INACTIVATED COVID-19 VACCINE CAN INDUCE REACTIVE POLYARTHRITIS IN OLDER PATIENTS: REPORT OF TWO CASES.

Reactive arthritis is an acute, sterile, non-suppurative and inflammatory arthropathy that usually follows infection process. Gastrointestinal, genitourinary and respiratory tract infections generally provoke reactive arthritis. Also, reactive arthritis can be seen after vaccination. Reactive arthritis cases have been reported after tetanus, combined diphteria-poliomyelitis-tetanus toxoid, hepatitis B or influenza vaccination. Although reactive arthritis is more common in youngs, healthcare workers should be aware of the development of post inactivated COVID-19 vaccine reactive arthritis in older patients. We present two cases with ReA induced by inactivated coronavirus 2019 (COVID-19) vaccination (CoronaVac, Sinovac). Both patients in our study were over 70 years old and presented with polyarthritis that developed after vaccination. Rheumatoid factor and anti-nucleer antibody were negative and patients responded well to short-term steroid therapy, arthritis were not resistant.

Antifungal activity of cinnamic acid derivatives involves inhibition of benzoate 4-hydroxylase (CYP53).

AIMS: CYP53A15, from the sorghum pathogen Cochliobolus lunatus, is involved in detoxification of benzoate, a key intermediate in aromatic compound metabolism in fungi. Because this enzyme is unique to fungi, it is a promising drug target in fungal pathogens of other eukaryotes. METHODS AND RESULTS: In our work, we showed high antifungal activity of seven cinnamic acid derivatives against C. lunatus and two other fungi, Aspergillus niger and Pleurotus ostreatus. To elucidate the mechanism of action of cinnamic acid derivatives with the most potent antifungal properties, we studied the interactions between these compounds and the active site of C. lunatus cytochrome P450, CYP53A15. CONCLUSION: We demonstrated that cinnamic acid and at least four of the 42 tested derivatives inhibit CYP53A15 enzymatic activity. SIGNIFICANCE AND IMPACT OF THE STUDY: By identifying selected derivatives of cinnamic acid as possible antifungal drugs, and CYP53 family enzymes as their targets, we revealed a potential inhibitor-target system for antifungal drug development.

Determining the disinfection of textiles in compressed carbon dioxide using various indicator microbes.

AIMS: This paper presents a research on the disinfection efficiency of inoculated textile swatches by compressed carbon dioxide, an environmental friendly way to disinfect textiles as opposed to the conventional laundering procedures using water. The disinfection efficiency was determined by using the following microbes inoculated on cotton test fabrics: Enterococcus faecium, Enterobacter aerogenes and Candida albicans. METHODS AND RESULTS: The experiments were performed using the high pressure extraction device with a maximum pressure of 50MPa and a small extraction vessel of 500ml. Pure CO(2) and CO(2) with added disinfection agent or commercial detergent were used. The chosen disinfecting agent was hydrogen peroxide, a widespread disinfecting chemical. It was found that treatment with CO(2) for 25min at 5MPa and 40°C (313K) and the addition of 4ml of specific detergent per litre of CO(2) assures at least a five log step reduction of Enterobacter aerogenes and C. albicans, whilst treatment at 50°C (323K) with CO(2) for 25min at 5MPa is sufficient for at least a five log step reduction for Enterococcus faecium. It was also found that a 15-min CO(2) treatment at 7MPa and 20°C (293K) was sufficient for the inactivation of the yeast C. albicans, whilst these conditions were not rigorous enough for the challenge bacteria. On the other hand, the labscale treatment with CO(2) for 25min at pressure 4 and 6MPa with the addition of detergent or hydrogen peroxide only yields a log step reduction of up to 4 log steps, thus proving the slightly disinfective properties of the CO(2) treatment with added agents, but not reaching efficient results as a 5 log step reduction was not reached. CONCLUSIONS: Addition of heat to the compressed CO(2) treatment of textiles inoculated with microorganisms proved more effective than the addition of detergent or disinfectant with compressed CO(2) treatment at temperature of 20°C. SIGNIFICANCE AND IMPACT OF THE STUDY: CO(2) treatment of textiles is a promising ecological alternative dry-cleaning method for the disinfection of medical textiles.

Evaluation of echocardiographic indices for the prediction of major adverse events during long-term follow-up in chronic hemodialysis patients with normal left ventricular ejection fraction.

BACKGROUND: Cardiovascular disease is the leading cause of mortality in endstage renal failure. Prognostic role of echocardiography has not been fully elucidated in chronic hemodialysis patients. AIM: To assess the ability of Doppler echocardiographic parameters of left ventricular (LV) diastolic function along with conventional echocardiographic indices to predict long-term adverse major events in chronic hemodialysis patients with normal LV ejection fraction (EF). PATIENTS AND METHODS: A total of 45 chronic hemodialysis patients (aged 49 +/- 15 years) were included to the study. All patients underwent complete standard and tissue Doppler imaging echocardiography before and immediately after hemodialysis session and were followed-up prospectively. Major outcome measure was the combination of all-cause death and hospitalization for any cardiovascular event. RESULTS: During the follow up period (52 +/- 26 months) 23 major events occured (17 all-cause deaths and 6 cardiovascular events requring hospitalization). Post-dialytic values of mean left atrial diameter, mitral E (peak early mitral inflow velocity), E/Vp [ratio of mitral E to flow propagation velocity (Vp)] and E/Ea [ratio of mitral E to peak early diastolic mitral annular velocity (Ea)] (average of 4 segments of mitral annulus) were significantly higher in patients who had major events. In Cox proportional hazard analysis only E/Ea ratio predicted combined endpoint of all-cause mortality and nonfatal cardiovascular events (hazard ratio: 1.20; confidence interval: 1.03-1.39; p = 0.018). The optimum cut-off value for E/Ea determined by ROC curve analysis revealed that E/Ea ratio higher than 9.8 predicted future events with sensitivity of 74% and specificity of 86%. CONCLUSIONS: E/Ea might be an accurate echocardiographic indice during long-term follow up for the prediction of major adverse events in chronic hemodialysis patients with normal LV EF.

Aspergillus thyroiditis in a renal transplant recipient mimicking subacute thyroiditis.

Fungal pathogens are increasingly encountered after renal transplantation. Aspergillus causes significant morbidity and mortality in transplant patients. Fungal thyroiditis is a rare occurrence owing to unique features of the thyroid gland. Most cases are caused by Aspergillus species and have been described in immunocompromised patients. Presentation may be identical with that of subacute thyroiditis, in which hyperthyroidism features and painful thyroid are the prominent findings. Diagnosis can be ascertained by fine-needle aspiration of thyroid showing branching hyphae of Aspergillus. We describe a renal transplant patient who developed Aspergillus thyroiditis as part of a disseminated infection successfully treated with voriconazole.

A novel multifunctional NCQDs-based injectable self-crosslinking and in situ forming hydrogel as an innovative stimuli responsive smart drug delivery system for cancer therapy.

In this work, we offer an easy approach to develop a novel injectable, pH sensitive and in situ smart drug delivery system for use in cancer treatments. The developed hydrogels containing nitrogen doped carbon quantum dots (NCQD), doxorubicin (Dox) and hydroxyapatite (HA) were obtained by in situ self-crosslinking. Characterization of the synthesized nanomaterials, interactions between NCQD/Dox/HA hydrogel structure were carried out by TEM, FESEM, EDS, FTIR, XPS, XRD, Zeta potential, DLS, UV-Vis, SEM, gelation time, injectability and DIST measurements. In addition, antibacterial evaluation which was performed against Staphylococcus aureus realized that HA compound significantly increased the antibacterial activity of the hybrid hydrogel. The anticancer drug release to the tumor cell microenvironment with a pH of 5.5 was found to be higher compared to the release in the normal physiological range of pH 6.5 and 7.4. MTT and live/dead assays were also performed using L929 fibroblastic cell lines to investigate the cytotoxic behavior of NCQDs, and NCQDs/Dox/HA hydrogels. Furthermore, the NCQDs/Dox/HA hydrogel could transport Dox within a MCF-7 cancerous cell at specifically acidic pH. Additionally, imaging of cell line was observed using NCQDs and their use in imaging applications and multicolor features in the living cell system were evaluated. The overall study showed that in situ formed NCQDs/Dox/HA hydrogel represented a novel and multifunctional smart injectable controlled-release drug delivery system with great potential, which may be considered as an attractive minimal invasive smart material for future intelligent delivery of chemotherapeutic drug and disease therapy applications.

Three critical clinicobiological phases of the human SARS-associated coronavirus infections.

OBJECTIVE: COVID-19 immune syndrome is a multi-systemic disorder induced by the COVID-19 infection. Pathobiological transitions and clinical stages of the COVID-19 syndrome following the attack of SARS-CoV-2 on the human body have not been fully explored. The aim of this review is to outline the three critical prominent phase regarding the clinicogenomics course of the COVID-19 immune syndrome. MATERIALS AND METHODS: In the clinical setting, the COVID-19 process presents as "asymptomatic/pre-symptomatic phase", "respiratory phase with mild/moderate/severe symptoms" and "multi-systemic clinical syndrome with impaired/disproportionate and/or defective immunity". The corresponding three genomic phases include the "ACE2, ANPEP transcripts in the initial phase", "EGFR and IGF2R transcripts in the propagating phase" and the "immune system related critical gene involvements of the complicating phase". RESULTS: The separation of the phases is important since the genomic features of each phase are different from each other and these different mechanisms lead to distinct clinical multi-systemic features. Comprehensive genomic profiling with next generation sequencing may play an important role in defining and clarifying these three unique separate phases for COVID-19. From our point of view, it is important to understand these unique phases of the syndrome in order to approach a COVID-19 patient bedside. CONCLUSIONS: This three-phase approach may be useful for future studies which will focus on the clinical management and development of the vaccines and/or specific drugs targeting the COVID-19 processes. ANPEP gene pathway may have a potential for the vaccine development. Regarding the specific disease treatments, MAS agonists, TXA127, Angiotensin (1-7) and soluble ACE2 could have therapeutic potential for the COVID-19 course. Moreover, future CRISPR technology can be utilized for the genomic editing and future management of the clinical course of the syndrome.

Distribution and relative frequency of immunohistochemically detected endocrine cells in the stomach of New Zealand White rabbit (Oryctolagus cuniculus).

BACKGROUND: Gastrointestinal (GI) endocrine cells produce many GI hormones that perform various physiological functions of the digestive system. AIMS: We aimed to investigate the presence and distribution of immunoreactive (IR) endocrine cells to glucagon, somatostatin, cholecystokinin-8 (CCK-8), serotonin, secretin and histamine in the stomach of adult male New Zealand White rabbit (Oryctolagus cuniculus). METHODS: For immunohistochemical staining, peroxidase anti-peroxidase (PAP) method was applied to stomach samples. RESULTS: Glucagon-IR cells of closed- and open type were found throughout all the stomach parts examined. Somatostatin-IR cells of closed- and open type in the cardiac and oxyntic glands were localized to deep portions of foveola gastrica. CCK-8 IR cells that were not observed in the cardia and fundus were mostly localized to the glands and lamina epithelialis in the pyloric part near the duodenum. Oval-shaped open and closed type serotonin-IR cells were mostly dispersed throughout the fundic and pyloric glands. Secretin-IR cells were rare in the pyloric and cardiac region although they were not observed in the fundic glands. Histamine-IR cells were rarely found in the cardia, fundus and pylorus. CONCLUSION: Our findings show that glucagon, histamine, somatostatin, secretin and serotonin might be produced by all the stomach regions while pyloric region had only CCK-8 IR. These distribution patterns also provide further evidence of species-specific differences, which might be important from the evolutionary aspect of the digestive tract in relation to evolutional niches and nutrient resources.

Biomimetic synthesis of Ag, Zn or Co doped HA and coating of Ag, Zn or Co doped HA/fMWCNT composite on functionalized Ti.

In this study a novel composite containing fMWCNT and Ag, Zn or Co doped hydroxyapatite (HA) and undoped HA successfully synthesized using biomimetic process and coated on the chemically silane functionalized Ti via facile surface functionalization and deposition approach through the covalent immobilization. In this approach, the properties of Ti pretreated with NaOH and APTES were examined before the composite coating. Functional groups with oxygen were created on the MWCNT surface by acidic treatment. The synthesized powders were characterized using SEM, XRD, EDS, TGA, FTIR, Raman. Surface energies were decreased from 46.03 mN/m (Ti) to 43,79 mN/m (Ti-/fMWCNT/Ag, Zn or Co doped HA). The contact angles of APTES, fMWCNT and undoped or doped HA containing Ti surfaces increased from 40.03 ±â€¯1.2 to 49.05 ±â€¯1.6°. The bond strengths between the coatings and Ti substrates were measured using an adhesive strength test which indicates that the crosslinking process increased the adhesion strength (from 14.6 ±â€¯0.9 to 19.8 ±â€¯1.2 MPa). The results indicated that the Ag+, Zn2+ or Co2+ added in prepared SBF medium have been located in the HA lattice structure at rate of 1.06, 1.86 and 1.78 at.%, respectively. The biocompatibility of the synthesized composites was evaluated using MTT assays in vitro and no negative effect was observed on cell viability. This work shows that the fMWCNT/Ag, Zn or Co doped HA coating promise for the potential implementation in biomaterial coating fields.

Oxidative stress in leucocytospermic prostatitis patients: preliminary results.

The aim of this study was to contribute to the knowledge concerning pathogenesis of inflammatory chronic prostatitis by revealing possible shifts in the balance of markers of oxidative stress and anti-oxidative activity in case of leucocytospermic prostatitis. We also attempted to identify possible relations between seminal micro-organisms and oxidative stress parameters. A many-sided complex of local (spermatozoa, seminal plasma) and general (blood, urine) markers in 21 prostatitis patients and nine controls was compared. In both spermatozoa and seminal plasma, the content of diene conjugates was significantly higher in prostatitis patients compared with healthy controls. At the same time total anti-oxidative status in spermatozoa and total anti-oxidative activity in seminal plasma were lower in prostatitis patients than in controls. In urine, the level of 8-isoprostanes was significantly higher in prostatitis patients than in healthy controls, correlating well with 8-hydroxy-2'-deoxyguanosine. The latter correlated with cellular Fe and Ni contents as well, confirming that these metals with varying valency may cause DNA damage. Reduced glutathione showed higher levels in blood of controls than in prostatitis patients. Coryneform bacteria appeared to be associated with prostatitis-related oxidative stress. In conclusion, leucocytospermic prostatitis patients are characterised by oxidative stress at all levels: systemic (general), seminal plasma and cellular.

Immunolocalization of natriuretic peptides and their receptors in goat (Capra hircus) heart.

Natriuretic peptides are structurally similar, but genetically distinct, hormones that participate in cardiovascular homeostasis by regulating blood and extracellular fluid volume and blood pressure. We investigated the distribution of natriuretic peptides and their receptors in goat (Capra hircus) heart tissue using the peroxidase-anti-peroxidase (PAP) immunohistochemical method. Strong staining of atrial natriuretic peptide (ANP) was observed in atrial cardiomyocytes, while strong staining for brain natriuretic peptide (BNP) was observed in ventricular cardiomyocytes. Slightly stronger cytoplasmic C-type natriuretic peptide (CNP) immunostaining was detected in the ventricles compared to the atria. Natriuretic peptide receptor-A (NPR-A) immunoreactivity was more prominent in the atria, while natriuretic peptide receptor-B (NPR-B) immunoreactivity was stronger in the ventricles. Cytoplasmic natriuretic peptide receptor-C (NPR-C) immunoreactivity was observed in both the atria and ventricles, although staining was more prominent in the ventricles. ANP immunoreactivity ranged from weak to strong in endothelial and vascular smooth muscle cells. Endothelial cells exhibited moderate to strong BNP immunoreactivity, while vascular smooth cells displayed weak to strong staining. Endothelial cells exhibited weak to strong cytoplasmic CNP immunoreactivity. Vascular smooth muscle cells were labeled moderately to strongly for CNP. Weak to strong cytoplasmic NPR-A immunoreactivity was found in the endothelial cells and vascular smooth muscle cells stained weakly to moderately for NPR-A. Endothelial and vascular smooth cells exhibited weak to strong cytoplasmic NPR-B immunoreactivity. Moderate to strong NPR-C immunoreactivity was observed in the endothelial and smooth muscle cells. Small gender differences in the immunohistochemical distribution of natriuretic peptides and receptors were observed. Our findings suggest that endothelial cells, vascular smooth cells and cardiomyocytes express both natriuretic peptides and their receptors.

3D porous collagen/functionalized multiwalled carbon nanotube/chitosan/hydroxyapatite composite scaffolds for bone tissue engineering.

In this study, we describe new collagen/functionalized multiwalled carbon nanotube/chitosan/hydroxyapatite (Col/f-MWCNT/CS/HA) composite scaffolds which were fabricated by freezing (-40 °C at 0.9 °C/min) and lyophilization (48 h, 0 °C and 200 mtorr). The compressive stresses (from 523 to 1112 kPa), swelling (from 513.9 ±â€¯27 to 481.05 ±â€¯25%), porosity (from 98 ±â€¯0.15 to 95.7 ±â€¯0.1%), contact angle (from 87.8 to 76.7°) properties examined before and after biomineralization for comparison 3D porous Col, CS, Col/f-MWCNT and Col/f-MWCNT/CS scaffolds. Biomineralization was performed by biomimetic method in concentrated SBF (10 × SBF, at 37 °C and 6.5 pH). XRD, SEM, EDS, FTIR, TGA, Optical microscopy and BET results showed that compared to Col, CS and Col/f-MWCNT scaffolds, Col/f-MWCNT/CS scaffolds had higher in vitro bioactivity, large surface area (11.746 m2/g) and a good pore volume (0.026 cc/g), interconnected porous microstructure (with 20-350 µm pore size) and incorporates the advantageous properties of both Col, f-MWCNT, CS and HA. Finally, the methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate scaffolds cytotoxicity which showed that Col/f-MWCNT/CS scaffolds have the best biocompatibility.

Impact of sexual debut on culturable human seminal microbiota.

Micro-organisms are tightly integrated into host-microbiota ecosystem. Microbiota of human semen has been studied so far mostly in case of infertility or prostatitis. We aimed to reveal possible impact of sexual debut on seminal microbiota in healthy young men. The study group included 68 young healthy men, of them 12 men without sexual experience, 11 men with single lifetime sexual partner and 45 men with multiple lifetime sexual partners. Basic semen parameters were similar for all subgroups, and no correlation between sexual experience and WBC counts in semen was found. A man could harbour one to nine different bacteria in his semen; the total concentration of bacteria ranged from 2.3 to 7.3 log10 CFU/mL of semen. Lower total bacterial concentration and lower bacterial diversity was observed in men without sexual experience than in sexually experienced men (p < 0.05), with significant positive correlation between these two parameters (r = 0.54; p < 0.0001). In conclusion, the sexual debut is associated with the enrichment of seminal microbiota but not with the influx of WBC or changes in basic seminal parameters.

Antimicrobial disinfection effect of a laundering procedure for hospital textiles against various indicator bacteria and fungi using different substrates for simulating human excrements.

Recent studies confirm the increase of nosocomial infections and microbial resistance. One of the possible causes is infected textiles due to inappropriate laundering procedures. Most Slovenian laundries use thermal laundering procedures with high energy and water consumption to disinfect hospital textiles. In addition to this fact, there is an increasing number of hospital textiles composed of cotton/polyester blends that cannot endure high temperatures of thermal disinfection. On the other hand, decreasing the temperature of laundering procedures enhances the possibility of pathogenic microorganisms to survive the laundering procedure. In our research, we determined the antimicrobic laundering effect by simulating a common laundering procedure for hospital textiles in the laboratory washing machine at different temperatures by the use of bioindicators. Enterococcus faecium, Staphylococcus aureus, Mycobacterium terrae, Enterobacter aerogenes, and Pseudomonas aeruginosa were used for determining the antibacterial laundering effect. Candida albicans was used for determining the antifungal laundering effect. Swine blood, artificial sweat, and swine fat were used as substrates for simulating human excrements and were inoculated together with the chosen microorganisms onto cotton pieces to simulate real laundering conditions. It was found that E. faecium, S. aureus, E. aerogenes, and P. aeruginosa survived at 60 degrees C, but no microorganisms were found at 75 degrees C.

Regional and mucosal distributions of some intestinal immunoreactive endocrine cells in New Zealand white rabbit (Oryctolagus cuniculus L.).

The aim of this study was to detect the regional and mucosal distribution of endocrine cells that secrete gulcagon, somatostatin, Chyholecystokinin-8 (CCK-8), serotonin, secretin, substance P (SP) and histamine in the small and large intestine of New Zealand white rabbit (Oryctolagus cuniculus L.) using immunohistochemical peroxidase-antiperoxidase (PAP) method. It was found that most of the immunoreactive (IR) endocrine cells, which are oval- or spindle-shaped, are spotted in the basal parts of the relevant glands. It was noticed that cells in the lamina epithelialis of small and large intestine is linked to the lumen and that the cells in their glands cannot reach the lumen. Immunoreactive cells for glucagon, somatostatin, serotonin, secretin and SP were identified in lamina epithelialis of the small and large intestine. It was seen that secretin, SP and histamine-IR cells are rarely deployed throughout the intestinal tract. It was determined that somatostatin-IR cells were identified throughout the intestinal tract. In conclusion, the immunohistochemical study shows that gastrointestinal tract of this species contained different types of endocrine cells similar to those found in other vertebrate species. However, some species-dependent unique distributions and frequencies of endocrine cells were also observed in the present study.

Microwave-assisted biomimetic synthesis of hydroxyapatite using different sources of calcium.

In this study, some properties of biomimetic synthesized hydroxyapatite by using different sources of calcium were investigated. Biomimetic synthesis of hydroxyapatite was carried out in microwave oven using 1.5 simulated body fluid (SBF) solution having different calcium sources with 800W power for 15min. As phosphorus source di-ammonium hydrogen phosphate ((NH4)2HPO4) while for each sample as a calcium sources calcium chloride (CaCl2), calcium nitrate tetra hydrate (Ca(NO3)2·4H2O) and calcium hydroxide (Ca(OH)2) were utilized, respectively. For comparison, precipitation process was also performed in only 1.5 SBF solution without calcium and phosphorus sources. The presence of phases in synthesized hydroxyapatite was confirmed by XRD. The crystallinity and crystalline size of the phases in as synthesized powders were also calculated by using XRD data. It was found that the unique phase is hydroxyapatite (HAp, Ca5(PO4)3(OH)) by using the calcium nitrate tetra hydrate and calcium hydroxide sources, while the dominant phases are tri-calcium phosphates (TCP) and HAp for CaCl2 source and 1.5SBF which does not contain any additional Ca source. SEM studies revealed that nano-hexagonal rods and nano-spherical hydroxyapatites could be synthesized by using this process. Energy-dispersive X-ray spectroscopy (EDS) analysis revealed that the Ca/P ratio near to be as 1.5 which is the value for HAp in bone. Raman and Fourier transform-infrared spectroscopy (FT-IR) results combined with the X-ray diffraction (XRD) indicates that dominantly the present of single phase is HAp. The crystal size and fraction crystallinity of as synthesized HAp powders were changed between 29.5 and 45.4nm and 0.53-2.37, respectively. Results showed that microwave assisted biomimetic synthesis is a promising method for obtaining HAp powders in shorter process time.

Treatment with antidepressive drugs improved quality of life in chronic hemodialysis patients.

BACKGROUND: Despite some improvements in dialysis therapies, depression still remains an important problem in chronic hemodialysis (HD) patients. In this study, we aimed to investigate the association of depression and its treatment with quality of life (QOL) in HD patients. PATIENTS AND METHODS: 97 HD patients (52 male, 45 female, mean age 55 +/- 16 years) were enrolled. All patients had been dialyzed for more than 6 months. In order to evaluate QOL of the patients, a short form of Medical Outcomes Study (SF-36) was used. Depression was assessed by using Beck Depression Inventory (BDI). Patients who had BDI score > or = 15 were diagnosed as to have depression. Patients with depression received antidepressive treatment (sertralin HCl, 50 mg/day) for an 8-week period. After 8-week antidepressive treatment, all biochemical analysis, SF-36 and BDI were performed again. RESULTS: 40 patients (20 male, 20 female, mean age 56 +/- 14 years) had depression. All parameters related to QOL were significantly decreased in patients with depression as compared to patients without depression. Severity of depression was correlated with QOL parameters. After 8 weeks of treatment, as parallel to changes in BDI, QOL parameters improved in patients with depression. CONCLUSION: Decrease in QOL, associated with depression and antidepressive treatment, improves QOL in HD patients. Hemodialysis patients should be followed-up closely for presence of depression. Treatment of depression with antidepressive drug regimen would lead to relieve the symptoms related to depression and improvement of QOL in these patients. Antidepressive treatment should be required more often than we prescribe in routine clinical practice now.

DNA polymerase in nuclei isolated from herpes simplex virus type-2-infected cells. Characterization of the reaction product and inhibition by substrate analogs.

Nuclei isolated from herpes simplex virus (HSV) type 2-infected KB cells were examined for their capacity to serve as an in situ source of herpes DNA polymerase. In contrast to purified enzymes with added template, approx. 80% of the DNA synthesized in isolated nuclei was viral. The average size of DNA fragments labeled in vitro was 3.2 X 10(6) Da. Based on an increase in DNA density when nuclei were incubated in the presence of BrdUTP rather than dTTP, 16% of the nucleotides were added during the in vitro reaction. Sucrose gradient analysis of DNA polymerase activity in extracts of isolated nuclei demonstrated the nearly exclusive presence of herpes DNA polymerase. Km concentrations for the four dNTPs were from 0.14 to 0.55 microM. DNA synthesis was inhibited competitively by the 5'-triphosphates of ara-A and ara-C (Ki = 0.03 and 0.22 microM, respectively) but not by the 5'-triphosphate of dideoxythymidine. aATP also served as a substrate (Km = 0.014 microM) for the reaction. We conclude that nuclei from HSV-infected cells have significant advantages for the detailed study of inhibitors of herpesvirus replication.

Implementing hygiene monitoring systems in hospital laundries in order to reduce microbial contamination of hospital textiles.

As textiles sent to hospital laundries contain many types of pathogenic organisms, it is important that laundering not only has an appropriate cleaning effect but also has a satisfactory disinfecting effect. Critical to this process is the maintenance of an appropriate hygiene level in the clean area of laundries in order to prevent recontamination of textiles from manual handling when ironing, folding, packing etc. The aims of this study were to evaluate the hygienic state of a hospital laundry, to introduce continuous sanitary measures, and to introduce a continuous hygiene monitoring system with an infection control programme. Two systems for evaluating hospital laundry hygiene were combined: HACCP principles (hazard analysis and critical control points) and RAL-GZ 992 standards (quality assurance standard for textile care of hospital laundry). Evaluation of the hygienic state of the hospital laundry was carried out by evaluating the number and types of micro-organisms present at the critical control points throughout the whole laundering process, using RODAC agar plates for surface sampling and the pour plate method for investigating water samples. The initial examination showed that the sanitary condition of the laundry did not reach the required hygiene level. Therefore, fundamental sanitation measures were instituted and the examination was repeated. Results were then satisfactory. The most important critical control point was the chemothermal laundering efficiency of the laundering process. To prevent micro-organisms spreading into the entire clean working area, it is important that, in addition to regular sanitary measures such as cleaning/disinfecting all working areas, technical equipment and storage shelves etc., regular education sessions for laundry employees on proper hand hygiene is undertaken and effective separation of the clean and dirty working areas is achieved.

Application of experimental design methodology in development and optimization of drug release method.

The aim of our research was to apply experimental design methodology in the development and optimization of drug release methods. Diclofenac sodium (2-[(2,6-dichlorophenyl)amino]benzeneacetic acid monosodium salt) was selected as a model drug and Naklofen retard prolonged release tablets, containing 100 mg of diclofenac sodium, were chosen as a model prolonged release system. On the basis of previous results, a three-level three-factorial Box-Behnken experimental design was used to characterize and optimize three physicochemical parameters, i.e. rotation speeds of the stirring elements, pH, and ionic strengths of the dissolution medium, affecting the release of diclofenac sodium from the tablets. The chosen dependent variables (responses) were a cumulative percentage of dissolved diclofenac sodium in 2, 6, 12 and 24 h. For estimation of coefficients in the approximating polynomial function, the least square regression method was applied. Afterwards, the information about the model reliability was verified by using the analysis of variance (ANOVA). The estimation of model factors' significance was performed by Student's t-test. For investigation of the shape of the predicted response surfaces and for model optimization, the canonical analysis was applied. Our study proved that experimental design methodology could efficiently be applied for characterization and optimization of analytical parameters affecting drug release and that it is an economical way of obtaining the maximum amount of information in a short period of time and with the fewest number of experiments.