Miracle-dz/UniFAR-FTUs huggingface dataset:
A Null Mutation in the Gene Encoding the Herpes Simplex Virus Type 1 UL37 Polypeptide Abrogates Virus Maturation

ABSTRACT The tegument is an integral and essential structural component of the herpes simplex virus type 1 (HSV-1) virion. The UL37 open reading frame of HSV-1 encodes a 120-kDa virion polypeptide which is a resident of the tegument. To analyze the function of the UL37-encoded polypeptide a null mutation was generated in the gene encoding this protein. In order to propagate this mutant virus, transformed cell lines that express the UL37 gene product in trans were produced. The null mutation was transferred into the virus genome using these complementing cell lines. A mutant virus designated KΔUL37 was isolated based on its ability to form plaques on the complementing cell line but not on nonpermissive (noncomplementing) Vero cells. This virus was unable to grow in Vero cells; therefore, UL37 encodes an essential function of the virus. The mutant virus KΔUL37 produced capsids containing DNA as judged by sedimentation analysis of extracts derived from infected Vero cells. Therefore, the UL37 gene product is not required for DNA cleavage or packaging. The UL37 mutant capsids were tagged with the smallest capsid protein, VP26, fused to green fluorescent protein. This fusion protein decorates the capsid shell and consequently the location of the capsid and the virus particle can be visualized in living cells. Late in infection, KΔUL37 capsids were observed to accumulate at the periphery of the nucleus as judged by the concentration of fluorescence around this organelle. Fluorescence was also observed in the cytoplasm in large puncta. Fluorescence at the plasma membrane, which indicated maturation and egress of virions, was observed in wild-type-infected cells but was absent in KΔUL37-infected cells. Ultrastructural analysis of thin sections of infected cells revealed clusters of DNA-containing capsids in the proximity of the inner nuclear membrane. Occasionally enveloped capsids were observed between the inner and outer nuclear membranes. Clusters of unenveloped capsids were also observed in the cytoplasm of KΔUL37-infected cells. Enveloped virions, which were observed in the cytoplasm of wild-type-infected cells, were never detected in the cytoplasm of KΔUL37-infected cells. Crude cell fractionation of infected cells using detergent lysis demonstrated that two-thirds of the UL37 mutant particles were associated with the nuclear fraction, unlike wild-type particles, which were predominantly in the cytoplasmic fraction. These data suggest that in the absence of UL37, the exit of capsids from the nucleus is slowed. UL37 mutant particles can participate in the initial envelopment at the nuclear membrane, although this process may be impaired in the absence of UL37. Furthermore, the naked capsids deposited in the cytoplasm are unable to progress further in the morphogenesis pathway, which suggests that UL37 is also required for egress and reenvelopment. Therefore, the UL37 gene product plays a key role in the early stages of the maturation pathway that give rise to an infectious virion.
Drinking Motives as Mediators of Social Anxiety and Hazardous Drinking Among College Students

Although social anxiety (SA) and alcohol use disorders commonly co-occur, the relationship between these variables in college populations has been inconsistent. The present study tested the hypothesized model that negatively reinforcing, but not positively reinforcing, drinking motives (or reasons for drinking) would mediate the association between SA and three aspects of hazardous drinking (quantity/frequency, consequences, and dependence symptoms) in an ethnically diverse sample of college drinkers (N = 817; mean age = 19.9 years, range = 18–29). Structural equation modeling (SEM) results using the asymmetrical distribution of products test indicated that coping motives partially mediated the relationship between SA and negative consequences and dependence symptoms but not the quantity/frequency outcome. Contrary to the hypothesized model, conformity motives did not mediate the association between SA and hazardous drinking. As expected, positive reinforcement motives did not mediate the SA–hazardous drinking association. Multigroup SEM analyses revealed that the mediation models did not differ for men (n = 215) and women (n = 602). Overall, the present findings support extant research and theoretical models regarding the mediating role of coping motives in the relationship between SA and problem drinking, suggesting a potential pathway for the development and maintenance of SA and alcohol use disorder comorbidity. Such findings could contribute to improved intervention programs by targeting coping drinking motives and building coping skills.
Respiratory Symptoms and Ventilatory Function in Never- Smoking Males Working in Dusty Occupations

Abstract AIM: To assess the effect of occupational exposure on respiratory symptoms and ventilatory capacity in never-smoking male workers exposed to mineral or organic dusts. MATERIAL AND METHODS: We performed a cross-sectional study including 138 never-smoking male workers exposed to mineral or organic dust (34 construction workers, 32 furniture manufacturers, 37 agricultural workers, and 35 bakers) and 35 unexposed controls (office workers). Evaluation of all study subjects included completion of a questionnaire and spirometric measurements. RESULTS: The prevalence of the overall respiratory symptoms in the last 12 months was higher in dusty occupation workers than its prevalence in office workers. Statistically significant difference was found between the prevalence of cough in construction workers, agricultural workers and bakers, as well as between the prevalence of phlegm in construction workers, furniture manufacturers and agricultural workers as compared to its prevalence in office workers. The mean values of spirometric parameters were lower in all groups of exposed workers as compared to their mean values in office workers with statistical significance for all measured parameters in construction workers and furniture manufacturers, as well as for small airways indices in agricultural workers and bakers. CONCLUSION: Our findings indicate significant effect of occupational exposure on respiratory symptoms and ventilatory capacity impairment in workers exposed to mineral or organic dusts.
Inhibition of the nitrosothiol production of cultured osteoarthritic chondrocytes by rhein, cortisol and diclofenac.

OBJECTIVE Nitric oxide (NO degrees ) is a free molecule produced by NO synthases which acts as a mediator in inflammatory processes. NO degrees can react with thiol groups of proteins to produce nitrosothiols. Increased concentrations of these bioactive compounds have been found in sera and synovial fluids from patients with osteoarthritis (OA). The aim of this study was to assess the ability of human osteoarthritic chondrocytes to synthesize nitrosothiols and to compare the in vitro effects of rhein, cortisol and diclofenac on nitrosothiol and nitrite production. METHODS Osteoarthritic chondrocytes were incubated for 24 h with 1 ng/ml of recombinant human interleukin-1beta (IL-1beta) in the presence or absence of rhein (1.3x10(-5) M, 6.5x10(-6) M, or 1.3x10(-6) M), cortisol (10(-5) M) or diclofenac (10(-5) M or 10(-6) M). Nitrite levels were measured in cell supernatants by the Griess method; nitrosothiol levels were determined in supernatants and cellular lysates by fluorimetry. RESULTS At the basal level, nitrosothiols represented 80% of the total of nitrite and nitrosothiol production. After IL-1beta stimulation, NO degrees production was highly increased in the supernatants (45-fold increase in nitrite, 60-fold increase in nitrosothiols) as well as in cell lysates (35-fold increase in nitrosothiols). Rhein caused a dose-dependent decrease in nitrosothiol and nitrite production. In comparison, diclofenac (10(-5) M) moderately decreased nitrite and nitrosothiol levels in the supernatants but had no effect on lysate nitrosothiol. Cortisol had no significant effect on NO degrees production. CONCLUSIONS The IL-1beta stimulation increased nitrosothiol production by osteoarthritic chondrocytes. These results demonstrate the need to measure nitrosothiol as well as nitrite production. Rhein inhibited the IL-1beta induced NO degrees production, and may be a suitable treatment for osteoarthritis.
Correspondence regarding SNX8 haploinsufficiency and its potential for cardiac anomalies including tetralogy of Fallot

Correspondence Regarding SNX8 Haploinsufficiency and its Potential for Cardiac Anomalies Including Tetralogy of Fallot Rena J. Vanzo,* Megan M. Martin, Mallory R. Sdano, Kathie Teta, and Sarah T. South Lineagen—Genetic Counseling, Salt Lake City, Utah Pediatric & Adolescent Neurodevelopmental Associates, Atlanta, Georgia ARUP Laboratories—Cytogenetics, Salt Lake City, Utah University of Utah, Department of Pediatrics, Salt Lake City, Utah University of Utah, Department of Pathology, Salt Lake City, Utah
Should production and trading activities be separated ?

Firms often supplement profits from production by trading in the item that they produce such as oil or electricity. We ask whether firms that use a commodity in production and also trade in that commodity should keep their production and trading activities together in one corporate entity, or divide them into separate entities. What is the connection between this choice and the capital structure decision? Results indicate that firms engaging in both production and trading of a commodity benefit from diversification, from flexibility in the allocation of capital that improves risk management, and from information spillovers between production and trading. Separation is preferred when trading revenues are risky, sensitive to contamination by losses in the production arm, and sensitive to the effects of leverage.
Identification and Validation of Small Molecules That Enhance Recombinant Adeno-associated Virus Transduction following High-Throughput Screens

ABSTRACT While the recent success of adeno-associated virus (AAV)-mediated gene therapy in clinical trials is promising, challenges still face the widespread applicability of recombinant AAV(rAAV). A major goal is to enhance the transduction efficiency of vectors in order to achieve therapeutic levels of gene expression at a vector dose that is below the immunological response threshold. In an attempt to identify novel compounds that enhance rAAV transduction, we performed two high-throughput screens comprising 2,396 compounds. We identified 13 compounds that were capable of enhancing transduction, of which 12 demonstrated vector-specific effects and 1 could also enhance vector-independent transgene expression. Many of these compounds had similar properties and could be categorized into five groups: epipodophyllotoxins (group 1), inducers of DNA damage (group 2), effectors of epigenetic modification (group 3), anthracyclines (group 4), and proteasome inhibitors (group 5). We optimized dosing for the identified compounds in several immortalized human cell lines as well as normal diploid cells. We found that the group 1 epipodophyllotoxins (teniposide and etoposide) consistently produced the greatest transduction enhancement. We also explored transduction enhancement among single-stranded, self-complementary, and fragment vectors and found that the compounds could impact fragmented rAAV2 transduction to an even greater extent than single-stranded vectors. In vivo analysis of rAAV2 and all of the clinically relevant compounds revealed that, consistent with our in vitro results, teniposide exhibited the greatest level of transduction enhancement. Finally, we explored the capability of teniposide to enhance transduction of fragment vectors in vivo using an AAV8 capsid that is known to exhibit robust liver tropism. Consistent with our in vitro results, teniposide coadministration greatly enhanced fragmented rAAV8 transduction at 48 h and 8 days. This study provides a foundation based on the rAAV small-molecule screen methodology, which is ideally used for more-diverse libraries of compounds that can be tested for potentiating rAAV transduction. IMPORTANCE This study seeks to enhance the capability of adeno-associated viral vectors for therapeutic gene delivery applicable to the treatment of diverse diseases. To do this, a comprehensive panel of FDA-approved drugs were tested in human cells and in animal models to determine if they increased adeno-associated virus gene delivery. The results demonstrate that particular groups of drugs enhance adeno-associated virus gene delivery by unknown mechanisms. In particular, the enhancement of gene delivery was approximately 50 to 100 times better with than without teniposide, a compound that is also used as chemotherapy for cancer. Collectively, these results highlight the potential for FDA-approved drug enhancement of adeno-associated virus gene therapy, which could result in safe and effective treatments for diverse acquired or genetic diseases.
Influence of subthalamic deep brain stimulation versus levodopa on motor perseverations in Parkinson's disease

Patients with Parkinson's disease (PD) show impairment in generating random motor sequences reflecting a higher order motor deficit in set‐shifting and suppression of perseverative behavior. The impact of deep brain stimulation (DBS) of the subthalamic nucleus (STN) on motor perseverations has not yet been elucidated. In 35 patients with PD, we evaluated the effect of STN‐DBS and levodopa on motor perseverations using the Vienna perseveration task. The task was performed 6 months after implantation of stimulation electrodes in the following three conditions: Stimulation off/medication off (Stim OFF/Med OFF), Stim ON/Med OFF, and Stim OFF/Med ON. Perseverations were measured by redundancy of second order (R2) with higher values indicating more severe perseverations. ANCOVA analysis revealed that influence of STN‐DBS on R2 significantly depended on R2 severity during Stim OFF/Med OFF (F = 4.69, P = 0.035). Accordingly, we classified patients with PD into two groups based on the R2 value during off treatment. In patients with mild perseveration (R2 < 35) neither STN‐DBS nor levodopa changed perseverations. By contrast, in patients with severe perseveration (R2 > 35), STN‐DBS significantly reduced R2 by 9.7 ± 2.6 (P < 0.001) whereas levodopa had no impact (R2 reduction 3.7 ± 1.6, P = 0.081). This demonstrates that STN‐DBS, by reducing motor perseveration, influences higher order aspects of motor behavior of patients with PD. © 2009 Movement Disorder Society
High HIV, hepatitis C and sexual risks among drug-using men who have sex with men in northern Thailand

Background:Men who have sex with men (MSM) and who use drugs have shown high HIV risks in Europe, and the Americas. We investigated MSM–drug user demographics, HIV sexual and drug use risks and behaviors in Chiang Mai, northern Thailand to identify prevention targets. Methods:A total of 2005 males aged 13 years and older were enrolled during inpatient drug treatment from 1999–2000 and assessed for HIV, hepatitis C virus (HCV), syphilis, and for demographics and risks by questionnaire. Data were analyzed using χ2 and multiple logistic regression to estimate odds ratios (ORs) and 95% confidence intervals (CIs). Results:Of 2005 males in treatment, 1752 (87.4%) had ever had sex, and 66 of 1752 (3.8%) reported ever having sex with another man; mostly Katoey (transgendered male) partners. MSM had higher HIV rates (OR, 2.32; 95% CI, 1.36–3.96) and were younger (P = 0.002); more likely to be Thai (P < 0.0001); better educated (P < 0.0001); had more lifetime sex partners (P < 0.0001), more female partners (P = 0.002), more female paid partners (P < 0.0001), and been paid for sex (P < 0.0001). MSM were more likely to have ever injected (P < 0.0001), sold drugs, been in prison, injected in prison, used heroin, and to have HCV (OR, 2.59; 95% CI, 1.55–4.34). Conclusions:Northern Thai MSM–drug users are at high HIV and HCV risk. In addition to sex risks with men, they have more sex with women and sex workers than other men, which fits Thai MSM patterns but not Western ones. Prevention must take into account their high rates of substance use and multiple partner types.
Phrenic nerve diabetic neuropathy in rats: unmyelinated fibers morphometry

We have demonstrated that phrenic nerves' large myelinated fibers in streptozotocin (STZ)‐induced diabetic rats show axonal atrophy, which is reversed by insulin treatment. However, studies on structural abnormalities of the small myelinated and the unmyelinated fibers in the STZ‐model of neuropathy are limited. Also, structural changes in the endoneural vasculature are not clearly described in this model and require detailed study. We have undertaken morphometric studies of the phrenic nerve in insulin‐treated and untreated STZ‐diabetic rats and non‐diabetic control animals over a 12‐week period. The presence of neuropathy was assessed by means of transmission electron microscopy, and morphometry of the unmyelinated fibers was performed. The most striking finding was the morphological evidence of small myelinated fiber neuropathy due to the STZ injection, which was not protected or reversed by conventional insulin treatment. This neuropathy was clearly associated with severe damage of the endoneural vessels present on both STZ groups, besides the insulin treatment. The STZ‐diabetes model is widely used to investigate experimental diabetic neuropathies, but few studies have performed a detailed assessment of either unmyelinated fibers or capillary morphology in this animal model. The present study adds useful information for further investigations on the ultrastructural basis of nerve function in diabetes.
A comparative analysis of pre- and post-dialysis albumin as indicators of nutritional and morbi-mortality risks in haemodialysis patients.

INTRODUCTION Pre-dialysis albumin is likely to be falsely low due to a dilution effect, making its usefulness in assessing protein status questionable. OBJEVTIVES: The purpose of this study was to assess whether post-dialysis albumin would be a better marker of malnutrition and risk of mortality, when compared to pre-dialysis albumin. METHODS We evaluated the correlation between pre- and post-dialysis albumin and the following parameters: body mass index (BMI), adequacy of muscle arm circumference (MAC) and tricipital skinfold (TS) to the 50th percentile (P50), C-reactive protein (CRP), phase angle (PA), protein equivalent of nitrogen appearance (PNA), the Kt/V index of dialysis adequacy, and the hydration status (Pearson's correlation coefficient). Agreement in the nutritional status according to pre- and post-dialysis (post-HD) albumin and PA was estimated according to the Kappa (K) coefficient (Bland-Altman). RESULTS A total of 58 haemodialysis (HD) patients were included in this study (30 female; mean age: 49 years). BMI, PA and CRP had a significant correlation with pre- and post-HD albumin, while MAC and PNA correlated only with post-HD albumin. Agreement in the diagnosis of malnutrition according to PA < 5 and pre- and post-HD albumin < 3.2 g/dL was regular (K = 0.432). When using an albumin cut-off value of 3.7 g/dL for malnutrition (mild malnutrition or risk of malnutrition), the diagnosis was concordant only in the post-HD period (K = 0.544). CONCLUSIONS Post-dialysis albumin levels may be a better marker of protein status and mortality risk in cases of mild malnutrition or risk of malnutrition and in patients with low/medium mortality risk. Pre-dialysis fluid overload may be a confounding factor when evaluating albumin levels.
Ultrafast real-time visualization of active site flexibility of flavoenzyme thymidylate synthase ThyX

In many bacteria the flavoenzyme thymidylate synthase ThyX produces the DNA nucleotide deoxythymidine monophosphate from dUMP, using methylenetetrahydrofolate as carbon donor and NADPH as hydride donor. Because all three substrates bind in close proximity to the catalytic flavin adenine dinucleotide group, substantial flexibility of the ThyX active site has been hypothesized. Using femtosecond time-resolved fluorescence spectroscopy, we have studied the conformational heterogeneity and the conformational interconversion dynamics in real time in ThyX from the hyperthermophilic bacterium Thermotoga maritima. The dynamics of electron transfer to excited flavin adenine dinucleotide from a neighboring tyrosine residue are used as a sensitive probe of the functional dynamics of the active site. The fluorescence decay spanned a full three orders of magnitude, demonstrating a very wide range of conformations. In particular, at physiological temperatures, multiple angstrom cofactor-residue displacements occur on the picoseconds timescale. These experimental findings are supported by molecular dynamics simulations. Binding of the dUMP substrate abolishes this flexibility and stabilizes the active site in a configuration where dUMP closely interacts with the flavin cofactor and very efficiently quenches fluorescence itself. Our results indicate a dynamic selected-fit mechanism where binding of the first substrate dUMP at high temperature stabilizes the enzyme in a configuration favorable for interaction with the second substrate NADPH, and more generally have important implications for the role of active site flexibility in enzymes interacting with multiple poly-atom substrates and products. Moreover, our data provide the basis for exploring the effect of inhibitor molecules on the active site dynamics of ThyX and other multisubstrate flavoenzymes.
Transcriptional regulation of CacyBP/SIP gene and the influence of increased CacyBP/SIP level on gene expression pattern in colorectal cancer HCT116 cells

The CacyBP/SIP protein is expressed at a particularly high level in brain, spleen, and various tumors. In this work, we have studied transcriptional regulation of the CacyBP/SIP gene and the influence of increased CacyBP/SIP level on gene expression in colorectal cancer HCT116 cells. We have shown that E2F1, EGR1, and CREB transcription factors bind to the CacyBP/SIP gene promoter and stimulate transcription of CacyBP/SIP gene. The role of CREB was further confirmed by the observation that forskolin, a strong activator of CREB phosphorylation/activity, increased CacyBP/SIP gene promoter activity. Moreover, we have shown that CREB dominant negative mutants, CREB133 and KCREB, inhibits CacyBP/SIP promoter activity. To check the biological significance of increased CacyBP/SIP expression/level we have applied RNA microarray analysis and have found that upregulation of CacyBP/SIP entails changes in mRNA level of many genes involved, among others, in immune processes. © 2017 IUBMB Life, 70(1):50–59, 2018