Deubiquitinases in muscle physiology and disorders.

In vivo, muscle and neuronal cells are post-mitotic, and their function is predominantly regulated by proteostasis, a multilayer molecular process that maintains a delicate balance of protein homeostasis. The ubiquitin-proteasome system (UPS) is a key regulator of proteostasis. A dysfunctional UPS is a hallmark of muscle ageing and is often impacted in neuromuscular disorders (NMDs). Malfunction of the UPS often results in aberrant protein accumulation which can lead to protein aggregation and/or mis-localization affecting its function. Deubiquitinating enzymes (DUBs) are key players in the UPS, controlling protein turnover and maintaining the free ubiquitin pool. Several mutations in DUB encoding genes are linked to human NMDs, such as ATXN3, OTUD7A, UCHL1 and USP14, whilst other NMDs are associated with dysregulation of DUB expression. USP5, USP9X and USP14 are implicated in synaptic transmission and remodeling at the neuromuscular junction. Mice lacking USP19 show increased maintenance of lean muscle mass. In this review, we highlight the involvement of DUBs in muscle physiology and NMDs, particularly in processes affecting muscle regeneration, degeneration and inflammation following muscle injury. DUBs have recently garnered much respect as promising drug targets, and their roles in muscle maturation, regeneration and degeneration may provide the framework for novel therapeutics to treat muscular disorders including NMDs, sarcopenia and cachexia.

Is Mean Platelet Volume a Predictive Marker for the Development of Thrombosis in Patients with COVID-19 Infection?

Mean platelet volume (MPV) can provide important information about the course and prognosis of many diseases. MPV is an early indicator of platelet activation, which has an important role in the pathogenesis of thrombosis. In this study, we aimed to investigate whether MPV was a predictive marker for the development of thrombosis in hospitalized patients with COVID-19 infection. Fifty-seven patients whose courses were followed after the diagnosis of COVID-19 infection using a polymerase chain reaction test during the pandemic were included in the study. Our results demonstrated that there was a negative correlation between platelet count and MPV (r=0.470, p≤ 0.01), and there was a positive correlation between Platelet Distribution Width (PDW) and MPV (r=0,933, p≤ 0.01), but no significant correlation was found between the other variables and MPV.

Impact of Ferroptosis Inducers on Chronic Radiation-exposed Survivor Glioblastoma Cells.

INTRODUCTION: The median survival of patients diagnosed with glioblastoma is very poor, despite efforts to improve the therapeutic effects of surgery, followed by treatment with temozolomide (TMZ) and ionizing radiation (IR). The utilization of TMZ or IR survivor cell models has enhanced the understanding of glioblastoma biology and the development of novel therapeutic strategies. In this present study, naïve U373 and clinically relevant U373 IRsurvivor (Surv) cells were used, as the IR-Surv cell model mimics the chronic long-term exposure to standardized radiotherapy for patients with glioblastoma in the clinic. As the role of ferroptosis in the IR survivor cell model has not previously been reported, we aimed to clarify its involvement in the clinically relevant IR-Surv glioblastoma model. METHODS: Transcriptomic alterations of ferroptosis-related genes were studied on naïve U373 and IR-Surv cell populations. To determine the effects of glutathione peroxidase inhibitors, ferroptosis-inducing agent 56 (FIN56) and Ras synthetic lethal 3 (RSL3), on the cells, several properties were assessed, including colony formation, cell viability and lipid peroxidation. RESULTS: Results from the transcriptomic analysis identified ferroptosis as a critical mechanism after radiation exposure in glioblastoma. Our findings also identified the role of ferroptosis inducers (FINs) in IR-survivor cells and suggested using FINs to treat glioblastoma. CONCLUSION: FINs serve an important role in radioresistant cells; thus, the results of the present study may contribute to improving survival in patients with glioblastoma.

Prospective analysis of febrile neutropenia patients with bacteraemia: the results of an international ID-IRI study.

OBJECTIVES: Bacteraemia during the course of neutropenia is often fatal. We aimed to identify factors predicting mortality to have an insight into better clinical management. METHODS: The study has a prospective, observational design using pooled data from febrile neutropenia patients with bacteraemia in 41 centres in 16 countries. Polymicrobial bacteraemias were excluded. It was performed through the Infectious Diseases-International Research Initiative platform between 17 March 2021 and June 2021. Univariate analysis followed by a multivariate binary logistic regression model was used to determine independent predictors of 30-d in-hospital mortality (sensitivity, 81.2%; specificity, 65%). RESULTS: A total of 431 patients were enrolled, and 85 (19.7%) died. Haematological malignancies were detected in 361 (83.7%) patients. Escherichia coli (n = 117, 27.1%), Klebsiellae (n = 95, 22% %), Pseudomonadaceae (n = 63, 14.6%), Coagulase-negative Staphylococci (n = 57, 13.2%), Staphylococcus aureus (n = 30, 7%), and Enterococci (n = 21, 4.9%) were the common pathogens. Meropenem and piperacillin-tazobactam susceptibility, among the isolated pathogens, were only 66.1% and 53.6%, respectively. Pulse rate (odds ratio [OR], 1.018; 95% confidence interval [CI], 1.002-1.034), quick SOFA score (OR, 2.857; 95% CI, 2.120-3.851), inappropriate antimicrobial treatment (OR, 1.774; 95% CI, 1.011-3.851), Gram-negative bacteraemia (OR, 2.894; 95% CI, 1.437-5.825), bacteraemia of non-urinary origin (OR, 11.262; 95% CI, 1.368-92.720), and advancing age (OR, 1.017; 95% CI, 1.001-1.034) were independent predictors of mortality. Bacteraemia in our neutropenic patient population had distinctive characteristics. The severity of infection and the way to control it with appropriate antimicrobials, and local epidemiological data, came forward. CONCLUSIONS: Local antibiotic susceptibility profiles should be integrated into therapeutic recommendations, and infection control and prevention measures should be prioritised in this era of rapidly increasing antibiotic resistance.

A case report of brain abscess caused by carbapenem-resistant Klebsiella pneumoniae.

The treatment of infections caused by carbapenem-resistant Klebsiella pneumoniae (CR-Kp) strains is difficult due to the limited antimicrobial options and high mortality. There are many reports on intracranial infections caused by CR-Kp, but only a few on brain abscesses caused by CR-Kp. Here, we present a case of brain abscess caused by CR-Kp successfully treated with combined antibiotics. A 26-year-old male patient was admitted to our hospital due to high fever and headache. His past medical history includes a surgical intervention due to an acute subdural hematoma, performed at an external healthcare center. After the current diagnosis of cerebral abscess, he underwent two surgeries. During the procedure, multiple cerebral abscesses were drained and capsulotomies were performed under ultrasound guidance. The combination of meropenem and vancomycin was started. The contents of the abscesses were sent to the microbiology and pathology laboratory. On the 3 rd day of treatment, the medical team was informed that CR-Kp grew in an abscess culture. The patient's treatment was changed to meropenem + colistin + tigecycline. The patient developed electrolyte disturbances during the follow-up and this was considered an adverse effect of colistin. On the 41 st day of treatment, colistin was discontinued, fosfomycin was added, and meropenem and tigecycline were maintained. Treatment was discontinued on the 68 th day, when the patient was discharged. The general condition of the patient, who has been followed up for two years, is satisfactory. The treatment of CR-Kp infections should be individualized, and the pharmacokinetics and pharmacodynamics of antibiotics should be considered in each case.

The possible effects of sodium fluoroscein to primary cell culture sampling in glioblastoma surgery.

•FL increases beta-galactosidase activity in GBM cell cultures.•FL cause a decrease in GBM cell numbers.•Sampling in GBM cell culture should be performed before using FL.

A case report of brain abscess caused by carbapenem-resistant Klebsiella pneumoniae

ABSTRACT The treatment of infections caused by carbapenem-resistant Klebsiella pneumoniae (CR-Kp) strains is difficult due to the limited antimicrobial options and high mortality. There are many reports on intracranial infections caused by CR-Kp, but only a few on brain abscesses caused by CR-Kp. Here, we present a case of brain abscess caused by CR-Kp successfully treated with combined antibiotics. A 26-year-old male patient was admitted to our hospital due to high fever and headache. His past medical history includes a surgical intervention due to an acute subdural hematoma, performed at an external healthcare center. After the current diagnosis of cerebral abscess, he underwent two surgeries. During the procedure, multiple cerebral abscesses were drained and capsulotomies were performed under ultrasound guidance. The combination of meropenem and vancomycin was started. The contents of the abscesses were sent to the microbiology and pathology laboratory. On the 3 rd day of treatment, the medical team was informed that CR-Kp grew in an abscess culture. The patient's treatment was changed to meropenem + colistin + tigecycline. The patient developed electrolyte disturbances during the follow-up and this was considered an adverse effect of colistin. On the 41 st day of treatment, colistin was discontinued, fosfomycin was added, and meropenem and tigecycline were maintained. Treatment was discontinued on the 68 th day, when the patient was discharged. The general condition of the patient, who has been followed up for two years, is satisfactory. The treatment of CR-Kp infections should be individualized, and the pharmacokinetics and pharmacodynamics of antibiotics should be considered in each case.

Comparison of antibiotic use in the COVID-19 pandemic with the pre-pandemic period in a university hospital.

Introduction: The appropriate use of antibiotics is an important strategy in slowing the development of antimicrobial resistance. This study aimed to evaluate antibiotic consumption and antibiotic use during the coronavirus disease 2019 (COVID-19) pre-pandemic period and pandemic period. Methods: Antibiotic consumption was evaluated with the antibiotic consumption index (ACI). Results: Antibiotics with the largest increase in ACI value during the pandemic period compared to the previous year increased from 0.4 to 1.8 DDI/100 bed days in moxifloxacin. Teicoplanin, linezolid, and clindamycin were not affected in terms of consumption. Conclusions: It was observed that the use of many intravenous antibiotics in our hospital increased during the pandemic period.

[Investigation of Factors Affecting Viral Clearance Time by Real-Time Polymerase Chain Reaction Method in Nasopharyngeal Swab Samples of COVID-19 Patients]. / COVID-19 Hastalarinin Nazofarengeal Sürüntü Örneklerinde Gerçek Zamanli Polimeraz Zincir Reaksiyonu ile Yöntemiyle Viral Klirens Süresine Etki Eden Faktörlerin Arastirilmasi.

The coronavirus disease-2019 (COVID-19) pandemic continues to threaten the lives of millions of people. Viral shedding through the respiratory tract is the main risk factor for the transmission of the severe acute respiratory syndrome-2 (SARS-CoV-2) virus from sick individuals to healthy individuals. In this study, we aimed to investigate the viral clearance (VC) time in PCR tests of COVID-19 patients and the possible factors affecting this time. Seventy patients older than 18 years of age whose presence of SARS-CoV-2 virus was proven by real-time polymerase chain reaction (Rt-PCR) in nasopharyngeal swab samples were included in the study. The presence of SARS-CoV-2 RNA was investigated by RT-PCR in nasopharyngeal swab samples at 48-72 hour intervals, five days after the initial diagnosis. Demographic , physical examination, laboratory test, computed tomography (CT) results, concomitant diseases, and duration of VC were recorded. Of the cases, 41 were female and 29 were male. The mean age was 45.8 ± 19.2 years. According to the CT results, in the group with no involvement, local involvement and widespread involvement, the duration of VC was 9.66 ± 5.91 days, 9.99 ± 4.68 days, and 10.94 ± 5.34 days, respectively (p> 0.05). While the duration of VC was determined as 8.93 ± 4.33 days in the group without comorbidity, this period was found to be 12.26 ± 5.69 days (p= 0.025) in the group with the comorbidity. It was determined that the duration of VC was 9.55 ± 6.37 days in women and 9.20 ± 7.22 days in men (p= 0.040). The duration of VC was found to be 10.18 ± 7.1 days in patients over 50 years of age and 8.87 ± 5.15 days under 50 years of age (p= 0.03). A significant correlation was found between the laboratory test lactate dehydrogenase level and VC duration (p= 0.007). However, a significant relationship could not be established between other laboratory test results and the duration of VC. In this retrospective observational study, the relationship between viral clearance duration in Rt-PCR and gender, age, CT results, comorbidities and laboratory results in nasopharyngeal swab samples was investigated and it was determined that the duration of VC was significantly prolonged in case of female gender, being over 50 years old and having a comorbid disease. The results obtained may contribute to predict the isolation times of the patients and to reveal the factors that may affect viral shedding.

Chronically Radiation-Exposed Survivor Glioblastoma Cells Display Poor Response to Chk1 Inhibition under Hypoxia.

Glioblastoma is the most malignant primary brain tumor, and a cornerstone in its treatment is radiotherapy. However, tumor cells surviving after irradiation indicates treatment failure; therefore, better understanding of the mechanisms regulating radiotherapy response is of utmost importance. In this study, we generated clinically relevant irradiation-exposed models by applying fractionated radiotherapy over a long time and selecting irradiation-survivor (IR-Surv) glioblastoma cells. We examined the transcriptomic alterations, cell cycle and growth rate changes and responses to secondary radiotherapy and DNA damage response (DDR) modulators. Accordingly, IR-Surv cells exhibited slower growth and partly retained their ability to resist secondary irradiation. Concomitantly, IR-Surv cells upregulated the expression of DDR-related genes, such as CHK1, ATM, ATR, and MGMT, and had better DNA repair capacity. IR-Surv cells displayed downregulation of hypoxic signature and lower induction of hypoxia target genes, compared to naïve glioblastoma cells. Moreover, Chk1 inhibition alone or in combination with irradiation significantly reduced cell viability in both naïve and IR-Surv cells. However, IR-Surv cells' response to Chk1 inhibition markedly decreased under hypoxic conditions. Taken together, we demonstrate the utility of combining DDR inhibitors and irradiation as a successful approach for both naïve and IR-Surv glioblastoma cells as long as cells are refrained from hypoxic conditions.

A 5-year surveillance of healthcare-associated infections in a university hospital: A retrospective analysis.

Objectives: "Nosocomial infections" or "healthcare-associated infections" are a significant public health problem around the world. This study aimed to assess the rate of laboratory-confirmed healthcare-associated infections, frequency of nosocomial pathogens, and the antimicrobial resistance patterns of bacterial isolates in a University Hospital. Methods: A retrospective evaluation of healthcare-associated infections in a University Hospital, between the years 2015 and 2019 in Tekirdag, Turkey. Results: During the 5 years, the incidence densities of healthcare-associated infections in intensive care units and clinics were 10.31 and 1.70/1000 patient-days, respectively. The rates of ventilator-associated pneumonia, central line-associated bloodstream infections, and catheter-associated urinary tract infections in intensive care units were 11.57, 4.02, and 1.99 per 1000 device-days, respectively. The most common healthcare-associated infections according to the primary sites were bloodstream infections (55.3%) and pneumonia (20.4%). 67.5% of the isolated microorganisms as nosocomial agents were Gram-negative bacteria, 24.9% of Gram-positive bacteria, and 7.6% of Candida. The most frequently isolated causative agents were Escherichia coli (16.7%) and Pseudomonas aeruginosa (15.7%). The rate of extended-spectrum beta-lactamase production among E. coli isolates was 51.1%. Carbapenem resistance was 29.8% among isolates of P. aeruginosa, 95.1% among isolates of Acinetobacter baumannii, and 18.2% among isolates of Klebsiella pneumoniae. Colistin resistance was 2.4% among isolates of A. baumannii. Vancomycin resistance was 5.3% among isolates of Enterococci. Conclusion: Our study results demonstrate that healthcare-associated infections are predominantly originated by intensive care units. The microorganisms isolated from intensive care units are highly resistant to many antimicrobial agents. The rising incidence of multidrug-resistant microorganisms indicates that more interventions are urgently needed to reduce healthcare-associated infections in our intensive care units.

Comparison of astrocytes and gap junction proteins in the white matter of genetic absence epileptic and control rats: an experimental study.

OBJECTIVES: The role of white matter astrocytes in absence epilepsy is unknown. The present study aims to quantify astrocytic markers glial fibrillary acidic protein (GFAP), gap junction's proteins connexin 30 (Cx30) and connexin 43 (Cx43) in the corpus callosum (CC) of genetic absence epileptic rats from Strasbourg (GAERS), Wistar albino glaxo rats from Rijswijk (WAG/Rij)and compare the results with control animals. METHODS: -The density of GFAP, Cx30 and Cx43 positive astrocytes in per unite area were quantified in the CC of GAERS, WAG/Rij and control animals using immunohistochemistry and real-time quantitative polymerase chain reaction (RT-qPCR). The quantifications were made from three regions of CC; below the primary somatosensory (S1BF), below the motor (M1) and below the retrosplenial (RSG) cortices. RESULTS: oThe number GFAP, Cx30 and Cx43 immunopositive astrocytes showed heterogeneous distribution within the CC. The GFAP immunopositive astrocytes was significantly high in the S1BF region of the three strains. The immunopositive GFAP and Cx43 showed significant decrease in the S1BF and M1 regions in GAERS and WAG/Rij compared to control animals, however, an increase in the immunopositive Cx30 was observed in the same regions in both GAERS and WAG/Rij compared to control Wistar animals but the increase was significant for GAERS but not for WAG/Rij. The RT-qPCR analysis was corroborated by GFAP immunohistochemistry results. CONCLUSION: The different expression pattern of the two Cx's in the CC of the epileptic strains compared to control animals may indicate a compensatory response or maybe the cause of generalization of absence seizures.

Does astrocyte gap junction protein expression differ during development in absence epileptic rats?

Intercellular communication via gap junctions (GJs) has a wide variety of complex and essential functions in the CNS. In the present developmental study, we aimed to quantify the number of astrocytic GJs protein connexin 30 (Cx30) of genetic model of absence epilepsy rats from Strasbourg (GAERS) at postnatal P10, P30, and P60 days in the epileptic focal areas involved in the cortico-thalamic circuit. We compared the results with Wistar rats using immunohistochemistry and western blotting. The number of Cx30 immunopositive astrocytes per unit area were quantified for the somatosensory cortex (SSCx), ventrobasal (VB), and lateral geniculate (LGN) thalamic nuclei of the two strains and Cx30 western blot was applied to the tissue samples from the same regions. Both immunohistochemical and western blot results revealed the presence of Cx30 in all regions studied at P10 in both Wistar and GAERS animals. The SSCx, VB, and LGN of Wistar animals showed progressive increase in the number of Cx30 immunopositive labeled astrocytes from P10 to P30 and reached a peak at P30; then a significant decline was observed from P30 to P60 for the SSCx and VB. However, in GAERS Cx30 immunopositive labeled astrocytes showed a progressive increase from P10 to P60 for all brain regions studied. The immunohistochemical data highly corresponded with western blotting results. We conclude that the developmental disproportional expression of Cx30 in the epileptic focal areas in GAERS may be related to the onset of absence seizures or may be related to the neurogenesis of absence epilepsy.

The incidence of acute kidney injury and its association with mortality in patients diagnosed with COVID-19 followed up in intensive care unit.

INTRODUCTION: The kidneys are some of the most frequently affected organs during coronavirus disease 2019 (COVID-19). This multicenter study evaluated the incidence of and risk factors for acute kidney injury (AKI) in COVID-19 patients followed up in intensive care unit (ICU) and its association with mortality. METHODS: Three hundred twenty-eight patients diagnosed with COVID-19 and hospitalized in ICU were included. Risk factors associated with AKI and mortality were evaluated. RESULTS: Eighty-eight patients (27.9%) were diagnosed with AKI. AKI was significantly associated with older age, higher baseline creatinine level, lower albumin level, and coexistence of cardiovascular disease and chronic obstructive pulmonary disease. Mortality in the entire study group was significantly associated with AKI, older age, requirement of invasive mechanical ventilation, higher neutrophil level, lower lymphocyte, and albumin levels. CONCLUSION: AKI is frequently seen during the course of COVID-19 and is associated with high mortality. Identifying AKI-related risk factors appears essential in the management of COVID-19 patients.

The place of infectious markers in predicting culture positivity in patients with renal failure hospitalized with suspected infection.

Infectious diseases are an important cause of mortality in patients with renal failure. The markers used to diagnose infection in patients with renal failure have various limitations. Culture positivity is an objective guide in that context. The purpose of this study was to examine the effectiveness of frequently used markers of bacterial infection in predicting culture positivity in renal failure patients with renal failure hospitalized with suspected bacterial infection over an approximately 1.5-year period were included in this prospective observational study. Patients' demographic and laboratory findings were recorded. Demographic and laboratory findings and mortality were compared between patients with and without culture-positivity. Parameters affecting culture positivity were also analyzed. Four hundred twenty-six patients (median age 67.50, 45.5% female) were included in the study. Culture positivity was determined in 54.5% of patients. Hospital stay was longer (p < 0.001) and leukocyte (p < 0.001), neutrophil percentage (p < 0.05) and CRP (p < 0.001) values were significantly higher in culture-positive patients. Mortality was also significantly higher in culture-positive patients than in culture-negative patients (p < 0.05). CRP was determined as a predictor of culture positivity at logistic regression analysis (p = 0.000, exp ß [1.004]). Culture positivity was determined in more than half of the patients hospitalized with suspected bacterial infection. CRP, a longstanding marker, was identified as a parameter predicting culture positivity. We think that the determination in further studies of a cut-off point for CRP in determining culture positivity may be a useful diagnostic guide.

A 24-generation-old founder mutation impairs splicing of RBBP8 in Pakistani families affected with Jawad syndrome.

Jawad syndrome is a multiple congenital anomaly and intellectual disability syndrome with mutation in RBBP8 reported only in two families. Here, we report on two new families from Pakistan and identified a previously reported variant in RBBP8, NM_002894.3:c.1808-1809delTA. We could show that this mutation impairs splicing resulting in two different abnormal transcripts. Finally, we could verify a shared haplotype among all four families and estimate the founder event to have occurred some 24 generations ago.

Comparing astrocytic gap junction of genetic absence epileptic rats with control rats: an experimental study.

The synchronization of astrocytes via gap junctions (GJ) is a crucial mechanism in epileptic conditions, contributing to the synchronization of the neuronal networks. Little is known about the endogenous response of GJ in genetic absence epileptic animal models. We evaluated and quantified astrocyte GJ protein connexin (Cx) 30 and 43 in the somatosensory cortex (SSCx), ventrobasal (VB), centromedian (CM), lateral geniculate (LGN) and thalamic reticular (TRN) nuclei of thalamus of genetic absence epilepsy rats from Strasbourg (GAERS), Wistar albino glaxo rats from Rijswijk (WAG/Rij) and control Wistar animals using immunohistochemistry and Western Blot. The Cx30 and Cx43 immunopositive astrocytes per unit area were quantified for each region of the three animal strains. Furthermore, Cx30 and Cx43 Western Blot was applied to the tissue samples from the same regions of the three strain. The number of Cx30 immunopositive astrocytes showed significant increase in both GAERS and WAG/Rij compared to control Wistar in all brain regions studied except LGN of WAG/Rij animals. Furthermore, Cx43 in both GAERS and WAG/Rij showed significant increase in SSCx, VB and TRN. The protein expression was increased in both Cx30 and Cx43 in the two epileptic strains compared to control Wistar animals. The significant increase in the astrocytic GJ proteins Cx30 and Cx43 and the differences in the co-expression of Cx30 and Cx43 in the genetically absence epileptic strains compared to control Wistar animals may suggest that astrocytic Cx's may be involved in the mechanism of absence epilepsy. Increased number of astrocytic Cx's in GAERS and WAG/Rij may represent a compensatory response of the thalamocortical circuitry to the absence seizures or may be related to the production and/or development of absence seizures.

Gemella morbillorum endocarditis in a patient with a bicuspid aortic valve.

Gemella morbillorum is one of the rare causative microorganisms of endocarditis. We herein report a case of infective endocarditis in a patient with bicuspid aortic valve caused by G. morbillorum. Infective endocarditis diagnosis was established based on the Modified Duke's criteria. The patient was successfully treated with medical-surgical management.