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Objective: Rural areas account for approximately 6% of AIDS cases in the United States. Many HIV-infected persons in rural areas live with elevated levels of psychiatric distress, suicidal ideation, and loneliness. This pilot study tested whether brief interpersonal psychotherapy delivered via telephone could reduce psychiatric distress among persons living with HIV-AIDS in rural areas in the United States.Methods: Seventy-nine participants were assigned randomly to a usual care control condition or to a six-session, telephone-delivered, interpersonal psychotherapy intervention (hereafter referred to as the teletherapy group); participants in the teletherapy group continued to receive standard services available to them in the community. Participants completed self-administered surveys pre- and postintervention that assessed depressive and psychiatric symptoms, perceptions of loneliness, and social support.Results: Participants in the teletherapy group evidenced greater reductions in depressive symptoms and in overall levels of psychiatric distress, compared with those in the control group. Nearly one-third of teletherapy participants reported clinically meaningful reductions in psychiatric distress from pre- to postintervention.Conclusions: The telephone-delivered interpersonal therapy intervention showed potential to reduce depressive and psychiatric symptoms among HIV-infected persons in rural areas. On the basis of these encouraging findings, additional research examining this intervention with this clinical population is warranted. [ABSTRACT FROM AUTHOR]

Synaptic vesicles are organelles with a precisely defined protein and lipid composition 1,2 , yet the molecular mechanisms for the biogenesis of synaptic vesicles are mainly unknown. Here we discovered a well-defined interface between the synaptic vesicle V-ATPase and synaptophysin by in situ cryo-electron tomography and single-particle cryo-electron microscopy of functional synaptic vesicles isolated from mouse brains 3 . The synaptic vesicle V-ATPase is an ATP-dependent proton pump that establishes the proton gradient across the synaptic vesicle, which in turn drives the uptake of neurotransmitters 4,5 . Synaptophysin 6 and its paralogues synaptoporin 7 and synaptogyrin 8 belong to a family of abundant synaptic vesicle proteins whose function is still unclear. We performed structural and functional studies of synaptophysin-knockout mice, confirming the identity of synaptophysin as an interaction partner with the V-ATPase. Although there is little change in the conformation of the V-ATPase upon interaction with synaptophysin, the presence of synaptophysin in synaptic vesicles profoundly affects the copy number of V-ATPases. This effect on the topography of synaptic vesicles suggests that synaptophysin assists in their biogenesis. In support of this model, we observed that synaptophysin-knockout mice exhibit severe seizure susceptibility, suggesting an imbalance of neurotransmitter release as a physiological consequence of the absence of synaptophysin., (© 2024. The Author(s).)

Objectives: Treatment of carious primary molars is always indicated, especially on young children; however, there are no clear guidelines that precisely explain the best treatment approach for Class II carious molars with marginal breakdown (International Caries Detection and Assessment System [ICDAS] 5). The objective of this prospective observational clinical study was to assess the efficacy of three restorative techniques in treating ICDAS 5 Class II lesions in primary molars: compomer fillings (CF), preformed metal crowns (PMC), and pulpotomy and conventional preformed metal crowns (PMC+P). The secondary goal was to evaluate the impact of some cofactors on the course of treatment., Method and Materials: Overall, 92 children (female, n = 50, 54.3%; male, n = 42, 45.7%) aged 2 to 9 years old (mean age = 5.9 ± 1.9 years) with 166 treated teeth were included. The average number of decayed, missing, or filled teeth (d3mft) of the whole sample was 8.0 ± 3.4. The distribution of the sample according to type of treatment was CF = 53 (31.9%), PMC = 64 (38.6%), and PMC+P = 49 (29.5%). Paired t test, nonparametric Friedman ANOVA test, and decision tree analysis were used as the basis for the statistics., Results: After 12 months, data from 75.8% (72/95) treated patients, corresponding to 62.0% (103/166) of the treated teeth (CF = 42/53, 79.2%; PMC = 38/64, 59.4%; PMC+P = 23/49, 46.9%) were available for analysis. The mean patients age was 6.8 ± 1.8 years; 32 (47.1%) boys and 36 (52.9%) girls. The mean d3mft of the remaining sample was 7.8 ± 3.35. PMC and PMC+P arms showed the highest success rates (> 91%) as compared to the CF arm, which showed the lowest success rates (61.9%), with 9/42 teeth of the CF group (21.4%) presenting with minor failures, and 7/42 teeth (16.7%) with major failures (P < .0001)., Conclusion: According to the decision tree analysis, PMC and PMC+P had a success rate of 99%, whereas CF had a success rate of only 69%. Some cofactors (treatment decision, Approximal Plaque Index, and tooth number) had a higher impact on the decision tree analysis than others (age, dmfs, and dmft values), especially when the treatment selection was CF. In future studies it is necessary to examine the impact of other cofactors on the outcomes of conventional fillings using a larger sample size.

TsaC/Sua5 family of enzymes catalyzes the first step in the synthesis of N6-threonyl-carbamoyl adenosine (t 6 A) one of few truly ubiquitous tRNA modifications important for translation accuracy. TsaC is a single domain protein while Sua5 proteins contains a TsaC-like domain and an additional SUA5 domain of unknown function. The emergence of these two proteins and their respective mechanisms for t 6 A synthesis remain poorly understood. Here, we performed phylogenetic and comparative sequence and structure analysis of TsaC and Sua5 proteins. We confirm that this family is ubiquitous but the co-occurrence of both variants in the same organism is rare and unstable. We further find that obligate symbionts are the only organisms lacking sua5 or tsaC genes. The data suggest that Sua5 was the ancestral version of the enzyme while TsaC arose via loss of the SUA5 domain that occurred multiple times in course of evolution. Multiple losses of one of the two variants in combination with horizontal gene transfers along a large range of phylogenetic distances explains the present day patchy distribution of Sua5 and TsaC. The loss of the SUA5 domain triggered adaptive mutations affecting the substrate binding in TsaC proteins. Finally, we identified atypical Sua5 proteins in Archaeoglobi archaea that seem to be in the process of losing the SUA5 domain through progressive gene erosion. Together, our study uncovers the evolutionary path for emergence of these homologous isofunctional enzymes and lays the groundwork for future experimental studies on the function of TsaC/Sua5 proteins in maintaining faithful translation., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Pichard-Kostuch, Da Cunha, Oberto, Sauguet and Basta.)

In Archaea and Eukaryotes, the synthesis of a universal tRNA modification, N 6 -threonyl-carbamoyl adenosine (t 6 A), is catalyzed by the KEOPS complex composed of Kae1, Bud32, Cgi121, and Pcc1. A fifth subunit, Gon7, is found only in Fungi and Metazoa. Here, we identify and characterize a fifth KEOPS subunit in Archaea. This protein, dubbed Pcc2, is a paralog of Pcc1 and is widely conserved in Archaea. Pcc1 and Pcc2 form a heterodimer in solution, and show modest sequence conservation but very high structural similarity. The five-subunit archaeal KEOPS does not form dimers but retains robust tRNA binding and t 6 A synthetic activity. Pcc2 can substitute for Pcc1 but the resulting KEOPS complex is inactive, suggesting a distinct function for the two paralogs. Comparative sequence and structure analyses point to a possible evolutionary link between archaeal Pcc2 and eukaryotic Gon7. Our work indicates that Pcc2 regulates the oligomeric state of the KEOPS complex, a feature that seems to be conserved from Archaea to Eukaryotes., (© 2023. The Author(s).)

DNA gyrase is a type II topoisomerase with the unique capacity to introduce negative supercoiling in DNA. In bacteria, DNA gyrase has an essential role in the homeostatic regulation of supercoiling. While ubiquitous in bacteria, DNA gyrase was previously reported to have a patchy distribution in Archaea but its emergent function and evolutionary history in this domain of life remains elusive. In this study, we used phylogenomic approaches and an up-to date sequence dataset to establish global and archaea-specific phylogenies of DNA gyrases. The most parsimonious evolutionary scenario infers that DNA gyrase was introduced into the lineage leading to Euryarchaeal group II via a single horizontal gene transfer from a bacterial donor which we identified as an ancestor of Gracilicutes and/or Terrabacteria. The archaea-focused trees indicate that DNA gyrase spread from Euryarchaeal group II to some DPANN and Asgard lineages via rare horizontal gene transfers. The analysis of successful recent transfers suggests a requirement for syntropic or symbiotic/parasitic relationship between donor and recipient organisms. We further show that the ubiquitous archaeal Topoisomerase VI may have co-evolved with DNA gyrase to allow the division of labor in the management of topological constraints. Collectively, our study reveals the evolutionary history of DNA gyrase in Archaea and provides testable hypotheses to understand the prerequisites for successful establishment of DNA gyrase in a naive archaeon and the associated adaptations in the management of topological constraints., (© The Author(s) 2022. Published by Oxford University Press on behalf of Society for Molecular Biology and Evolution.)

The proteasome system allows the elimination of functional or structurally impaired proteins. This includes the degradation of nascent peptides. In Archaea, how the proteasome complex interacts with the translational machinery remains to be described. Here, we characterized a small orphan protein, Q9UZY3 (UniProt ID), conserved in Thermococcales. The protein was identified in native pull-down experiments using the proteasome regulatory complex (proteasome-activating nucleotidase [PAN]) as bait. X-ray crystallography and small-angle X-ray scattering experiments revealed that the protein is monomeric and adopts a β-barrel core structure with an oligonucleotide/oligosaccharide-binding (OB)-fold, typically found in translation elongation factors. Mobility shift experiment showed that Q9UZY3 displays transfer ribonucleic acid (tRNA)-binding properties. Pull-downs, co-immunoprecipitation and isothermal titration calorimetry (ITC) studies revealed that Q9UZY3 interacts in vitro with PAN. Native pull-downs and proteomic analysis using different versions of Q9UZY3 showed that the protein interacts with the assembled PAN-20S proteasome machinery in Pyrococcus abyssi (Pa) cellular extracts. The protein was therefore named Pbp11, for Proteasome-Binding Protein of 11 kDa. Interestingly, the interaction network of Pbp11 also includes ribosomal proteins, tRNA-processing enzymes and exosome subunits dependent on Pbp11's N-terminal domain that was found to be essential for tRNA binding. Together these data suggest that Pbp11 participates in an interface between the proteasome and the translational machinery., (© 2022 The Authors. Molecular Microbiology published by John Wiley & Sons Ltd.)

Binding of the neurotransmitter acetylcholine to its receptors on muscle fibers depolarizes the membrane and thereby triggers muscle contraction. We sought to understand at the level of three-dimensional structure how agonists and antagonists alter nicotinic acetylcholine receptor conformation. We used the muscle-type receptor from the Torpedo ray to first define the structure of the receptor in a resting, activatable state. We then determined the receptor structure bound to the agonist carbachol, which stabilizes an asymmetric, closed channel desensitized state. We find conformational changes in a peripheral membrane helix are tied to recovery from desensitization. To probe mechanisms of antagonism, we obtained receptor structures with the active component of curare, a poison arrow toxin and precursor to modern muscle relaxants. d-Tubocurarine stabilizes the receptor in a desensitized-like state in the presence and absence of agonist. These findings define the transitions between resting and desensitized states and reveal divergent means by which antagonists block channel activity of the muscle-type nicotinic receptor., (© 2022. The Author(s), under exclusive licence to Springer Nature America, Inc.)

Electron tomography of the chemical synapse provides important architectural information regarding the organization of synaptic organelles including synaptic vesicles, Nissl bodies, and early endosomes. Here, we describe methods for the preparation of select murine brain regions for high-pressure freezing, freeze substitution, and EM tomographic analysis of synaptic structures. The method uses fresh brain slices prepared using a vibratome and biopsy punches to collect specific brain regions of interest suitable for subsequent preservation and EM tomographic imaging., (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

In all cells, DNA topoisomerases dynamically regulate DNA supercoiling allowing essential DNA processes such as transcription and replication to occur. How this complex system emerged in the course of evolution is poorly understood. Intriguingly, a single horizontal gene transfer event led to the successful establishment of bacterial gyrase in Archaea, but its emergent function remains a mystery. To better understand the challenges associated with the establishment of pervasive negative supercoiling activity, we expressed the gyrase of the bacterium Thermotoga maritima in a naïve archaeon Thermococcus kodakarensis which naturally has positively supercoiled DNA. We found that the gyrase was catalytically active in T. kodakarensis leading to strong negative supercoiling of plasmid DNA which was stably maintained over at least eighty generations. An increased sensitivity of gyrase-expressing T. kodakarensis to ciprofloxacin suggested that gyrase also modulated chromosomal topology. Accordingly, global transcriptome analyses revealed large scale gene expression deregulation and identified a subset of genes responding to the negative supercoiling activity of gyrase. Surprisingly, the artificially introduced dominant negative supercoiling activity did not have a measurable effect on T. kodakarensis growth rate. Our data suggest that gyrase can become established in Thermococcales archaea without critically interfering with DNA transaction processes., (© The Author(s) 2021. Published by Oxford University Press on behalf of Nucleic Acids Research.)

Compromised protein homeostasis underlies accumulation of plaques and tangles in Alzheimer's disease (AD). To observe protein turnover at early stages of amyloid beta (Aβ) proteotoxicity, we performed pulse-chase proteomics on mouse brains in three genetic models of AD that knock in alleles of amyloid precursor protein (APP) prior to the accumulation of plaques and during disease progression. At initial stages of Aβ accumulation, the turnover of proteins associated with presynaptic terminals is selectively impaired. Presynaptic proteins with impaired turnover, particularly synaptic vesicle (SV)-associated proteins, have elevated levels, misfold in both a plaque-dependent and -independent manner, and interact with APP and Aβ. Concurrent with elevated levels of SV-associated proteins, we found an enlargement of the SV pool as well as enhancement of presynaptic potentiation. Together, our findings reveal that the presynaptic terminal is particularly vulnerable and represents a critical site for manifestation of initial AD etiology. A record of this paper's transparent peer review process is included in the Supplemental Information., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)

The synaptotagmin (syt) proteins have been widely studied for their role in regulating fusion of intracellular vesicles with the plasma membrane. Here we report that syt-17, an unusual isoform of unknown function, plays no role in exocytosis, and instead plays multiple roles in intracellular membrane trafficking. Syt-17 is localized to the Golgi complex in hippocampal neurons, where it coordinates import of vesicles from the endoplasmic reticulum to support neurite outgrowth and facilitate axon regrowth after injury. Further, we discovered a second pool of syt-17 on early endosomes in neurites. Loss of syt-17 disrupts endocytic trafficking, resulting in the accumulation of excess postsynaptic AMPA receptors and defective synaptic plasticity. Two distinct pools of syt-17 thus control two crucial, independent membrane trafficking pathways in neurons. Function of syt-17 appears to be one mechanism by which neurons have specialized their secretory and endosomal systems to support the demands of synaptic communication over sprawling neurite arbors.

Objectives: To assess the survival of patients with cervical cancer (CC). Since the recommendations concerning cervical cancer management adopted by Polish medical societies do not differ significantly from the ESGO or non-European guidelines, and the fact that evaluation of the system for CC treatment in Poland, as well as the mortality rate of Polish women with CC, which is 70% higher than the average for European Union (EU) countries, justifies the hypothesis that treatment of CC in Poland deviates from the Polish and international recommendations. This article puts forward the current management of cervical cancer in Poland and discusses it in the context of ASCO guidelines., Material and Methods: A survey retrospective multicenter analysis of the medical records of 1247 patients with cervical cancer who underwent treatment for disease and who had completed at least two years of follow-up., Results: Although concurrent radiotherapy and chemotherapy is a standard treatment of FIGO IB to IVA cervical cancer patients in enhanced- and maximum-resources settings, in our analysis, we found that the percentage of women subjected to chemotherapy was lower than in countries where total survival rates were lower., Conclusion: Within the IA to II A cervical cancer patients studied group, the methods of treatment remained in line with ASCO guidelines for countries with the highest standard of care. Although concurrent radiotherapy and chemotherapy is a standard treatment of FIGO IB to IVA cervical cancer patients in enhanced- and maximum-resources settings, in our analysis, we found that the percentage of women subjected to chemotherapy was lower than in countries where total survival rates were lower. Our findings, together with the inconsistencies within the cervical cancer screening program, may be one of the explanations of poorer survival rate of women with cervical cancer in Poland., Competing Interests: The authors have declared that no competing interests exist.

N -threonyl-carbamoyl adenosine (t 6 A) is a universal tRNA modification found at position 37, next to the anticodon, in almost all tRNAs decoding ANN codons (where N = A, U, G, or C). t 6 A stabilizes the codon-anticodon interaction and hence promotes translation fidelity. The first step of the biosynthesis of t 6 A stabilizes the codon-anticodon interaction and hence promotes translation fidelity. The first step of the biosynthesis of t 6 A, the production of threonyl-carbamoyl adenylate (TC-AMP), is catalyzed by the Sua5/TsaC family of enzymes. While TsaC is a single domain protein, Sua5 enzymes are composed of the TsaC-like domain, a linker and an extra domain called SUA5 of unknown function. In the present study, we report structure-function analysis of Pyrococcus abyssi Sua5 ( Pa -Sua5). Crystallographic data revealed binding sites for bicarbonate substrate and pyrophosphate product. The linker of Pa -Sua5 forms a loop structure that folds into the active site gorge and closes it. Using structure-guided mutational analysis, we established that the conserved sequence motifs in the linker and the domain-domain interface are essential for the function of Pa -Sua5. We propose that the linker participates actively in the biosynthesis of TC-AMP by binding to ATP/PPi and by stabilizing the N -carboxy-l-threonine intermediate. Hence, TsaC orthologs which lack such a linker and SUA5 domain use a different mechanism for TC-AMP synthesis., (© 2018 Pichard-Kostuch et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.)

Galloway-Mowat syndrome (GAMOS) is an autosomal-recessive disease characterized by the combination of early-onset nephrotic syndrome (SRNS) and microcephaly with brain anomalies. Here we identified recessive mutations in OSGEP, TP53RK, TPRKB, and LAGE3, genes encoding the four subunits of the KEOPS complex, in 37 individuals from 32 families with GAMOS. CRISPR-Cas9 knockout in zebrafish and mice recapitulated the human phenotype of primary microcephaly and resulted in early lethality. Knockdown of OSGEP, TP53RK, or TPRKB inhibited cell proliferation, which human mutations did not rescue. Furthermore, knockdown of these genes impaired protein translation, caused endoplasmic reticulum stress, activated DNA-damage-response signaling, and ultimately induced apoptosis. Knockdown of OSGEP or TP53RK induced defects in the actin cytoskeleton and decreased the migration rate of human podocytes, an established intermediate phenotype of SRNS. We thus identified four new monogenic causes of GAMOS, describe a link between KEOPS function and human disease, and delineate potential pathogenic mechanisms.

In general, U.S. college students have low perceived susceptibility of acquiring HIV infection while 15-25 percent of youth have had negative perceptions towards HIV positive individuals. Factors associated with HIV stigma among college students were examined in a convenience sample of 200 students. Descriptive and inferential statistics were utilized to summarize the data. Only four percent of participants responded correctly to HIV transmission knowledge items. HIV transmission knowledge scores were significantly higher for participants who were single with partner and those who resided outside university residential dorms ( p < 0.05). There was a significant negative correlation between composite HIV knowledge scores and stigma scores r = -0.18 ( p < 0.05). After adjusting for confounders, a marginal significant negative linear relationship emerged ( β = -0.09, p = 0.06) between HIV knowledge and stigma. HIV prevention education among college students needs to be addressed with nuance to minimize HIV knowledge gaps, stigma and student risk perception that impacts HIV prevention and stigma against those living with HIV., Competing Interests: Conflict of Interest: The authors declare no conflict of interest.

Introduction: Cervical cancer (CC) is the fourth most common, in terms of incidence of new cases, cancer in women and the third leading cause of cancer deaths in women worldwide. Survival of patients with CC depends on many factors, including the type of cancer, grading, FIGO staging and treatment., Material and Methods: Analysis of survival of 524 patients diagnosed with invasive and non-invasive CC depending on histopathologic diagnosis, clinical staging, tumor grading and combination of therapy., Results: The 2-fold increase in the risk of death at diagnosis in order of HSIL> ca planoepitheliale> adenocarcinoma> sarcoma was noted. Grading 2 and 3 significantly reduces the average survival in patients diagnosed with CC. The higher staging, the shorter the average survival. Each pass by one FIGO stage was shown to increase the risk of death by 46%. The risk of death increases by 4% with every year of woman’s life. The longest average survival, 72 months, characterized a group of women undergoing curettage, followed by radical hysterectomy/ trachelectomy and lymphadenectomy without adiuvant radio-/ chemotherapy. The shortest survival, 26.9 months, was observed in the group treated with curettage followed by chemoradiation., Conclusions: Histopathology, clinical staging, grading, age and combination of treatment proved to be significant factors affecting survival in women with CC.

The onset and the course of autoimmune diseases is influenced among other factors by the sex hormones. Hormonal contraception might affect the course of the autoimmune disease. The paper summarises the manner of save application of hormonal contraception in patients with autoimmune disease.

Unlabelled: Ovarian cancer (OC) affects over 3 000 women in Poland annually The efficacy of the therapy remains relatively low due to challenges of systematic improvement in the early detection OC rates. International comparisons indicate a positive correlation between health expenditures and 5-year survival rates of cancer patients. To the best of our knowledge, our study has been the first to present a correlation between the 5-year survival rates (SRs) and the cost of ovarian cancer therapy in particular regions of Poland., Material and Methods: The study was based on the National Health Fund (NHF) data, available in the Disease Treatment Registry The analysis included approximately 13,000 OC patients who started their treatment between 2005 and 2008 to allow for the evaluation of long-term therapy results. The 5-year survival rates were analyzed in relation to average NHF expenditures in various regions of Poland, distinguishing the population of patients aged 45-64 years., Results: The 5-year survival rate in the cohorts diagnosed in 2005 and 2008 changed marginally from 42% to 43%, maintaining relatively large differences between the regions (from 35% to 53% in patients diagnosed in 2008). The NHF expenditures in particular regions differed significantly: mean cost for the entire treatment cycle ranged from 31.600 PLN do 58.000 PLNperperson among patients diagnosed in 2008. No significant correlation between the survival and the cost was found., Conclusions: SRs of OC patients in particular regions of Poland are not correlated with average treatment cost. Thus, the differences in SRs between various regions of Poland have their source in other factors, e.g., clinical stage at diagnosis, or prevailing treatment patterns in the given region. Further studies may decrease regional discrepancies in patient care and SRs in OC subjects.

Human Papilloma Virus (HPV) infection is the most common sexually transmitted disease. Chronic HPV infection is indispensable for development of cervical intraepithelial neoplasia and cervical cancer. First data stating that HPV transmission among heterosexual partners is very common appeared in last years. The aim of this study was to estimate the HPV infection prevalence among male sexual partners of women diagnosed with subclinical HPV infection, cervical intraepithelial neoplasia and early invasive cervical cancer. The study was conducted among 289 women aged 25-60 diagnosed with CIN and cervical cancer stage IA; control group consisted of 44 women aged 28-56 HPV testing using the material from retroglandular sulcus was conducted among male sexual partners of women from the study group. Testing was performed with HC2 method. In the study group, HPV infection was stated in 218 (75.43%) women and in 6 (13.63%) in control group. HPV DNA was present in 148 (51.21%) men--sexual partners of women from the study group and only 1 (2.27%) from control group. Additionally, HPV types of high and low oncogenic potential were analyzed with regard to histological diagnosis (SPI, CIN, early invasive cervical cancer). As the analysis shows, HPV infection of male sexual partners of women diagnosed with SPI and CIN is relatively high (9.09-93.33%).

Purpose of Study: The aim of the study was to assess the ongoing changes of the type of regression, progression and steady state among patients diagnosed with subclinical changes of an HPV infection as well as changes in the type of CIN 1 and CIN 2., Materials and Methods: The study was conducted in a group of 289 women between the ages of 25-60 with abnormal cytology taking part in the CIN cervical cancer prevention program., Results: The patients were observed over a period of 6 years; no detectable differences were discovered in the frequency of regression between patients with SPI in comparision to patients with CIN1. In addition, no differences were identified in the frequency of regression between groups of patients with CIN1 and CIN2. In contrast, regression was more common in patients with SPI than in patients with CIN2. Steady state was more frequent in patients with CIN1 and CIN 2 than in patients with SPI. The results illustrated no differences in the progression of SPI and the CIN1 to CIN2. The group of patients with CIN2 were frequently associated with progression to CIN3 more than in the group of patients with SPI. The group of patients with CIN2 were frequently associated with progression to CIN3 more than in the group of patients with CIN1. Further investigation of cervical changes associated with SPI, CIN1 and CIN2 were dependent on the presence of transcription genes E6 and E7 of HPV. In 138 cases, the presence of these transcription genes lead to progression in 19.56% of women; more specifically in the introduction of mE6 and E7 RNA. There were changes typical of remission in 56,52% of cases primarily in the absence of transcriptor genes HPV E6 and E7., Conclusion: 1. The histological changes of the cervix observed in subclinical HPV infection, CIN1 and CIN2 may be subject to a higher degree of progression of CIN. In addition, these changes may progress to cervical cancer, remain stationary in a steady state, or decline into remission. 2. The types of HPV infection with high oncogenic potential are not only important in initiation of cerival changes but also in the developmental process of carcinogenesis in the cervix by several independent mechanisms.

Introduction: Human papillomavirus (HPV) is the most common sexually transmitted infection. Data reporting vertical transmission of HPV from the mother to the fetus are inconsistent and scant. Vertical transmission may occur by hematogenic route (transplacental), or by ascending contamination, or through the birth canal, which may result in the dreaded and rare laryngeal papillomatosis. Infected sperm at fertilization is a potential route of infection, too., Objective: The objective of the study was to evaluate the rate of vertical transmission of HPV in HPV-positive pregnant women to their newborn infants, as well as the risk factors of HPV vertical transmission., Material and Methods: The clinical material was provided by 136 pregnant women, aged 18-45 years. Out of this group, 30 (22.05%) women with abnormal Pap test and positive DNA HPV test were prospectively observed Neonatal status, i.e. DNA HPV from the nasopharyngeal smear was recorded in all infants during the perinatal period. The conventional Pap test was performed with the cervix brush in all women. The Bethesda 2011 classification system was applied., Results: An average C Reactive Protein (CRP) concentration in the studied pregnant women was 711.6083 (Std Dev--12.93). The most frequent cytological findings in the cervical smears from the examined women were ASCUS, n = 13 (43.3%), then--LSIL, n = 10 (33.3%), HSIL--n = 5 (16.7%) and AGC--n = 2 (6.7%). In the neonates, the presence of LR HPV DNA was detected in 9 cases (30.0%) and HR HPV DNA in 7 cases (23.3%). Fourteen neonates (46.7%) tested HPV DNA negative in the perinatal period., Conclusions: HPV infection (incidental or chronic) is observed in approximately 22% of pregnant women from the Matopolska province. Neonatal HPV infection in HPV-positive women was observed in 53.3% of the subjects. CRP concentration > 10 mg/dl in the serum of pregnant women statistically significantly (p 0.001) reduces the risk of vertical transmission of HPV from the mother to the fetus.

Threonylcarbamoyladenosine (t(6)A) is a universal modification located in the anticodon stem-loop of tRNAs. In yeast, both cytoplasmic and mitochondrial tRNAs are modified. The cytoplasmic t(6)A synthesis pathway was elucidated and requires Sua5p, Kae1p, and four other KEOPS complex proteins. Recent in vitro work suggested that the mitochondrial t(6)A machinery of Saccharomyces cerevisiae is composed of only two proteins, Sua5p and Qri7p, a member of the Kae1p/TsaD family (L. C. K. Wan et al., Nucleic Acids Res. 41:6332-6346, 2013, http://dx.doi.org/10.1093/nar/gkt322). Sua5p catalyzes the first step leading to the threonyl-carbamoyl-AMP intermediate (TC-AMP), while Qri7 transfers the threonyl-carbamoyl moiety from TC-AMP to tRNA to form t(6)A. Qri7p localizes to the mitochondria, but Sua5p was reported to be cytoplasmic. We show that Sua5p is targeted to both the cytoplasm and the mitochondria through the use of alternative start sites. The import of Sua5p into the mitochondria is required for this organelle to be functional, since the TC-AMP intermediate produced by Sua5p in the cytoplasm is not transported into the mitochondria in sufficient amounts. This minimal t(6)A pathway was characterized in vitro and, for the first time, in vivo by heterologous complementation studies in Escherichia coli. The data revealed a potential for TC-AMP channeling in the t(6)A pathway, as the coexpression of Qri7p and Sua5p is required to complement the essentiality of the E. coli tsaD mutant. Our results firmly established that Qri7p and Sua5p constitute the mitochondrial pathway for the biosynthesis of t(6)A and bring additional advancement in our understanding of the reaction mechanism., (Copyright © 2014, American Society for Microbiology. All Rights Reserved.)

The exceptional genomic content and genome organization of the Acidianus filamentous virus 1 (AFV1) that infects the hyperthermophilic archaeon Acidianus hospitalis suggest that this virus might exploit an unusual mechanism of genome replication. An analysis of replicative intermediates of the viral genome by two-dimensional (2D) agarose gel electrophoresis revealed that viral genome replication starts by the formation of a D-loop and proceeds via strand displacement replication. Characterization of replicative intermediates using dark-field electron microscopy, in combination with the 2D agarose gel electrophoresis data, suggests that recombination plays a key role in the termination of AFV1 genome replication through the formation of terminal loops. A terminal protein was found to be attached to the ends of the viral genome. The results allow us to postulate a model of genome replication that relies on recombination events for initiation and termination., (© 2014 John Wiley & Sons Ltd.)

We demonstrate that membrane proteins and phospholipids can self-assemble into polyhedral arrangements suitable for structural analysis. Using the Escherichia coli mechanosensitive channel of small conductance (MscS) as a model protein, we prepared membrane protein polyhedral nanoparticles (MPPNs) with uniform radii of ∼ 20 nm. Electron cryotomographic analysis established that these MPPNs contain 24 MscS heptamers related by octahedral symmetry. Subsequent single-particle electron cryomicroscopy yielded a reconstruction at ∼ 1-nm resolution, revealing a conformation closely resembling the nonconducting state. The generality of this approach has been addressed by the successful preparation of MPPNs for two unrelated proteins, the mechanosensitive channel of large conductance and the connexon Cx26, using a recently devised microfluidics-based free interface diffusion system. MPPNs provide not only a starting point for the structural analysis of membrane proteins in a phospholipid environment, but their closed surfaces should facilitate studies in the presence of physiological transmembrane gradients, in addition to potential applications as drug delivery carriers or as templates for inorganic nanoparticle formation.

Unlabelled: Cervical intraepithelial neoplasia--CIN affects women in their repro- ductive life period. CIN may proceed squamous cervical cancer. CIN is divided into: CIN1, CIN2, CIN3. CIN3 comprises cervical cancer in situ- CIS which is the true precancer state within the cervix. CIN, depending on grade may progress, regress or persist for many years. According to a few publication vitamins C, E and A may protect against carcinogenesis within the cervix. The aim of this paper was evalua- tion of vitamins A and E serum concentration of cervical intraepithelial neoplasia patients. The study material consisted of 289 women aged 25-60 years diagnosed with CIN and early invasive cervical cancer IA. The subjects of the study were selected amongst participants of National Cervical Cancer Screen- ing Program attending Department of Gynecology and Oncology of Jagiellonian University Medical College in Krakow. The control group consisted of 44 women aged 28-56 years diagnosed and treated in the same centre and period due to a non oncologic gynecologic conditions. Serum vitamin A and E was measured with HPLC method with ultraviolet detector (UV) (254 nm)., Results: Medium serum vitamin A concentration in the study group was 2.67 ± 1.15 mg/l and was significantly (p < 0.001) lower than in control group -3.81 ± 1.62 mg/l. Mean serum vitamin E concentration in the study group was 3.95 ± 1.93 mg/l and was also significantly (p < 0.001) lower than in control group (8.63 ± 2.84 mg/l). To conclude, the observed significantly lower vitamins A and E serum concentrations may be related to the cervical neoplasia process. The normal vitamin A and E serum levels may have a protective effect against cervical carcinogenesis.