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Abstract The mosquito-borne flaviviruses West Nile virus (WNV) and Usutu virus (USUV) pose a significant threat to the health of humans and animals. Both viruses co-circulate in numerous European countries including Germany. Due to their overlapping host and vector ranges, there is a high risk of co-infections. However, it is largely unknown if WNV and USUV interact and how this might influence their epidemiology. Therefore, in-vitro infection experiments in mammalian (Vero B4), goose (GN-R) and mosquito cell lines (C6/36, CT) were performed to investigate potential effects of co-infections in vectors and vertebrate hosts. The growth kinetics of German and other European WNV and USUV strains were determined and compared. Subsequently, simultaneous co-infections were performed with selected WNV and USUV strains. The results show that the growth of USUV was suppressed by WNV in all cell lines. This effect was independent of the virus lineage but depended on the set WNV titre. The replication of WNV also decreased in co-infection scenarios on vertebrate cells. Overall, co-infections might lead to a decreased growth of USUV in mosquitoes and of both viruses in vertebrate hosts. These interactions can strongly affect the epidemiology of USUV and WNV in areas where they co-circulate.

The arenavirus nucleoprotein (NP) plays an important role in the virus’ ability to block interferon (IFN) production, and its exonuclease function appears to contribute to this activity. However, efforts to analyze this contribution are complicated by the functional overlap between the exonuclease active site and a neighboring region involved in IKKε-binding and subsequent inhibition of IRF3 activation, which also plays an important role in IFN production. To circumvent this issue, we mutated a residue located away from the active site that is involved in binding of the dsRNA substrate being targeted for exonuclease digestion, i.e. H426A. We found that expression of Tacaribe virus (TCRV) NP containing this RNA-binding H426A mutation was still able to efficiently block IFN-β promoter activity in response to Sendai virus infection, despite being strongly impaired in its exonuclease activity. This was in contrast to a conventional exonuclease active site mutant (E388A), which was impaired with respect to both exonuclease activity and IFN antagonism. Importantly, growth of a recombinant virus encoding the RNA-binding mutation (rTCRV-H426A) was similar to wild-type in IFN-deficient cells, unlike the active site mutant (rTCRV-E388A), which was already markedly impaired in these cells. Further, in IFN-competent cells, the TCRV-H426A RNA-binding mutant showed more robust growth and delayed IFN-β mRNA upregulation compared to the TCRV-E388A active site mutant. Taken together, this novel mutational approach, which allows us to now dissect the different contributions of the NP exonuclease activity and IKKε-binding/IRF3 inhibition to IFN antagonism, clearly suggests that conventional exonuclease mutants targeting the active site overestimate the contribution of the exonuclease function, and that rather other IFN antagonistic functions of NP play the dominant role in IFN-antagonism. Author summary The ability to suppress interferon production plays an important role in the successful establishment of viral infection, and for arenaviruses, the exonuclease domain of the nucleoprotein is suggested to play an important role in this process. However, analyzing its contribution is challenging because mutations that disrupt the exonuclease active site also block antagonism of IRF3 activation, which is also important for interferon production. Here we report a mutation that instead targets binding of the exonuclease’s dsRNA substrate. As expected, this mutation strongly impaired exonuclease activity; however, unlike classical mutations targeting the exonuclease active site, it was still able to efficiently block interferon production. This suggests that data based on active site mutants overestimate the contribution of the NP exonuclease function to interferon antagonism. This advance in our understanding regarding the specific contributions of different NP functions to interferon antagonism has important implications for understanding arenavirus pathogenesis and the development of future treatment strategies.

The arenavirus nucleoprotein (NP) plays an important role in the virus' ability to block interferon (IFN) production, and its exonuclease function appears to contribute to this activity. However, efforts to analyze this contribution are complicated by the functional overlap between the exonuclease active site and a neighboring region involved in IKKε-binding and subsequent inhibition of IRF3 activation, which also plays an important role in IFN production. To circumvent this issue, we mutated a residue located away from the active site that is involved in binding of the dsRNA substrate being targeted for exonuclease digestion, i.e. H426A. We found that expression of Tacaribe virus (TCRV) NP containing this RNA-binding H426A mutation was still able to efficiently block IFN-β promoter activity in response to Sendai virus infection, despite being strongly impaired in its exonuclease activity. This was in contrast to a conventional exonuclease active site mutant (E388A), which was impaired with respect to both exonuclease activity and IFN antagonism. Importantly, growth of a recombinant virus encoding the RNA-binding mutation (rTCRV-H426A) was similar to wild-type in IFN-deficient cells, unlike the active site mutant (rTCRV-E388A), which was already markedly impaired in these cells. Further, in IFN-competent cells, the TCRV-H426A RNA-binding mutant showed more robust growth and delayed IFN-β mRNA upregulation compared to the TCRV-E388A active site mutant. Taken together, this novel mutational approach, which allows us to now dissect the different contributions of the NP exonuclease activity and IKKε-binding/IRF3 inhibition to IFN antagonism, clearly suggests that conventional exonuclease mutants targeting the active site overestimate the contribution of the exonuclease function, and that rather other IFN antagonistic functions of NP play the dominant role in IFN-antagonism.

Design work was done on a broadband and wide-angle selective pyramid metamaterial emitter. COMSOL Multiphysics software was used to study the emitter, which was made of tungsten and aluminum nitride. The width of the unit cell and the tungsten ground thickness were fixed while other geometric parameters, such as the base length of the pyramid, the height of the pyramid, and the thickness of the dielectric, were tuned to produce the emitter’s desired broadband emission. A high average emissivity over 0.96 below the cutoff wavelength (0.1 μm-2.2 μm) was seen in the numerical simulation. The developed metamaterial emitter also had good emissivity across a broad range of incidence angles, from 0° to 60°, and was polarization independent. In addition, the planned emitter for the InGaAs cell has a better spectral efficiency than the blackbody other designed emitter. In general, the planned selective nanopyramid emitter was realized with 75% spectrum efficiency for InGaAs band gap energy (0.55 eV) at 1200 K, which was greater than blackbody and prior papers.

Due to globalisation and climate change, mosquito-borne pathogens are emerging in new areas on all continents, including Europe, which has recently faced outbreaks of dengue, chikungunya and West Nile fever. The present study complements previous investigations to evaluate the circulation of mosquito-borne viruses in Germany, with the aim of identifying potential vector species and risk areas. Mosquitoes collected from 2019 to 2021 and identified to species or species group level were screened for viruses of the families Flaviviridae, Peribunyaviridae and the genus Alphavirus of the family Togaviridae. In total, 22,528 mosquitoes were examined, thus providing the most comprehensive study on West Nile virus (WNV) circulation so far in the German mosquito population. Usutu virus (USUV) RNA was detected in six samples, Sindbis virus (SINV) RNA in 21 samples and WNV RNA in 11 samples. Samples containing RNA of USUV and WNV consisted of mosquitoes collected in the East German federal states of Brandenburg, Saxony and Saxony-Anhalt, while samples with RNA of SINV originated from more widespread locations. Although minimum infection rates have remained relatively low, the intensity of virus circulation appears to be increasing compared to previous studies. Continuous mosquito screening contributes to the early detection of the introduction and spread of mosquito-borne pathogens.

The increasing threat of arboviruses such as West Nile virus (WNV) and Usutu virus (USUV) requires the fast and efficient surveillance of these viruses. The examination of mosquitoes takes up an important part; however, these investigations are usually very time-consuming. An alternative sample type for arbovirus surveillance might be mosquito excreta. In order to determine the excretion dynamics under laboratory conditions, laboratory colonies of Aedes vexans and Culex pipiens biotype molestus were infected with WNV, USUV or tick-borne encephalitis virus (TBEV). After infection, the excreta were sampled and investigated for viral RNA. Excretion of viral RNA together with infectious blood meal could be detected up to five days after infection. Further excretion seemed to correlate with a disseminated infection in mosquitoes, at least after USUV infection. In addition, it could be determined that the amount of viral RNA in the excretions correlated positively with the viral load in the mosquito bodies. Overall, this study shows that the usage of mosquito excreta as a sample type for surveillance enables the detection of endemic viruses (WNV, USUV) as well as non-mosquito-borne viruses (TBEV). In addition, examination of viral shedding during vector competence studies can provide insights into the course of infection without sacrificing animals.

West Nile virus (WNV) is a zoonotic flavivirus transmitted by mosquitoes as a biological vector. Because of its biting behavior, the widespread snow-melt mosquito Aedes punctor could be a potential bridge vector for WNV to humans and nonhuman mammals. However, little is known on its role in transmission of WNV. The aim of this study was to determine the vector competence of German Ae. punctor for WNV lineages 1 and 2. Field-collected larvae and pupae were reared to adults and offered infectious blood containing either an Italian WNV lineage 1 or a German WNV lineage 2 strain via cotton stick feeding. Engorged females were incubated for 14/15 or 21 days at 18 °C. After incubation; surviving mosquitoes were dissected and forced to salivate. Mosquito bodies with abdomens, thoraces and heads, legs plus wings and saliva samples were investigated for WNV RNA by RT-qPCR. Altogether, 2/70 (2.86%) and 5/85 (5.88%) mosquito bodies were found infected with WNV lineage 1 or 2, respectively. In two mosquitoes, viral RNA was also detected in legs and wings. No saliva sample contained viral RNA. Based on these results, we conclude that Ae. punctor does not play an important role in WNV transmission in Germany.

Abstract Background Depression and anxiety are among the common mental health problems among medical students and are associated with poor academic performance, disability and poor quality of life. A better understanding of the magnitude and correlates of depression and anxiety is essential for planning appropriate intervention for those population groups. However, research into depression and anxiety and the potential contributing factors is limited in low and middle-income countries including Ethiopia. Therefore, this study aimed to determine the prevalence and associated factors of depression and anxiety among medical students. Methods A cross-sectional study was conducted among 273 medical students selected by systematic sampling technique. Hospital anxiety and depression scale (HADS) was used to assess anxiety and depression. Binary and multivariable logistic regression models were fitted, adjusting for the potential confounding factors. Odds ratios (OR) with the corresponding 95% confidence interval (95% CI) was computed to assess the strength of association. Result The prevalence of co-morbid depression and anxiety was found to be 21.20% (16.35% to 26.05%) and prevalence of depression and anxiety was 51.30% (45.37% to 57.23%) and (30.1% 24.66% to 35.54%), respectively. Multivariate analysis showed that being female [AOR 2.56, 95% CI (1.32, 4.95)], first-year educational level [AOR 12.06, 95% CI (2.18, 66.72)], second-year educational level [AOR 8.99, 95% CI (1.67, 48.45)] and those who had poor/low social support [AOR 5.36, 95% CI (2.08, 13.76)] were significantly association with anxiety. Students who were in the age interval of 18–21 years [AOR 2.42, 95% CI (1.64, 9.22)], first-year educational level [AOR 1.63, 95% CI (1.43, 6.26)], second-year educational level [AOR 1.39, 95% CI (1.17, 5.18)] and who had stressful life events [AOR 1.61, 95% CI (1.14, 2.76)] were significantly associated with depression among medical students. Conclusion The current study demonstrated that a remarkable proportion of medical students are suffering from depression (51.30%), anxiety (30.10%) as well as comorbid depression and anxiety (21.20%). There was strong evidence of association between anxiety and female sex, first-year educational level, second-year educational level and having poor/low social support. Whereas a significant association was observed between depression and younger age (18–21 years old), first-year educational level, second-year educational level and having one or more stressful life events in the last 6 months. Screening of depression and anxiety among medical students was recommended. Future studies focusing on better ways of preventing and treating depression and anxiety among medical students are warranted.

In the context of climate change, globalization, and enhanced human traveling, arboviruses continue to represent a threat to public health. West Nile and Usutu viruses are mosquito-borne viruses belonging to the Flaviviridae family (Flavivirus genus) and members of the Japanese encephalitis virus serocomplex. Included in the Togaviridae family (Alphavirus genus), the Sindbis virus is also vectored by mosquitoes. In the present study, we aimed to analyze the presence of antibodies concerning the abovementioned viruses in migratory and resident birds in the South-Eastern region of Romania, as avian hosts represent the main reservoir for human infection. Blood samples were collected from wild birds between May 2018 and October 2019 in nine locations from three counties. All the samples were serologically tested by ELISA and a serum neutralization test. Overall, a seroprevalence of 8.72% was registered for the West Nile virus, 2.71% for the Usutu virus, and 0% for the Sindbis virus. To our best knowledge, this is the first large-scale comprehensive study to assess the West Nile virus seropositivity in wild birds and the first serological confirmation of the Usutu virus in wild birds in Romania. Moreover, this is the only follow-up study reviewing the current seroprevalence of the Sindbis virus in Romania since 1975.

Mosquitoes collected from mid-December 2020 to early March 2021 from hibernacula in northeastern Germany, a region of West Nile virus (WNV) activity since 2018, were examined for WNV-RNA. Among the 6101 mosquitoes tested in 722 pools of up to 12 specimens, one pool of 10 Culex pipiens complex mosquitoes collected in early March 2021 in the cellar of a medieval castle in Rosslau, federal state of Saxony-Anhalt, tested positive. Subsequent mosquito DNA analysis produced Culex pipiens biotype pipiens. The pool homogenate remaining after nucleic acid extraction failed to grow the virus on Vero and C6/36 cells. Sequencing of the viral NS2B-NS3 coding region, however, demonstrated high homology with virus strains previously collected in Germany, e.g., from humans, birds, and mosquitoes, which have been designated the East German WNV clade. The finding confirms the expectation that WNV can overwinter in mosquitoes in Germany, facilitating an early start to the natural transmission season in the subsequent year. On the other hand, the calculated low infection prevalence of 0.016–0.20%, depending on whether one or twelve of the mosquitoes in the positive pool was/were infected, indicates a slow epidemic progress and mirrors the still-hypoendemic situation in Germany. In any case, local overwintering of the virus in mosquitoes suggests its long-term persistence and an enduring public health issue.

Abstract Chlamydiae are bacterial pathogens that grow in vacuolar inclusions. Dendritic cells (DCs) disintegrate these compartments, thereby eliminating the microbes, through auto/xenophagy, which also promotes chlamydial antigen presentation via MHC I. Here, we show that TNF-α controls this pathway by driving cytosolic phospholipase (cPLA)2-mediated arachidonic acid (AA) production. AA then impairs mitochondrial function, which disturbs the development and integrity of these energy-dependent parasitic inclusions, while a simultaneous metabolic switch towards aerobic glycolysis promotes DC survival. Tubulin deacetylase/autophagy regulator HDAC6 associates with disintegrated inclusions, thereby further disrupting their subcellular localisation and stability. Bacterial remnants are decorated with defective mitochondria, mito-aggresomal structures, and components of the ubiquitin/autophagy machinery before they are degraded via mito-xenophagy. The mechanism depends on cytoprotective HSP25/27, the E3 ubiquitin ligase Parkin and HDAC6 and promotes chlamydial antigen generation for presentation on MHC I. We propose that this novel mito-xenophagic pathway linking innate and adaptive immunity is critical for effective DC-mediated anti-bacterial resistance.

Pestiviruses express the unique essential envelope protein Erns, which exhibits RNase activity, is attached to membranes by a long amphipathic helix, and is partially secreted from infected cells. The RNase activity of Erns is directly connected with pestivirus virulence. Formation of homodimers and secretion of the protein are hypothesized to be important for its role as a virulence factor, which impairs the host’s innate immune response to pestivirus infection. The unusual membrane anchor of Erns raises questions with regard to proteolytic processing of the viral polyprotein at the Erns carboxy-terminus. Moreover, the membrane anchor is crucial for establishing the critical equilibrium between retention and secretion and ensures intracellular accumulation of the protein at the site of virus budding so that it is available to serve both as structural component of the virion and factor controlling host immune reactions. In the present manuscript, we summarize published as well as new data on the molecular features of Erns including aspects of its interplay with the other two envelope proteins with a special focus on the biochemistry of the Erns membrane anchor.

West Nile virus (WNV) is a mosquito-borne agent that has also been isolated from several tick species. Vector competence of Ixodes ricinus, one of the most common tick species in Europe, has been poorly investigated for WNV to date. As such, to evaluate the vector competence, laboratory reared Ixodes ricinus nymphs were in vitro fed with WNV lineage 1 infectious blood, allowed to molt, and the resulting females artificially fed to study the virus transmission. Furthermore, we studied the kinetics of WNV replication in ticks after infecting nymphs using an automatic injector. Active replication of WNV was detected in injected nymphs from day 7 post-infection until 28 dpi. In the nymphs infected by artificial feeding, the transstadial transmission of WNV was confirmed molecularly in 46.7% of males, while virus transmission during in vitro feeding of I. ricinus females originating from infected nymphs was not registered. The long persistence of WNV in I. ricinus ticks did not correlate with the transmission of the virus and it is unlikely that I. ricinus represents a competent vector. However, there is a potential reservoir role that this tick species can play, with hosts potentially acquiring the viral agent after ingesting the infected ticks.

One year after the first autochthonous transmission of West Nile virus (WNV) to birds and horses in Germany, an epizootic emergence of WNV was again observed in 2019. The number of infected birds and horses was considerably higher compared to 2018 (12 birds, two horses), resulting in the observation of the first WNV epidemy in Germany: 76 cases in birds, 36 in horses and five confirmed mosquito-borne, autochthonous human cases. We demonstrated that Germany experienced several WNV introduction events and that strains of a distinct group (Eastern German WNV clade), which was introduced to Germany as a single introduction event, dominated mosquito, birds, horse and human-related virus variants in 2018 and 2019. Virus strains in this clade are characterized by a specific-Lys2114Arg mutation, which might lead to an increase in viral fitness. Extraordinary high temperatures in 2018/2019 allowed a low extrinsic incubation period (EIP), which drove the epizootic emergence and, in the end, most likely triggered the 2019 epidemic. Spatiotemporal EIP values correlated with the geographical WNV incidence. This study highlights the risk of a further spread in Germany in the next years with additional human WNV infections. Thus, surveillance of birds is essential to provide an early epidemic warning and thus, initiate targeted control measures.

In 2018, West Nile virus (WNV) broke out for the first time in Germany, with continuation of the epidemic in 2019, involving birds, horses and humans. To identify vectors and characterize the virus, mosquitoes were collected in both years in zoological gardens and on a horse meadow immediately following the diagnosis of disease cases in birds and horses. Mosquitoes were identified and screened for WNV by qRT-PCR, with virus-positive samples being sequenced for the viral envelope protein gene. While no positive mosquitoes were found in 2018, seven mosquito pools tested positive for WNV in 2019 in the Tierpark (Wildlife Park) Berlin. The pools consisted of Cx. pipiens biotype pipiens (n = 5), and a mixture of Cx. p. biotype pipiens and Cx. p. biotype molestus (n = 2), or hybrids of these, and were collected between 13 August and 24 September 2019. The virus strain turned out to be nearly identical to two WNV strains isolated from birds diseased in 2018 in eastern Germany. The findings represent the first demonstration of WNV in mosquitoes in Germany and include the possibility of local overwintering of the virus.

Tick-borne encephalitis is the most important tick-transmitted zoonotic virus infection in Eurasia, causing severe neurological symptoms in humans. The causative agent, the tick-borne encephalitis virus (TBEV), circulates between ticks and a variety of mammalian hosts. To study the interaction between TBEV and one of its suspected reservoir hosts, bank voles of the Western evolutionary lineage were inoculated subcutaneously with either one of eight TBEV strains or the related attenuated Langat virus, and were euthanized after 28 days. In addition, a subset of four strains was characterized in bank voles of the Carpathian linage. Six bank voles were inoculated per strain, and were housed together in groups of three with one uninfected in-contact animal each. Generally, most bank voles did not show any clinical signs over the course of infection. However, one infected bank vole died and three had to be euthanized prematurely, all of which had been inoculated with the identical TBEV strain (Battaune 17-H9, isolated in 2017 in Germany from a bank vole). All inoculated animals seroconverted, while none of the in-contact animals did. Viral RNA was detected via real-time RT-PCR in the whole blood samples of 31 out of 74 inoculated and surviving bank voles. The corresponding serum sample remained PCR-negative in nearly all cases (29/31). In addition, brain and/or spine samples tested positive in 11 cases, mostly correlating with a positive whole blood sample. Our findings suggest a good adaption of TBEV to bank voles, combining in most cases a low virulence phenotype with detectable virus replication and hinting at a reservoir host function of bank voles for TBEV.

West Nile virus (WNV) is a zoonotic flavivirus whose transmission cycle in nature includes wild birds as amplifying hosts and ornithophilic mosquito vectors. Bridge vectors can transmit WNV to mammal species potentially causing West Nile Fever. Wild bird migration is a mode of WNV introduction into new areas. The Danube Delta Biosphere Reserve (DDBR) is a major stopover of wild birds migrating between Europe and Africa. The aim of this study was to investigate the presence of WNV in the DDBR during the 2016 transmission season in wild birds and mosquitoes. Blood from 68 wild birds (nine different species) trapped at four different locations was analyzed by competitive ELISA and Virus Neutralization Test (VNT), revealing positive results in 8/68 (11.8%) of the wild birds by ELISA of which six samples (three from juvenile birds) were confirmed seropositive by VNT. Mosquitoes (n = 6523, 5 genera) were trapped with CDC Mini Light traps at two locations and in one location resting mosquitoes were caught. The presence of WNV RNA was tested in 134 pools by reverse transcription quantitative PCR (RT-qPCR). None of the pools was positive for WNV-specific RNA. Based on the obtained results, WNV was circulating in the DDBR during 2016.

Chronic Hepatitis C infection is a global health problem. While primary infection is often inapparent, it becomes chronic in most cases. Chronic infection with Hepatitis C virus (HCV) frequently leads to liver cirrhosis or liver cancer. Consequently, HCV infection is one of the leading causes for liver transplantation in industrialized countries. Current treatment is not HCV specific and is only effective in about half of the infected patients. This situation underlines the need for new antivirals against HCV. To develop new and more efficient drugs, it is essential to specifically target the different steps of the viral life cycle. Of those steps, the targeting of HCV assembly has the potential to abolish virus production. This review summarizes the advances in our understanding of HCV particle assembly and the identification of new antiviral targets of potential interest in this late step of the HCV life cycle.

Abstract Musicians create sound that is enjoyable to themselves and their audience, but this same sound also threatens their hearing health due to high sound pressure levels generated by their own and their fellow musicians’ instruments. Here we seek to identify musicians’ hearing health awareness in relation to their coping strategies. 370 professional and 401 amateur musicians in total responded to a questionnaire on hearing health, which included quantitative elements and open-ended questions. Findings reveal that musicians are generally aware of hearing health risks but also indicate a lack of discussion on that topic. However, inasmuch as respondents consider hearing protection as necessary, they articulate justified concerns about its potential impact on quality of performance and musical communication. In sum, musicians show literacy regarding their hearing health, but more guidance and training is needed to ensure effective protective measures.

Abstract Background The occurrence of potential drug‒drug interactions (pDDIs) is a serious global issue that affects all age groups, with the elderly population being the most vulnerable. This is due to their relatively high rates of comorbidity and polypharmacy, as well as physiological changes that can increase the potential for DDIs and the likelihood of adverse drug reactions. The aim of this study was to estimate the prevalence of pDDIs and associated factors among elderly patients in Ethiopia. Methods A comprehensive literature search using the preferred reporting items for systematic review and meta-analysis statement was conducted on HINARI, Science Direct, Embase, PubMed/MEDLINE, Google Scholar, and Research Gate. Data were extracted via a Microsoft Excel spreadsheet and analyzed via STATA version 11.0. Egger regression tests and funnel plot analysis were used to check publication bias, and the I2 statistic was used to evaluate statistical heterogeneity. Sensitivity and subgroup analyses were also conducted to identify potential causes of heterogeneity. Results Seven articles were analyzed, and a total of 1897 pDDIs were identified in 970 patients, resulting in an average of 1.97 DDIs per patient. The number of DDIs per patient ranged from 0.18 to 5.86. The overall prevalence of pDDIs among elderly patients was 50.69% (95% CI 18.77–82.63%). However, the prevalence of pDDIs ranged widely from 2.80 to 90.1%. When the severity of the interactions was considered, the prevalence of potential DDIs was found to be 28.74%, 70.68%, and 34.20% for major, moderate, and minor pDDIs, respectively. Polypharmacy and long hospital stays were identified as factors associated with pDDIs among elderly patients in Ethiopia. Conclusions The overall prevalence of pDDIs among elderly patients was high, with a wide range of prevalence rates. Moderate-severity interactions were the most prevalent. Polypharmacy and long hospital stays were identified as factors associated with pDDIs among elderly patients. The study suggests that DDIs identification database itself could have modified the DDIs prevalence rate. As a result, a single DDIs identification database needs to be authorized; otherwise, clinical knowledge should be taken into account when interpreting the information obtained.

Abstract Hematological changes is one of the most common complications occurred in cancer patients. Therefore, the current study aimed to assess the hematological toxicity of cervical cancer patients before and after the initiation of treatment. The retrospective cohort study was conducted from 2015 to 2022 at the University of Gondar Comprehensive Specialized Hospital. The hematological profile and sociodemographic and clinical data of the cervical cancer patients were collected using data extraction sheets. The Epidata version 3.1 and SPSS version 25 were used for data entry and analysis, respectively. Descriptive statistics were employed to summarize the data. To compare the median differences in hematological parameters before and after treatment, the Wilcoxon rank test was used. In addition, to assess the presence of an independent association between hematological abnormalities and the independent variables, logistic regression models were used. A p value less than 0.05 was considered to indicate statistical significance for all tests. In current study, the median (Interquartile range) of hemoglobin levels, white blood cell counts, and platelet counts, before treatment were 13.2 (12.1, 15) g/dl, 7.5 (5.8, 11.8) *109/L, and 330 (252, 383) *109/L, respectively. However, after treatment the median (Interquartile range) value of hemoglobin levels, white blood cell counts, and platelet counts were significantly lowered. On the other hand, red cell distribution width was significantly greater after treatment. At baseline, the magnitude of leucocytosis, anemia, and thrombocytosis were 27.9%, 24.6%, and 18.7%, respectively. After the treatment, anemia increased to 44.3%, but leucocytosis and thrombocytosis were replaced by leucopenia 18.3% and thrombocytopenia 17.8%, respectively. Hematological abnormalities such as anemia, leucopenia and thrombocytopenia were high after chemo-radiotherapy, and surgery. As the stage of cancer advances, the risk of developing anemia, leucocytosis, and thrombocytosis increased in 7.6, 6.9 and 9 times, respectively. Furthermore, being HIV patients and rural resident increased the risk of developing anemia about twofold. In conclusion hematological abnormalities were observed before and after cervical cancer treatment, with significance increment after chemo-radiotherapy and surgery. As the stage of cancer advances, the risk of developing hematological abnormalities increases. Therefore, routine monitoring of hematological changes before and after treatment and screening for major risk factors are important for improved patients’ management.

Societal Impact Statement Protecting diversity is critical to maintaining functional and adaptive ecosystems. However, research groups that explore these relationships tend to lack diversity themselves. To address the challenges of our changing world, it is crucial to consider a wide range of perspectives and knowledge. We propose that just as greater diversity in ecosystems relates to a variety of functions and, consequently, enhances resilience, stability, and efficiency, a comparable diversity within research teams results in a broader spectrum of creative endeavors. Thus, it is important to re‐evaluate our scientific communities' openness toward greater diversity to meet the diverse problems of our changing world. Summary Ecosystems face multiple threats, jeopardizing natural and human well‐being. Biodiversity loss can increase ecosystem vulnerability to global change, risking abrupt and irreversible changes. Protecting biodiversity can prevent ecosystems from crossing tipping points, preserving their functions and resilience, which sustain life on our planet. In addressing these challenges, society demands creativity, innovation, and diverse perspectives. Yet, our societal structures often neglect the full range of voices and experiences, limiting problem‐solving. Understanding how research teams with diverse demographics may stimulate creative solutions to global change challenges may help. In this context, we emphasize examples from plant ecology, where the advantages of diverse ecosystems are well recognized, and we draw an analogy of these features to the innovation of research teams. In ecosystems, biodiversity can maximize productivity and function but can also serve as a protective “insurance” against disruptions. Similarly, in a time of multifaceted environmental changes, diverse research groups play a critical and analogous role, resembling the biodiversity within ecosystems. Diverse research teams not only bring complementary strengths, fostering creative problem‐solving by embracing various perspectives, but also drive scientific innovation by integrating a wide range of research topics (e.g., ecosystems, taxa, and conservation priorities). These human‐centered dynamics are congruent with the mechanisms in ecosystems, with a diversity of solutions promoting resilience, stability, and efficiency, all of which ultimately provide a counterbalancing effect on global change. Much like biodiverse ecosystems, a diverse spectrum of knowledge, and expertise within the scientific community can enrich a dynamic landscape of innovation.

Abstract Background Hepatitis virus infection remains a serious public health concern worldwide. Percutaneous, mucosal, or non-intact skin exposure to infectious blood, semen, and other body fluids are the major routes of infection in endemic areas, including Ethiopia, where little is known about the community’s knowledge, attitudes, and practice toward hepatitis infection. Thus, this study aimed to determine the knowledge, attitude, and practices towards hepatitis infection among adults in Gondar town. Methods A community-based cross-sectional study was conducted from July to August 2021 on a total of 390 participants. Proportional sample allocation was used in 3 randomly selected sub-cities. Then, study participants from randomly selected households were recruited by using a systematic random sampling technique. Socio-demographic data and questions regarding KAP were collected via a self-administered and interviewer-administered questionnaire. Statistical analysis was performed using SPSS version 20. Logistic regression analyses were used to identify the associated factors and a p-value

Abstract Background The level of experienced sociocultural pressure to have an idealized body can vary depending on a person’s gender identity and sexual orientation. The current study explored whether differences in levels of body appreciation among people with different sexual orientations and gender identities vary because of differing levels of experienced pressure by in-group members and varying levels of experienced hostile behaviors because of their looks or body. Thereby, the study tests the social cure model, according to which high levels of identity centrality are associated with better mental health. Methods An online cross-sectional questionnaire study was conducted with 1,587 people (51.3% cisgender women, 39.3% cisgender men, 9.5% non-binary; 52.9% identified as heterosexual, 27.7% identified as bisexual/pansexual, 11.2% identified as gay/lesbian, 8.2% identified as asexual/demi/queer; M age = 32.9, SD = 12.6) from German-speaking countries. Variables were assessed with German-language versions of the Multidimensional and Multicomponent Measure of Social Identification, Body Appreciation Scale-2, the Perceived Stigmatization Questionnaire, and the Sociocultural Attitudes Towards Appearance Questionnaire-4, revised. A manifest-path model was calculated. Results Non-binary persons reported lower levels of body appreciation than did cisgender men and sexual minority persons reported lower levels of body appreciation than did heterosexual persons. Furthermore, sexual minority persons experienced more hostile behaviors directed towards them because of their looks or body than did heterosexual persons. Similarly, non-binary persons experienced more hostile behaviors than did men. Non-binary persons were subjected to lower levels of in-group pressure than were men. Gay/lesbian persons and asexual persons were subjected to lower levels of in-group pressure than were heterosexual persons. More hostile behaviors and stronger in-group pressure were related to lower body appreciation. In cisgender women and men indirect links revealed associations between strong identity centrality and low levels of body appreciation through the mediator of high in-group pressure. Conclusions Data in sexual minority individuals or non-binary persons supported the social cure model, according to which persons can find support and validation for their looks and body from in-group members. In cisgender women and men, strong identification as a woman or man can be related to stronger in-group pressure and in turn to lower body appreciation.

Abstract Vitamin D deficiency is an emerging public health problem globally, with devastating health consequences. Some studies suggest that exogenous sex hormones, found in hormonal contraceptives, may enhance vitamin D levels. However, the reasons for this association are not fully understood, as women using hormonal contraception may have different lifestyle habits affecting their vitamin D status. Therefore, this study seeks to explore the relationship between hormonal contraceptive use and vitamin D levels. A Facility based comparative cross-sectional study was conducted in Gondar town from February to April 2023, involving a total of 162 women using three types of hormonal contraceptives (Norplant, DMPA, and COC) and 162 age and BMI-matched non-users as controls in a 1:1 ratio. Participants were selected using systematic random sampling. A semi-structured questionnaire was used to collected data regarding the socio-demographic, economic, obstetric, lifestyle, and clinical information. 5 milliliters of blood samples were collected from each participant for Laboratory analysis of serum vitamin D, calcium, and alkaline phosphates using a Beckman Coulter chemistry analyzer. Independent t-tests, ANOVA with post hoc Bonferroni test was used to compare statistics between the two groups, and logistic regression models to identify factors associated with Vitamin D deficiency. The mean serum Vitamin D levels of Norplant, DMPA, and COC users were 24.08 (± 5.17), 24.83 (± 5.52), and 31.90 (± 6.94) respectively; whereas control group has mean Vitamin D level of 22.00 (± 7.97). On the current study the prevalence of Vitamin D deficiency (

Background: Blood biomarkers offers an independent insight for the pathophysiology of hyperbilirubinemia. However, they are not practically used for the differential diagnosis of the hyperbilirubinemia severity. Therefore, the current study aimed to assess the differential diagnostic value of peripheral blood biomarkers with disease severity as an alternative. Methods: A cross-sectional study was done on conveniently selected neonates admitted with hyperbilirubinemia during study period. A 4 ml of venous blood was collected for laboratory analysis. The Sysmex KX-21 cellular analysis and Mindray BS-240 automated chemistry analyzer was used for complete blood count and biochemical analysis, respectively. The data were entered into Epi-data (4.6.0) and analyzed by STATA (14) software. The summary statistics were used. The Kruskal Wallis H tests were utilized to compare median differences between groups. To ascertain the diagnostic value, receiver operator characteristic (ROC) curve analysis was performed. Blood biomarkers score area under curve >.7 was selected as the best discriminative marker. The accepted threshold for statistical significance was set at P

Abstract This study aimed to assess the magnitude of hematological toxicity and associated factors in newborns with hyperbilirubinemia. A cross-sectional study was conducted from April to December 2023. A total of 247 newborns were included. The data were collected using questionnaires and a data extraction sheet. Four 4 ml of blood was collected. A Sysmex KX-21 analyzer was used for blood analysis, and a Mindray BS-240 analyzer was used for bilirubin measurement. The data were entered into Epi-data and analyzed by SPSS. The logistic regression was used. The P value was set at 0.05. Before phototherapy, the hematological toxicities, such as anemia, leucopenia, and thrombocytopenia, were 45.7%, 22.2%, and 6.1%, respectively, whereas after phototherapy, anemia and thrombocytopenia, significantly increased, but the leucopenia, significantly decreased. The risk of developing anemia increased, 3.5, 2.7, and 2.1-fold among newborns with bilirubin > 18 mg/dl, with Rh blood group incompatibility, and treated with intensive phototherapy, respectively. Both low birth weight and intensive phototherapy increased the incidence of thrombocytopenia by 2 and 3.4-fold, respectively. Hematological toxicity was found to be a severe public health issue in newborns. Thus, strict follow-up and early detection of toxicity by considering aggravation factors are necessary.

Fast charging is a critical factor for the widespread adoption of electric vehicles (EVs), yet it poses challenges to the durability of lithium-ion batteries (LiBs). This study develops an aging model for a 10 Ah LiB cell using COMSOL Multiphysics, integrating a one-dimensional electrochemical model with a zero-dimensional thermal model. The model is constructed using cell design and electrochemical parameters derived from existing literature, tailored to the specific electrode and electrolyte materials utilized in this study. Key aging phenomena, such as the Solid Electrolyte Interface (SEI) growth and lithium plating, are embedded within the model to simulate the aging patterns and the interplay between these factors across various C-rates and temperatures. Simulation results suggest that while higher charging rates can reduce charging time, they also lead to increased degradation, with lithium plating becoming more severe at these elevated rates. The model also indicates that temperature management during cell operation can influence cycle life, with strategic temperature increases potentially mitigating some negative effects of fast charging. The study also explores the concept of total charging time as an alternative metric for battery wear, examining its relationship with the charging current profile and temperature. Results indicate that while fast charging offers convenience, it significantly impacts cycle life, underscoring the need for optimized charging strategies. The insights from this work enhance the understanding of Li-ion cell aging mechanisms, shedding light on the complex trade-offs between charging speed, temperature management, and lifespan. This contributes to the development of more sophisticated battery management systems and charging protocols that aim to extend the operational life of LiBs while accommodating the demands of fast charging.

Abstract Proficiency testing (PT) is an impartial laboratory performance-evaluating system using an independent body. It is a mandatory accreditation requirement and means for improving the laboratory’s performance. The study aimed to evaluate the performance of PT, with a focus on identifying and discussing determinants that influence PT performance at comprehensive specialized hospitals in northwest Ethiopia. A retrospective cross-sectional study was carried out from 2020 to 2022. Using a convenient sampling technique, laboratory tests with recorded PT results in each hospital laboratory were included. A data collection template and customized checklists were used to collect the data. Epi Data Version 3.1 for data entry and STATA Version 14.1 for cleaning and analysis were used. Binary logistic regression analyses were used. Variables with p

Abstract Neurodegenerative disorders are characterized by the progressive loss of structure and function of neurons, often including the death of the neuron. Previously, we reported that, by removing the cell death stimulus, dying/injured neurons could survive and recover from the process of regulated cell death, even if the cells already displayed various signs of cellular damage. Now we investigated the role of mitochondrial dynamics (fission/fusion, biogenesis, mitophagy) in both degeneration and in recovery of neuronal cells. In neuronal PC12 cells, exposure to ethanol (EtOH) induced massive neurite loss along with widespread mitochondrial fragmentation, mitochondrial membrane potential loss, reduced ATP production, and decreased total mitochondrial volume. By removing EtOH timely all these mitochondrial parameters recovered to normal levels. Meanwhile, cells regrew neurites and survived. Study of the mitochondrial dynamics showed that autophagy was activated only during the cellular degeneration phase (EtOH treatment) but not in the recovery phase (EtOH removed), and it was not dependent on the Parkin/PINK1 mediated mitophagy pathway. Protein expression of key regulators of mitochondrial fission, phospho-Drp1Ser616 and S-OPA1, increased during EtOH treatment and recovered to normal levels after removing EtOH. In addition, the critical role of PGC-1α mediated mitochondrial biogenesis in cellular recovery was revealed: inhibition of PGC-1α using SR-18292 after EtOH removal significantly impeded recovery of mitochondrial damage, regeneration of neurites, and cell survival in a concentration-dependent manner. Taken together, our study showed reversibility of mitochondrial morphological and functional damage in stressed neuronal cells and revealed that PGC-1α mediated mitochondrial biogenesis played a critical role in the cellular recovery. This molecular mechanism could be a target for neuroprotection and neurorescue in neurodegenerative diseases.

Abstract Aim To systematically review longitudinal studies investigating the impact of dental visiting patterns on oral health across the life course. Methods Five databases (MEDLINE, Embase, Scopus, Web of Science, CINAHL) were searched up to March 2023. Results were screened based on eligibility criteria in a two-stage process: title and abstract, and full-text review. A backward search of reference lists and a forward search of citations of the included papers was also conducted. The quality of the included papers was assessed using the Newcastle-Ottawa Scale. Key study information was extracted and a narrative synthesis of the findings was performed. Results Eleven papers from five longitudinal studies in five countries (Australia, Brazil, China, New Zealand, Sweden) met the inclusion criteria. Studies of moderate to high quality consistently reported that regular dental attendance was associated with having less dental caries experience, fewer missing teeth and better oral health-related quality of life. Inconsistent findings were observed for decayed teeth, and no association was found for periodontal condition. Conclusions This review highlights an association between regular dental visiting pattern and improved oral health, notably less dental caries experience and better oral health-related quality of life. Dental attendance emerges as an important predictor of oral health across the life course, underscoring the importance of routine dental care. Registration information The PROSPERO registration number is CRD42023396380.

Cyclophilin (Cyp) inhibitors are of clinical interest in respect to their antiviral activities in the context of many viral infections including chronic hepatitis B and C. Cyps are a group of enzymes with peptidyl-prolyl isomerase activity (PPIase), known to be required for replication of diverse viruses including hepatitis B and C viruses (HBV and HCV). Amongst the Cyp family, the molecular mechanisms underlying the antiviral effects of CypA have been investigated in detail, but potential roles of other Cyps are less well studied in the context of viral hepatitis. Furthermore, most studies investigating the role of Cyps in viral hepatitis did not investigate the potential therapeutic effects of their inhibition in already-established infections but have rather been performed in the context of neo-infections. Here, we investigated the effects of genetically silencing Cyps on persistent HCV and HBV infections. We confirm antiviral effects of CypA and CypD knock down and demonstrate novel roles for CypG and CypH in HCV replication. We show, furthermore, that CypA silencing has a modest but reproducible impact on persistent HBV infections in cultured human hepatocytes.

Sector coupling plays a crucial role in reducing CO2 emissions. The usage of renewable energies in Power-to-Liquid (PtL) processes is one option to link the energy, transport and chemical sectors. In addition to the sectors mentioned, other industries such as the cement industry need to be coupled for decarbonization. The BlueFire research project focuses on the investigation of an innovative process, plasma-based calcium looping. This process has the potential to serve as a fundamental component of a PtL plant by absorbing CO2 from the environment and converting it into the syngas component carbon monoxide. It is furthermore an option for electrification of the important process of calcination in the cement industry. In this study, a techno-economic analysis is carried out to evaluate the potential of the plasma-based calcium looping. Integrated in a PtL plant to produce marine diesel, scenarios for the years 2020 and 2050 as well as different process schemes are defined in order to investigate the effects of different optimizations. In 2050, the integration of the plasma-based calcium looping is estimated to increase the PtL efficiency from 25 % to 32 %. This leads to net production costs (NPC) of 2.5 EUR/L marine diesel. By illustrating the techno-economic potential, further development goals can be set to achieve NPC below 2.0 EUR/L. This can be achieved in the presented process setup with plasma efficiencies of 45 % at conversions of 65 %. The investigations allow statements regarding the system integration and the degree of optimization of the plasma-based calcium looping.

Background: Antipsychotics are commonly administered to nursing home residents with dementia, despite the associated risk of severe adverse events. Objective: This study aimed to explore healthcare professionals' experiences in caring for nursing home residents with dementia, with a focus on rationales behind the use of antipsychotics. Method: Twelve semi-structured interviews with healthcare professionals' from Danish nursing homes were conducted and analyzed using the method Systematic Text Condensation. Results: Nonpharmacological interventions were reported as the primary approach to care and the first-choice treatment for behavioral and psychological symptoms of dementia (BPSD). Use of antipsychotics was considered to serve as a last resort, reserved for residents with severe symptoms. However, most informants preferred a more limited use. The study identified four main barriers to reduce the use of antipsychotics: “Scarcity of resources”, “Perceiving antipsychotic use to provide relieve”, “Reluctance towards deprescribing” and “Limited access to medical counseling”, and three potential enablers: “Updating knowledge and nonpharmacological competencies”, “Management support and clear procedures” and “Regularity in interdisciplinary collaboration”. Conclusion: The treatment and care were reported as primarily following guidelines in BPSD. Several barriers were perceived to challenge the healthcare professionals' preference of limited use of antipsychotics. To further reduce the use, this study highlights the importance of understanding the adverse effects caused by limited resources, enhancing employee knowledge and competencies and ensuring regular interprofessional collaboration for assessing and reassessing the need to use antipsychotics.

Abstract Abnormal mitochondria have been observed in bronchial- and alveolar epithelial cells of patients with chronic obstructive pulmonary disease (COPD). However, it is unknown if alterations in the molecular pathways regulating mitochondrial turnover (mitochondrial biogenesis vs mitophagy) are involved. Therefore, in this study, the abundance of key molecules controlling mitochondrial turnover were assessed in peripheral lung tissue from non-COPD patients (n = 6) and COPD patients (n = 11; GOLDII n = 4/11; GOLDIV n = 7/11) and in both undifferentiated and differentiated human primary bronchial epithelial cells (PBEC) from non-COPD patients and COPD patients (n = 4–7 patients/group). We observed significantly decreased transcript levels of key molecules controlling mitochondrial biogenesis (PPARGC1B, PPRC1, PPARD) in peripheral lung tissue from severe COPD patients. Interestingly, mRNA levels of the transcription factor TFAM (mitochondrial biogenesis) and BNIP3L (mitophagy) were increased in these patients. In general, these alterations were not recapitulated in undifferentiated and differentiated PBECs with the exception of decreased PPARGC1B expression in both PBEC models. Although these findings provide valuable insight in these pathways in bronchial epithelial cells and peripheral lung tissue of COPD patients, whether or not these alterations contribute to COPD pathogenesis, underlie changes in mitochondrial function or may represent compensatory mechanisms remains to be established.

Abstract Background Retinal ganglion cell (RGC) degeneration and death cause vision loss in patients with glaucoma. Regulated cell death, once initiated, is generally considered to be an irreversible process. Recently, we showed that, by timely removing the cell death stimulus, stressed neuronal PC12 cells can recover from phosphatidylserine (PS) exposure, nuclear shrinkage, DNA damage, mitochondrial fragmentation, mitochondrial membrane potential loss, and retraction of neurites, all hallmarks of an activated cell death program. Whether the cell death process can be reversed in neurons of the central nervous system, like RGCs, is still unknown. Here, we studied reversibility of the activated cell death program in primary rat RGCs (prRGCs). Methods prRGCs were exposed to ethanol (5%, vol/vol) to induce cell death. At different stages of the cell death process, ethanol was removed by washing and injured prRGCs were further cultured in fresh medium to see whether they recovered. The dynamics of single cells were monitored by high-resolution live-cell spinning disk microscopy. PS exposure, mitochondrial structure, membrane potential, and intracellular Ca2+ were revealed by annexin A5-FITC, Mito-tracker, TMRM, and Fluo 8-AM staining, respectively. The distribution of cytochrome c was investigated by immunofluorescence. The ultrastructure of mitochondria was studied by electron microscopy. Results Analysis of temporal relationships between mitochondrial changes and PS exposure showed that fragmentation of the mitochondrial network and loss of mitochondrial membrane potential occurred before PS exposure. Mitochondrial changes proceeded caspase-independently, while PS exposure was caspase dependent. Interestingly, prRGCs recovered quickly from these mitochondrial changes but not from PS exposure at the plasma membrane. Correlative light and electron microscopy showed that stress-induced decrease in mitochondrial area, length and cristae number was reversible. Intracellular Ca2+ was elevated during this stage of reversible mitochondrial injury, but there was no sign of mitochondrial cytochrome c release. Conclusions Our study demonstrates that RGCs with impaired mitochondrial structure and function can fully recover if there is no mitochondrial cytochrome c release yet, and no PS is exposed at the plasma membrane. This finding indicates that there is a time window for rescuing dying or injured RGCs, by simply removing the cell death stimulus. Video Abstract

Abstract The goal of affirmative action programs is to establish equal opportunities for women and men. Past research has focused on one type of affirmative action, namely quotas, and found that the implementation of quotas is often met by a backlash from employees. The current study adds to the literature by investigating fairness and importance perceptions of career development programs offered only to women at Austrian universities or the Austrian Science Fund. Using the model of attitudes toward affirmative action programs the current experimental study tested whether providing participants with information about the benefits (gain-message condition) or costs (loss-message condition) of the implementation of affirmative action programs influenced participants’ perceptions of affirmative action programs. In the current online study, the 510 participants (52.5% cisgender women and 47.5% cisgender men; M age = 29.5, SD = 9.5) from German-speaking countries in Europe gave on average higher fairness and importance ratings to career development programs offered to all employees than to such programs offered specifically to women. Men in the gain-message condition and loss-message condition gave higher fairness ratings to affirmative action programs than did men in the control condition (i.e., men who read a text that gave no justification for the implementation of affirmative action programs). Men in the gain-message condition also gave higher importance ratings to affirmative action programs than did men in the control condition. Women were not influenced in their ratings by the justification of the implementation of affirmative action programs. Nevertheless, women’s perceptions of affirmative action programs were more favorable than men’s. When implementing affirmative action programs in organizations, providing information that explains why affirmative action is needed and how all employees benefit therefrom can increase support from men who, as seen from past research, are known to be most opposed to affirmative action.

Thermal propagation events are characterized by fire and thick black smoke, leading to propagation methods with a focus on preventing heat transfer and optimizing gas flow. Yet little attention is being paid to the electric conductivity of the gas, leading to possibly unexpected battery casing openings due to lightning arcs as well as potentially providing the minimum ignition energy. This gas composition (omitting particles) was used at different temperatures and pressures in a lightning arc test bench, leading to the Paschen curve. Using a mini-module cell setup, filtered venting gas was flowed through another lightning arc test bench, allowing for in situ measurements.

The integrity of the 12Vdc power distribution system on a vehicle is essential to guarantee continuous power supply to safety-relevant consumers. Safety-relevant consumers are critical loads, for example, electric power steering, braking systems with functionalities like Anti-Lock Braking or Electronic Stability Control, and autonomous drive systems. To prevent insufficient power supply for safety-relevant consumers due to an increased wiring harness resistance, a novel diagnostic approach is developed to determine the condition of the power distribution, especially the electrical resistance. The influence of measurement errors and bus commutation on the estimation is investigated by using a simulation. By using the diagnostic, a resistance determination in the milliohm range with a standard deviation of σ=0.3mΩ can be achieved under realistic conditions. This ensures that failures in the wiring harness can be identified, avoiding cascading effects and minimizing recalls. Compared to the state of the art, redundancies, costs, and weight can be saved with the proposed diagnostic system based on electrical resistance estimation.

Abstract Lower respiratory tract infections caused by Streptococcus pneumoniae (Spn) are a leading cause of death globally. Here we investigate the bronchial epithelial cellular response to Spn infection on a transcriptomic, proteomic and metabolic level. We found the NAD+ salvage pathway to be dysregulated upon infection in a cell line model, primary human lung tissue and in vivo in rodents, leading to a reduced production of NAD+. Knockdown of NAD+ salvage enzymes (NAMPT, NMNAT1) increased bacterial replication. NAD+ treatment of Spn inhibited its growth while growth of other respiratory pathogens improved. Boosting NAD+ production increased NAD+ levels in immortalized and primary cells and decreased bacterial replication upon infection. NAD+ treatment of Spn dysregulated the bacterial metabolism and reduced intrabacterial ATP. Enhancing the bacterial ATP metabolism abolished the antibacterial effect of NAD+. Thus, we identified the NAD+ salvage pathway as an antibacterial pathway in Spn infections, predicting an antibacterial mechanism of NAD+.

Dereje Mengesha Berta,1 Mulugeta Melku,1,2 Tiruneh Adane,1 Mekonnen Girma,3 Zewudu Mulatie,4 Elias Chane,5 Bisrat Birke Teketelew1 1Department of Hematology and Immunohematology, School of Biomedical and Laboratory Sciences, College of Medicine and Health Sciences, University of Gondar, Gondar, Ethiopia; 2College of Medicine and Public Health, Flinders University, Adelaide, SA, Australia; 3Department of Quality Assurance and Laboratory Management, School of Biomedical and Laboratory Sciences, College of Medicine and Health Sciences, University of Gondar, Gondar, Ethiopia; 4Department of Medical Laboratory Science, Wollo University, Wollo, Ethiopia; 5Department of Clinical Chemistry, School of Biomedical and Laboratory Sciences, College of Medicine and Health Sciences, University of Gondar, Gondar, EthiopiaCorrespondence: Dereje Mengesha Berta, Department of Hematology and Immunohematology, School of Biomedical and Laboratory Sciences, College of Medicine and Health Sciences, University of Gondar, PO Box 196, Gondar, Ethiopia, Tel +251910594497, Email mengeshad826@gmail.comIntroduction: In clinical laboratory, the performance of the hematology analyzer should be checked routinely to ensure the desired quality. Therefore, the aim of the study was to assess the analytical performance of hematology analyzer using sigma metrics.Methods: The study included all daily internal quality control (IQC) data of hematology analyzer prospectively from August to October 2022. Data was collected using record formats by trained laboratory professionals. The sigma values of each CBC parameter were calculated using the formula: Sigma = (TEa – Bias) / CV. The TEa data were adopted from five different guidelines to calculate sigma value of the laboratory based on different specification. The bias of all complete blood count (CBC) parameters was calculated from the laboratory mean of the daily IQC data and the target value of the manufacturer in the insert kit. A coefficient of variations was also calculated using IQC data.Results: The current study found that sigma value of the analyzer varied based on source of TEa. Except HCT out 5 parameters included based on CLIA guideline, except MCV, MCHC, RDW, MPV and Basophil out of 15 parameters included based on EFLM 2022 minimum guideline, except Hb and PLT out of 9 parameters included based on SOTA guideline other parameters meets minimum specification (< 3 sigma value). On the other hand, all parameters included in Rilibak and Standards of Spanish guideline achieved minimum specification (> 3 sigma value).Conclusion: Sigma values of the CBC parameters have significantly varied depends on the TEa sources. So, it is recommended laboratory to use alternative sigma value based on its preference. Additionally, it is suggested that the laboratory to design local Westgard rules for each parameter based on sigma value.Keywords: analytical performance, hematology, laboratory, sigma metrics

Abstract Open science practices such as posting data or code and pre-registering analyses are increasingly prescribed and debated in the applied sciences, but the actual popularity and lifetime usage of these practices remain unknown. This study provides an assessment of attitudes toward, use of, and perceived norms regarding open science practices from a sample of authors published in top-10 (most-cited) journals and PhD students in top-20 ranked North American departments from four major social science disciplines: economics, political science, psychology, and sociology. We observe largely favorable private attitudes toward widespread lifetime usage (meaning that a researcher has used a particular practice at least once) of open science practices. As of 2020, nearly 90% of scholars had ever used at least one such practice. Support for posting data or code online is higher (88% overall support and nearly at the ceiling in some fields) than support for pre-registration (58% overall). With respect to norms, there is evidence that the scholars in our sample appear to underestimate the use of open science practices in their field. We also document that the reported lifetime prevalence of open science practices increased from 49% in 2010 to 87% a decade later.

Introduction Illness perception refers to patients’ subjective representations and appraisals of somatic and mental symptoms. These are relevant for self-management and outcome. In clinical practice, patients with functional neurological disorder (FND) often encounter a fragmented biomedical attitude, which leaves them without clear concepts. In this context, illness perception is relevant.Methods Illness perception was assessed in FND patients and compared with samples of psychosomatic patients (PSM) as well as poststroke patients (STR). The three samples (FND, n=87; PSM, n=97 and STR, n=92) were almost all in inpatient treatment or rehabilitation. Illness perception was assessed with the revised German version of the Illness Perception Questionnaire (IPQ-R). For assessments of correlations, depressive symptoms were tested with the Patient Health Questionnaire-9, dissociative and functional neurological symptoms by the German adaption of the Dissociative Experiences Scale and biopsychosocial complexity by the INTERMED Self-Assessment questionnaire.Results Apart from the chronicity subscale, all dimensions of the IPQ-R differed between groups. FND patients perceived lower illness coherence and personal control than both other groups and attributed their illness more to chance than to behavioural risk factors. PSM patients had the strongest emotional representations. There were only few correlations with dissociative scores and biopsychosocial complexity.Conclusion Illness perception is an important issue in patients with FND with particular emphasis on low illness coherence and personal control. Missing associations with biopsychosocial complexity suggest that subjective illness perception is an important complementary but separate issue, which likely influences therapeutic alliance and self-management in FND. Future studies should assess its influences on outcome.Trial registration number DRKS00024685; German Clinical Trials Register; www.drks.de.

Introduction Sexual and gender minority (SGM) people are often found to have lower levels of body appreciation than do cis-heteronormative people. Objectives The current study utilizes the social cure model and the intragroup status and health model to investigate whether identification with a SGM social group and identity centrality (i.e., the degree to which a specific social identity is important to an individual) is linked to experiences of hostile behaviors because of a person’s looks or body and consequently, to body appreciation. Methods A cross-sectional online-questionnaire study was conducted with 1,680 German-speaking participants (49.2% cisgender women, 37.7% cisgender men, 9.0% non-binary, 4.1% transgender; M age = 32.7, SD = 12.5). The Multidimensional and Multicomponent Measure of Social Identification, the Body Appreciation Scale-2, the Perceived Stigmatization Questionnaire and the Sociocultural Attitudes Towards Appearance Questionnaire-4, revised were used. A manifest path model was calculated. Results People who identified as gender minority (GM) people and LGBTQ* people reported lower levels of body appreciation. Sexual minority (SM) individuals who identified with a social group other than LGBTQ* people reported levels of body appreciation similar to those of individuals who identified as women. Individuals who identified as GM people experienced fewer instances of hostile behaviors because of their looks or body the higher their level of identity centrality was. On the other hand, individuals who identified as LGBTQ* people more frequently encountered hostile behaviors because of their looks or body when their identity centrality was strong. Frequent experiences of hostile behaviors because of a person’s looks or body was linked to poor body appreciation in all social groups. Conclusions Identity centrality might help alleviate experiences of discrimination, especially in people who identify as GM people, as the social cure model suggests. In line with the intragroup status and health model, individuals who strongly identify as LGBTQ* people might be more visible as SM people and experience more discrimination than do SM people who identify with another social group. Disclosure of Interest None Declared

Abstract Background Lung infections caused by Streptococcus pneumonia are a global leading cause of death. The reactive oxygen species H2O2 is one of the virulence factors of Streptococcus pneumoniae. The Golgi apparatus is essential for the inflammatory response of a eukaryotic cell. Golgi fragmentation was previously shown to be induced by bacterial pathogens and in response to H2O2 treatment. This led us to investigate whether the Golgi apparatus is actively involved and targeted in host–pathogen interactions during pneumococcal infections. Methods Following in vitro infection of BEAS-2B bronchial epithelial cells with Streptococcus pneumoniae for 16 h, the structure of the Golgi apparatus was assessed by fluorescence staining of the Golgi-associated protein, Golgin-97. To investigate the effect of H2O2 production on Golgi structure, BEAS-2B cells were treated with H2O2 or the H2O2 degrading enzyme Catalase, prior to Golgi staining. Artificial disruption of the Golgi apparatus was induced by treatment of cells with the GBF1 inhibitor, Golgicide A. A proinflammatory cellular response was induced by treatment of cells with the bacterial cell wall component and TLR4 ligand lipoteichoic acid. Results In vitro infection of bronchial epithelial cells with wild type Streptococcus pneumoniae led to a disruption of normal Golgi structure. Golgi fragmentation was not observed after deletion of the pneumococcal H2O2-producing gene, spxB, or neutralization of H2O2 by catalase treatment, but could be induced by H2O2 treatment. Streptococcus pneumoniae infection significantly reduced host cell protein glycosylation and artificial disruption of Golgi structure significantly reduced bacterial adherence, but increased bacterial counts in the supernatant. To understand if this effect depended on cell-contact or soluble factors, pneumococci were treated with cell-supernatant of cells treated with Golgicide A and/or lipoteichoic acid. This approach revealed that lipoteichoic acid conditioned medium inhibits bacterial replication in presence of host cells. In contrast, artificial Golgi fragmentation by Golgicide A treatment prior to lipoteichoic acid treatment rescued bacterial replication. This effect was associated with an increase of IL-6 and IL-8 in the supernatant of lipoteichoic acid treated cells. The increased cytokine release was abolished if cells were treated with Golgicide A prior to lipoteichoic acid treatment. Conclusion Streptococcus pneumoniae disrupts the Golgi apparatus in an H2O2-dependent manner, thereby inhibiting paracrine anti-infective mechanisms. Video Abstract

Abstract Loss of neurons in chronic neurodegenerative diseases may occur over a period of many years. Once initiated, neuronal cell death is accompanied by distinct phenotypic changes including cell shrinkage, neurite retraction, mitochondrial fragmentation, nuclear condensation, membrane blebbing and phosphatidylserine (PS) exposure at the plasma membrane. It is still poorly understood which events mark the point of no return for dying neurons. Here we analyzed the neuronal cell line SH-SY5Y expressing cytochrome C (Cyto.C)-GFP. Cells were exposed temporarily to ethanol (EtOH) and tracked longitudinally in time by light and fluorescent microscopy. Exposure to EtOH induced elevation of intracellular Ca2+ and reactive oxygen species, cell shrinkage, neurite retraction, mitochondrial fragmentation, nuclear condensation, membrane blebbing, PS exposure and Cyto.C release into the cytosol. Removing EtOH at predetermined time points revealed that all phenomena except Cyto.C release occurred in a phase of neuronal cell death in which full recovery to a neurite-bearing cell was still possible. Our findings underscore a strategy of treating chronic neurodegenerative diseases by removing stressors from neurons and harnessing intracellular targets that delay or prevent trespassing the point of no return.

Dereje Berta,1 Mekonnen Girma,2 Mulugeta Melku,1,3 Tiruneh Adane,1 Bisrat Birke,1 Aregawi Yalew1 1Department of Hematology and Immunohematology, School of Biomedical and Laboratory Sciences, College of Medicine and Health Sciences, University of Gondar, Gondar, Ethiopia; 2Department of Quality Assurance and Laboratory Management, School of Biomedical and Laboratory Sciences, College of Medicine and Health Sciences, University of Gondar, Gondar, Ethiopia; 3College of Medicine and Public Health, Flinders University, Adelaide, SA, AustraliaCorrespondence: Dereje Berta, Email mengeshad826@gmail.comAbstract: Ribonucleic acid splicing is a crucial process to create a mature mRNA molecule by removing introns and ligating exons. This is a highly regulated process, but any alteration in splicing factors, splicing sites, or auxiliary components affects the final products of the gene. In diffuse large B-cell lymphoma, splicing mutations such as mutant splice sites, aberrant alternative splicing, exon skipping, and intron retention are detected. The alteration affects tumor suppression, DNA repair, cell cycle, cell differentiation, cell proliferation, and apoptosis. As a result, malignant transformation, cancer progression, and metastasis occurred in B cells at the germinal center. B-cell lymphoma 7 protein family member A (BCL7A), cluster of differentiation 79B (CD79B), myeloid differentiation primary response gene 88 (MYD88), tumor protein P53 (TP53), signal transducer and activator of transcription (STAT), serum- and glucose-regulated kinase 1 (SGK1), Pou class 2 associating factor 1 (POU2AF1), and neurogenic locus notch homolog protein 1 (NOTCH) are the most common genes affected by splicing mutations in diffuse large B cell lymphoma.Keywords: RNA-splicing, aberrant splicing, alternative splicing, diffuse large B-cell lymphoma

Biogenic polyamines are ubiquitous compounds. Dysregulation of their metabolism is associated with the development of various pathologies, including cancer, hyperproliferative diseases, and infections. The canonical pathway of polyamine catabolism includes acetylation of spermine and spermidine and subsequent acetylpolyamine oxidase (PAOX)-mediated oxidation of acetylpolyamines (back-conversion) or their direct efflux from the cell. PAOX is considered to catalyze a non-rate-limiting catabolic step. Here, we show that PAOX transcription levels are extremely low in various tumor- and non-tumor cell lines and, in most cases, do not change in response to altered polyamine metabolism. Its enzymatic activity is undetectable in the majority of cell lines except for neuroblastoma and low passage glioblastoma cell lines. Treatment of A549 cells with N1,N11-diethylnorspermine leads to PAOX induction, but its contribution to polyamine catabolism remains moderate. We also describe two alternative enzyme isoforms and show that isoform 4 has diminished oxidase activity and isoform 2 is inactive. PAOX overexpression correlates with the resistance of cancer cells to genotoxic antitumor drugs, indicating that PAOX may be a useful therapeutic target. Finally, PAOX is dispensable for the replication of various viruses. These data suggest that a decrease in polyamine levels is achieved predominantly by the secretion of acetylated spermine and spermidine rather than by back-conversion.

Hepatitis C virus (HCV) is an oncogenic virus that causes chronic liver disease in more than 80% of patients. During the last decade, efficient direct-acting antivirals were introduced into clinical practice. However, clearance of the virus does not reduce the risk of end-stage liver diseases to the level observed in patients who have never been infected. So, investigation of HCV pathogenesis is still warranted. Virus-induced changes in cell metabolism contribute to the development of HCV-associated liver pathologies. Here, we studied the impact of the virus on the metabolism of polyamines and proline as well as on the urea cycle, which plays a crucial role in liver function. It was found that HCV strongly suppresses the expression of arginase, a key enzyme of the urea cycle, leading to the accumulation of arginine, and up-regulates proline oxidase with a concomitant decrease in proline concentrations. The addition of exogenous proline moderately suppressed viral replication. HCV up-regulated transcription but suppressed protein levels of polyamine-metabolizing enzymes. This resulted in a decrease in polyamine content in infected cells. Finally, compounds targeting polyamine metabolism demonstrated pronounced antiviral activity, pointing to spermine and spermidine as compounds affecting HCV replication. These data expand our understanding of HCV’s imprint on cell metabolism.