Antimicrobial Therapy as a Risk Factor of Multidrug-Resistant Acinetobacter Infection in COVID-19 Patients Admitted to the Intensive Care Unit.

Background: Multidrug-resistant Acinetobacter (MDR-Ab) is one of the most important pathogens causing superinfections in COVID-19 patients hospitalised in the intensive care unit (ICU). The occurrence of MDR-Ab superinfection significantly impairs the prognosis of patients in the ICU. Overuse of antibiotics in COVID-19 patients might contribute to the risk of developing MDR-Ab infection. Objective: The objective was to assess the role of prior antibiotic exposure as an independent predictor of MDR-Ab infection in COVID-19 patients admitted to the ICU. Methods: We conducted a retrospective cohort study in 90 patients admitted to the ICU of the Department of Infectology and Geographical Medicine, University Hospital in Bratislava, for respiratory failure due to COVID-19 between 1 September 2021 and 31 January 2022 (delta variant predominance). Patients underwent regular microbial screening. Superinfection was defined as infection occurring ≥48 h after admission. We assessed the role of prior antibiotic exposure and other factors as independent predictors of MDR-Ab isolation. Results: Fifty-eight male and 32 female patients were included in the analysis. Multidrug-resistant bacteria were cultured in 43 patients (47.8%), and MDR-Ab was isolated in 37 patients. Thirty-three (36.7%) patients had superinfection caused by MDR-Ab. Fifty-four (60%) patients were exposed to antibiotics prior to MDR-Ab isolation; of those, 35 (64.8%) patients received ceftriaxone. Prior exposure to ceftriaxone (odds ratio (OR) 4.1; 95% confidence interval (CI) 1.4-11.9; P < 0.05), tocilizumab therapy (OR 4.7; 95% CI 1.3-15.0; P < 0.05), and ICU length of stay exceeding 11 days (OR 3.7; 95% CI 1.3-10.3; P < 0.05) were independent predictors of MDR-Ab infection. Conclusions: Prior exposure to ceftriaxone increases the risk of MDR-Ab infection in COVID-19 patients admitted to the ICU. Our findings suggest that antibiotic use in COVID-19 patients admitted to the ICU should be restricted to patients with documented bacterial superinfection.

Early dynamics of glial fibrillary acidic protein and extracellular DNA in plasma of mice after closed head traumatic brain injury.

BACKGROUND: Glial fibrillary acidic protein (GFAP) in plasma is an established biomarker of traumatic brain injury (TBI) in humans. Plasma extracellular DNA (ecDNA) is a very sensitive, although nonspecific marker of tissue damage including TBI. Whether plasma GFAP or ecDNA could be used as an early non-invasive biomarker in the mouse model of closed head injury is unknown. The aim of this paper was to describe the early dynamics of plasma GFAP and ecDNA in the animal model of closed head TBI. METHODS: Closed head TBI was induced using the weight-drop method in 40 adult CD1 mice and blood was collected in different time points (1, 2 or 3h) after TBI in different groups of mice. Plasma GFAP and ecDNA and ecDNA fragmentation from the experimental groups were compared to healthy controls. In the surviving mice, a static rods test was performed 30 days after TBI to assess the neurological outcome of TBI. RESULTS: Despite a trend of higher plasma GFAP after TBI the differences between the groups were not statistically significant. Plasma ecDNA was higher by 50% after 1h (P<0.05) and 2h (P<0.05) after TBI and was highly variable after 3h. Plasma ecDNA, but not GFAP, was partially predictive of the neurological impairment of the mice. CONCLUSION: In this study, we have described the early dynamics of plasma GFAP and ecDNA after TBI in mice. According to our results, ecDNA in plasma is a more sensitive early marker of TBI than GFAP. Analysis of tissue-specific ecDNA might improve its predictive value regarding the survival and neurobehavioral outcome.

Sex steroid hormones in depressive disorders as a basis for new potential treatment strategies.

The sex steroid hormones (SSHs) such as testosterone, estradiol, progesterone, and their metabolites have important organizational and activational impacts on the brain during critical periods of brain development and in adulthood. A variety of slow and rapid mechanisms mediate both organizational and activational processes via intracellular or membrane receptors for SSHs. Physiological concentrations and distribution of SSHs in the brain result in normal brain development. Nevertheless, dysregulation of hormonal equilibrium may result in several mood disorders, including depressive disorders, later in adolescence or adulthood. Gender differences in cognitive abilities, emotions as well as the 2-3 times higher prevalence of depressive disorders in females, were already described. This implies that SSHs may play a role in the development of depressive disorders. In this review, we discuss preclinical and clinical studies linked to SSHs and development of depressive disorders. Our secondary aim includes a review of up-to-date knowledge about molecular mechanisms in the pathogenesis of depressive disorders. Understanding these molecular mechanisms might lead to significant treatment adjustments for patients with depressive disorders and to an amelioration of clinical outcomes for these patients. Nevertheless, the impact of SSHs on the brain in the context of the development of depressive disorders, progression, and treatment responsiveness is complex in nature, and depends upon several factors in concert such as gender, age, comorbidities, and general health conditions.

Sex Difference in Plasma Deoxyribonuclease Activity in Rats.

Extracellular DNA (ecDNA) activates immune cells and is involved in the pathogenesis of diseases associated with inflammation such as sepsis, rheumatoid arthritis or metabolic syndrome. DNA can be cleaved by deoxyribonucleases (DNases), some of which are secreted out of cells. The aim of this experiment was to describe plasma DNase activity in relation to extracellular DNA in adult rats, to analyse potential sex differences and to prove whether they are related to endogenous testosterone. Adult Lewis rats (n=28) of both sexes were included in the experiment. Male rats were gonadectomized or sham-operated and compared to intact female rats. Plasma ecDNA and DNase activity were measured using fluorometry and single radial enzyme diffusion assay, respectively. Concentrations of nuclear ecDNA and mitochondrial ecDNA were determined using real-time PCR. Females had 60% higher plasma DNase activity than males ( p=0.03). Gonadectomy did not affect plasma DNase in males. Neither the concentration of total ecDNA, nor nuclear or mitochondrial DNA in plasma differed between the groups. No significant correlations between DNase and ecDNA were found. From previous studies on mice, it was expected, that male rats will have higher DNase activity. In contrast, our study in rats showed the opposite sex difference. This sex difference seems not to be caused by endogenous testosterone. Interestingly, no sex differences were observed in plasma ecDNA suggesting a complex or missing association between plasma ecDNA and DNase. The observed sex difference in plasma DNase should be taken into account in animal models of ecDNA-associated diseases.

Testosterone, personality traits and aggressive driving among young male drivers.

INTRODUCTION: Psychological testing to examine potentially aggressive behaviour is a gold standard, but it is not sufficient. Testosterone might increase an aggressive behaviour. AIM: The aim of this study was to evaluate whether testosterone along with psychological assessment of fitness to drive could help to identify aggressive drivers. METHODS: Male participants (n=150) aged from 20 to 25, who possessed a driving license and drive at least 100 km per week, were evaluated in this study using an Inventory of traffic-relevant personality characteristics, the Sensation Seeking Scale and the Buss-Durkee Aggression Inventory. Saliva was collected for testosterone and cortisol measurements. The five binomial logistic models with dependent variables Caused an accident, Driving license taken away, Court trial, Intoxicated driving and Sporty self-report were tested in this study. RESULTS: The 'Intoxicated driving' model, was found to be statistically highly significant, explaining 48.8 % of the dependent variable's variance (χ2(16)=36.145, p<0.01). In this model with sensation seeking, actual testosterone and their interaction was highly significant and explained 20.4 % of intoxicated driving variability (χ2(3)=14.283, p<0.01). This was higher than sensation seeking scores only. CONCLUSION: To conclude, salivary testosterone might prove a biological marker that improves the identification of those with a high probability of aggressive driving or its subtypes (Tab. 3, Ref. 53).

Acute effect of cola and caffeine on locomotor activity in drosophila and rat.

Caffeine is well known for reducing fatigue and its effect on behavior is widely studied. Usually, caffeine is not ingested in its pure form but rather in sugar-sweetened beverages such as cola. Our aim was to compare the acute effect of cola and caffeine on locomotor activity. Rats and flies ingested cola or caffeine solution for 24 hours. The open field test revealed higher locomotor activity in cola groups for both flies and rats. Surprisingly, no differences have been observed between caffeineand control group. We conclude that caffeine itself does not explain the effect of cola on locomotor activity. Effect of cola cannot be generalized and interpreted for any caffeinated drink with other contents. Rather, the observed effect on locomotor activity may be caused by interaction of caffeine with other substances present in cola.

Role of interleukin 6 as a predictive factor for a severe course of Covid-19: retrospective data analysis of patients from a long-term care facility during Covid-19 outbreak.

BACKGROUND: Covid-19 is a disease with high morbidity and mortality among elderly residents of long-term care facilities (LTCF). During an outbreak of SARS-CoV-2 infection in the LTCF an effective screening tool is essential to identify the patients at risk for severe disease. We explored the role of interleukin 6 (IL-6) as a predictor for severe disease during the outbreak of Covid-19 in one LTCF in Slovakia. METHODS: We conducted a retrospective data analysis of cases of COVID-19, diagnosed during the outbreak in one LTCF in Slovakia between April 11, 2020, and May 5, 2020. Within 24 h after the diagnosis of Covid-19, clinical and laboratory screening was performed in the LTCF to identify patients in need of hospitalization. Patients with oxygen saturation below 90% were immediately referred to the hospital. Patients staying in the LFTC were monitored daily and those that developed hypoxemia were transferred to the hospital. We analyzed the association between the IL-6 at the initial assessment and development of hypoxemia during follow up and determined the cut-off of the IL-6 able to predict the development of hypoxemia requiring oxygen therapy. RESULTS: Fifty-three patients (11 men, 42 women) with diagnosed Covid-19 were included in the analysis. 19 (53%) patients developed hypoxemia during the disease. Patients with hypoxemia had significantly higher concentrations of IL-6, C-reactive protein, procalcitonin, fibrinogen, total bilirubin, aspartate aminotransferase and alanine aminotransferase at initial screening. ROC analyses identified IL-6 as the most robust predictor of hypoxemia. The concentration of IL-6 > 24 pg/mL predicted the development of hypoxemia with the sensitivity of 100% and specificity of 88.9%. The positive and negative predictive values were 76.9, and 100% respectively. CONCLUSIONS: The concentration of IL-6 > 24 pg/mL at initial assessment predicted the development of hypoxemia requiring hospitalization with excellent sensitivity and good specificity. IL-6 appears as a potential predictor for the development of the severe Covid-19 and might serve for early identification of patients in need of hospitalization. Further studies are needed to evaluate the robustness of the use of IL-6 as an effective screening tool for the severe course of Covid-19.

Measurement of Urea in the Saliva of Healthy Mice - a Pilot Study.

Salivary urea is studied as a non-invasive alternative for screening and monitoring of renal diseases. Its high variability prevents a wider clinical use. Animal experiments are needed to identify factors affecting this marker. The aim of this study was to describe the inter-individual variability of salivary urea in healthy mice, establish reference intervals, and analyse the effects of sex, age and body weight. Plasma and saliva samples were obtained from 37 male and 41 female healthy adult CD1 mice aged 13-69 weeks (body weight 22-51 g). The reference interval for salivary urea in heathy mice based on our results is 2.7-8.4 mmol/l (CV = 23 %). Multivariate analysis did not show any significant effect of age, sex, or body weight. In addition, salivary urea did not correlate with its plasma concentrations. The high variability of the promising salivary marker of kidney function in healthy mice requires further research before its use to diagnose or monitor renal failure in animal models of kidney diseases. Other potential confounders should be analysed, including intra-individual and pre-analytical variability. In addition, a normalization factor such as total salivary proteins or salivation rate is likely needed.

Testosterone and the brain: from cognition to autism.

Sex and gender matter in all aspects of life. Humans exhibit sexual dimorphism in anatomy, physiology, but also pathology. Many of the differences are due to sex chromosomes and, thus, genetics, other due to endocrine factors such as sex hormones, some are of social origin. Over the past decades, huge number of scientific studies have revealed striking sex differences of the human brain with remarkable behavioral and cognitive consequences. Prenatal and postnatal testosterone influence brain structures and functions, respectively. Cognitive sex differences include especially certain spatial and language tasks, but they also affect many other aspects of the neurotypical brain. Sex differences of the brain are also relevant for the pathogenesis of neuropsychiatric disorders such as autism spectrum disorders, which are much more prevalent in the male population. Structural dimorphism in the human brain was well-described, but recent controversies now question its importance. On the other hand, solid evidence exists regarding gender differences in several brain functions. This review tries to summarize the current understanding of the complexity of the effects of testosterone on brain with special focus on their role in the known sex differences in healthy individuals and people in the autism spectrum.

Does the 2nd and 4th digit ratio reflect prenatal androgen exposure?

OBJECTIVES:  The aim of our study was to describe the effect of prenatal testosterone exposure on 2D:4D in both sexes, and to determine whether this effect is mediated via the androgen receptor. In addition, the sex differences in lengths of 2D, 4D, and 2D:4D ratio were analyzed. BACKGROUND:  Clinical studies suggest a negative correlation between prenatal testosterone exposure and ratio of the lengths of the second and fourth digits (2D:4D). However, less is known about the underlying molecular mechanisms. METHODS:  Pregnant rats were treated with olive oil, testosterone, flutamide or testosterone with flutamide daily from the fourteenth day of pregnancy until delivery. The finger lengths of adult offspring were measured using both, digital scanning of the paws and µCT analysis of the phalanges. RESULTS:  None of the aforementioned methods revealed any effect of testosterone on 2D:4D. µCT measurements showed that prenatal hyperandrogenism in both sexes leads to shorter 2D compared to controls. Moreover, the testosterone treatment in males resulted in the shortening of 4D when compared to controls. CONCLUSION:  Prenatal hyperandrogenism leads to shorter lengths of 2D and 4D; however, it does not affect 2D:4D ratio. Whether other steroid hormones and/or testosterone metabolites affect the 2D:4D ratio requires further investigation (Tab. 5, Fig. 3, Ref. 32).

Sex differences in the effect of prenatal testosterone exposure on steroid hormone production in adult rats.

Maternal hyperandrogenism during pregnancy might have metabolic and endocrine consequences on the offspring as shown for the polycystic ovary syndrome. Despite numerous experiments, the impact of prenatal hyperandrogenic environment on postnatal sex steroid milieu is not yet clear. In this study, we investigated the effect of prenatal testosterone excess on postnatal concentrations of luteinizing hormone, corticosterone and steroid hormones including testosterone, pregnenolone, progesterone, estradiol and 7beta-hydroxy-epiandrosterone in the offspring of both sexes. Pregnant rats were injected daily with either testosterone propionate or vehicle from gestational day 14 until parturition. The hormones were evaluated in plasma of the adult offspring. As expected, females had lower testosterone and higher pregnenolone, progesterone and estradiol in comparison to males. In addition, corticosterone was higher in females than in males, and it was further elevated by prenatal testosterone treatment. In males, prenatal testosterone exposure resulted in higher 7beta-hydroxy-epiandrosterone in comparison to control group. None of the other analyzed hormones were affected by prenatal testosterone. In conclusion, our results did not show major effects on sex hormone production or luteinizing hormone release in adult rats resulting from testosterone excess during their fetal development. However, maternal hyperandrogenism seems to partially affect steroid biosynthesis in sex-specific manner.

Body proportions as possible predictors for free gracilis one-stage facial reanimation.

BACKGROUND: Gracilis muscle and its motor nerve belongs to most commonly used flap for facial reanimation. However, it is performed in two steps, which is time consuming. One stage technique can be also performed, but the length of the motor nerve cannot be currently determined before surgery. AIM: The present study was conducted in order to evaluate the body composition on the length and suitability of the motor nerve of gracilis muscle for one stage facial reanimation. METHODS: The gracilis flaps along with the motoric nerve were dissected from 20 fresh cadavers (6 females, 14 males). The length of the lower extremity from superior iliac anterior spine to the bottom of the heel and BMI were measured. Regression analysis of lower extremity length and BMI to the actual length of the motor nerve of gracilis flap was performed. RESULTS: The linear regression analysis showed a positive correlation between the length of the lower limb and the size of the motor nerve length (r = 0.5060, p < 0.05), as well as between the BMI and the size of the motor nerve length (r = 0.5073, p < 0.05). Also, the males had longer motor nerve when compared to females by 13 % (p < 0.05). No difference between females and males in BMI was observed. CONCLUSION: The length from the superior iliac anterior spine, BMI and gender seemed to be potential factors that could help to predict the length of the gracilis flap motor nerve for the one stage facial reanimation. However, further studies evaluating other anatomical factors and validating the possible prediction rule for one stage reanimation success are needed (Fig. 3, Ref. 14).

Effect of ivabradine, captopril and melatonin on the behaviour of rats in L-nitro-arginine methyl ester-induced hypertension.

Cardiovascular diseases including hypertension are often associated with behavioural alterations. The aim of this study was to show, whether ivabradine, the blocker of If-channel in sinoatrial node, is able to modify the behaviour of rats in L-nitro-arginine methyl ester (L-NAME)-induced hypertension and to compare the effect of ivabradine with captopril and melatonin. 12-week-old male Wistar rats were divided into the following groups: controls, ivabradine (10 mg/kg/24 h), L-NAME (40 mg/kg/24 h), L-NAME + ivabradine, L-NAME + captopril (100 mg/kg/24 h), L-NAME + melatonin (10 mg/kg/24 h). Systolic blood pressure (SBP) and heart rate (HR) were measured by tail-cuff method once a week. The behaviour of rats was investigated during 23-hours in the phenotyper after four weeks of the treatment. Chronic administration of L-NAME induced hypertension without a change in HR. All tested substances partly prevented the increase of SBP, while ivabradine and melatonin also reduced HR. Ivabradine, captopril and melatonin reduced daily food intake, slightly decreased daily water intake and attenuated body weight gain. In L-NAME group, locomotor activity was enhanced by ivabradine, whereas exploratory behaviour was increased by melatonin and captopril. In conclusion, ivabradine, besides its potentially protective hemodynamic actions, does not seem to exert any disturbing effects on behaviour in L-NAME-induced hypertension in rats, while some of its effects were similar to captopril or melatonin. It is suggested that ivabradine used in cardiovascular indications is harmless regarding the effect on behaviour.

Testosterone and estradiol are not affected in male and female patients with obstructive sleep apnea treated with continuous positive airway pressure.

BACKGROUND: Continuous positive airway pressure (CPAP) improves symptoms in patients with obstructive sleep apnea syndrome (OSAS). It is currently unclear, whether CPAP also alters endocrine parameters such as sex hormone levels. In a previous study, we have found no changes in sex hormones in patients with OSAS after one night with CPAP. AIM: The aim of this study was to prove long-term effects of CPAP on sex hormone concentrations in patients with OSAS. METHODS: Twenty-two women and 67 men with severe OSAS (respiratory distress index > 30/h) were enrolled in the study. Fasting blood venous samples were taken before CPAP therapy and after 1 and 6 months of CPAP treatment. Testosterone and estradiol were measured in all samples using commercially available ELISA kits. RESULTS: No effects of long-term CPAP treatment were found on testosterone or estradiol levels in OSAS patients of either gender. CONCLUSIONS: The results are in line with previous smaller studies. However, our study is larger and longer than previously published studies. In addition, this is the first study analyzing the effects of CPAP on testosterone and estradiol and in both genders. Positive effects of CPAP on sexual functions reported in other studies might, thus, be mediated by other than endocrine effects.

Short-term effects of continuous positive airway pressure on sex hormones in men and women with sleep apnoea syndrome.

Patients with sleep apnoea syndrome (SAS) suffer from symptoms of hypogonadism. Besides surgical interventions, in some cases, the standard care of SAS for most patients is continuous positive airway pressure (CPAP). Studies focusing on the long-term effects of CPAP on testosterone levels revealed conflicting results. None of the studies included female patients with SAS. The aim of our study was to analyse and compare sex hormone levels in saliva before and after a night without and with CPAP in women and men with SAS. The results were negative. One night with CPAP did not affect the dynamics of sex hormones, neither in men nor in women. Future studies should focus on long-term effects of CPAP in both genders.

Reference values of skin autofluorescence as an estimation of tissue accumulation of advanced glycation end products in a general Slovak population.

AIMS: For decades, Slovakia has maintained a prominent place in mortality rates from cardiovascular diseases among European Union (EU-27) countries. Determination of skin autofluorescence serves as an estimate of tissue accumulation of advanced glycation end products--substances accumulating in tissues and body fluids that play a pathophysiological role in age-related diseases and their complications, such as diabetes. METHODS: In 1385 apparently healthy Slovakian subjects aged from a few days old to 77 years, skin autofluorescence was determined using an advanced glycation end product reader and compared with reference data from Dutch Caucasians. The impact of the weekly frequency of recreational physical exercise on skin autofluorescence was investigated in the adults, and the impact of feeding regimen in the infants. RESULTS: With the exception of 10- to 19-year-olds, Slovaks had lower skin autofluorescence values in comparison with the Dutch Caucasians. In healthy non-smokers, physical exercise for > 30 min/day performed ≥ 3 times/week was associated with lower skin autofluorescence levels. In infants, breastfeeding (advanced glycation end product-poor diet) was associated with lower skin autofluorescence levels in comparison with consumption of infant formulas (advanced glycation end product-rich diet). CONCLUSIONS: Reference ranges of skin autofluorescence in Slovak Caucasians, detailed for paediatric age groups, are provided. Our data show that, in healthy adults, regular physical exercise associates with lower skin autofluorescence. Infants fed or weaned from infant formulas (advanced glycation end product-rich diet) have higher skin autofluorescence than their breast milk-consuming counterparts. It is unclear why Slovaks have lower skin autofluorescence compared with a Dutch population with lower cardiovascular mortality rates. Reference data on skin autofluorescence from diverse populations are needed for the precise clinical interpretation of skin autofluorescence measurements.

Effect of exogenous testosterone on oxidative status of the testes in adult male rats.

The aim of this study was to investigate the testosterone-induced changes in the oxidative status of testes in adult male rats treated either with testosterone or after blockade of androgen receptors with cyproterone acetate. A total of 40 intact rats were divided into four groups: a control group receiving sterile oil, the testosterone group receiving testosterone isobutyras, the cyproterone group receiving cyproterone acetate and the combination group receiving both testosterone isobutyras and cyproterone acetate. Treatments were carried out for 2 days by intramuscular application. Parameters of oxidative stress and the expression levels of the steroidogenic acute regulatory protein (StAR) gene were measured in testes. Significantly increased TBARS and advanced glycation end products (AGEs) levels were found in the testosterone group when compared to the control group. The °1 ferric-reducing ability of the tissue and total antioxidative capacity were lower in the testosterone group in comparison with the control group. Gene expression analysis revealed significant downregulation of the StAR gene in the testes of rats in the testosterone and combination groups with respect to control animals. In conclusion, administration of exogenous testosterone influences the lipid peroxidation and carbonyl stress and decreases the antioxidant defence in the testes. These data might have implications for male fertility in humans.

Bacteria in gene therapy: bactofection versus alternative gene therapy.

Recent advances in gene therapy can be attributed to improvements of gene delivery vectors. New viral and nonviral transport vehicles that considerably increase the efficiency of transfection have been prepared. However, these vectors still have many disadvantages that are difficult to overcome, thus, a new approach is needed. The approach of bacterial delivery could in the future be important for gene therapy applications. In this article we try to summarize the most important modifications that are used for the preparation of applied strains, difficulties that are related with bacterial gene delivery and the current use of bactofection in animal experiments and clinical trials. Important differences to the alternative gene therapy (AGT) are discussed. AGT resembles bacteria-mediated protein delivery, as the therapeutical proteins are produced not by host cells but by the bacteria in situ and the expression can be regulated exogenously. Although the procedure of bacterial gene delivery is far from being definitely solved, bactofection remains a promising technique for transfection in human gene therapy.

On the origin of cardiovascular complications of sleep apnea syndrome by the means of molecular interactions.

In this article we present a novel hypothesis of the pathogenesis of cardiovascular complications of sleep apnea syndrome (SAS). Chronic intermittent hypoxia occurring in association with SAS represents a variation of chronic ischaemia reperfusion injury of the heart. In the hypoxic cells hypoxia inducible factor induces adaptation processes, including production of vascular endothelial growth factor and suppression of antioxidative mechanisms. Resulting oxidative and carbonyl stress are responsible for endothelial dysfunction leading to the development of systemic hypertension. Metabolic and vascular changes stimulate the atherogenic process. Besides the pathogenetic pathway of cardiovascular complications of SAS, we also present the latest concluding results from experimental observations and epidemiological studies concerning sleep disordered breathing and diseases of heart and vessels. Our theoretical assumption should be further proved.