated to the exposure level and duration for all conditions examined. These findings support hemoglobin as a potential preneurotoxic biomarker of effect for CS2 possessing several practical advantages relative to the use of CS2-mediated spectrin cross-linking.


[Mutagenic substances in amino acid and protein pyrolysates and in heat-treated food].
Information on the recent knowledge of occurrence and formation of heterocyclic substances as results of the pyrolysis of amino acids and protein as well as of heat-treatment of food is given. Positive results obtained by means of the Ames-test point to the possibility that the tested substances might also have mutagenic or carcinogenic effect on man. Up to now a corroboration of possible delayed lesions could not be definitely proved in animal experiments. It may be taken for granted that the preparation and especially the heat treatment of foods could induce the formation of products being injurious to health. Chemical substances being known in this respect are specified with reference to their occurrence.


Hierarchical nanostructures derived from cellulose for lithium-ion batteries.
Introducing the sophisticated morphologies and structures of natural substances into artificial materials provides a promising strategy for the fabrication of functional materials with tailored structures and functionalities. Herein, our recent advances achieved in the fabrication of hierarchically structured functional materials derived from natural cellulose substances (e.g., ordinary laboratory filter paper) for lithium-ion batteries are reviewed. The surface sol-gel process was employed to fabricate metal oxides (Sn, Ti, Fe, etc.), silica and silicon based nanocomposites by using the corresponding metal alkoxides as the precursors, which faithfully inherited the unique porous network structures of the initial cellulose substances. However, the nanomaterials synthesized as such are limited to the above-mentioned species due to the limitation of specific metal alkoxide precursors used. To break this limitation, specific polyoxometalate clusters were employed as the building blocks to be assembled onto the surfaces of cellulose nanofibres and eventually to obtain the corresponding molybdenum based nanomaterials with three-dimensional porous network structures and specific properties. These cellulose derived nanocomposites exhibit improved lithium storage capacities and cycling stabilities when applied as anodic materials for lithium-ion batteries, resulting from the unique hierarchically porous network structures inherited from the initial cellulose substance and the strong synergistic interactions among the components contained therein.


Evidence for increased somatic cell mutations at the glycophorin A locus in atomic bomb survivors.
A recently developed assay for somatic cell mutations was used to study survivors of the atomic bomb at Hiroshima. This assay measures the frequency of variant erythrocytes produced by erythroid precursor cells with mutations that result in a loss of gene expression at the polymorphic glycophorin A (GPA) locus. Significant linear relations between variant frequency (VF) and radiation exposure were observed for three different variant cell phenotypes. The spontaneous and induced VFs agree with previous measurements of radiation-induced mutagenesis in other systems; this evidence supports a mutational origin for variant cells characterized by a loss of GPA expression and suggests that the GPA assay system may provide a cumulative dosimeter of past radiation exposures. VFs for some survivors differ dramatically from the calculated dose response, and these deviations appear to result primarily from statistical fluctuations in the number of mutations in the stem-cell pool. These fluctuations allow one to estimate the number of long-lived hemopoietic stem cells in humans.


Real world data on the demographic and clinicopathological profile and management of patients with early-stage HER2-positive breast cancer and residual disease treated with adjuvant trastuzumab emtansine (KARMA study).
Trastuzumab emtansine (T-DM1) significantly improves invasive disease-free survival and reduces the risk of recurrence in patients with HER2-positive early breast cancer (EBC) with residual disease (RD). The KARMA study aimed to describe the characteristics and management of these patients in clinical practice in Spain. We conducted a multicentre retrospective study in patients with HER2-positive EBC with RD following neoadjuvant treatment (NeoT) and who had received ≥1 dose of T-DM1 as adjuvant treatment. The primary endpoint was the evaluation of sociodemographic and clinicopathological characteristics of these patients. A total of 114 patients were included (March-July 2020). At diagnosis, most tumours were infiltrating ductal carcinoma (IDC) (93.9 %), grade 2 (56.1 %), and hormone receptor (HR)-positive (79.8 %). Over 75 % of patients had disease in operable clinical stages (T1-3 N0-1). In the neoadjuvant setting, 86.8 % of patients received trastuzumab plus pertuzumab, and 23.6 % achieved radiological complete response. Breast-conserving surgery was performed in 55.8 % of patients. Surgical specimens showed that 89.5 % of patients had IDC, 49.1 % grade 2, 84.1 % HR-positive, and 8.3 % HER2-negative disease. Most patients had RD classified as RCB-II and Miller/Payne grade 3/4. Grade 3 treatment-related adverse events (trAEs) occurred in 5.3 % of patients. No grade 4/5 AEs occurred. Over 95 % of patients were free of invasive-disease during T-DM1 adjuvant treatment. The KARMA study describes the characteristics of patients with HER2-positive EBC with RD after NeoT and the real-life management of a T-DM1 adjuvant regimen, which showed a manageable safety profile in line with the KATHERINE trial data.


MS-Based Allotype-Specific Analysis of Polyclonal IgG-Fc N-Glycosylation.
Current approaches to study glycosylation of polyclonal human immunoglobulins G (IgG) usually imply protein digestion or glycan release. While these approaches allow in-depth characterization, they also result in a loss of valuable information regarding certain subclasses, allotypes and co-occuring post-translational modifications (PTMs). Unfortunately, the high variability of polyclonal IgGs makes their intact mass spectrometry (MS) analysis extremely challenging. We propose here a middle-up strategy for the analysis of the intact fragment crystallizable (Fc) region of human plasma IgGs, with the aim of acquiring integrated information of the N-glycosylation and other PTMs of subclasses and allotypes. Human plasma IgG was isolated using Fc-specific beads followed by an on-bead C H 2 domain digestion with the enzyme IdeS. The obtained mixture of Fc subunits was analyzed by capillary electrophoresis (CE) and hydrophilic interaction liquid chromatography (HILIC) hyphenated with MS. CE-MS provided separation of different IgG-subclasses and allotypes, while HILIC-MS allowed resolution of the different glycoforms and their oxidized variants. The orthogonality of these techniques was key to reliably assign Fc allotypes. Five individual donors were analyzed using this approach. Heterozygosis was observed in all the analyzed donors resulting in a total of 12 allotypes identified. The assignments were further confirmed using recombinant monoclonal IgG allotypes as standards. While the glycosylation patterns were similar within allotypes of the same subclass, clear differences were observed between IgG subclasses and donors, highlighting the relevance of the proposed approach. In a single analysis, glycosylation levels specific for each allotype, relative abundances of subclasses and information on co-occurring modifications are obtained. This middle-up method represents an important step toward a comprehensive analysis of immunoglobulin G-Fc variants.


Expression of proopiomelanocortin-related peptides in human follicular fluid.
In order to evaluate the expression of the opioid precursor proopiomelanocortin (POMC) in the ovarian follicle, we measured 6 of its main end-products in 23 follicular fluids. We coupled high performance liquid chromatography (HPLC) to specific radioimmunoassays. Seven follicles were immature (diameter less than 9 mm), 10 were obtained from superovulated patients during an in vitro fertilization-embryo transfer program (greater than 22 mm) and six were persistent follicles, collected during the luteal phase [15-31 mm, luteinized unruptured follicles (LUF)]. Follicular fluids were extracted by mean of Sep-pak cartridges and then purified by HPLC with a reverse-phase C-18 column eluted in a linear gradient with acetonitrile/0.01 M hydrochloric acid (from 18:82 to 40:60). Fractions were tested with specific antisera for ACTH (1-39), alpha-MSH, beta-lipotropin (beta-LPH), beta-endorphin (beta-EP) and gamma-endorphin (gamma-EP) immunoreactivities. No presence of beta-LPH, beta-EP and ACTH was confirmed, while gamma-EP, alpha-MSH and des-alpha-MSH were detected for the first time in follicular fluid. In every class of follicles shorter chain peptides predominate over their longer chain precursor. Immature follicles are characterized by the highest amounts of gamma-EP, ACTH, alpha-MSH and des-alpha-MSH if compared to superovulated and LUF. On the contrary, beta-EP amount was highest after superovulation. Apart from this finding, peptide levels in superovulated patients and LUF are similar.(ABSTRACT TRUNCATED AT 250 WORDS)


The relationship between school lunch attendance and the food intakes of French schoolchildren aged 3-17 years.
Recently, school meal composition regulations have been implemented in France in order to improve the nutritional status of children. The present study investigated the link between school lunch attendance and the food intakes of schoolchildren aged 3-17 years. Second French cross-sectional dietary survey (2006-2007). Eating frequencies were assessed for twenty-four food groups with a 7 d food record. Eating locations were recorded for main meals. Food group intakes at weekday lunches were compared for the school canteen and for other locations. The children's overall dietary intake was compared based on school lunch attendance. Mainland France. Schoolchildren aged 3-17 years (n 1068). Lunchtime food intake differed between the school canteen and other locations. Some intakes at school canteens were more in accordance with the regulations (more fruit and vegetables, fish and dairy products, and less sandwiches, soft drinks, chocolate and confectionery), whereas others highlighted needs for improvement (more sweet biscuits and pastries, ice cream and dairy desserts, pizzas and salty pastries). Many of these differences were also observed in the children's overall diet: children regularly attending school lunches ate more mashed fruit, fish and sweet biscuits or pastries, and less sandwiches and soft drinks. The link between school lunch attendance and overall diet was less pronounced in secondary-school children. School canteen attendance is associated with both potentially beneficial and deleterious differences in the lunchtime and overall diets of French children. These findings are important to consider when setting national regulations for school meal composition.


[Emergency medicine and anesthesiology aspects of neurofibromatosis in childhood].
In infancy mediastinal and major airway associated tumours can cause respiratory emergency situations requiring resuscitation and special anaesthetic management. The case report of an infant with a cervicothoracal neurofibroma and resulting acute respiratory distress underlines these problems. In anaesthetic and surgical procedures these patients can experience respiratory difficulties with severe consequences. Preoperative possibilities to evaluate and reduce these risks are presented and discussed.


Nuclear HIF1A expression is strongly prognostic in sporadic but not familial male breast cancer.
Male breast cancer is poorly understood with a large proportion arising in the familial context particularly with the BRCA2 germline mutation. As phenotypic and genotypic differences between sporadic and familial male breast cancers have been noted, we investigated the importance of a hypoxic drive in these cancers as this pathway has been shown to be of importance in familial female breast cancer. Expression of two major hypoxia-induced proteins, the hypoxia-inducible factor-1α (HIF1A) and the carbonic anhydrase IX (CA9), examined within a large cohort including 61 familial (3 BRCA1, 28 BRCA2, 30 BRCAX) and 225 sporadic male breast cancers showed that 31% of all male breast cancers expressed either HIF1A (25%) and/or CA9 (8%) in the combined cohort. Expression of HIF1A correlated with an increased incidence of a second-major malignancy (P=0.04), histological tumor type (P=0.005) and basal phenotype (P=0.02). Expression of CA9 correlated with age (P=0.004) in sporadic cases and an increased tumor size (P=0.003). Expression of HIF1A was prognostic for disease-specific survival in sporadic male breast cancers (HR: 3.8, 95% CI: 1.5-9.8, P=0.006) but not within familial male breast cancer, whereas CA9 was only prognostic in familial male breast cancers (HR: 358.0, 95% CI: 9.3-13781.7, P=0.002) and not in sporadic male breast cancer. This study found that hypoxic drive is less prevalent in male breast cancer compared with female breast cancer, possibly due to a different breast microenvironment. The prognostic impact of HIF1A is greatest in sporadic male breast cancers with an alternate dominant mechanism for the oncogenic drivers suggested in high risk familial male breast cancers.


Comparing different non-invasive brain stimulation interventions for bipolar depression treatment: A network meta-analysis of randomized controlled trials.
Non-invasive brain stimulation (NIBS) is a promising treatment for bipolar depression. We systematically searched for randomized controlled trials on NIBS for treating bipolar depression (INPLASY No: 202340019). Eighteen articles (N = 617) were eligible for network meta-analysis. Effect sizes were reported as standardized mean differences (SMDs) or odds ratios (ORs) with 95% confidence intervals (CIs). Anodal transcranial direct current stimulation over F3 plus cathodal transcranial direct current stimulation over F4 (a-tDCS-F3 +c-tDCS-F4; SMD = -1.18, 95%CIs = -1.66 to -0.69, N = 77), high-definition tDCS over F3 (HD-tDCS-F3; -1.17, -2.00 to -0.35, 25), high frequency deep transcranial magnetic stimulation (HF-dTMS; -0.81, -1.62 to -0.001, 25), and high frequency repetitive TMS over F3 plus low frequency repetitive TMS over F4 (HF-rTMS-F3 +LF-rTMS-F4; -0.77, -1.43 to -0.11, 38) significantly improved depressive symptoms compared to sham controls. Only a-tDCS-F3 +c-tDCS-F4 (OR = 4.53, 95%CIs = 1.51-13.65) and HF-rTMS-F3 +LF-rTMS-F4 (4.69, 1.02-21.56) showed higher response rates. No active NIBS interventions exhibited significant differences in dropout or side effect rates, compared with sham controls.


Hepatitis e vaccine to prevent morbidity and mortality during epidemics.
Recurrent, large, waterborne epidemics of hepatitis E virus (HEV) occur regularly after monsoon rains contaminate water supplies in Asia or during humanitarian crises in Africa. These epidemics commonly affect thousands of persons, and it has a high mortality in pregnant women who become infected. Although a subunit HEV vaccine has been developed by Chinese investigators and was found to be highly effective and safe in a large clinical trial, this vaccine is only available in China. Until it is prequalified by the World Health Organization, the vaccine may not be available for use outside of China in low-income countries that lack national vaccine regulatory agencies. In this manuscript, we explore possible strategies for providing access to this potentially important vaccine for international use in responding to epidemics of HEV in low-resource countries.


Reduced dopamine function within the medial shell of the nucleus accumbens enhances latent inhibition.
Latent inhibition (LI) manifests as poorer conditioning to a CS that has previously been presented without consequence. There is some evidence that LI can be potentiated by reduced mesoaccumbal dopamine (DA) function but the locus within the nucleus accumbens of this effect is as yet not firmly established. Experiment 1 tested whether 6-hydroxydopamine (6-OHDA)-induced lesions of DA terminals within the core and medial shell subregions of the nucleus accumbens (NAc) would enhance LI under conditions that normally disrupt LI in controls (weak pre-exposure). LI was measured in a thirst motivated conditioned emotional response procedure with 10 pre-exposures (to a noise CS) and 2 conditioning trials. The vehicle-injected and core-lesioned animals did not show LI and conditioned to the pre-exposed CS at comparable levels to the non-pre-exposed controls. 6-OHDA lesions to the medial shell, however, produced potentiation of LI, demonstrated across two extinction tests. In a subsequent experiment, haloperidol microinjected into the medial shell prior to conditioning similarly enhanced LI. These results underscore the dissociable roles of core and shell subregions of the NAc in mediating the expression of LI and indicate that reduced DA function within the medial shell leads to enhanced LI.


Mechanism of recovery from acute virus infection. IV. Questionable role of mononuclear phagocytes in the clearance of lymphocytic choriomeningitis virus from spleens of mice.
After intravenous infection of mice with 10(3) infectious units (IU) the WE strain lymphocytic choriomeningitis (LCM) virus multiplied in the spleens (as in all other major organs), reaching more than 10(8) IU/g of tissue on days 4 to 5. Subsequently, the virus was quickly eliminated, being below detectability usually by day 10. During the time of virus clearance, the mononuclear phagocytes (MNP) of the spleen were activated as revealed by suppression of growth of Listeria monocytogenes and increase of cell-associated hydrolytic enzymes. In athymic nude mice, in whom the MNP system is assumed to be permanently activated, the virus replicated slightly but reproducibly less than in their euthymic counterparts. However, when the MNP were activated by Corynebacterium parvum, virus in spleens attained higher concentrations than in mice not so treated, and the rate of elimination was not altered. In mice whose MNP had been damaged by injection of dextran sulfate 500, the spleen virus titers were also increased, but the subsequent immune elimination was slightly delayed. Activation of spleen MNP was not evident at the time virus was rapidly cleared as a result of transfusion of LCM-immune T lymphocytes. Adoptive immunization was as successful in mice that had been pretreated with gamma-rays or cyclophosphamide, suggesting that replicating cells or their descendants, in particular monocytes, did not participate measurably in the process of elimination. Pretreatments of recipients with dextran sulfate 500 reduced the efficacy of transfused LCM-immune T lymphocytes, but this compound probably directly affected the cells. We interpret these findings to mean that the LCM virus in the mouse's spleen is controlled by a mechanism in which MNP do not play an essential role.


Development and validation of a multiplex UHPLC-MS/MS method for the determination of the investigational antibiotic against multi-resistant tuberculosis macozinone (PBTZ169) and five active metabolites in human plasma.
The emergence of Mycobacterium tuberculosis strains resistant to current first-line antibiotic regimens constitutes a major global health threat. New treatments against multidrug-resistant tuberculosis (MDR-TB) are thus eagerly needed in particular in countries with a high MDR-TB prevalence. In this context, macozinone (PBTZ169), a promising drug candidate with an unique mode of action and highly potent in vitro tuberculocidal properties against MDR Mycobacterium strains, has now reached the clinical phase and has been notably tested in healthy male volunteers in Switzerland. To that endeavor, a multiplex UHPLC-MS/MS method has been developed for the sensitive and accurate human plasma levels determination of PBTZ169 along with five metabolites retaining in vitro anti-TB activity. Plasma protein precipitation with methanol was carried out as a simplified sample clean-up procedure followed by direct injection of the undiluted supernatant for the bioanalysis of the six analytes within 5 min, using 1.8 μm reversed-phase chromatography coupled to triple quadrupole mass spectrometry employing electrospray ionization in the positive mode. Stable isotopically-labelled PBTZ169 was used as internal standard (ISTD), while metabolites could be reliably quantified using two unlabeled chemical analogues selected as ISTD from a large in-house analogous compounds library. The overall methodology was fully validated according to current recommendations (FDA, EMEA) for bioanalytical methods, which include selectivity, carryover, qualitative and quantitative matrix effect, extraction recovery, process efficiency, trueness, precision, accuracy profiles, method and instrument detection limits, integrity to dilution, anticoagulant comparison and short- and long-term stabilities. Stability studies on the reduced metabolite H2-PBTZ169 have shown no significant impact on the actual PBTZ169 concentrations determined with the proposed assay. This simplified, rapid, sensitive and robust methodology has been applied to the bioanalysis of human plasma samples collected within the frame of a phase I clinical study in healthy volunteers receiving PBTZ169.


Illegal alcohol sales and use of alcohol control policies at community festivals.
The primary goals of this study were to assess the propensity for alcohol sales to underage customers and obviously intoxicated customers at community festivals, and to assess the prevalence of alcohol control policies at these events. A secondary goal was to identify server and festival characteristics and festival policies related to the likelihood of illegal alcohol sales. We conducted pseudo-underage purchase attempts at 43 festivals and pseudo-intoxicated purchase attempts at 50 festivals to assess the likelihood of illegal sales. Research staff made observations at festivals and contacted festival planners by telephone following each event to assess which alcohol policies were implemented. We conducted backwards stepwise multivariate analyses for each purchase attempt outcome to identify policies and characteristics related to likelihood of illegal alcohol sales. Pseudo-intoxicated buyers purchased beer in 89% of 95 attempts (standard deviation [SD]=0.31) and pseudo-underage buyers were able to purchase beer in 50% of 82 attempts (SD=0.50). All festival planners reported having at least two of the 10 alcohol policies we assessed, but no festival had implemented all 10 policies. Server characteristics were not related to either purchase attempt outcome. In the multivariate analyses, having more alcohol control policies was related to a greater likelihood of illegal sales to intoxicated customers; however, having more alcohol control policies was associated with a lesser likelihood of alcohol sales to underage customers. Restricting the number of servings per person was also associated with a lesser likelihood of alcohol sales to underage customers. Propensity for illegal alcohol sales at festivals is very high. Research is needed to identify interventions to prevent illegal alcohol sales at these events.


Replacement of the V3 region of gp120 with SDF-1 preserves the infectivity of T-cell line-tropic human immunodeficiency virus type 1.
Interaction between the human immunodeficiency virus type 1 (HIV-1) envelope and the relevant chemokine receptors is crucial for subsequent membrane fusion and viral entry. Although the V3 region of gp120 is known to determine the cell tropism as well as the coreceptor usage, the significance of the binding of the V3 region to the chemokine receptor has not been fully understood. To address this issue, we adopted the pseudotyped virus infection assay in which the V3 region of the T-cell line-tropic (T-tropic) NL4-3 envelope was replaced with a portion of stromal cell-derived factor 1 (SDF-1), the ligand of CXCR4. The V3 region of the NL4-3 envelope expression vector was replaced with three different stretches of SDF-1 cDNA. Expression of each chimeric envelope protein was confirmed by immunoprecipitation and Western blotting. Luciferase reporter viruses were prepared by cotransfection of the pNL4-3.Luc.E(-)R(-) vector and each chimeric envelope expression vector, and the infection assay was then carried out. We showed that pseudotyped viruses with one of the chimeric envelopes, NL4-3/SDF1-51, could infect U87.CD4.CXCR4 but not U87.CD4 or U87.CXCR4 cells and that this infection was inhibited by the ligand of CXCR4, SDF-1beta, by anti-human SDF-1 antibody, or by an anti-CD4 antibody, Leu3a, in a dose-dependent manner. Furthermore, chimeric NL4-3/SDF1-51 gp120 significantly inhibited binding of labeled SDF-1 to CXCR4. It was suggested that replacement of the V3 region of the NL4-3 envelope with SDF-1 preserved the CD4-dependent infectivity of T-tropic HIV-1. These results indicate that binding between the V3 region and the relevant coreceptor is important for viral entry, whether its amino acid sequence is indigenous to the virus or not.


Is appropriate management of atrial fibrillation in primary care a utopia?
The authors aimed at reporting on whether or not primary care doctors follow atrial fibrillation (AF) treatment protocols, and on the mental distress of such patients. A total of 138 patients with first detected or recurrent AF were examined in a health center. Demographic data were collected and their lifestyle and medical history for rhythm-related pathologies and chronic medication were investigated. Physical examination, electrocardiogram (EKG), and in selected cases, lab analysis were carried-out. CHADS2 index was used for assessing the stroke risk in patients with AF, while the General Health Questionnaire-12 (GHQ-12) for personal health perception was performed in all patients. According to CHADS2 the majority of the patients had at least 1 risk factor and half of those receiving oral vitamin K antagonists presented an out-of-range international normalized ratio (INR). In 24 cases, patients used both aspirin and oral anticoagulants, while in 41 cases, medication was corrected according to index. GHQ-12 seemed to be significantly worse in paroxysmal and persistent cases, as well as in women with recurrent AF. Many paroxysmal AF patients under 75 years continued caffeine intake, whereas an extensive use of benzodiazepines was noticed in the majority of patients. Shortages and limitations of the peripheral or rural units and health centers and inadequate knowledge and application of the guidelines, seemed to be major factors responsible for mismanaging AF patients. More education in prehospital cardiology may contribute in improving management of arrhythmias in primary care.


Ritonavir-boosted lopinavir plus nucleoside or nucleotide reverse transcriptase inhibitors versus ritonavir-boosted lopinavir plus raltegravir for treatment of HIV-1 infection in adults with virological failure of a standard first-line ART regimen (SECOND-LINE): a randomised, open-label, non-inferiority study.
Uncertainty exists about the best treatment for people with HIV-1 who have virological failure with first-line combination antiretroviral therapy of a non-nucleoside analogue (NNRTI) plus two nucleoside or nucleotide analogue reverse transcriptase inhibitors (NtRTI). We compared a second-line regimen combining two new classes of drug with a WHO-recommended regimen. We did this 96-week, phase 3b/4, randomised, open-label non-inferiority trial at 37 sites worldwide. Adults with HIV-1 who had confirmed virological failure (plasma viral load >500 copies per mL) after 24 weeks or more of first-line treatment were randomly assigned (1:1) to receive ritonavir-boosted lopinavir plus two or three NtRTIs (control group) or ritonavir-boosted lopinavir plus raltegravir (raltegravir group). The randomisation sequence was computer generated with block randomisation (block size four). Neither participants nor investigators were masked to allocation. The primary endpoint was the proportion of participants with plasma viral load less than 200 copies per mL at 48 weeks in the modified intention-to-treat population, with a non-inferiority margin of 12%. This study is registered with ClinicalTrials.gov, number NCT00931463. We enrolled 558 patients, of whom 541 (271 in the control group, 270 in the raltegravir group) were included in the primary analysis. At 48 weeks, 219 (81%) patients in the control group compared with 223 (83%) in the raltegravir group met the primary endpoint (difference 1·8%, 95% CI -4·7 to 8·3), fulfilling the criterion for non-inferiority. 993 adverse events occurred in 271 participants in the control group versus 895 in 270 participants in the raltegravir group, the most common being gastrointestinal. The raltegravir regimen was no less efficacious than the standard of care and was safe and well tolerated. This simple NtRTI-free treatment strategy might extend the successful public health approach to management of HIV by providing simple, easy to administer, effective, safe, and tolerable second-line combination antiretroviral therapy. University of New South Wales, Merck, AbbVie, the Foundation for AIDS Research.


[The effect of alpha- and beta-ecdysone on differentiation of imaginal disks of Drosophila melanogaster cultivated in vitro].
The effect of alpha- and beta-ecdysone was studied upon differentiation of eye and leg imaginal discs of Drosophila cultivated together with neural ganglia in the medium C-39. In the medium without hormone, eye and leg imaginal discs from larvae of the early 3rd instar are not capable of differentiation. In these conditions, eye imaginal discs from larvae of the late 3rd instar become pigmented in 5 weeks and leg imaginal discs form sometimes segmented limbs on the 4th week of cultivation. In the medium with alpha- and beta-ecdysone, eye and leg imaginal discs from larvae of the early 3rd instar reveal only initial stages of differentiation. The effect of beta-ecdysone concentration was studied upon differentiation of leg and eye imaginal discs from larvae of the late 3rd instar. On decrease of beta-ecdysone concentration from 0.05 down to 0.005 mug/ml, initial stages of differentiation of leg imaginal discs proceed at a slower rate but, then, more complete differentiation with the formation of chitin and neuromuscular tissue is observed. In eye imaginal discs alpha-ecdysone at a concentration of 0.5 mug/ml and beta-ecdysone at a concentration of 0.005-0.5 mug/ml induce the pigment accumulation already at the 1st week of cultivation and beta-ecdysone at a concentration of 5 mug/ml fully inhibits the formation and accumulation of pigment.


Effects of Dihydroartemisinin against Cystic Echinococcosis In Vitro and In Vivo.
Cystic echinococcosis (CE) is a disease caused by the infection of Echinococcus granulosus. We sought to investigate the effects of dihydroartemisinin (DHA) against CE under in vitro and in vivo conditions. Protoscoleces (PSCs) from E. granulosus were divided into control, DMSO, ABZ, DHA-L, DHA-M, and DHA-H groups. PSC viability after DHA treatment was determined based on the eosin dye exclusion test, alkaline phosphatase content detection, and ultrastructure observation. DNA oxidative damage inducer hydrogen peroxide (H2O2), reactive oxygen species (ROS) scavenger mannitol, and the DNA damage repair inhibitor velparib were used to explore the anti-CE mechanism of DHA. The anti-CE effects and CE-induced liver injury and oxidative stress of DHA at different doses (50, 100, and 200 mg/kg) were assessed in CE mice. DHA showed antiparasitic effects on CE in both in vivo and in vitro experiments. DHA could elevate the ROS level and induce oxidative DNA damage in PSCs, thereby destroying hydatid cysts. DHA could inhibit the growth of cysts in a dose-dependent manner and reduce the content of biochemical parameters associated with liver injury in CE mice. It also significantly reversed oxidative stress in CE mice, which was characterized as the decreased tumor necrosis factor alpha and H2O2 content, as well as the increase of the ratio of glutathione/oxidized glutathione and total superoxide dismutase content. DHA showed antiparasitic effects. DNA damages induced by oxidative stress played important roles in this process.


FTO is a relevant factor for the development of the metabolic syndrome in mice.
The metabolic syndrome is a worldwide problem mainly caused by obesity. FTO was found to be a obesity-risk gene in humans and FTO deficiency in mice led to reduction in adipose tissue. Thus, FTO is an important factor for the development of obesity. Leptin-deficient mice are a well characterized model for analysing the metabolic syndrome. To determine the relevance of FTO for the development of the metabolic syndrome we analysed different parameters in combined homozygous deficient mice (Lep(ob/ob);Fto(-/-)). Lep(ob/ob);Fto(-/-) mice showed an improvement in analysed hallmarks of the metabolic syndrome in comparison to leptin-deficient mice wild type or heterozygous for Fto. Lep(ob/ob);Fto(-/-) mice did not develop hyperglycaemia and showed an improved glucose tolerance. Furthermore, extension of beta-cell mass was prevented in Lep(ob/ob);Fto(-/-)mice and accumulation of ectopic fat in the liver was reduced. In conclusion this study demonstrates that FTO deficiency has a protective effect not only on the development of obesity but also on the metabolic syndrome. Thus, FTO plays an important role in the development of metabolic disorders and is an interesting target for therapeutic agents.


Physical activity in people age 80 years and older as a means of counteracting disability, balanced in relation to frailty.
The aim of this study was to describe experiences of physical activity, perceived meaning, and the importance of and motives and barriers for participation in physical activity in people 80 years of age and older. A qualitative design with focus-group methodology was used. The sample consisted of 20 community-living people age 80-91 yr. Data analyses revealed 4 themes: physical activity as a part of everything else in life, joie de vivre, fear of disease and dependence, and perceptions of frailty. Our results suggest that physical activity was not seen as a separate activity but rather as a part of activities often rated as more important than the physical activity itself. Thus, when designing physical activity interventions for elderly people, health care providers should consider including time for social interaction and possibilities to be outdoors. Moreover, assessment of physical activity levels among elderly people should include the physical activity in everyday activities.


Evidence for syntactic alignment in children with autism.
We report an experiment that examined whether children with Autistic Spectrum Disorder (ASD) spontaneously converge, or align, syntactic structure with a conversational partner. Children with ASD were more likely to produce a passive structure to describe a picture after hearing their interlocutor use a passive structure to describe an unrelated picture when playing a card game. Furthermore, they converged syntactic structure with their interlocutor to the same extent as did both chronological and verbal age-matched controls. These results suggest that the linguistic impairment that is characteristic of children with ASD, and in particular their difficulty with interactive language usage, cannot be explained in terms of a general deficit in linguistic imitation.


Cognitive function following traumatic brain injury: effects of injury severity and recovery period in a parasagittal fluid-percussive injury model.
Previous work in this laboratory has demonstrated that rats show substantial deficits on the cued and hidden versions of the Morris water maze, as well as an apparent time-dependent recovery over a period of months, following moderate parasagittal fluid-percussion (FP) injury. However, the longitudinal nature of those studies precluded definitive statements regarding recovery because of the possible confound of practice-dependent improvements in performance. The present experiments were undertaken to address this issue and to investigate more closely the relationship between impact severity and posttraumatic learning/memory deficits, which have not been examined thoroughly in this model. Separate groups of rats were subjected to mild (1.1 to 1.4 atm), moderate (1.8 to 2.1 atm), or severe (2.2 to 2.7 atm) FP injury (or sham surgery) and tested on several water maze tasks at either 5 days or 8 weeks after injury. Moderately and severely injured animals showed impairment in acquisition of the hidden platform task at both time points. Cued platform task performance was impaired significantly in severely injured animals 8 weeks after insult. Mildly injured animals exhibited no significant deficits on either task at either time point. The results indicate that deficits on the hidden platform task are more robust than those on the cued platform task, and that performance on both tasks is dependent on injury severity. They also indicate that the learning/memory deficits in this model are relatively enduring, suggesting that the model is a reasonable one for assessing potential treatment regimens.


[Effect of vitamins A, E and K on the indices of the glutathione antiperoxide system in gingival tissues in periodontosis].
Glutathione reductase is activated and the content of glutathione sulfhydryl groups is increased in the gingival tissue of patients afficted with parodontosis. The degree of alterations depends on the degree of the development and character of the disease. Application of antioxidant vitamin therapy (vitamins A, E and K) locally and per os normalizes the parameters under study and improves the status of the parodontium.


Branched-Selective Cross-Electrophile Coupling of 2-Alkyl Aziridines and (Hetero)aryl Iodides Using Ti/Ni Catalysis.
The arylation of 2-alkyl aziridines by nucleophilic ring-opening or transition-metal-catalyzed cross-coupling enables facile access to biologically relevant β-phenethylamine derivatives. However, both approaches largely favor C-C bond formation at the less-substituted carbon of the aziridine, thus enabling access to only linear products. Consequently, despite the attractive bond disconnection that it poses, the synthesis of branched arylated products from 2-alkyl aziridines has remained inaccessible. Herein, we address this long-standing challenge and report the first branched-selective cross-coupling of 2-alkyl aziridines with aryl iodides. This unique selectivity is enabled by a Ti/Ni dual-catalytic system. We demonstrate the robustness of the method by a twofold approach: an additive screening campaign to probe functional group tolerance and a feature-driven substrate scope to study the effect of the local steric and electronic profile of each coupling partner on reactivity. Furthermore, the diversity of this feature-driven substrate scope enabled the generation of predictive reactivity models that guided mechanistic understanding. Mechanistic studies demonstrated that the branched selectivity arises from a TiIII-induced radical ring-opening of the aziridine.


Molecular determination of H antigens of Salmonella by use of a microsphere-based liquid array.
Serotyping of Salmonella has been an invaluable subtyping method for epidemiologic studies for more than 70 years. The technical difficulties of serotyping, primarily in antiserum production and quality control, can be overcome with modern molecular methods. We developed a DNA-based assay targeting the genes encoding the flagellar antigens (fliC and fljB) of the Kauffmann-White serotyping scheme. Fifteen H antigens (H:a, -b, -c, -d, -d/j, -e,h, -i, -k, -r, -y, -z, -z(10), -z(29), -z(35), and -z(6)), 5 complex major antigens (H:G, -EN, -Z4, -1, and -L) and 16 complex secondary antigens (H:2, -5, -6, -7, -f, -m/g,m, -m/m,t, -p, -s, -t/m,t, -v, -x, -z(15), -z(24), -z(28), and -z(51)) were targeted in the assay. DNA probes targeting these antigens were designed and evaluated on 500 isolates tested in parallel with traditional serotyping methods. The assay correctly identified 461 (92.2%) isolates based on the 36 antigens detected in the assay. Among the isolates considered correctly identified, 47 (9.4%) were partially serotyped because probes corresponding to some antigens in the strains were not in the assay, and 13 (2.6%) were monophasic or nonmotile strains that possessed flagellar antigen genes that were not expressed but were detected in the assay. The 39 (7.8%) strains that were not correctly identified possessed an antigen that should have been detected by the assay but was not. Apparent false-negative results may be attributed to allelic divergence. The molecular assay provided results that paralleled traditional methods with a much greater throughput, while maintaining the integrity of the Kauffmann-White serotyping scheme, thus providing backwards-compatible epidemiologic data. This assay should greatly enhance the ability of clinical and public health laboratories to serotype Salmonella.


In vitro quantification of botulinum neurotoxin type A1 using immobilized nerve cell-mimicking nanoreactors in a microfluidic platform.
The bacterial toxin botulinum neurotoxin A (BoNT/A) is not only an extremely toxic substance but also a potent pharmaceutical compound that is used in a wide spectrum of neurological disorders and cosmetic applications. The quantification of the toxin is extremely challenging due to its extraordinary high physiological potency and is further complicated by the toxin's three key functionalities that are necessary for its activity: receptor binding, internalization-translocation, and catalytic activity. So far, the industrial standard to measure the active toxin has been the mouse bioassay (MBA) that is considered today as outdated due to ethical issues. Therefore, recent introductions of cell-based assays were highly anticipated; their impact however remains limited due to their labor-intensive implementation. This report describes a new in vitro approach that combines a nanosensor based on the use of nerve cell-mimicking nanoreactors (NMN) with microfluidic technology. The nanosensor was able to measure all three key functionalities, and therefore suitable to quantify the amount of physiologically active BoNT/A. The integration of such a sensor in a microfluidic device allowed the detection and quantification of BoNT/A amounts in a much shorter time than the MBA (<10 h vs. 2-4 days). Lastly, the system was also able to reliably quantify physiologically active BoNT/A within a simple final pharmaceutical formulation. This complete in vitro testing system and its unique combination of a highly sensitive nanosensor and microfluidic technology represent a significant ethical advancement over in vivo measures and a possible alternative to cell-based in vitro detection methods.


Myocardial infarction complexity: A multi-omics approach.
Myocardial infarction (MI), a prevalent cardiovascular disease, is fundamentally precipitated by thrombus formation in the coronary arteries, which subsequently decreases myocardial perfusion and leads to cellular necrosis. The intricacy of MI pathogenesis necessitates extensive research to elucidate the disease's root cause, thereby addressing the limitations present in its diagnosis and prognosis. With the continuous advancement of genomics technology, genomics, proteomics, metabolomics and transcriptomics are widely used in the study of MI, which provides an excellent way to identify new biomarkers that elucidate the complex mechanisms of MI. This paper provides a detailed review of various genomics studies of MI, including genomics, proteomics, transcriptomics, metabolomics and multi-omics studies. The metabolites and proteins involved in the pathogenesis of MI are investigated through integrated protein-protein interactions and multi-omics analysis by STRING and Metascape platforms. In conclusion, the future of omics research in myocardial infarction offers significant promise.


In vivo imaging of the glucagonlike peptide 1 receptor in the pancreas with 68Ga-labeled DO3A-exendin-4.
The glucagonlike peptide 1 receptor (GLP-1R) is mainly expressed on β-cells in the islets of Langerhans and is therefore an attractive target for imaging of the β-cell mass. In the present study, (68)Ga-labeled exendin-4 was evaluated for PET imaging and quantification of GLP-1R in the pancreas. Dose escalation studies of (68)Ga-labeled 1,4,7-tris(carboxymethylaza)cyclododecane-10-azaacetyl (DO3A)-exendin-4 were performed in rats (organ distribution) and cynomolgus monkeys (PET/CT imaging) to determine the GLP-1R-specific tissue uptake in vivo. Pancreatic uptake (as determined by organ distribution) in healthy rats was compared with that in diabetic rats. GLP-1R occupancy in the cynomolgus pancreas was quantified with a 1-tissue-compartment model. In rodents, uptake in the pancreas was decreased from the baseline by up to 90% (P < 0.0001) by coadministration of DO3A-exendin-4 at 100 μg/kg. Pancreatic uptake in diabetic animals was decreased by more than 80% (P < 0.001) compared with that in healthy controls, as measured by organ distribution. GLP-1R occupancy in the cynomolgus pancreas after coinjection of DO3A-exendin-4 at 0.15-20 μg/kg ranged from 49% to 97%, as estimated by compartment modeling. These results strongly support the notion that (68)Ga-DO3A-exendin-4 uptake in the pancreas is mediated by specific receptor binding. In addition, pancreatic uptake was decreased by selective destruction of β-cells. This result suggests that GLP-1R can be quantified in vivo, which has major implications for the prospect of imaging of native β-cells.


[Nosocomial fungal infections in surgical patients. Older age as factor of risk and early presumptive and systemic therapy versus prophylactic therapy].
Objective of our study has been to verify if the age is a factor of additional risk for the opportunistic fungal infections and if prophylactic therapy cause real advantages in terms of reduction of morbidity, mortality and times of hospitalization. To such we report the experience of the Operational Unity of Emergency Surgery of the Hospital of Relief National and High Specialization of Caserta relatively to the patients submitted to intervention of great abdominal surgery. In the year 2000 antifungal therapy has been administered to 20 patient (of which 18 over 65 years) on the escort of an laboratory diagnosis; in the year 2001 we have administered prophylactic therapy to 53 high-risk patient (of which 48 over 65). In the group of patients essays with preventive therapy the middle hospitalization has been slightly inferior with a reduction of around 2 days of refuge for the elderly patients and of around 1.5 days in the youngest patients. Mortality in the elderly patients has been reduced from 38% of the year 2000 at 23% of the year 2001. Our data confirm that the age is an important factor of risk for the fungal infections in the surgical departments and we believe to a prophylactic therapy can be of benefit in well selected patient, although an ordinary application of antifunfal therapy can not have recommended without an laboratory diagnosis because to emerge of new resistances to the medicines.


A peripheral primitive neuroectodermal tumor with generalized bone metastases in a puppy.
A peripheral primitive neuroectodermal tumor (pPNET), most consistent with a human Ewing's sarcoma, is described in a 5-month-old male Australian Shepherd puppy. The first tumor site detected was in the left frontal bone of the skull with apparent subsequent rapid metastases to multiple sites in the axial and appendicular skeleton and bone marrow, kidneys, and perihyphophyseal meninges. Radiographically, all bone lesions were lytic and there was also a humeral bone fracture. Histologically, the tumor was diagnosed as a small round blue cell tumor. At this stage, the differential diagnosis included a lymphoma, rhabdomyosarcoma, and a PNET of the peripheral nervous system. However, the cells had positive expression of triple neurofilament antigens as detected immunocytochemically. The cells were negative for a broad panel of canine-specific leucocyte cell marker antigens for desmin, smooth muscle actin, synaptophysin, and CD99. Ultrastructurally, the cells contained occasional dense core neurosecretory granules and intermediate filaments with intercellular desmosomal-like junctions and abundant glycogen clusters. Based on the age of the dog, the clinical history, the distribution of gross lesions, histologic characteristics of a small round blue cell tumor, and immunocytochemical and ultrastructural evidence of neuroectodermal differentiation, a diagnosis of a pPNET similar to a human Ewing's sarcoma was made.


[Occupational hypoacusis in transport workers].
The article concerns prevalence and causes of occupational neurosensory hypoacusis (ONH) in transport workers exposed to intensive industrial noise. Most often ONH is diagnosed in civil aviation air crews and railway locomotive crews aged 45-50 years with service history 15 years and longer. Because ONH morbidity has a trend to rise, it is necessary to update medical criteria of professional fitness and rules of medical examination of transport workers, especially those engaged in safety control. International classifications and scientific approaches may be considered. Innovations in noise reduction, modern hearing protection measures, early diagnosis and rehabilitation must be promoted.


Disordered insulin secretion in the development of insulin resistance and Type 2 diabetes.
For many years, the development of insulin resistance has been seen as the core defect responsible for the development of Type 2 diabetes. However, despite extensive research, the initial factors responsible for insulin resistance development have not been elucidated. If insulin resistance can be overcome by enhanced insulin secretion, then hyperglycaemia will never develop. Therefore, a β-cell defect is clearly required for the development of diabetes. There is a wealth of evidence to suggest that disorders in insulin secretion can lead to the development of decreased insulin sensitivity. In this review, we describe the potential initiating defects in Type 2 diabetes, normal pulsatile insulin secretion and the effects that disordered secretion may have on both β-cell function and hepatic insulin sensitivity. We go on to examine evidence from physiological and epidemiological studies describing β-cell dysfunction in the development of insulin resistance. Finally, we describe how disordered insulin secretion may cause intracellular insulin resistance and the implications this concept has for diabetes therapy. In summary, disordered insulin secretion may contribute to development of insulin resistance and hence represent an initiating factor in the progression to Type 2 diabetes.


Gene duplication and deletion caused by over-replication at a fork barrier.
Replication fork stalling can provoke fork reversal to form a four-way DNA junction. This remodelling of the replication fork can facilitate repair, aid bypass of DNA lesions, and enable replication restart, but may also pose a risk of over-replication during fork convergence. We show that replication fork stalling at a site-specific barrier in fission yeast can induce gene duplication-deletion rearrangements that are independent of replication restart-associated template switching and Rad51-dependent multi-invasion. Instead, they resemble targeted gene replacements (TGRs), requiring the DNA annealing activity of Rad52, the 3'-flap nuclease Rad16-Swi10, and mismatch repair protein Msh2. We propose that excess DNA, generated during the merging of a canonical fork with a reversed fork, can be liberated by a nuclease and integrated at an ectopic site via a TGR-like mechanism. This highlights how over-replication at replication termination sites can threaten genome stability in eukaryotes.


Cancer treatment-induced bone loss in premenopausal women: a need for therapeutic intervention?
Current clinical treatment guidelines recommend cytotoxic chemotherapy, endocrine therapy, or both (with targeted therapy if indicated) for premenopausal women with early-stage breast cancer, depending on the biologic characteristics of the primary tumor. Some of these therapies can induce premature menopause or are specifically designed to suppress ovarian function and reduce circulating estrogen levels. In addition to bone loss associated with low estrogen levels, cytotoxic chemotherapy may have a direct negative effect on bone metabolism. As a result, cancer treatment-induced bone loss poses a significant threat to bone health in premenopausal women with breast cancer. Clinical trials of antiresorptive therapies, such as bisphosphonates, have demonstrated the ability to slow or prevent bone loss in this setting. Current fracture risk assessment tools are based on data from healthy postmenopausal women and do not adequately address the risks associated with breast cancer therapy, especially in younger premenopausal women. We therefore recommend that all premenopausal women with breast cancer be informed about the potential risk of bone loss prior to beginning anticancer therapy. Women who experience amenorrhea should have bone mineral density assessed by dual-energy X-ray absorptiometry and receive regular follow-up to monitor bone health. Regular exercise and daily calcium and vitamin D supplementation are recommended. Women with a Z-score <-2.0 or Z-score ≤-1.0 and/or a 5-10% annual decrease in bone mineral density should be considered for bisphosphonate therapy in addition to calcium and vitamin D supplements.


Gender equality or patriarchal dividend: Structural change in Turkish nursing.
Turkey is attempting to join the European Union and is facing pressure to eliminate many forms of discrimination, including discrimination based on gender. In keeping with these aims, in early 2007 the Turkish government changed the law to permit Turkish men to become nurses. Given that Turkey is a highly patriarchal society and that occupational segregation by sex is a persistent feature, it is important to examine the potential outcome of the legislative changes. The aim of this paper is to explore the paradoxical potential for Turkish female nurses to experience increased discrimination in a system that is restructured by legislation to be non-discriminatory.


A near-infrared responsive germanium complex of Ge/GeO2 for targeted tumor phototherapy.
The development of photoactive nanomaterials with high biocompatibility for targeted tumor phototherapy is of great significance for antitumor applications; this study presents a novel phototherapeutic agent, the Ge/GeO2 complex, which shows broad photoabsorption in the near infrared (NIR) region. As a result, it can synchronously produce reactive oxygen species (ROS) and heat under NIR irradiation. After being loaded onto macrophages, Ge/GeO2 could be delivered to tumors in a targeted fashion. Combining the abovementioned merits together, macrophage-loaded Ge/GeO2 realized in vivo synergetic photothermal and photodynamic outcomes to completely remove solid tumors in mice via intravenous administration. In this study, B-ultrasonography was also employed to monitor the tumor evolution after phototherapy, revealing a sequential process of tumor necrosis, liquefaction/softening, and finally disappearance. In addition, Ge/GeO2 proposed in this study shows negligible cytotoxicity and hematotoxicity, especially after being loaded onto macrophages.


Inducing transient language disruptions by mapping of Broca's area with modified patterned repetitive transcranial magnetic stimulation protocol.
Until now there has been no reliable stimulation protocol for inducing transient language disruptions while mapping Broca's area. Despite the promising data of only a few studies in which speech arrest and language disturbances have been induced, certain concerns have been raised. The purpose of this study was to map Broca's area by using event-related navigated transcranial magnetic stimulation (nTMS) to generate a modified patterned nTMS protocol. Eleven right-handed subjects underwent nTMS to Broca's area while engaged in a visual object-naming task. Navigated TMS was triggered 300 msec after picture presentation. The modified patterned nTMS protocol consists of 4 stimuli with an interstimulus interval of 6 msec; 8 or 16 of those bursts were repeated with a burst repetition rate of 12 Hz. Prior to mapping of Broca's area, the primary motor cortices (M1) for hand and laryngeal muscles were mapped. The Euclidian distance on MRI was measured between cortical points eliciting transient language disruptions and M1 for the laryngeal muscle. On stimulating Broca's area, transient language disruptions were induced in all subjects. The mean Euclidian distance between cortical spots inducing transient language disruptions and M1 for the laryngeal muscle was 17.23 ± 4.73 mm. The stimulation paradigm with the modified patterned nTMS protocol was shown to be promising and might gain more widespread use in speech localization in clinical and research applications.


Chitosan-genipin microspheres for the controlled release of drugs: clarithromycin, tramadol and heparin.
The aim of this study was to first evaluate whether the chitosan hydrochloride-genipin crosslinking reaction is influenced by factors such as time, and polymer/genipin concentration, and second, to develop crosslinked drug loaded microspheres to improve the control over drug release. Once the crosslinking process was characterized as a function of the factors mentioned above, drug loaded hydrochloride chitosan microspheres with different degrees of crosslinking were obtained. Microspheres were characterized in terms of size, morphology, drug content, surface charge and capacity to control in vitro drug release. Clarithromycin, tramadol hydrochloride, and low molecular weight heparin (LMWH) were used as model drugs. The obtained particles were spherical, positively charged, with a diameter of 1-10 microm. X-Ray diffraction showed that there was an interaction of genipin and each drug with chitosan in the microspheres. In relation to the release profiles, a higher degree of crosslinking led to more control of drug release in the case of clarithromycin and tramadol. For these drugs, optimal release profiles were obtained for microspheres crosslinked with 1 mM genipin at 50 °C for 5 h and with 5 mM genipin at 50 °C for 5 h, respectively. In LMWH microspheres, the best release profile corresponded to 0.5 mM genipin, 50 °C, 5 h. In conclusion, genipin showed to be eligible as a chemical-crosslinking agent delaying the outflow of drugs from the microspheres. However, more studies in vitro and in vivo must be carried out to determine adequate crosslinking conditions for different drugs.


Direct relationship between radiobiological hypoxia in tumors and monoclonal antibody detection of EF5 cellular adducts.
While the potential importance of hypoxia in limiting the sensitivity of tumor cells to ionizing radiation has long been appreciated, methods for accurately quantifying the number of radiation-resistant hypoxic cells within tumors have been lacking. We have used the pentafluorinated derivative [2-(2-nitro-1H-imidazol-1-yl)-N-(2,2,3,3,3-pentafluoropropyl)-acet amide] of etanidazole (EF5), which binds selectively to hypoxic cells. The adducts formed between EF5 and cellular proteins in the hypoxic cells were detected using the specific monoclonal antibody (MAb), ELK3-51 conjugated to the flurochrome Cy3, and the number of hypoxic cells was quantified by flow cytometry. To verify the validity of this technique for the detection of hypoxic cells, mice bearing KHT sarcomas were treated with various agents to alter tumor oxygenation and hence vary the fraction of radiobiologically hypoxic tumor cells. The percentage of EF5 binding cells was then compared directly with the clonogenic survival of the tumor cells following radiation treatment under the various pretreatment conditions. The results showed that allowing the mice to breathe carbogen (5% CO2/95% O2) prior to irradiation reduced clonogenic cell survival approx. 6-fold and led to an absence of cells binding high levels of EF5. In contrast, pretreating the tumor-bearing animals with either hydralazine, which decreased tumor blood flow, or phenylhydrazine hydrochloride, which made the mice anemic, increased tumor cell survival following irradiation 2- to 4-fold, indicative of an increase in the fraction of hypoxic tumor cells. EF5 measurements made under identical conditions illustrated a shift in the cells in the tumor to high EF5 binding. Our results demonstrate that flow cytometric measurement by fluorescent MAb binding to EF5 adducts may relate directly to radiobiological hypoxia in KHT tumors measured by conventional methods.


The effect of single tooth implant restorations on the survival, morbidity, pulpal, and periapical health of adjacent teeth: A chart review.
To determine whether the placement and restoration of a single tooth posterior implant affects the survival, morbidity, pulpal, and periapical health of adjacent natural teeth. A retrospective chart review identified patients who received single posterior tooth implants between August 2004 and July 2015 at the UNC SOD and met the study inclusion criteria. Preoperative and postoperative records were reviewed; survival and changes in coronal, pulpal, and periapical status of teeth adjacent to the implant and contralateral tooth were recorded. Dichotomous survival, restoration, and retreatment outcomes were analyzed using conditional logistic regression with patient as strata and implant versus control as the predictor. Exact odds ratio estimates and the 95% confidence intervals were obtained for the relationship of implant versus control side and outcomes. Five hundred and fifty-five sites with follow-up time averaging 5 years ± 30.8 months were reviewed. Teeth adjacent to implants had 1.75 (95% CI: 1.17, 2.64) times the odds of restorative retreatments as compared to controls (P = .005). On the implant side, 48 adjacent teeth (4.5%) were more heavily restored at follow up, while 84 (7.9%) experienced retreatment with comparable number of surfaces restored. On the contralateral side, 54 adjacent teeth (5.0%) were more heavily restored, and 56 (5.2%) experienced comparable levels of retreatment. In addition, 17 (1.7%) implant adjacent teeth required root canal treatment, compared to 12 (1.2%) on the contralateral side; 1 implant adjacent tooth required root canal retreatment. Forty-two teeth (3.8%) adjacent to implants were lost, compared to 35 (3.2%) adjacent to natural teeth. The incidence for restorative retreatment was significantly higher on teeth adjacent to implant restorations as compared to the contralateral controls. There were no significant differences in the survival, morbidity, pulpal, or periapical health of teeth adjacent to single tooth implants compared to those adjacent to the contralateral natural tooth.


The role of natriuretic peptides in volume assessment and mortality prediction in Haemodialysis patients.
Maintaining optimal fluid balance is essential in haemodialysis (HD) patients but clinical evaluation remains problematic. Other technologies such as bioimpedance are emerging as valuable adjuncts. This study was undertaken to explore the potential utility of the natriuretic peptides - atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP) in the assessment of fluid status and cardiovascular risk in this setting. This was a cross-sectional study carried out in an unselected cohort of 170 prevalent HD patients. Volume status was assessed by clinical parameters - the presence or absence of peripheral oedema, raised jugular venous pressure and basal lung crepitations; by extracellular fluid volume (ECFV) status determined by whole body bioimpedance; and by serum levels of BNP and ANP (pre- and post -dialysis). The relationships of ANP and BNP levels to clinical and bioimpedance parameters of volume status was determined. Patients were followed up for 5 years to assess the relationship of natriuretic peptide levels to mortality. Bioimpedance estimates of ECFV expansion (>105 % of ideal ECFV) was present in 52 % of patients pre-dialysis. A significant proportion (21 %) of pre-dialysis patients had a depleted ECFV (<95 % of ideal ECFV) pre-dialysis. The situation was reversed post-dialysis. A raised JVP >3 cm was the most reliable clinical sign of ECFV expansion inferred from bioimpedance measurements and natriuretic peptide levels. The vast majority of patients with this sign also had lung crepitations or peripheral oedema or both. BNP was a stronger predictor of ECFV expansion than either pre- or post-dialysis ANP. BNP was also a stronger predictor of five-year survival. Serum levels of BNP have a strong relationship to both volume status and survival in HD patients. We found no clear role for measurement of ANP, though changes in blood levels may be a sensitive indicator of acute changes in volume status. Whether monitoring levels of these peptides has a role in the management of volume status and cardiovascular risk requires further study.


Use of biochemical profile in children's hospital: results of two controlled trials.
Two controlled trials of the use of a biochemical profile were conducted in a childern's hospital to see whether the profile led to diagnoses which would not otherwise [have been made and to see what effect it had on the number of extra requests for pathololgical investigations and the length of stay in hospital]. Altogether 2816 children were examined and 13 new diagnoses made. There was a significant increases in the total number of pathorequests but the profile did not alter the length of stay in hospital. We conclude that the profile mad only a small contribtuion to the overall care of the pateints.


Parental perspectives about information and deferred versus two-stage consent in studies of neonatal asphyxia.
The ALBINO Trial (NCT03162653) investigates effects of very early postnatal allopurinol on neurocognitive outcome following perinatal asphyxia where prenatal informed consent (IC) is impossible. Ethically and legally, waiver of consent and/or deferred consent (DC) is acceptable in such an emergency. Short oral/two-step consent (SOC, brief information and oral consent followed by IC) has recently been investigated. Mixed-methods analysis of parental opinions on DC versus SOC in the context of neonatal asphyxia in a survey at two German centres. Prospective parents (ProP), parents of healthy newborns (PNeo) and parents of asphyxiated infants (PAx) born between 2006 and 2016 were invited. 108 of 422 parents participated (ProP:43; PNeo:35; PAx:30). Most parents trusted physicians, wanted preinterventional information and agreed that in emergencies interventions should begin immediately. Intergroup and intragroup variability existed for questions about DC and SOC. In the ALBINO Trial situation, 55% preferred SOC, and 26% reported DC without information might adversely affect their trust. Only 3% reported to potentially take legal action after DC. PAx were significantly more likely to support DC. PAx more frequently expressed positive emotions and appreciation for neonatal research. In open-ended questions, parents gave many constructive recommendations. In this survey, parents expressed diverse opinions on consent, but the majority preferred SOC over DC. Parents who had experienced emergency admission of their asphyxiated neonates were more trusting. Obtaining parental perspectives is essential when designing studies, while being cognisant that these groups of parents may not represent the opinion of all parents.


Ubiquitin Modulates Liquid-Liquid Phase Separation of UBQLN2 via Disruption of Multivalent Interactions.
Under stress, certain eukaryotic proteins and RNA assemble to form membraneless organelles known as stress granules. The most well-studied stress granule components are RNA-binding proteins that undergo liquid-liquid phase separation (LLPS) into protein-rich droplets mediated by intrinsically disordered low-complexity domains (LCDs). Here we show that stress granules include proteasomal shuttle factor UBQLN2, an LCD-containing protein structurally and functionally distinct from RNA-binding proteins. In vitro, UBQLN2 exhibits LLPS at physiological conditions. Deletion studies correlate oligomerization with UBQLN2's ability to phase-separate and form stress-induced cytoplasmic puncta in cells. Using nuclear magnetic resonance (NMR) spectroscopy, we mapped weak, multivalent interactions that promote UBQLN2 oligomerization and LLPS. Ubiquitin or polyubiquitin binding, obligatory for UBQLN2's biological functions, eliminates UBQLN2 LLPS, thus serving as a switch between droplet and disperse phases. We postulate that UBQLN2 LLPS enables its recruitment to stress granules, where its interactions with ubiquitinated substrates reverse LLPS to enable shuttling of clients out of stress granules.


A systematic review and meta-analysis of radon risk exposure from drinking water resources in Nigeria.
Elevated radon concentrations in drinking water pose an increased risk of cancer among nonsmokers. A Monte-Carlo Simulation was employed to assess the effective dose and cancer risk associated with radon exposure in humans, utilizing a systematic review and meta-analysis of related studies. These studies were sourced from databases including PubMed, Web of Science, Scopus, Science Direct, and Google Scholar, focusing on drinking water from Nigeria's six geopolitical zones. The random effects models revealed a 222Rn concentration in drinking water of Nigeria at 25.01, with 95% confidence intervals (CI) of 7.62 and 82.09, indicating significant heterogeneity of (I2 = 100%; p < 0.001). The probabilistic risk of effective dose revealed a best-scenario (P 5%) at Kundiga and Magiro that exceeded the World Health Organization's (WHO) recommended effective dose limit of 200 µSv/y. Conversely, the worst-case scenario (P 95%) indicated concentrations surpassing the recommended limit at Kundiga, Edbe, Magiro, Ekiti, and Abeokuta. Excess Life Cancer Risk for infants, children, and adults attributed to the ingestion and inhalation of radon from various drinking water sources exceeded the recommended values of 0.2 x 10-3 established by the International Commission on Radiological Protection (ICRP) and the United Nations Scientific Committee on the Effect of Atomic Radiation (UNSCEAR). It underscores the necessity for treating radon-polluted water, employing methos such as aeration and granular activated carbon (GAC) processes.


Concomitant appearance of pisa syndrome and striatal hand in Parkinson's disease.
Pisa syndrome is (PS) usually seen in patients receiving antipsychotic drugs and characterised by lateral flexion of trunk and axial dystonia. It is believed that antipsychotic drugs lead to dopamine blockage causing PS. We describe a Parkinson's disease patient who was doing well with levodopa/carbidopa for 3 years and developed lateral flexion of trunk. His abnormal posture used to completely improve upon lying down position. He also had striatal hand deformity suggestive of focal dystonia.


Effect of aliskiren on postdischarge mortality and heart failure readmissions among patients hospitalized for heart failure: the ASTRONAUT randomized trial.
Hospitalizations for heart failure (HHF) represent a major health burden, with high rates of early postdischarge rehospitalization and mortality. To investigate whether aliskiren, a direct renin inhibitor, when added to standard therapy, would reduce the rate of cardiovascular (CV) death or HF rehospitalization among HHF patients. International, double-blind, placebo-controlled study that randomized hemodynamically stable HHF patients a median 5 days after admission. Eligible patients were 18 years or older with left ventricular ejection fraction (LVEF) 40% or less, elevated natriuretic peptides (brain natriuretic peptide [BNP] ≥ 400 pg/mL or N -terminal pro-BNP [NT-proBNP] ≥ 1600 pg/mL), and signs and symptoms of fluid overload. Patients were recruited from 316 sites across North and South America, Europe, and Asia between May 2009 and December 2011. The follow-up period ended in July 2012. All patients received 150 mg (increased to 300 mg as tolerated) of aliskiren or placebo daily, in addition to standard therapy. The study drug was continued after discharge for a median 11.3 months. MAIN OUTCOME MEASURES Cardiovascular death or HF rehospitalization at 6 months and 12 months. In total, 1639 patients were randomized, with 1615 patients included in the final efficacy analysis cohort (808 aliskiren, 807 placebo). Mean age was 65 years; mean LVEF, 28%; 41% of patients had diabetes mellitus, mean estimated glomerular filtration rate, 67 mL/min/1.73 m2. At admission and randomization, median NT-proBNP levels were 4239 pg/mL and 2718 pg/mL, respectively. At randomization, patients were receiving diuretics (95.9%), β-blockers (82.5%), angiotensin-converting enzyme inhibitors or angiotensin II receptor blockers (84.2%), and mineralocorticoid receptor antagonists (57.0%). In total, 24.9% of patients receiving aliskiren (77 CV deaths, 153 HF rehospitalizations) and 26.5% of patients receiving placebo (85 CV deaths, 166 HF rehospitalizations) experienced the primary end point at 6 months (hazard ratio [HR], 0.92; 95% CI, 0.76-1.12; P = .41). At 12 months, the event rates were 35.0% for the aliskiren group (126 CV deaths, 212 HF rehospitalizations) and 37.3% for the placebo group (137 CV deaths, 224 HF rehospitalizations; HR, 0.93; 95% CI, 0.79-1.09; P = .36). The rates of hyperkalemia, hypotension, and renal impairment/renal failure were higher in the aliskiren group compared with placebo. Among patients hospitalized for HF with reduced LVEF, initiation of aliskiren in addition to standard therapy did not reduce CV death or HF rehospitalization at 6 months or 12 months after discharge. clinicaltrials.gov Identifier: NCT00894387.


VHH Structural Modelling Approaches: A Critical Review.
VHH, i.e., VH domains of camelid single-chain antibodies, are very promising therapeutic agents due to their significant physicochemical advantages compared to classical mammalian antibodies. The number of experimentally solved VHH structures has significantly improved recently, which is of great help, because it offers the ability to directly work on 3D structures to humanise or improve them. Unfortunately, most VHHs do not have 3D structures. Thus, it is essential to find alternative ways to get structural information. The methods of structure prediction from the primary amino acid sequence appear essential to bypass this limitation. This review presents the most extensive overview of structure prediction methods applied for the 3D modelling of a given VHH sequence (a total of 21). Besides the historical overview, it aims at showing how model software programs have been shaping the structural predictions of VHHs. A brief explanation of each methodology is supplied, and pertinent examples of their usage are provided. Finally, we present a structure prediction case study of a recently solved VHH structure. According to some recent studies and the present analysis, AlphaFold 2 and NanoNet appear to be the best tools to predict a structural model of VHH from its sequence.


Individual differences in eye blink rate predict both transient and tonic pupil responses during reversal learning.
The pupil response under constant illumination can be used as a marker of cognitive processes. In the past, pupillary responses have been studied in the context of arousal and decision-making. However, recent work involving Parkinson's patients suggested that pupillary responses are additionally affected by reward sensitivity. Here, we build on these findings by examining how pupil responses are modulated by reward and loss while participants (N = 30) performed a Pavlovian reversal learning task. In fast (transient) pupil responses, we observed arousal-based influences on pupil size both during the expectation of upcoming value and the evaluation of unexpected monetary outcomes. Importantly, after incorporating eye blink rate (EBR), a behavioral correlate of striatal dopamine levels, we observed that participants with lower EBR showed stronger pupil dilation during the expectation of upcoming reward. Subsequently, when reward expectations were violated, participants with lower EBR showed stronger pupil responses after experiencing unexpected loss. Across trials, the detection of a reward contingency reversal was reflected in a slow (tonic) dilatory pupil response observed already several trials prior to the behavioral report. Interestingly, EBR correlated positively with this tonic detection response, suggesting that variability in the arousal-based detection response may reflect individual differences in striatal dopaminergic tone. Our results provide evidence that a behavioral marker of baseline striatal dopamine level (EBR) can potentially be used to describe the differential effects of value-based learning in the arousal-based pupil response.


The D-JH complex is an intermediate to the complete immunoglobulin heavy-chain V-region gene.
We have examined the organization of the immunoglobulin JH segments in three clones derived from a single Abelson murine leukemia virus-transformed cell. Cloning and nucleotide sequence analyses of the JH-containing fragments have revealed the rearrangement from the preformed D-JH complex to the complete VH-D-JH gene, which was accompanied by the expression of the intra-cytoplasmic mu chain. In one case a JH segment downstream to the preformed D-JH was used to create a new VH-D-JH gene. Upon the D-JH and VH-D-JH rearrangements the intervening D segments were deleted from the chromosome. One of the expressed VH genes suffered from a large deletion of the 3' portion (including the 95th cysteine residue) of the VH segment. We discuss the possible mechanism of the allelic exclusion.


Effects of telemedicine interventions on essential hypertension: a protocol for a systematic review and meta-analysis.
Essential hypertension is a major preventable risk factor for early cardiovascular disease, premature death and disability. It has been reported that telemedicine interventions can provide an innovative solution to essential hypertension to overcome the barriers that exist in traditional treatment or control. Nevertheless, this subject has not been thoroughly investigated. The goal of this study is to systematically evaluate and describe the impact of telemedicine interventions on essential hypertension. To find relevant research, we will conduct a systematic literature search of three databases (PubMed, Embase and Cochrane Library), with no language limitations, in addition to researching grey literature. Two reviewers will extract the data individually, and any disagreements will be resolved by discussion or by a third reviewer. The randomised controlled trials will be chosen based on predetermined inclusion criteria. Primary outcomes will include systolic blood pressure and diastolic blood pressure after the telemedicine intervention. Secondary outcomes will include medication adherence (eg, the Morisky Medication Adherence Questionnaire), quality of life (eg, the MOS item scale of the Health Survey Short Form 36 questionnaire), blood pressure control rate and adverse events (eg, stroke, chronic renal failure, aortic dissection, myocardial infarction and heart failure). The quality of the included studies will be assessed using the Cochrane risk-of-bias method. The data will be analysed using RevMan V.5.3.5 software and STATA V.16.0 software. If heterogeneity testing reveals little or no statistical heterogeneity, a fixed effect model will be used for data synthesis; otherwise, a random effect model would be employed. We will synthesise the available evidence to perform a high-quality meta-analysis. This project does not require ethical approval because it will be conducted using publicly available documents. The review's findings will be published in peer-reviewed journals and publications. CRD42021293539.


Interaction of Vitamin D with Peptide Hormones with Emphasis on Parathyroid Hormone, FGF23, and the Renin-Angiotensin-Aldosterone System.
The seminal discoveries that parathyroid hormone (PTH) and fibroblast growth factor 23 (FGF23) are major endocrine regulators of vitamin D metabolism led to a significant improvement in our understanding of the pivotal roles of peptide hormones and small proteohormones in the crosstalk between different organs, regulating vitamin D metabolism. The interaction of vitamin D, FGF23 and PTH in the kidney is essential for maintaining mineral homeostasis. The proteohormone FGF23 is mainly secreted from osteoblasts and osteoclasts in the bone. FGF23 acts on proximal renal tubules to decrease production of the active form of vitamin D (1,25(OH)2D) by downregulating transcription of 1α-hydroxylase (CYP27B1), and by activating transcription of the key enzyme responsible for vitamin D degradation, 24-hydroxylase (CYP24A1). Conversely, the peptide hormone PTH stimulates 1,25(OH)2D renal production by upregulating the expression of 1α-hydroxylase and downregulating that of 24-hydroxylase. The circulating concentration of 1,25(OH)2D is a positive regulator of FGF23 secretion in the bone, and a negative regulator of PTH secretion from the parathyroid gland, forming feedback loops between kidney and bone, and between kidney and parathyroid gland, respectively. In recent years, it has become clear that vitamin D signaling has important functions beyond mineral metabolism. Observation of seasonal variations in blood pressure and the subsequent identification of vitamin D receptor (VDR) and 1α-hydroxylase in non-renal tissues such as cardiomyocytes, endothelial and smooth muscle cells, suggested that vitamin D may play a role in maintaining cardiovascular health. Indeed, observational studies in humans have found an association between vitamin D deficiency and hypertension, left ventricular hypertrophy and heart failure, and experimental studies provided strong evidence for a role of vitamin D signaling in the regulation of cardiovascular function. One of the proposed mechanisms of action of vitamin D is that it functions as a negative regulator of the renin-angiotensin-aldosterone system (RAAS). This finding established a novel link between vitamin D and RAAS that was unexplored until then. During recent years, major progress has been made towards a more complete understanding of the mechanisms by which FGF23, PTH, and RAAS regulate vitamin D metabolism, especially at the genomic level. However, there are still major gaps in our knowledge that need to be filled by future research. The purpose of this review is to highlight our current understanding of the molecular mechanisms underlying the interaction between vitamin D, FGF23, PTH, and RAAS, and to discuss the role of these mechanisms in physiology and pathophysiology.


The developmental evaluation center as a clinical setting for mental health nursing students.
The DEC is seen as a viable option to the more traditional clinical settings used for psychiatric/mental health nursing students. Such a placement affords students the opportunities for direct involvement with individuals, families, and communities in the application of theoretical concepts within the framework of the nursing process. Ongoing client contacts over the course of the semester facilitate continuity in the implementation of intervention strategies and allow opportunities for evaluation of both short-term and long-term goals. The interdisciplinary nature of the DEC assists students in more clearly identifing their role and function as nurses within the larger health care team. Additionally, students gain first-hand experience in developing and utilizing colleagueal relationships to more effectively meet the needs of clients. Since the DEC concept is found not only in North Carolina, but throughout the U.S., it is thought that the DEC's offer a rich and rewarding alternative to the more traditional clinical placement sites.