Effect of postpartum drenching on plasma parameters of cows at a large-scale dairy farm.

The aim of the present study was to explore the influence of postpartum drenching with a feed additive on the plasma concentration of biochemical parameters while factoring in prepartum rumination times (RT). One hundred and sixty-one cows were fitted with a Ruminact© HR-Tag approximately 5 days before calving. Drenching and control groups were established based on calving dates. Animals in the drenched group were treated three times (Day 1/day of calving/, Day 2, and Day 3 postpartum) using a feed additive containing calcium propionate, magnesium sulphate, yeast, potassium chloride and sodium chloride mixed in approximately 25 L of lukewarm tap water. Blood samples were collected on Days 1, 2, 3, 7 and 12. Cows with below the average RT were categorised as "low rumination" and those above it as "high rumination" animals. Drenching decreased the plasma concentrations of total protein, urea and creatinine and increased the levels of alanine aminotransferase (ALT), gamma-glutamyl transferase (GGT) and chloride. Low rumination time prepartum resulted in higher concentrations of beta-hydroxybutyrate, total protein and activities of alkaline phosphatase and GGT, while it decreased the activity of ALT and the concentrations of calcium, magnesium, sodium and potassium. The day of lactation had an effect on all parameters except for potassium.

Study protocol of the Hungarian Longitudinal Study of Healthy Brain Aging (HuBA).

Background and purpose:

Neuro­cog­nitive aging and the associated brain diseases impose a major social and economic burden. Therefore, substantial efforts have been put into revealing the lifestyle, the neurobiological and the genetic underpinnings of healthy neurocognitive aging. However, these studies take place almost exclusively in a limited number of highly-developed countries. Thus, it is an important open question to what extent their findings may generalize to neurocognitive aging in other, not yet investigated regions. The purpose of the Hungarian Longitudinal Study of Healthy Brain Aging (HuBA) is to collect multi-modal longitudinal data on healthy neurocognitive aging to address the data gap in this field in Central and Eastern Europe.

We adapted the Australian Ima­ging, Biomarkers and Lifestyle (AIBL) study of aging study protocol to local circumstances and collected demographic, lifestyle, men­tal and physical health, medication and medical history related information as well as re­cor­ded a series of magnetic resonance imaging (MRI) data. In addition, participants were al­so offered to participate in the collection of blood samples to assess circulating in­flam­matory biomarkers as well as a sleep study aimed at evaluating the general sleep quality based on multi-day collection of subjective sleep questionnaires and whole-night elec­troencephalographic (EEG) data.

Baseline data collection has al­ready been accomplished for more than a hundred participants and data collection in the se­cond
session is on the way. The collected data might reveal specific local trends or could also indicate the generalizability of previous findings. Moreover, as the HuBA protocol al­so offers a sleep study designed for tho­rough characterization of participants’ sleep quality and related factors, our extended multi-modal dataset might provide a base for incorporating these measures into healthy and clinical aging research. 

Besides its straightforward na­tional benefits in terms of health ex­pen­di­ture, we hope that this Hungarian initiative could provide results valid for the whole Cent­ral and Eastern European region and could also promote aging and Alzheimer’s disease research in these countries.

Stereo-Controlled Liquid Phase Synthesis of Phosphorothioate Oligonucleotides on a Soluble Support.

5'-O-(2-Methoxyisopropyl) (MIP)-protected 2'-deoxynucleosides as chiral P(V)-building blocks, based on the limonene-derived oxathiaphospholane sulfide, were synthesized and used for the assembly of di-, tri-, and tetranucleotide phosphorothioates on a tetrapodal pentaerythritol-derived soluble support. The synthesis cycle consisted of two reactions and two precipitations: (1) the coupling under basic conditions, followed by neutralization and precipitation and (2) an acid catalyzed 5'-O-deacetalization, followed by neutralization and precipitation. The simple P(V) chemistry together with the facile 5'-O-MIP deprotection proved efficient in the liquid phase oligonucleotide synthesis (LPOS). Ammonolysis released nearly homogeneous Rp or Sp phosphorothioate diastereomers in ca. 80% yield/synthesis cycle.

West Nile virus infections in Hungary: Epidemiological update and phylogenetic analysis of the Hungarian virus strains between 2015 and 2022.

Following the introduction of the West Nile virus (WNV) into Hungary in 2004, it has shortly become one of the most important human arbovirus infections, with a gradually increasing number of cases. The study aimed to summarize the current epidemiological situation in Hungary and sequence the WNV PCR-positive clinical specimens and virus isolates by next-generation whole genome sequencing (NGS) to obtain a detailed phylogenetic analysis of the circulating virus strains. Whole blood and urine samples from confirmed WNV-infected patients and WNV isolates were investigated by reverse transcription PCR assays. Genome sequencing was carried out by Sanger-method, followed by NGS on the Illumina MiSeq platform. Altogether 499 human infections were diagnosed between 2004 and 2022. A particularly remarkable increase in human WNV infections was observed in 2018, while the number of reported cases significantly decreased during the COVID-19 pandemic. Between 2015 and 2022, 15 WNV isolates, and 10 PCR-positive clinical specimens were investigated by NGS. Phylogenetic analysis revealed that the major European WNV lineage 2 clades, namely the Eastern European (or Russian) and the Central European (or Hungarian) clades, are presented in Hungary. Strains of the Balkan and other European clusters within the Central European clade are co-circulating in the country, following a characteristic geographical distribution. In Hungary, the presence and co-circulation of multiple lineage 2 WNV strains could be identified in the last few years. Therefore, in light of the 2018 WNV outbreak, sequence-based typing of the currently circulating strains could highly support outbreak investigations.

Comparative evaluation of Duragen® and skim milk extenders to enhance ram semen quality and fertility: A promising alternative in ovine artificial insemination.

This study aimed to investigate the effectiveness of Duragen® and skimmed milk (SM) extenders on the quality parameters, bacterial load and fertilization ability of stored ram semen. A total of 50 ejaculates from Sardi rams (n = 5) aged 2.5-3 years, were collected and stored in Duragen® and SM at 15°C. The motilities and velocity parameters generated by the CASA system were then evaluated at 0, 8 and 24 h of storage. Afterward, bacterial loads of sperm extended in Duragen® and SM were determined at 0, 5 and 24 h of incubation. In addition, ewes (n = 100) aged 2 years, have been chosen in the same herd. The selected ewes were then synchronized and inseminated using semen extended in Duragen® and SM and stored for 5 h at 15°C. The results revealed that total and progressive motilities, straight velocity (VSL), straightness (SRT), lateral head displacement (ALH) and beat cross frequency (BCF) were not affected by the extender type after 24 h of storage (p > .05). However, curvilinear velocity (VCL), velocity average path (VAP), linearity (LIN) and wobble (WOB) showed higher values in Duragen® compared with SM extender after 24 h of storage (p < .05). Bacterial loads were observed mainly in sperm stored in SM at 5 h (183 UFC/mL) and at 24 h (357 UFC/mL) of incubation. However, the only case showing a bacterial load in Duragen® is when the storage time attains 24 h (199 UFC/mL). Concerning fertility, sperm diluted in both extenders resulting in high fertility rates which reaches 66% and 73% for Duragen® and SM, respectively, with no statistical difference (p > .05). In summary, Duragen® extender decreased bacterial load in stored semen and maintained high ram sperm quality and fertility. These findings suggest that Duragen® extender could be used as SM alternative in ovine artificial insemination (OAI).

Effect of postpartum drenching on rumination time and reticuloruminal pH at a Hungarian dairy farm.

The aim of this research was to investigate the effect of drenching with a feed additive on rumination time (RT) and reticuloruminal pH post-partum at a Hungarian large-scale dairy farm. One hundred and sixty-one cows were fitted with a Ruminact© HR-Tag and from these 20 also received SmaXtec© ruminal boli approximately 5 days before calving. Drenching and control groups were established based on calving dates. Animals in the drenching group were dosed three times (Day 0/day of calving/, Day 1, and Day 2 after calving) using a feed additive containing calcium propionate, magnesium sulphate, yeast, potassium chloride and sodium chloride mixed in approximately 25 L of lukewarm water. RT before calving and sensitivity to subacute ruminal acidosis (SARA) were considered in the final analysis. There was a significant decrease in RT in the drenched groups compared to the controls after drenching. Reticuloruminal pH was significantly higher and time below reticuloruminal pH 5.8 remained significantly lower in SARA-tolerant drenched animals on the days of the first and the second drenchings. Drenching temporarily decreased RT in both drenched groups compared to controls. The feed additive had a positive effect on reticuloruminal pH and time below reticuloruminal pH 5.8 in tolerant drenched animals.

Efficient link prediction in the protein-protein interaction network using topological information in a generative adversarial network machine learning model.

BACKGROUND: The investigation of possible interactions between two proteins in intracellular signaling is an expensive and laborious procedure in the wet-lab, therefore, several in silico approaches have been implemented to narrow down the candidates for future experimental validations. Reformulating the problem in the field of network theory, the set of proteins can be represented as the nodes of a network, while the interactions between them as the edges. The resulting protein-protein interaction (PPI) network enables the use of link prediction techniques in order to discover new probable connections. Therefore, here we aimed to offer a novel approach to the link prediction task in PPI networks, utilizing a generative machine learning model. RESULTS: We created a tool that consists of two modules, the data processing framework and the machine learning model. As data processing, we used a modified breadth-first search algorithm to traverse the network and extract induced subgraphs, which served as image-like input data for our model. As machine learning, an image-to-image translation inspired conditional generative adversarial network (cGAN) model utilizing Wasserstein distance-based loss improved with gradient penalty was used, taking the combined representation from the data processing as input, and training the generator to predict the probable unknown edges in the provided induced subgraphs. Our link prediction tool was evaluated on the protein-protein interaction networks of five different species from the STRING database by calculating the area under the receiver operating characteristic, the precision-recall curves and the normalized discounted cumulative gain (AUROC, AUPRC, NDCG, respectively). Test runs yielded the averaged results of AUROC = 0.915, AUPRC = 0.176 and NDCG = 0.763 on all investigated species. CONCLUSION: We developed a software for the purpose of link prediction in PPI networks utilizing machine learning. The evaluation of our software serves as the first demonstration that a cGAN model, conditioned on raw topological features of the PPI network, is an applicable solution for the PPI prediction problem without requiring often unavailable molecular node attributes. The corresponding scripts are available at https://github.com/semmelweis-pharmacology/ppi_pred .

Synthesis of Phosphorodiamidate Oligonucleotide Dimers.

Azido nucleosides couple with phosphoramidites via an initial iminophosphorane, which eliminates acrylonitrile to generate the coupled dimer P(V) product. The vulnerable phosphite triester intermediate is bypassed entirely, making the methodology very suitable to solution-phase synthesis. This new coupling protocol requires no protection of the 5'-OH function and provides a new method of installing internucleosidic phosphorodiamidate bonds with near quantitative yields.

Drug Delivery through the Psoriatic Epidermal Barrier-A "Skin-On-A-Chip" Permeability Study and Ex Vivo Optical Imaging.

Psoriasis is a chronic inflammatory disease with unmet medical needs. To clarify potential therapeutic targets, different animal models have been developed. In the current study, imiquimod-induced psoriasiform dermatitis was used for monitoring the changes in skin thickness, transepidermal water loss, body weight, blood perfusion and drug permeability for a topical cream formulation of caffeine, both in wild type and in knock out mice. Morphological characterization of control and diseased tissues was performed by scanning electron microscopy and two-photon microscopy. The chemically induced psoriatic group showed increased skin permeability for the model drug during disease progression. In wild type and TRPA1 KO mice, however, enhanced skin thickness and hyperkeratosis blocked further increase of drug penetration at the late phase (96 h). These results indicate that topical drug therapy can be more effective in early phases of plaque development, when skin thickness is lower. Although paracellular connections (tight junctions) are looser in the advanced phase, hyperkeratosis blocks drug delivery through the transappendageal routes. Novel drug formulations may have the potency for effective drug delivery across the epidermal barrier even in the advanced phase. For development of more effective topical drugs, further research is proposed to explore drug penetration both in healthy and diseased conditions.

[Can we influence the negative drug attitude? Interpretation of the rejection of COVID-19 vaccine in the light of results of a pilot study].

Background and purpose: Vaccination refusal is a serious obstacle to minimizing the spread of COVID-19. Nevertheless, the rejection of vaccine can be considered the result of a negative attitude towards medical treatment, and according to our previously published data, it can be influenced by the underlying affective state. Increased incidence of affective disorders and anxiety could be observed globally during the pandemic, which may have a significant impact on vaccination acceptance. The aim of our pilot study was to determine the association between clinical improvement of affective and neurocognitive symptoms and change of drug attitude and health control beliefs in a sample of psychiatric patients. Methods: A data set of 85 patients with psychiatric disorder has been analysed with the use of Patient's Health Belief Questionnaire on Psychiatric Treatment (PHBQPT) with 5 subscales (Negative Aspect of Medication - NA; Positive Aspect of Medication - PA; Doctor health locus of control- Doctor HLOC; Internal HLOC; Psychological Reactance - PR); Hospital Anxiety Depression Scale (HADS-Anx; HADS-Dep) and neurocognitive tests, such as the Stroop test and the Trail Making Tests. All the tests were performed before and after a 14 days treatment. Paired t-tests and generalized linear models were performed to assess the associations between the variables. Results: The baseline scores of NA and HADS-Anx correlated significantly (p=0.001) and after two weeks of treatment NA decreased (p=0.001), while Doctor HLOC and Internal HLOC increased (p=0.001 and p=0.006). The patients performance of the neurocognitive tests improved (all p<0.05). The reduction of HADS-Anx (p=0.002) and HADS-Dep (p=0.006) scores showed significant associations with the decrease of NA. Increase of the PA score was associated with reduction of HADS-Dep (p=0.028). Improvement of neurocognitive functions had no effect on PHBQPT scores. Conclusion: Important conclusions can be drawn regarding the rejection of the COVID-19 vaccine based on the associations found between the intensity of affective and anxiety symptoms and the attitude towards treatment. Our findings suggest that affective symptoms have a negative influence on the attitude towards treatment and that the improvement of these symptoms can facilitate the acceptance of the therapy, regardless of diagnosis. The modest effect of the improvement of neurocognitive functioning on the attitude towards drugs and the significant role of affective-emotional factors suggest that the accep-tance of vaccination probably cannot be facilitated solely with the aid of educational programs. Considering the increasing incidence of affective disorders during the COVID-19 pandemic, the screening of affective and anxiety symptoms and treatment of these disorders could be an important step towards the acceptance of the vaccine. Although psychiatry is not considered as a frontline care unit of the COVID cases, more attention is needed to pay on the availability of mental health services because refuse of vaccine can develop due to affective disorders too.

Isoprostanes evoke contraction of the murine and human detrusor muscle via activation of the thromboxane prostanoid TP receptor and Rho kinase.

Local or systemic inflammation can severely impair urinary bladder functions and contribute to the development of voiding disorders in millions of people worldwide. Isoprostanes are inflammatory lipid mediators that are upregulated in the blood and urine by oxidative stress and may potentially induce detrusor overactivity. The aim of the present study was to investigate the effects and signal transduction of isoprostanes in human and murine urinary bladders in order to provide potential pharmacological targets in detrusor overactivity. Contraction force was measured with a myograph in murine and human urinary bladder smooth muscle (UBSM) ex vivo. Isoprostane 8-iso-PGE2 and 8-iso-PGF2α evoked dose-dependent contraction in the murine UBSM, which was abolished in mice deficient in the thromboxane prostanoid (TP) receptor. The responses remained unaltered after removal of the mucosa or incubation with tetrodotoxin. Smooth muscle-specific deletion of Gα12/13 protein or inhibition of Rho kinase by Y-27632 decreased the contractions. In Gαq/11-knockout mice, responses were reduced and in the presence of Y-27632 abolished completely. In human UBSM, the TP agonist U-46619 evoked dose-dependent contractions. Neither atropine nor the purinergic receptor antagonist pyridoxalphosphate-6-azophenyl-2',4'-disulfonic acid decreased the effect, indicating that TP receptors directly mediate detrusor muscle contraction. 8-iso-PGE2 and 8-iso-PGF2α evoked dose-dependent contraction in the human UBSM, and these responses were abolished by the TP antagonist SQ-29548 and were decreased by Y-27632. Our results indicate that isoprostanes evoke contraction in murine and human urinary bladders, an effect mediated by the TP receptor. The G12/13-Rho-Rho kinase pathway plays a significant role in mediating the contraction and therefore may be a potential therapeutic target in detrusor overactivity.NEW & NOTEWORTHY Voiding disorders affect millions of people worldwide. Inflammation can impair urinary bladder functions and contribute to the development of detrusor overactivity. The effects and signal transduction of inflammatory lipid mediator isoprostanes were studied in human and murine urinary bladders ex vivo. We found that isoprostanes evoke contraction, an effect mediated by thromboxane prostanoid receptors. The G12/13-Rho-Rho kinase signaling pathway plays a significant role in mediating the contraction and therefore may be a potential therapeutic target.

A field study using different technologies to detect calving at a large-scale hungarian dairy farm.

The objective of this study was to investigate three different calving detection systems in order to assess and compare their efficiency. The study was conducted at a large-scale dairy farm involving 54 Holstein-Friesian dairy cows and heifers. Animals were fitted with multiple devices: a rumination measuring device (Ruminact® (RA)), an intravaginal thermometer (Vel'Phone® (VP)) and a tail movement sensor (Moocall® (MC)) 5 to 7 days before expected calving and were removed after parturition. RA detects rumination time (RT) and calculates it in 2-hr intervals. VP detects a decrease in vaginal temperature that might indicate calving within 48 hr and the drop in temperature resulting from the expulsion of the device at calving (EXP message). MC detected increased tail movements and if they persisted for one hour, 1HA message was sent. If they continued during the subsequent hour, then 2HA message was sent. Messages sent by MC within 4 hr before calving (C4) were selected retrospectively as true positives for the prediction of calving, using the significant changes in RT as a baseline. All other messages were categorized as false positive. The mean value of RT decreased in a non-significant manner between interval -22 and -4 before calving. Significant decrease of RT was detectable between the two intervals of -4 and -2 before calving (24.7 ± 18.6 min/2 hr and 14.0 ± 13.0 min/2 hr, respectively). There was no significant difference between RT of primiparous and multiparous animals. EXP messages were accurate (positive predictive value 100%) indicators of the onset of calving. We received on average 12.7 ± 15.2 messages/animal (11.0 ± 10.1 and 16.6 ± 22.2 for cows and heifers, respectively). Positive predictive value was 12.6%. The number of false-positive messages was significantly higher in heifers. All three automatic systems could be used in a large-scale farm environment.

[The first lockdown due to COVID-19 pandemic from the psychiatric patients' perspective: an ambulatory care client experience survey]. / A COVID-19 járvány miatt elrendelt elso veszélyhelyzet a pszichiátriai betegek szemszögébol: gondozói felmérés.

INTRODUCTION: The coronavirus pandemic (COVID-19) required the declaration of a state of emergency in Hungary from 11 March 2020 to 18 June 2020. These governmental actions led to changes in everyday life, implementation of new rules, and reduced access to healthcare. Hospital beds were reserved for emergency use, face-to-face ambulatory care was mainly replaced by telemedicine. In our study we assessed opinion of the patients in two outpatient psychiatric care units in Budapest regarding the state of emergency. METHODS: We enrolled 438 patients in the survey (305 women and 133 men, mean age: 51.9±16.2 years). The patients completed a short questionnaire on a voluntary and anonymous basis following verbal informed consent. The questionnaire was comprised of 10 items and a 12-item "Problem Evaluation Scale" (fear, isolation and healthcare subscales). The comparison of groups was done using general linear models (GLM), pairwise comparison was performed using Tukey's test for post hoc analysis. The data set was analyzed with SPSS software, version 24.0. RESULTS: Up to 34% of enrolled patients believed that their condition worsened during the state of emergency, but 12% of these patients thought that this worsening was not related to the state of emergency. Twice as many patients (12.8%) were concerned about their financial situation than about their health status (6.1%). Loneliness and the implementation of specific regulations didn't cause relevant distress in almost half of the patients, isolation was the most frequently (55.2%) reported difficulty. The worsening of health status was reported more frequently (p=0.001) by the patients younger than 50 years, the sensation of fear was stronger (p=0.045), and they reported more serious adaptation difficulties (p=0.003) than subjects older than 50 years. Isolation caused significantly (p=0.003) more serious distress among women. The abundance of pandemic-related information caused more distress in the case of patients treated for anxiety than participants treated for psychotic disorders (p=0.024). Patients suffering from affective disorders perceived more pronounced feelings of vulnerability compared to patients treated for psychotic disorders (p=0.004). CONCLUSION: Approximately half of the enrolled psychiatric patients was able to adapt to this situation without major difficulties, the other half of this sample was more or less distressed by these circumstances. Unfortunately, depletion of emotional, psychological, social and material resources can be expected during the next stage of the pandemic. As a result, we can expect further worsening of the above presented aspects.

Visual Mismatch and Predictive Coding: A Computational Single-Trial ERP Study.

Predictive coding (PC) posits that the brain uses a generative model to infer the environmental causes of its sensory data and uses precision-weighted prediction errors (pwPEs) to continuously update this model. While supported by much circumstantial evidence, experimental tests grounded in formal trial-by-trial predictions are rare. One partial exception is event-related potential (ERP) studies of the auditory mismatch negativity (MMN), where computational models have found signatures of pwPEs and related model-updating processes. Here, we tested this hypothesis in the visual domain, examining possible links between visual mismatch responses and pwPEs. We used a novel visual "roving standard" paradigm to elicit mismatch responses in humans (of both sexes) by unexpected changes in either color or emotional expression of faces. Using a hierarchical Bayesian model, we simulated pwPE trajectories of a Bayes-optimal observer and used these to conduct a comprehensive trial-by-trial analysis across the time × sensor space. We found significant modulation of brain activity by both color and emotion pwPEs. The scalp distribution and timing of these single-trial pwPE responses were in agreement with visual mismatch responses obtained by traditional averaging and subtraction (deviant-minus-standard) approaches. Finally, we compared the Bayesian model to a more classical change model of MMN. Model comparison revealed that trial-wise pwPEs explained the observed mismatch responses better than categorical change detection. Our results suggest that visual mismatch responses reflect trial-wise pwPEs, as postulated by PC. These findings go beyond classical ERP analyses of visual mismatch and illustrate the utility of computational analyses for studying automatic perceptual processes.SIGNIFICANCE STATEMENT Human perception is thought to rely on a predictive model of the environment that is updated via precision-weighted prediction errors (pwPEs) when events violate expectations. This "predictive coding" view is supported by studies of the auditory mismatch negativity brain potential. However, it is less well known whether visual perception of mismatch relies on similar processes. Here we combined computational modeling and electroencephalography to test whether visual mismatch responses reflected trial-by-trial pwPEs. Applying a Bayesian model to series of face stimuli that violated expectations about color or emotional expression, we found significant modulation of brain activity by both color and emotion pwPEs. A categorical change detection model performed less convincingly. Our findings support the predictive coding interpretation of visual mismatch responses.

Circulating and tissue IMP3 levels are correlated with poor survival in renal cell carcinoma.

Tissue protein expression of IMP3 is emerging as a promising prognostic factor in renal cell carcinoma (RCC). The most commonly used immunohistochemical (IHC) antibody has been criticized for its low specificity. In addition, blood levels of IMP3 have not yet been analyzed in RCC. Therefore, we compared the prognostic performance of two different IMP3 IHC antibodies and assessed the prognostic relevance of IMP3 plasma levels in RCC. IMP3 levels were assessed in an overall number of 425 RCC (344× clear cell [ccRCC], 63× papillary [pRCC], 18× chromophobe [chRCC]) patients in three partly overlapping cohorts. Plasma IMP3 concentrations were determined by ELISA in 98 RCC (79× ccRCC, 15× pRCC, 4× chRCC) patients and 20 controls. IMP3 mRNA expression levels were analyzed in 73 frozen tissue samples (55× ccRCC, 12× pRCC, 6× chRCC), while protein expressions were assessed in 366 FFPE samples (294× ccRCC, 56× pRCC, 16× chRCC) using the M3626 and N-19 antibodies. IMP3 plasma and mRNA expression levels were significantly higher in patients compared to controls and in high-grade compared to low-grade tumors. In addition, IMP3 plasma and tissue protein levels (by M3626) were higher and IMP3 mRNA expression levels tended to be higher in patients with distant metastasis. Multivariate analyses in clear cell RCC revealed high IMP3 plasma concentration and mRNA expression as independent predictors of disease-specific survival. IMP3 immunostainings by M3626 but not by N-19 were independently associated with poor overall and disease-specific survival. High plasma and tissue levels of IMP3 are independently associated with poor RCC prognosis. The applied antibody significantly impacts the prognostic performance of analysis. IMP3 analysis may improve risk-stratification of RCC patients and therefore could help to optimize therapeutic and follow-up decisions.

Ring-Closing Metathesis on Commercial Scale: Synthesis of HCV Protease Inhibitor Simeprevir.

The key macrocyclization step in the synthesis of simeprevir, a hepatitis C virus (HCV) antiviral drug, was studied. N-Boc substitution on the diene precursor changes the site of insertion of the metathesis catalyst and, consequently, the kinetic model of the ring closing metathesis (RCM), enabling a further increase in the macrocyclization efficiency under simulated high dilution (SHD) conditions. NMR of the inserted species of both first and second generation RCM catalysts are reported and discussed.

Investigation of hydrostatic pressure-induced stress preconditioning of boar semen using modified cryopreservation.

The aim of this study was to investigate the effect of varying hydrostatic pressure treatments (HP) on the boar semen quality during the modified cryopreservation. In Experiment I, combinations of pressure level (20/40/80 MPa) and duration of application (40/80/120 min) were used. Before freezing, only the magnitude but not the duration influenced the total (TM%) and progressive motilities (PM%). The 20/40 MPa levels yielded a significant (p < 0.05) improvement compared to control samples (atmospheric), but the 80 MPa was detrimental. The post-freezing-thawing (FT) motilities were influenced significantly by both the HP level and its duration. For TM%, the 40 MPa:120 min gave the highest post-FT result (54.8% ± 3.3%); however, the 40 MPa:80 min (41.0% ± 3.1%) application showed the largest and significant improvement (18.4% ± 3.1%) compared to its control (22.6% ± 3.1%) and compared to the improvement (12.9% ± 3.6%) achieved by 40 MPa:120 min. For PM%, the improvement with the 40 MPa:120 min application was slightly larger than with the 40 MPa:80 min one (15.2% ± 4.2% vs. 13.8% ± 3.3%); furthermore, the difference was not significant. In Experiment II, the 40 MPa:80 min combination was tested at four different stages of the semen handling. By pressurization after dilution with the freezing extender without glycerol, significantly higher post-FT values (TM%, intact acrosome% and head membrane%) were obtained. The two experiments demonstrated possible improvement in post-FT semen quality achievable through the appropriate application of HP to boar semen during cryopreservation.

Validation of a protein panel for the noninvasive detection of recurrent non-muscle invasive bladder cancer.

CONTEXT: About 50-70% of patients with non-muscle invasive bladder cancer (NMIBC) experience relapse of disease. OBJECTIVE: To establish a panel of protein biomarkers incorporated in a multiplexed microarray (BCa chip) and a classifier for diagnosing recurrent NMIBC. MATERIALS AND METHODS: Urine samples from 45 patients were tested. Diagnostic performance was evaluated by receiver operating characteristic (ROC) analysis. RESULTS: A multi biomarker panel (ECadh, IL8, MMP9, EN2, VEGF, past recurrences, BCG therapies and stage at diagnosis) was identified yielding an area under the curve of 0.96. DISCUSSION AND CONCLUSION: This biomarker panel represents a potential diagnostic tool for noninvasive diagnosis of recurrent NMIBC.

The nucleotidohydrolases DCTPP1 and dUTPase are involved in the cellular response to decitabine.

Decitabine (5-aza-2'-deoxycytidine, aza-dCyd) is an anti-cancer drug used clinically for the treatment of myelodysplastic syndromes and acute myeloid leukaemia that can act as a DNA-demethylating or genotoxic agent in a dose-dependent manner. On the other hand, DCTPP1 (dCTP pyrophosphatase 1) and dUTPase are two 'house-cleaning' nucleotidohydrolases involved in the elimination of non-canonical nucleotides. In the present study, we show that exposure of HeLa cells to decitabine up-regulates the expression of several pyrimidine metabolic enzymes including DCTPP1, dUTPase, dCMP deaminase and thymidylate synthase, thus suggesting their contribution to the cellular response to this anti-cancer nucleoside. We present several lines of evidence supporting that, in addition to the formation of aza-dCTP (5-aza-2'-deoxycytidine-5'-triphosphate), an alternative cytotoxic mechanism for decitabine may involve the formation of aza-dUMP, a potential thymidylate synthase inhibitor. Indeed, dUTPase or DCTPP1 down-regulation enhanced the cytotoxic effect of decitabine producing an accumulation of nucleoside triphosphates containing uracil as well as uracil misincorporation and double-strand breaks in genomic DNA. Moreover, DCTPP1 hydrolyses the triphosphate form of decitabine with similar kinetic efficiency to its natural substrate dCTP and prevents decitabine-induced global DNA demethylation. The data suggest that the nucleotidohydrolases DCTPP1 and dUTPase are factors involved in the mode of action of decitabine with potential value as enzymatic targets to improve decitabine-based chemotherapy.

[Psychiatric disorders in epilepsy: clinical phenomena]. / Pszichiátriai zavarok epilepsziában: klinikai jelenségek.

The frequent psychiatric comorbidity in epilepsy compromises the quality of life of those affected, further increasing their suicide risk and the high burden of stigmatisation. Out of adulthood epilepsies, mesio-temporal lobe epilepsy carries the highest cognitive and psychiatric risk, making mental health comorbidities its inherent features. The pathomechanism of the epilepsy-related mental health symptoms and conditions appears to be related to undetected subclinical seizures, postictal exhaustion and inhibition, and the non-REM sleep related interictal epileptic activity. In our work we try to present some of the typical psychiatric syndromes and conditions seen in epilepsy. We aim to highlight the difficulties of treatment on the borderline of neurology and psychiatry.