High Prevalence of Clostridioides difficile Ribotype 176 in the University Hospital in Kosice.

Dysbiosis of the gut microbiota, caused by antibiotics, plays a key role in the establishment of Clostridioides difficile CD). Toxin-producing strains are involved in the pathogenesis of Clostridioides difficile infection (CDI), one of the most common hospital-acquired infections. We cultured a total of 84 C. difficile isolates from stool samples of patients hospitalized at Louis Pasteur University Hospital in Kosice, Slovakia, that were suspected of CDI and further characterized by molecular methods. The presence of genes encoding toxin A, toxin B, and binary toxin was assessed by toxin-specific PCR. CD ribotypes were detected using capillary-based electrophoresis ribotyping. A total of 96.4% of CD isolates carried genes encoding toxins A and B, and 54.8% of them were positive for the binary toxin. PCR ribotyping showed the presence of three major ribotypes: RT 176 (n = 40, 47.6%); RT 001 (n = 23, 27.4%); and RT 014 (n = 7, 8.3%). Ribotype 176 predominated among clinical CD isolates in our hospital. The proportion of RT 176 and RT 001 in four hospital departments with the highest incidence of CDI cases was very specific, pointing to local CDI outbreaks. Based on our data, previous use of antibiotics represents a significant risk factor for the development of CDI in patients over 65 years of age.

A Comprehensive Survey on the Non-Invasive Passive Side-Channel Analysis.

Side-channel analysis has become a widely recognized threat to the security of cryptographic implementations. Different side-channel attacks, as well as countermeasures, have been proposed in the literature. Such attacks pose a severe threat to both hardware and software cryptographic implementations, especially in the IoT environment where the attacker may easily gain physical access to a device, leaving it vulnerable to tampering. In this paper, we provide a comprehensive survey regarding the non-invasive passive side-channel analysis. We describe both non-profiled and profiled attacks, related security metrics, countermeasures against such attacks, and leakage-assessment methodologies, as available in the literature of more than twenty years of research.

Psychological Responses of Health Care Workers Are Strongly Associated With Pandemic Management.

Background: Globally, governments adopted protective measures to stabilize the worsening COVID-19 pandemic situation. These measures had a large impact on health care workers (HCWs) and could make the work environment more stressful for them. Therefore, we explored the associations of the psychological responses of HCWs and pandemic management among Slovak health care workers during the second wave of the pandemic. Materials and Methods: We obtained data about psychological responses, COVID-19 experiences, information overload, non-adherence of the public, work stress and barriers and facilitators of health care provision from HCWs at the COVID-19-related departments of one hospital that mainly covered eastern Slovakia (215 health care professionals, 77.7% females, M/SD = 44.4). Data were analyzed using logistic regression models adjusted for gender. Results: Pandemic management was most strongly associated with psychological responses, such as irritability, sadness, anxiety, dizziness, and back pain, in health care workers. The non-adherence of the public had the strongest associated psychological impact on HCWs, mostly affecting anxiety and irritability (odds ratios, ORs varying from 3.9 to 7.0). From the barriers and facilitators of health care provision, the use of personal protective equipment had the most impact psychologically, whereas efficient department management was most protective, with ORs around 0.3. Conclusion: Pandemic management has a large impact on health care workers, and promoting resilience may seriously decrease job-related stress and associated psychological responses of HCWs.

Changes in the Quality Attributes of Selected Long-Life Food at Four Different Temperatures over Prolonged Storage.

This study reports the development of selected indicators affecting changes in food quality and safety of selected long-life canned (Szeged goulash, canned chicken meat, pork pâté, canned tuna fish) and dehydrated (instant goulash soup) food during a two-year storage experiment at four different temperatures. The storage temperatures were selected to represent Arctic (−18 °C), temperate (5 °C), subtropical (25 °C) and tropical (40 °C) climatic zones where such food is likely to be stored during, for example, humanitarian and military missions. Microorganism amounts below the detection limit (p < 0.05), regardless of the storage temperature (p ≥ 0.05), were monitored in canned samples. The contents of dry matter, fat and proteins did not change during storage, regardless of the storage temperature (p ≥ 0.05). During the 24-month storage, all food showed an increase in the level of ammonia (p < 0.05) and the TBARS-value (p < 0.05), whereas the rate of increase in both parameters was significantly higher at higher storage temperatures (p < 0.05). The losses of individual amino acids during storage ranged from 5% rel. calculated on the amino acid contents in Month "0" up to 15% rel. (p < 0.05). With storage temperatures above the freezing point, the hardness values decreased with the increase in the storage temperature (p < 0.05) and prolongation of the storage period (p < 0.05). Moreover, with temperatures of −18 °C, the development of hardness, measured as the "decrease rate", was significantly higher compared to the absolute values.

Outbreak of Alimentary Tick-Borne Encephalitis in Eastern Slovakia: An Analysis of Affected Patients and Long-Term Outcomes.

OBJECTIVE: Tick-borne encephalitis (TBE) is an endemic zoonotic viral disease in many European countries and in the central and eastern parts of Asia. Slovakia reports the highest occurrence of alimentary tick-borne encephalitis in Europe, after the consumption of unpasteurized milk and cheese from domestic ruminants. In May 2016, an outbreak of tick-borne encephalitis that emerged after the consumption of sheep cheese occurred in eastern Slovakia. In total, 44 people were ill and 36 were hospitalized. METHODS: Data from the 36 hospitalized patients at the Department of Infectology and Travel Medicine in Kosice with TBE were retrospectively analysed from the medical documentation. The patients were contacted 3 years after discharge. RESULTS: Twenty of the hospitalized patients had meningoencephalitis and 16 had meningitis. The main symptoms that occurred in all patients were fever and headache. Nuchal rigidity was seen in 50% of the patients. Three patients developed late systemic complications and another six patients had psychiatric complications. None of the patients died. Three years after the disease onset, 52% of contacted patients reported persistent discomfort. CONCLUSIONS: TBE is an infection with a wide range of clinical courses. Our findings suggest that alimentary-acquired TBE lead to severe disease and persistent discomfort.

Hemorrhagic Epidermoid Cyst in Cerebellar Vermis: Case Report and Review of the Literature.

Intracranial epidermoid cysts are slow growing congenital avascular neoplasms that spread across the basal surface of the brain. They most commonly occur in the paramedial region in the cerebellopontine angle and the parasellar region. Despite its generally benign nature, sporadically they can be accompanied with hemorrhage or very rarely undergo malignant transformation. The authors present a case report of a patient with a hemorrhagic vermian epidermoid cyst and a review of all published similar cases.

Human galectin­3: Molecular switch of gene expression in dermal fibroblasts in vitro and of skin collagen organization in open wounds and tensile strength in incisions in vivo.

Understanding the molecular and cellular processes in skin wound healing can pave the way for devising innovative concepts by turning the identified natural effectors into therapeutic tools. Based on the concept of broad­scale engagement of members of the family of galactoside­binding lectins (galectins) in pathophysiological processes, such as cancer or tissue repair/regeneration, the present study investigated the potential of galectins­1 (Gal­1) and ­3 (Gal­3) in wound healing. Human dermal fibroblasts, which are key cells involved in skin wound healing, responded to galectin exposure (Gal­1 at 300 or Gal­3 at 600 ng/ml) with selective changes in gene expression among a panel of 84 wound­healing­related genes, as well as remodeling of the extracellular matrix. In the case of Gal­3, positive expression of Ki67 and cell number increased when using a decellularized matrix produced by Gal­3­treated fibroblasts as substrate for culture of interfollicular keratinocytes. In vivo wounds were topically treated with 20 ng/ml Gal­1 or ­3, and collagen score was found to be elevated in excisional wound repair in rats treated with Gal­3. The tensile strength measured in incisions was significantly increased from 79.5±17.5 g/mm2 in controls to 103.1±21.4 g/mm2 after 21 days of healing. These data warrant further testing mixtures of galectins and other types of compounds, for example a combination of galectins and TGF­ß1.

Seroprevalence of hepatitis E among hospitalized patients in Slovakia: first report.

OBJECTIVE: Hepatitis E infection is one of the most frequent acute hepatitis in the world. Currently five human genotypes with different geographical distributions and distinct epidemiologic patterns are identified. In Slovakia, only rare cases of hepatitis E have been reported in recent years. Therefore, the aim of the study was to evaluate the prevalence of anti-HEV total antibodies and the main risk factors for HEV in the general population in Eastern Slovakia. METHODS: Detection of anti-HEV total antibodies samples was done by a commercial enzyme-linked immunosorbent assay (ELISA) kit. RESULTS: Of 175 hospitalized patients included in the study, 76 (43.5%) showed positivity for anti-HEV total antibodies. No statistically significant differences were found in anti-HEV positivity between men and women or in the groups of different living areas (town/village - urban/rural). CONCLUSION: Prevalence of anti-HEV total antibodies of hospitalised patients was high. The risk factor significantly associated with antibody positivity was eating raw meat. Other factors, such as sex, age, living area and contact with animals were not associated with antibody positivity.

[Dalbavancin and its use in the treatment of methicillin-resistant Staphylococcus aureus - induced upper limb phlegmon].

The article discusses dalbavancin, a relatively new lipoglycopeptide antibiotic brought to market. It briefly describes the spectrum and mechanism of its antibacterial effect and dosing regimens that can be used in the treatment of skin and soft tissue infections. The authors present a case of a patient with shoulder phlegmon caused by MRSA who was successfully treated with this antibiotic.

Genistein Improves Skin Flap Viability in Rats: A Preliminary In Vivo and In Vitro Investigation.

Selective estrogen receptor modulators (SERMs) have been developed to achieve beneficial effects of estrogens while minimizing their side effects. In this context, we decided to evaluate the protective effect of genistein, a natural SERM, on skin flap viability in rats and in a series of in vitro experiments on endothelial cells (migration, proliferation, antioxidant properties, and gene expression profiling following genistein treatment). Our results showed that administration of genistein increased skin flap viability, but importantly, the difference is only significant when treatment is started 3 days prior the flap surgery. Based on our in vitro experiments, it may be hypothesized that the underlying mechanism may rather by mediated by increasing SOD activity and Bcl-2 expression. The gene expression profiling further revealed 9 up-regulated genes (angiogenesis/inflammation promoting: CTGF, CXCL5, IL-6, ITGB3, MMP-14, and VEGF-A; angiogenesis inhibiting: COL18A1, TIMP-2, and TIMP-3). In conclusion, we observed a protective effect of genistein on skin flap viability which could be potentially applied in plastic surgery to women undergoing a reconstructive and/or plastic intervention. Nevertheless, further research is needed to explain the exact underlying mechanism and to find the optimal treatment protocol.

Single centre clinical experience with fidaxomicin in the treatment of Clostridium difficile infection in Slovakia.

OBJECTIVE: Clostridium difficile infection (CDI) has become one of the most common causes of hospital-acquired infections. Fidaxomicin is one of the latest antibiotics used in the treatment of CDI, however, treatment cost affects recommendations for its use in several countries. We have analysed the treatment of our patients with CDI, treated by fidaxomicin since it was introduced to the market in 2018 and became available in the second biggest Slovak hospital, University Hospital of L. Pasteur. Our aim was to determine efficacy and safety of fidaxomicin in the treatment of CDI in Slovak patients. METHODS: We reviewed all courses of fidaxomicin use in our hospital (n = 60). Fidaxomicin was used for first recurrence (12 times), second recurrence (4 times), third recurrence (2 times), and fifth recurrence (1 patient). 41 patients received fidaxomicin first-line. RESULTS: Success of fidaxomicin treatment was recorded at 86.7% within the whole cohort. In the recurrent Clostridium difficile infection (rCDI) subgroup, fidaxomicin was 63% effective with three patients dying (15.7%) and two patients developing subsequent rCDI. During the duration of the study, 6 patients in total died. Only one of three patients, with three or more recurrences of CDI, had no further presentations after eight weeks of completion of treatment. CONCLUSIONS: The biggest benefit from fidaxomicin treatment was shown in a cohort of patients with primary CDI infection demonstrating a low recurrence rate and significant reduction of fidaxomicin effectiveness in preventing a recurrence when treating patients with multiple rCDI.

Pharmacological activation of estrogen receptors-α and -ß differentially modulates keratinocyte differentiation with functional impact on wound healing.

Estrogen deprivation is considered responsible for many age-related processes, including poor wound healing. Guided by previous observations that estradiol accelerates re­epithelialization through estrogen receptor (ER)­ß, in the present study, we examined whether selective ER agonists [4,4',4''-(4-propyl [1H] pyrazole-1,3,5-triyl)­trisphenol (PPT), ER­α agonist; 2,3-bis(4-hydroxyphenyl)-propionitrile (DPN), ER­ß agonist] affect the expression of basic proliferation and differentiation markers (Ki­67, keratin­10, ­14 and ­19, galectin­1 and Sox­2) of keratinocytes using HaCaT cells. In parallel, ovariectomized rats were treated daily with an ER modulator, and wound tissue was removed 21 days after wounding and routinely processed for basic histological analysis. Our results revealed that the HaCaT keratinocytes expressed both ER­α and ­ß, and thus are well-suited for studying the effects of ER agonists on epidermal regeneration. The activation of ER­α produced a protein expression pattern similar to that observed in the control culture, with a moderate expression of Ki­67 being observed. However, the activation of ER­ß led to an increase in cell proliferation and keratin­19 expression, as well as a decrease in galectin­1 expression. Fittingly, in rat wounds treated with the ER­ß agonist (DPN), epidermal regeneration was accelerated. In the present study, we provide information on the mechanisms through which estrogens affect the expression patterns of selected markers, thus modulating keratinocyte proliferation and differentiation; in addition, we demonstrate that the pharmacological activation of ER-α and -ß has a direct impact on wound healing.

Agrimonia eupatoria L. and Cynara cardunculus L. Water Infusions: Phenolic Profile and Comparison of Antioxidant Activities.

Reactive oxygen species (ROS) are highly considered in the ethiopathogenesis of different pathological conditions because they may cause significant damage to cells and tissues. In this paper, we focused on potential antioxidant properties of two medical plants such as the Agrimonia eupatoria L. and Cynara cardunculus L. Both plants have previously been studied for their pharmacological activities, especially as hepatoprotective and hypoglycemic activities. It has been suggested, that their effects are related to the antioxidant properties of polyphenols, which are dominant compounds of the plants' extracts. In the present study HPLC-MS analysis of water infusion was performed allowing the identification of several phenolic constituents. Furthermore, antioxidant effects of the two extracts were compared showing higher effects for agrimony extract compared to artichoke. Thus, agrimony was selected for the in vivo study using the skin flap viability model. In conclusion, our results provide evidence that the A. eupatoria extract may be a valuable source of polyphenols to be studied for the future development of supplements useful in the prevention of diseases linked to oxidative stress.

Regional changes in cardiac and stellate ganglion norepinephrine transporter in DOCA-salt hypertension.

Uptake of norepinephrine via the neuronal norepinephrine transporter is reduced in the heart during deoxycorticosterone (DOCA)-salt hypertension. We hypothesized that this was due to reduced norepinephrine transporter mRNA and/or protein expression in the stellate ganglia and heart. After 4 weeks of DOCA-salt treatment there was no change in norepinephrine transporter mRNA in either the right or the left stellate ganglia from hypertensive rats (n=5-7, p>0.05). Norepinephrine transporter immunoreactivity in the left stellate ganglion was significantly increased (n=4, p<0.05) while the right stellate ganglion was unchanged (n=4, p>0.05). Whole heart norepinephrine content was significantly reduced in DOCA rats consistent with reduced uptake function; however, when norepinephrine was assessed by chamber, a significant decrease was noted only in the right atrium and right ventricle (n=6, p<0.05). Cardiac norepinephrine transport binding by chamber revealed that it was only reduced in the left atrium (n=5-7, p>0.05). Therefore, 1) contrary to our hypothesis reduced reuptake in the hypertensive heart is not exclusively due to an overall reduction in norepinephrine transporter mRNA or protein in the stellate ganglion or heart, and 2) norepinephrine transporter regulation occurs regionally in the heart and stellate ganglion in the hypertensive rat heart.

Pre- and/or postsurgical administration of estradiol benzoate increases skin flap viability in female rats.

BACKGROUND: It has been shown that estrogens have a protective effect with regard to tissue ischemia. Therefore, in this macroscopic and histological investigation, the effect of estradiol benzoate on skin flap viability was studied in sham-operated and ovariectomized Sprague-Dawley rats. METHODS: Three months prior to flap surgery a group of rats underwent ovariectomy, while the remaining animals underwent a sham operation. Subsequently, all rats had a 2 × 8-cm skin flap created on the dorsum. Rats were randomly divided into estradiol- or saline-treated groups. Treatment started either on the day of flap excision or 3 days prior to the surgery. RESULTS: Our results showed that administration of estradiol benzoate prior to and after flap surgery significantly decreases skin flap necrosis in both sham-operated and ovariectomized rats, with the highest survival rate in animals where treatment started 3 days prior to flap surgery. CONCLUSION: In conclusion, the observed protective effect of estradiol on skin flap viability could potentially be applied to plastic and reconstructive surgery in postmenopausal women. Nevertheless, further research is needed to explain the exact underlying mechanism and to find the optimal treatment protocol for human clinical practice.

ER-α agonist induces conversion of fibroblasts into myofibroblasts, while ER-ß agonist increases ECM production and wound tensile strength of healing skin wounds in ovariectomised rats.

Oestrogen deprivation is one of the major factors responsible for many age-related processes, including poor wound healing in women. Previously, it has been shown that oestrogens have a modulatory effect in different wound-healing models. Therefore, in this study, the effect of selective oestrogen receptor (ER) agonists (PPT - ER-α agonist, DPN - ER-ß agonist) on excisional and incisional wound-healing models was compared in ovariectomised rats in vivo as well as on human dermal fibroblasts (HDF) and human umbilical endothelial cells (HUVEC) in vitro. In the in vivo study, 4 months after either ovariectomy or sham ovariectomy, Sprague-Dawley rats were randomly divided into four groups and subjected to two incisional and excisional wounds: (i) control - sham operated, vehicle-treated; (ii) ovariectomised, vehicle-treated; (iii) ovariectomised, PPT treated; (iv) ovariectomised, DPN treated. In the in vitro study, HDFs and HUVECs were used. After treatment with ER agonists, cells were processed for immunocytochemistry and gelatin zymography. Our study shows that stimulation of ER-α leads to the differentiation of fibroblasts into myofibroblasts both in vivo and in vitro. On the other hand, the formation of extracellular matrix was more prominent, and wound tensile strength (TS) was increased when ER-ß was stimulated. In contrast, stimulation of ER-α led to a more prominent increase in the expression of MMP-2 and decrease in wound TS. New information is presented in this investigation concerning oestrogen replacement therapy (ERT) in different wound-healing models. This study demonstrates that the ERT should be both wound and receptor-type specific.

Effects of equal daily doses delivered by different power densities of low-level laser therapy at 670 nm on open skin wound healing in normal and corticosteroid-treated rats: a brief report.

The optimal parameters for low-level laser therapy (LLLT) for wound healing are still open to discussion. Hence, our study was aimed at comparing the effects of different power densities of LLLT at 670 nm in rats. Four round full-thickness skin wounds were placed on the backs of 16 rats which were divided into two groups (non-steroid and steroid-treated). Three wounds were stimulated daily with a diode laser (daily dose 5 J/cm(2)) at different power densities (5, 15 and 40 mW/cm(2), respectively), and the fourth wound served as a control. Six days after surgery all animals were killed and samples removed for histological evaluation. Significant acceleration of fibroblast proliferation and new vessel formation was observed in wounds treated at the selected power densities. No significant differences were found in corticosteroid-treated rats. In conclusion, LLLT with the methodology used improved wound healing in non-steroid rats, but was not effective after corticosteroid-treatment.

Decrease in wound tensile strength following post-surgical estrogen replacement therapy in ovariectomized rats during the early phase of healing is mediated via ER-alpha rather than ER-beta: a preliminary report.

BACKGROUND: In cases of acute surgery or trauma, the most effective method of increasing the level of estrogen in postmenopausal women is its administration immediately pre- or postsurgery. However, in our previous study (J Surg Res 2008; 147:117-122) we showed that postsurgical administration of nonspecific estrogen receptor (ER) agonist decreases wound tensile strength. Therefore, the aim of this study was to evaluate whether this effect is mediated via the alpha or beta ER. MATERIALS AND METHODS: Three months prior to the wound healing experiment, 18 rats were anesthetized and underwent ovariectomy (OVX), while another six rats were sham operated. Two parallel full thickness skin incisions were performed on the back of each rat. Doses of 1mg/kg of either PPT (ER-alpha agonist) or DPN (ER-beta agonist) were administered to 12 OVX rats for 6 d postoperatively, whereas all other animals received vehicle. After 6 d, all animals were sacrificed and samples removed for wound tensile strength measurement and histologic evaluation. RESULTS: The mean wound tensile strength of PPT-treated rats (6.8+/-1.9 g/mm2) was significantly lower compared with all other groups (P<0.05). No significant differences were observed between DPN-treated (8.9+/-2.2 g/mm2), non-OVX vehicle-treated (8.7+/-2.0 g/mm2), and OVX vehicle-treated (9.1+/-1.7 g/mm2) rats. Nevertheless, no remarkable differences were found between groups during histologic evaluation. CONCLUSION: Our results indicate that the wound tensile strength decrease is mediated through the alpha rather than beta ER.

Effect of stellate ganglionectomy on basal cardiovascular function and responses to beta1-adrenoceptor blockade in the rat.

Cardiac sympathetic nerve activity is an important short-term controller of cardiac function and arterial pressure. Studies also suggest that long-term increases in cardiac sympathetic nerve activity may contribute to hypertension, coronary artery disease, and cardiac remodeling in heart failure. However, our understanding of the role of cardiac sympathetic nerves in chronic models of cardiovascular disease has been limited by inadequate experimental approaches. The present study was conducted to develop a surgical method to surgically denervate the sympathetic nerves of the rat heart for long-term cardiovascular studies. We characterized the effect of cardiac sympathetic denervation on basal levels of mean arterial pressure (MAP) and heart rate (HR) and the responses to a chronic administration of atenolol, a beta1-adrenoceptor antagonist. Rats were instrumented with telemetry transmitters for continuous recording of MAP and HR. After a 4-day baseline period, the rats were subjected to bilateral stellate ganglionectomy (SGX; n=9) or sham surgery (Sham; n=8). Seven days following SGX or Sham, the rats were administered atenolol for 5 days, followed by a 7-day recovery period. Following a transient decrease, SGX had no effect on basal MAP but decreased HR compared with baseline and Sham rats. Five days of atenolol treatment decreased MAP similarly in SGX and Sham rats. Atenolol resulted in a marked bradycardia in Sham rats but had a neglible effects on HR in SGX rats. The measurement of the content of cardiac catecholamines in all cardiac chambers at the end of the study verified a successful sympathetic denervation. This study confirms that bilateral SGX is a useful method to study the contribution of cardiac sympathetic nerves on the regulation of cardiac function. Moreover, these results suggest that cardiac sympathetic nerves are relatively unimportant in maintaining the basal level of MAP or the depressor response to atenolol in conscious, unrestrained rats.

Cardiac norepinephrine transporter protein expression is inversely correlated to chamber norepinephrine content.

The cardiac neuronal norepinephrine (NE) transporter (NET) in sympathetic neurons is responsible for uptake of released NE from the neuroeffector junction. The purpose of this study was to assess the chamber distribution of cardiac NET protein measured using [(3)H]nisoxetine binding in rat heart membranes and to correlate NE content to NET amount. In whole mounts of atria, NET was colocalized in nerve fibers with tyrosine hydroxylase (TH) immunoreactivity. NE content expressed as micrograms NE per gram tissue was lowest in the ventricles; however, NET binding was significantly higher in the left ventricle than the right ventricle and atria (P < 0.05), resulting in a significant negative correlation (r(2) = 0.922; P < 0.05) of NET to NE content. The neurotoxin 6-hydroxydopamine, an NET substrate, reduced NE content more in the ventricles than the atria, demonstrating functional significance of high ventricular NET binding. In summary, there is a ventricular predominance of NET binding that corresponds to a high NE reuptake capacity in the ventricles, yet negatively correlates to tissue NE content.