Search

Background: The aim of this study was to assess the epidemiological profile of unintentional opioid overdoses, the prevalence and number of psychotropic substances involved in opioid overdoses.Methods: This was a descriptive study, in which 180 participants were enrolled, and covered a nine-years-period. For collecting data was used the National patient electronic system “My term”. The variables as gender, age, duration of opioid dependence, number of overdoses, type of substance, number of antidote ampoules, duration of hospitalization were analyzed. Severity of poisoning was made by using the Poison severity score. Results: Opioid overdose cases were significantly higher among males than females. Mean age with standard deviation (SD) was 32.23 ± 6.71 years. Mean years (±SD) of duration of opioid use disorder was 11.60 ± 5.89 years. The most commonly used primary substance was methadone in 68.89% and heroin in 31.11% cases. Twenty patients were treated with mechanical ventilation because of the severe respiratory depression. Poison severity score was moderate in 51.11%, severe in 45.56% and fatal in 3.33% of the cases. Conclusion: Most of the cases, predominantly males used one or two substances. The combination of methadone and benzodiazepine was most frequently used and the most common way was by injecting the abused substances. In most of the subjects PSS score was moderate and severe with no differences between genders.

Leptin, as a major adipokine, positively correlates with the body’s fat, while atopy is an important feature in the development of childhood asthma. We aimed to evaluate the relationship between leptin, parameters of obesity, and atopy in children with asthma. The study included 112 children (73 boys, 39 girls, mean age 11.1±2.4). 41 were overweight, 38 had asthma and a normal body mass index (BMI), and 33 were overweight asthmatics. Serum leptin levels, BMI, waist circumference (WC), and waist to hips ratio (WHR) were measured. Skin prick test (SPT)/CAP, total serum IgE, fractional exhaled nitric oxide (FeNO), and pulmonary function tests were performed. In asthmatic children, serum leptin median level was 9.2±16.2 ng/ml, in overweight children was 30.6±21.6 ng/ml, and in overweight asthmatics was 31.1±20.3 ng/ml with a significant difference between the groups (p=0.0374), yet with a significantly lower median level in the group of children with asthma compared to the overweight children: with asthma (p=0.00001) and without asthma (p=0.00001). In the three groups of patients, BMI and WC displayed a significant positive correlation with leptin (for BMI r=0.652 vs. r=0.530 vs. r=0.563, respectively and for WC r=0.508 vs. r=0.426 vs. r=0.527, respectively). No significant correlations of leptin within atopy parameters (Eo, IgE, SPT/CAP, FeNO) in all three analyzed groups (p>0.05) was detected. Conclusion: Atopy was not confirmed as an underlying mechanism of the association between asthma and being overweight. Leptin had a significant linear correlation as a parameter of central obesity with BMI and WC in all three groups, but not with WHR.

Objective: We have set as objective to analyze epidemiological data of diagnosed thyroid carcinoma (TC) cases, incidence and prevalence rate by gender, age, histopathological type, and statistical regions in R. of N. Macedonia during the period 1999 to 2015. Material and Methods: Retrospective analysis of medical data collected from the 2 state thyroid departments. Inclusion criteria included newly diagnosed cases of TC in appropriate years for the period 1999 to 2015. We have evaluated: yearly incidence rate, incidence and prevalence by gender, age, the distribution in 8 statistical state regions and histopathological types and subtypes representation. Results: A total number of 422 TC patients were detected, average incidence rate of 1.22/105, with most prevalent papillary TCs79.5%, followed by follicular 10.9%, medullar 4.1%, anaplastic 3.1%, and other rare types with 2.3%. The highest incidence rate was detected in Skopje region, while the lowest in Southeast and the Polog region. The total prevalence rate for the female gender was 32.61/104 and for male 9.27/104 (f/m ratio = 3.52:1), with lowest female/male difference found in the elderly > 65 years (f/m = 2.21/1). Conclusion: Compared with regional epidemiological data we can conclude that Republic of N. Macedonia has very low incidence and prevalence rate, while female/male ratio was similar to that described in the literature. Our low incidence and prevalence rate may be due to 2 possible reasons, 1 would be insufficient diagnosis of only small portion of the real cases in the population, or the second reason may be a real low incidence resulting of specific etiopathogenetic circumstances.

Copyright: © 2021 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/), Self-management of health requires skills to obtain, process, understand, and use health-related information. Assessment of adolescents’ functional health literacy requires valid, reliable, and low-burden tools. The main objective of this study was to adapt and study the psychometric properties of the Newest Vital Sign for the Portuguese adolescents’ population (NVS-PTeen). Classic psychometric indicators of reliability and validity were combined with item response theory (IRT) analyses in a cross-sectional survey, complemented with a 3-month test-retest assessment. The NVS-PTeen was self-administered to students enrolled in grades 8 to 12 (12 to 17 years old) in a school setting. Overall, 386 students (191 girls) from 16 classes of the same school participated in the study (mean age = 14.5; SD = 1.5). Internal reliability of the NVS-PTeen was α = 0.60. The NVS-PTeen total score was positively and significantly correlated with Portuguese (r = 0.28) and mathematics scores (r = 0.31), school years (r = 0.31), and age (r = 0.19). Similar to the original scale (for the U.S.), the NVS-PTeen is composed of two dimensions, reading-related literacy and numeracy. Temporal reliability is adequate, though with a learning effect. IRT analyses revealed differences in difficulty and discriminative capacity among items, all with adequate outfit and infit values. Results showed that the NVS-PTeen is valid and reliable, sensible to inter-individual educational differences, and adequate for regular screening of functional health literacy in adolescents., This research was funded by the Fundação para a Ciência e a Tecnologia (FCT), under the project Health Information of Portuguese Population: Health Knowledge & Perceived Quality and Accessibility of Health Information Sources (Reference: HMSP-IISE/SAU-ICT/0004/2009). The writing of the paper also benefited from funding from the FCT under the grants UIDB/04295/2020 and UIDP/04295/2020. This funding entity had no role in the design or writing of this artic

Background: Sand flies (Diptera: Psychodidae) are medically important vectors of human and veterinary disease-causing agents. Among these, the genus Leishmania (Kinetoplastida: Trypanosomatidae), and phleboviruses are of utmost importance. Despite such significance, updated information about sand fly fauna is missing for Balkan countries where both sand flies and autochtonous leishmaniases are historically present and recently re-emerging. Therefore, a review of historical data on sand fly species composition and distribution in the region was followed by a large-scale entomological survey in eight Balkan countries to provide a recent update on local sand fly fauna. Methods: The literature search involved the period 1910–2019. The entomological survey was conducted at 1189 sampling stations in eight countries (Bulgaria, Bosnia and Herzegovina, Croatia, Kosovo, Montenegro, North Macedonia, Serbia and Slovenia), covering 49 settlements and 358 sampling sites between June and October in the years 2014 and 2016, accumulating 130 sampling days. We performed a total of 1189 trapping nights at these stations using two types of traps (light and CO2 attraction traps) in each location. Sampling was performed with a minimal duration of 6 (Montenegro) and a maximal of 47 days (Serbia) between 0–1000 m.a.s.l. Collected sand flies were morphologically identified. Results: In total, 8490 sand fly specimens were collected. Morphological identification showed presence of 14 species belonging to genera Phlebotomus and Sergentomyia. Historical data were critically reviewed and updated with our recent findings. Six species were identified in Bosnia and Herzegovina (2 new records), 5 in Montenegro (2 new records), 5 in Croatia (2 new records), 9 in Bulgaria (5 new records), 11 in North Macedonia (1 new record), 10 in Serbia (no new records), 9 in Kosovo (3 new records) and 4 in Slovenia (no new records). Conclusions: This study presents results of the first integrated sand fly fauna survey of such scale for the Balkan region, providing first data on sand fly populations for four countries in the study area and presenting new species records for six countries and updated species lists for all surveyed countries. Our findings demonstrate presence of proven and suspected vectors of several Leishmania species. [Figure not available: see fulltext.] © 2020, The Author(s)., European Food Safety Authority, EFSA European Centre for Disease Prevention and Control, ECDC, The work was carried out under the VectorNet project, a European network for sharing data on the geographic distribution of arthropod vectors, transmitting human and animal disease agents (Contract OC/EFSA/AHAW/2013/02-FWC1) funded by the European Food Safety Authority (EFSA) and the European Centre for Disease Prevention and Control (ECDC). Acknowledgements

Abnormal cerebrospinal fluid (CSF) pulsatility has been implicated in patients suffering from various diseases, including multiple sclerosis and hypertension. CSF pulsatility results in subarachnoid space (SAS) width changes, which can be measured with near-infrared transillumination backscattering sounding (NIR-T/BSS). The aim of this study was to combine NIR-T/BSS and wavelet analysis methods to characterise the dynamics of the SAS width within a wide range of frequencies from 0.005 to 2 Hz, with low frequencies studied in detail for the first time. From recordings in the resting state, we also demonstrate the relationships between SAS width in both hemispheres of the brain, and investigate how the SAS width dynamics is related to the blood pressure (BP). These investigations also revealed influences of age and SAS correlation on the dynamics of SAS width and its similarity with the BP. Combination of NIR-T/BSS and time-frequency analysis may open up new frontiers in the understanding and diagnosis of various neurodegenerative and ageing related diseases to improve diagnostic procedures and patient prognosis.

The Roma population is a European ethnic minority characterized by recent and multiple dispersals and founder effects. After their origin in South Asia around 1,500 years ago, they migrated West. In Europe, they diverged into ethnolinguistically distinct migrant groups that spread across the continent. Previous genetic studies based on genome-wide data and uniparental markers detected Roma founder events and West-Eurasian gene flow. However, to the best of our knowledge, it has not been assessed whether these demographic processes have equally affected both sexes in the population. The present study uses the largest and most comprehensive dataset of complete mitochondrial and Y chromosome Roma sequences to unravel the sex-biased patterns that have shaped their genetic history. The results show that the Roma maternal genetic pool carries a higher lineage diversity from South Asia, as opposed to a single paternal South Asian lineage. Nonetheless, the European gene flow events mainly occurred through the maternal lineages; however, a signal of this gene flow is also traceable in the paternal lineages. We also detect a higher female migration rate among European Roma groups. Altogether, these results suggest that sociocultural factors influenced the emergence of sex-biased genetic patterns at global and local scales in the Roma population through time., Tis work was supported by the Spanish Ministry of Economy and Competitiveness (grant numbers CGL2016- 75389-P (MINEICO/FEDER, UE), PID2019-106485GB-I00 (MINEICO), and “Unidad María de Maeztu” (MDM-2014-0370) to DC and FC; and Agència de Gestió d’Ajuts Universitaris i de la Recerca (Generalitat de Catalunya, grant 2017SGR00702). NF-P was supported by a FPU17/03501 fellowship

© The Author(s). 2019 Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated., Background: The existing knowledge on the interplay between reproductive and sexual health, migration and acculturation is recent and inconsistent, particularly on the sociocultural motives and constraints regarding fertility. Therefore, sexual and reproductive health (SRH) surveys are needed to provide accurate and comparable indicators to identify and address SRH inequalities, with specific focus on under researched aspects, such as the interrelation between migration and gender. FEMINA (FErtility, MIgratioN and Acculturation) aims to investigate intersectional SRH inequalities among Cape Verdean immigrant and Portuguese native families and how they impact on fertility in Portugal. This study will use a comprehensive approach exploring simultaneously the components of SRH, namely regarding identities, perceptions and practices of both women and men among lay people and relevant experts and stakeholders. The project has three main goals: 1) to identify social determinants of SRH among Cape Verdean immigrant and Portuguese native men and women of reproductive age; 2) to gain understanding of the diversity of the sexual and reproductive experiences and expectations of Cape Verdean immigrant and Portuguese native men and women of reproductive age, considering the singularities of their migratory, social and family dynamics; and 3) to produce recommendations for policy makers, employers and service providers on how to better address the SRH needs of Portuguese-born and immigrant populations. Methods: The study will address these goals using a mixed methods approach, including: a cross-sectional telephone survey with a probabilistic sample of 600 Cape Verdean immigrant and 600 Portuguese native women and men (women aged 18 to 49 and men aged 18 to 54), residents of the Greater Lisbon Area; a qualitative research through in-depth interviews with a subsample of 30 Cape Verdean immigrants and 30 Portuguese native men and women; and a Delphi technique for finding consensus on good practices in SRH for the entire population with a special emphasis on immigrants, namely extra-EU migrants. Discussion: Data will be used to produce a comprehensive set of indicators to monitor SRH in Portugal, to foster a greater understanding of its specificities and challenges to policy and decision makers, and to provide targeted recommendations to promote inclusive and migrant sensitive SRH services., The FEMINA project (PTDC/SOC-SOC/30025/2017) was granted by Fundação para a Ciência e a Tecnologia, I.P. with national funding. This publication is supported by Fundação para a Ciência e a Tecnologia project grant and ISCTE-IUL funds.

Aim To assess the association between azoospermia factor c microrearrangements and semen quality, and between Y-chromosome background with distinct azoospermia factor c microrearrangements and semen quality impairment. Methods This retrospective study, carried out in the Research Center for Genetic Engineering and Biotechnology “Georgi D. Efremov,” involved 486 men from different ethnic backgrounds referred for couple infertility from 2002- 2017: 338 were azoospermic/oligozoospermic and 148 were normozoospermic. The azoospermia factor c microrearrangements were analyzed with sequence tagged site and sequence family variant markers, quantitative fluorescent polymerase chain reaction, and multiplex ligation probe amplification analysis. The Y-haplogroups of all participants were determined with direct single nucleotide polymorphism typing and indirect prediction with short tandem repeat markers.Results Our participants had two types of microdeletions: gr/gr and b2/b3; three microduplications: b2/b4, gr/gr, and b2/b3; and one complex rearrangement gr/gr deletion + b2/b4 duplication. Impaired semen quality was not associated with microrearrangements, but b2/b4 and gr/ gr duplications were significantly associated with haplogroup R1a (P < 0.001 and P = 0.003, respectively) and b2/b3 deletions with haplogroup E (P = 0.005). There were significantly more b2/b4 duplication carriers in Albanians than in Macedonians with haplogroup R1a (P = 0.031). Conclusion Even though azoospermia factor c partial deletions/duplications and Y-haplogroups were not associated with impaired semen quality, specific deletions/ duplications were significantly associated with distinct haplogroups, implying that the Y chromosome background may confer susceptibility to azoospermia factor c microrearrangements.

Objectives The aim of our study was to evaluate the survival rate of all thyroid carcinomas (TCs) diagnosed in the 1999–2015 period in the Republic of North Macedonia and to analyze the prognostic influence of several characteristics on development of distant metastases, as well as to analyze the prognostic effect of seven clinical and constitutional features on mortality. Material and methods A retrospective analysis of medical data from all TCs diagnosed in 1999–2015 was performed. The survival rate of all types of TCs was estimated using the Kaplan Meier method. Univariate and multivariate logistic regression analysis was applied for evaluation of the predictive role of seven clinical and constitutional characteristics for development of distant metastases, and the univariate Cox-proportional model was applied for evaluation of the predictors for mortality. Results A total of 422 TC cases were diagnosed in the 17-year period, with an average survival time of 212.99 months. Results of the univariate regression analysis showed that dimension at initial ultrasound and histopathological type of tumor were significantly predictive variables for distant metastases. Multifocal tumors vs. unifocal tumors < 15 mm significantly increased the probability of distant metastases by 7.401 (p = 0.005, 95% CI = 1.817–30.190) times. Age, initial lymph node involvement, number of radioiodine therapies, and histopathology of the tumor were selected as independent significant predictors for mortality. Conclusion Our results showed an excellent overall prognosis of thyroid tumors in the Macedonian population. The dimension of the tumor, multifocality, and histopathological type were the most relevant prognostic predictive features for development of distant metastases.

Recently, Balkan virus (BALKV, family Phenuiviridae, genus Phlebovirus) was discovered in sand flies collected in Albania and genetically characterised as a member of the Sandfly fever Naples species complex. To gain knowledge concerning the geographical area where exposure to BALKV exists, entomological surveys were conducted in 2014 and 2015, in Croatia, Bosnia and Herzegovina (BH), Kosovo, Republic of Macedonia and Serbia. A total of 2830 sand flies were trapped during 2014 and 2015 campaigns, and organised as 263 pools. BALKV RNA was detected in four pools from Croatia and in one pool from BH. Phylogenetic relationships were examined using sequences in the S and L RNA segments. Study of the diversity between BALKV sequences from Albania, Croatia and BH showed that Albanian sequences were the most divergent (9–11% [NP]) from the others and that Croatian and BH sequences were grouped (0.9–5.4% [NP]; 0.7–5% [L]). The sand fly infection rate of BALKV was 0.26% in BH and 0.27% in Croatia. Identification of the species content of pools using cox1 and cytb partial regions showed that the five BALKV positive pools contained Phlebotomus neglectus DNA; in four pools, P neglectus was the unique species, whereas P. tobbi DNA was also detected in one pool. We report here (i) the first direct evidence that the Balkan virus initially described in coastal Albania has a much wider dissemination area than originally believed, (ii) two real-time RT-PCR assays that may be useful for further screening of patients presenting with fever of unknown origin that may be caused by Balkan virus infection, (iii) entomological results suggesting that Balkan virus is likely transmitted by Phlebotomus neglectus, and possibly other sand fly species of the subgenus Larroussius. So far, BALKV has been detected only in sand flies. Whether BALKV can cause disease in humans is unknown and remains to be investigated.

Introduction. Patients satisfaction is a very important part of any clinical practice both for evaluation and improvement of healthcare services. Objective. The aim of this study was to determine patient satisfaction with public outpatient healthcare services at secondary and tertiary level and to assess possible differences between the two levels. Methods. In a quantitative cross-sectional study, a convenient sample of 646 patients who experienced public outpatient healthcare services at the secondary and tertiary level during the last two months were interviewed. Patient satisfaction questionnaires, with statements regarding various aspects of satisfaction, were completed during face-to-face interviews (response rate 84.6%). The research instrument was tested for internal consistency using the Cronbach?s coefficient alpha estimate. Results. The patients were significantly more satisfied in tertiary than in secondary outpatient healthcare facilities in almost all aspects of assessment related to general settings, nurse/administrative staff performance and physician performance (p

Cystinuria is an autosomal recessive disorder caused by defective transport of cystine and dibasic amino acids in the proximal renal tubules and small intestine. So far, more than 128 mutations in SLC3A1 gene, and 93 in SLC7A9 gene have been described as a cause of cystinuria. We present a molecular characterization of the cystinuria in 47 unrelated south-east European families. The molecular methodology included direct sequencing, single strand conformational polymorphism, and restriction fragment length polymorphism. A total of 93 (94.9 %) out of 98 unrelated cystinuric chromosomes have been characterized. Mutations in SLC3A1 gene account for 64.3 % and in SLC7A9 gene for 30.6 % of the cystinuric chromosomes. Ten different mutations in SLC3A1 gene were found, and two of them were novel (C242R and L573X), while in SLC7A9 gene seven mutations were found, of which three were novel (G73R, V375I and c. 1048_1051delACTC). The most common mutations in this study were T216M (24.5 %), M467T (16.3 %) and R365L (11.2 %) in SLC3A1 and G105R (21.4 %) in SLC7A9 gene. A population specificity of cystinuria mutations was observed; T216M mutation was the only mutation present among Gypsies, G105R was the most common mutation among Albanians and Macedonians, and R365L among Serbs. The results of this study allowed introduction of rapid, simple and cost-effective genetic diagnosis of cystinuria that enables an early preventive care of affected patients and a prenatal diagnosis in affected families.

In order to evaluate whether arterial hypertension (AH) affects skin microcirculation, 46 newly diagnosed, nevertreated, hypertensive patients and 20 healthy normotensive controls underwent a forearm skin postocclusive reactive hyperaemia (PORH) test, using laser-Doppler flowmetry (LDF). Their resting skin blood flow oscillations (SBFOs) were also investigated using wavelet spectral analyses of skin LDF tracings within six frequency subintervals in the 0.005‐2Hz spectral range. To evaluate whether antihypertensive treatment affects skinmicrocirculation,the same measurements were repeated in 22 of the recruited hypertensive patients after 8W2 weeks of antihypertensive treatment. Significantly reduced PORH, together with significantly higher spectral amplitudes within the majority of the investigated SBFO subintervals, was found in untreated hypertensive patients compared with controls. In the 22 hypertensive patients who completed the follow-up, there was a significant increase in PORH after antihypertensive treatment compared with before (357W178vs.284W214%,respectively,P

Background Although age-related loss of chromosome Y (LOY) in normal hematopoietic cells is a well-known phenomenon, the phenotypic consequences of LOY have been elusive. However, LOY has been found in association with smoking, shorter survival and higher risk of cancer. It was suggested that LOY in blood cells could become a predictive biomarker of male carcinogenesis. Aims, Methods & Findings To investigate the association of LOY in blood cells with the risk for development of colorectal (CC) and prostate cancers (PC), we have analyzed DNA samples from peripheral blood of 101 CC male patients (mean age 60.5±11.9 yrs), 70 PC patients (mean age 68.8±8.0 yrs) and 93 healthy control males (mean age 65.8±16.6 yrs). The methodology included co-amplification of homologous sequences on chromosome Y and other chromosomes using multiplex quantitative fluorescent (QF) PCR followed by automatic detection and analysis on ABI 3500 Genetic Analyzer. The mean Y/X ratio was significantly lower in the whole group of cancer patients (0.907±0.12; p = 1.17x10-9) in comparison to the controls (1.015±0.15), as well as in CC (0.884±0.15; p = 3.76x10-9) and PC patients (0.941±0.06; p = 0.00012), when analyzed separately. Multivariate logistic regression analysis adjusting for LOY and age showed that LOY is a more significant predictor of cancer presence than age, and that age probably does not contribute to the increased number of subjects with detectable LOY in cancer patients cohort. Conclusion In conclusion, our results support the recent findings of association of LOY in blood cells with carcinogenesis in males.

BACKGROUND: Time-frequency analysis of the laser Doppler flowmetry signal, using wavelet transform, shows periodic oscillations at five characteristic frequencies related to the heart (0.6–2 Hz), respiration (0.15– 0.6 Hz), myogenic activity in the vessel wall (0.052– 0.15 Hz), sympathetic activity (0.021– 0.052 Hz), and very slow oscillations (0.0095– 0.021), which can be modulated by the endothelium-dependent vasodilator acetylcholine. We hypothesized that wavelet transform of laser Doppler flowmetry signals could detect changes in the microcirculation induced by general anesthesia, such as alterations in vasomotion and sympathetic activity. METHODS: Eleven patients undergoing faciomaxillary surgery were included. Skin microcirculation was measured on the lower forearm with laser Doppler flowmetry and iontophoresis with acetylcholine and sodium nitroprusside before and during general anesthesia with propofol, fentanyl, and midazolam. The laser Doppler flowmetry signals were analyzed using wavelet transform. RESULTS: There were significant reductions in spectral amplitudes in the 0.0095–0.021 (P < 0.01), the 0.021–0.052 (P < 0.001), and the 0.052–0.15 Hz frequency interval (P < 0.01) and a significant increase in the 0.15–0.6 Hz frequency interval. General anesthesia had no effect on the difference between acetylcholine and sodium nitroprusside on relative amplitudes in the 0.0095–0.021 Hz frequency interval (P < 0.001). CONCLUSION: General anesthesia reduces the oscillatory components of the perfusion signal related to sympathetic, myogenic activity and the component modulated by the endothelium. However, the iontophoretic data did not reveal a specific effect on the endothelium. The increase in the 0.15–0.6 Hz interval is related to the effect of mechanical ventilation.

Common European mole cricket (CEML) Grillotalpa grillotalpa L causes damage to field, vegetable crops, and small fruits growing at commercial plantations and nurseries. Chemical control if insecticides are used in poison bates, soil application or seedling/bulbs treatment is not environmentally friendly. Inundative and innoculative release of CTVL biocontrol agents, in particularly, Entomopathogenic nematode is a reliable alternative to chemical control. At the laboratory study the comparison of the ability of commercial strain (Nemastar) and local Ukrainian isolate of Steinernema carpocapsae in various concentrations to parasite in last instar nymph and adults of G. grillotalpa was investigated. Grillotalpa grillotalpa was found as a susceptible host for both commercial and local strains of S. carpocapsae. The life cycle of S. carpocapsae both strains in the adults of G. grillotapla with concentration 50 IJs per larva has been completed 12-15 days at t=25 C. Two generations of S. carpocapsae were able to develop in mole cricket for both strains. Two strains of S. carpocapsae nematode species tested were pathogenic to adults of G. grillotalpa. The mortalities of G. grillotapla last last instars' larva caused by S. carpocapsae were recorded in every concentration tested at least 20 to 150 IJs per larva. Mean larval mortality ranged from 48% to 95% depending upon nematode strain and rate of application. Larval mortality generally increased with increasing of nematode rates. It was significant for both S. carpocapsae strains (Ukr. Isolate F = 262,86) and commercial strain (Nemastar F = 102,952,86). Ukrainian local isolate caused a greater percentage of mortality of G. grillotapla adult than commercial strain of S. carpocapsae tested but interactions between nematode strains, application rates were not significant. This study presents new data on effect of S. carpocapsae isolated for Ukraine to key agricultural polyphagous pest G. grillotalpa susceptibility screening under controlled conditions, as presented in this paper is the first step towards the development of a biological control program of G. grillotaplpa by using of entomopathogenic nematodes.

Recurrent aphthous stomatitis (RAS) is a quite frequent, painful, ulcerative disease that affects the lining of the oral cavity and has an unknown etiology. The aim of this article is to examine the impact of the medication proaftol on epithelization speed and severity of pain in patients with RAS. In this randomized, double-blind, placebo-controlled study respondents were divided into two groups where one group was treated with proaftol spray and the other with a placebo. Aphthae considered for treatment had a diameter of 5-6 mm. The participants were given instruction on the use of the spray, two sprayings on the place of the aphtae 3-4 times a day. We examined two parameters in the symptomatology of RAS-lesion size (mm) and pain intensity (noted on four subjective levels: 0-no pain, 1-discomfort, 2-moderate pain, 3-severe pain). These parameters were noted on the baseline, the third, the fifth and the eighth days of examination.A significant faster reduction of the dimension of aphthous ulcers in patients treated with proaftol 3rd day p0.001, 5th day p0.0006, 8th day full epithelization in the control group. The magnitude of pain in the experimental compared with the control group on 3rd, 5th and 8th day was also significantly reduced: p0.0001, p0.007, p0.007 respectively.The use of proaftol positively influences the rate of epithelization and reduction of subjective feeling of pain in patients with RAS. The action of propolis should be the goal of studies with a larger number of subjects.

Background: The skin microcirculation may be evaluated noninvasively by laser Doppler flowmetry and iontophoresis with acetylcholine and sodium nitroprusside. Wavelet transform of the perfusion signal shows periodic oscillations of five characteristic frequencies in the interval 0.0095‐1.6 Hz. The aim of the current study was to investigate alterations in skin microcirculation induced by brachial plexus block, with emphasis on the periodic oscillations. Methods: Healthy nonsmokers undergoing hand surgery (n 13) were anesthetized with brachial plexus block, using bupivacaine, lidocaine, and epinephrine. Skin microcirculation was evaluated by laser Doppler flowmetry and iontophoresis with acetylcholine and sodium nitroprusside before and after brachial plexus block. Wavelet transform of the perfusion signal was performed. As a control group, 10 healthy nonsmokers were included. Results: In the anesthetized arm, skin perfusion after brachial plexus block increased from 19 (12‐30) to 24 (14‐39) arbitrary units (P < 0.01). A significant increase was also seen in the contralateral arm from 17 (14‐32) to 20 (14‐42) arbitrary units (P < 0.01). After brachial plexus block, spectral analysis revealed a significant reduction in relative amplitude of the oscillatory components within the 0.0095- to 0.021- (P < 0.001) and 0.021- to 0.052-Hz (P < 0.001) intervals in the anesthetized arm. Conclusion: Alterations in skin microcirculation induced by brachial plexus block can be evaluated by wavelet transform of the laser Doppler flowmetry signal. Brachial plexus block reduces the oscillatory components within the 0.0095- to 0.021and 0.021- to 0.052-Hz intervals of the perfusion signal. These alterations are related to inhibition of sympathetic activity and a possible impairment of endothelial function. THE human microcirculation shows rhythmical variations produced by central and local oscillators. The most obvious are the heart and respiration, which produce rhythmical variations in blood flow around 1.0 and 0.3 Hz, respectively. Guyton and Harris 1 in 1951 proposed the term vasomotor waves for slower oscillations than those related to respiration. Studies performed on humans and animals have concluded that these slow oscillations are influenced by the sympathetic nervous system and the microvascular wall. However, there have been conflicting results regarding the mechanisms for the different frequency intervals. 2 A relatively new approach to the analysis of nonstationary signals, such as human biologic signals, is wavelet analysis. 3 Wavelet analysis of laser Doppler flowmetry (LDF) signals from human cutaneous circulation has revealed five oscillations, including those from the heart and the respiration. Data indicate that the 0.1-, 0.03-, and 0.01-Hz oscillations are related to intrinsic myogenic activity of vascular smooth muscle cells, neurogenic activity on the vessel wall, and the vascular endothelium, respectively. 4,5 Brachial plexus block induces changes in peripheral hemodynamics. In addition, sympathetic impairment is a key finding and has been measured by stimulation of cutaneous vasoconstrictor reflexes, which are attenuated after brachial plexus block. 6,7 It has also been reported that local anesthetics impair endothelium-mediated vasodilatation. 8 –10 As far as we are aware, no reports exists on alterations in the periodic oscillations induced by brachial plexus block. We hypothesized that changes related to brachial plexus block, primarily the sympathetic block, could be detected by LDF, iontophoresis, and wavelet analysis of the human skin microcirculation.

Impaired blood supply is a significant risk factor for diabetic foot ulceration and gangrene. A possible relationship between peripheral macroangiopathy and the spectral components of microvascular skin blood flow in the lower extremities was tested in diabetic patients (DP) and non-diabetic subjects (C). Basal skin blood flow (BSBF) was recorded for 30 min at the right and left medial malleolus (predominantly nutritive capillary circulation) by laser Doppler flowmetry in 64 DP and 31 C. Its oscillatory components were analyzed using wavelet transform. Peripheral arterial obliterative disease (PAOD) was defined according to ankle/brachial index (ABI): PAOD+ (ABI < 0.9: 21 DP, 12 C), PAOD− (ABI 0.91–1.3: 43 DP, 19 C). No statistically significant differences in BSBF and its oscillatory components were observed between PAOD+ and PAOD−, neither in DP nor in C. In DP, the spectral component of microvascular flow associated with endothelial activity was in significant positive correlation with systolic pressures on brachial and dorsal pedal artery (p = 0.001 and 0.010, respectively). These results indicate that mean BSBF and its oscillatory components do not change with diabetic PAOD; however there is a strong correlation between systolic pressure and the oscillatory components of BSBF related to endothelial activity manifested in the frequency interval 0.0095–0.02 Hz.

Functional alterations of vascular endothelial cells may be evaluated by analysing differences in effects of endothelium-dependent [acetylcholine (ACh)] and endothelium-independent [sodium nitroprusside (SNP)] vasodilators. We evaluated whether a dynamic approach using spectral analysis of the blood flow signal, resulting from the cutaneous red cell flux and recorded by the technique of laser Doppler flowmetry (LDF), can detect higher endothelial responsiveness in trained versus less trained individuals. There was a 1.6 times higher ACh-induced cutaneous perfusion in athletes than in controls ( P

Hereditary multiple exostosis (HME) is a hereditary autosomal dominant disease in which multiple exostoses occur. Typically, the exostoses are primarily located at the metaphysis and migrate with continued growth towards the diaphysis. Clinical problems are caused by local pain, impingement of muscle tendons and neurovascular structures, malformation - especially in the forearm - and malignant transformation - especially exostoses at the trunc and pelvic girdle.A causal therapy is currently not available. Mechanical irritation is an indication for resection of the exostosis. Axial deviation of the lower extremity is treated according to the same principles as primary malalignments (temporary hemiepiphysiodesis/corrective osteotomy).The indication for correction of axial deviation at the upper extremity depends on age, extent as well as functional and cosmetic impairment. This should be discussed with the patient in detail. The patient has to be informed about the risk of malignant transformation after cessation of growth. Growing mass or new occurrence of symptoms after end of growth are suspicious and require further diagnostic examinations.

The aim of the study is to assess the trends of overdose and drug related fatalities in the Republic of Macedonia during the 11 years.Cross-sectional retrospective survey and reviewed of postmortem toxicological analyses which examined fatal poisonings with illegal drugs in years 2002-2013. Information about gender, age, drug consumption, reported years were analyzed. Narcotics were confirmed with toxicological semi quantitative fluorescence polarization immunoassay (FPIA) in urine (range 250-4000 ng/ml).Total of 165 deaths were observed. Out of them 145 (87.9%) were male. There is statistical significant differences between male and female DRD due to age (Mann-Whitney U Test = 925, Z = -2626, p = 0.0087). For p0.05 there is significant differences between genders due to cause of overdose (Pearson Chi-square = 9743, df = 4, p = 0.0449). DRD among male were mainly because of overdose due to heroin in 80 (51.17%) cases followed by DRD due to combination of methadone and BZD in 25 (11.72%) cases. Out of all DRD cases 50 (30.3%) are related to polydrug use. For p0.01 there is a significant differences between analyzed age groups due to cause of overdose (Pearson Chi-square = 33,886, df = 12, p = 0.0007).Death cause analysis reveals the difficulties in determining the role of substitution drugs, as many other factors may be involved. The findings also highlight the importance of further enhancing treatment interventions for benzodiazepine misuse among patients on methadone substitution treatment.

The wavelet transform technique, a time-frequency method with logarithmic frequency resolution, was used to analyze oscillations in human peripheral blood flow measured by laser Doppler flowmetry. The oscillations extended over a wide frequency scale and their periods varied in time. Within the frequency range studied, 0.0095-1.6 Hz, five characteristic oscillations were revealed, arising from both local and central regulatory mechanisms. After the insertion of endothelium-dependent and endothelium-independent vasodilators the spectra of blood flow markedly differed in the frequency interval 0.0095-0.02 Hz. In this way it was demonstrated that endothelial activity is a rhythmic process that contributes to oscillations in blood flow with a characteristic frequency of around 0.01 Hz. The study illustrates the potential of laser Doppler flowmetry combined with dynamical systems analysis for studies of both the micro- and macroscopic mechanisms of blood flow regulation in vivo.

Functional alterations to the endothelial cells of the vascular system may contribute to the improved circulatory performance induced by physical conditioning. We evaluated microvascular reactivity to iontophoretic application of acetylcholine (ACh) and sodium nitroprusside (SNP) through the skin and blood perfusion measurements in the same area using laser Doppler flowmetry. Whereas ACh acts on smooth muscle cells of the vascular system via the production of vasodilator substances from the endothelium, SNP is an endothelium-independent vasodilator acting on vascular smooth muscle cells directly. The study was performed using two groups of subjects with different levels of aerobic endurance, long distance runners competing at national level (n = 9) and controls (n = 9). The subjects were tested for 40 min on a treadmill before and after an exercise test at 80% of their maximal oxygen uptake. During stimulation by ACh cutaneous perfusion increased to a higher level in the athletes than in the controls (overall P0.05), whereas an acute period of exercise abolished this difference (overall P0.6). There was no significant difference between the athletes and the controls with respect to the SNP-induced increase in cutaneous perfusion either before (P0.9) or after (P0.9) exercise. The higher cutaneous perfusion responses to stimulation with ACh in the athletes than in the controls may support the hypothesis that regular exercise modifies the responsiveness of the cutaneous endothelium. The difference in ACh-induced perfusion and in unstimulated forearm perfusion between the two groups was present only at rest. This finding indicated that mechanisms were introduced during exercise, which compensated for the lower endothelial sensitivity to stimulation in the controls at rest.

Facial blood flow and temperature were significantly higher on the right side of the forehead compared to the left. This asymmetry implies that the hemispheric autonomic control of the face differs and could influence the expression of emotion.

Early laparoscopic cholecystectomy imposes itself as an option of choice for treatment of acute cholecystitis, while the rate of conversions to open procedure represents a key parameter for evaluating the outcome.The aim of this paper was to evaluate the results of the laparoscopic cholecystectomy in acute versus chronic cholecystitis through determining the conversion rate in open method, as well as to analye some predictive factors that may impact the decision to convert to open.An analytical case control study was conducted at the University Clinic for Digestive Surgery in Skopje within a period of 27 months. The first group included 62 patients with acute cholecystitis on whom the laparoscopic cholecystectomy was performed in the period from zero to the seventh day from the onset of symptoms. The second group included 62 patients with chronic cholecystitis who underwent laparoscopic cholecystectomy. Gender, age, history of cholecystitis, the time passed from the first symptoms till laparoscopic cholecystectomy and the duration of the intervention were analysed as factors that can possibly act on conversion.There were no significant differences (p0.05) between the group of patients with acute and the one with chronic cholecystitis due to conversion rate. In both groups, there was a significant difference in conversion due to the duration of the laparoscopic cholecystectomy (p0.05), and in the group with acute cholecystitis also due to the time passed from the first symptoms till the laparoscopic intervention (p0.01).The rate of conversions can be reduced with a prompt approach to predictive factors.

We experimentally altered the timing of respiratory motoneuron activity as a means to modulate and better understand otherwise hidden human central neural and hemodynamic oscillatory mechanisms. We recorded the electrocardiogram, finger photoplethysmographic arterial pressure, tidal carbon dioxide concentrations, and muscle sympathetic nerve activity in 13 healthy supine young men who gradually increased or decreased their breathing frequencies between 0.05 and 0.25 Hz over 9-min periods. We analyzed results with traditional time- and frequency-domain methods, and also with time-frequency methods (wavelet transform, wavelet phase coherence, and directional coupling). We determined statistical significance and identified frequency boundaries by comparing measurements with randomly generated surrogates. Our results support several major conclusions. First, respiration causally modulates both sympathetic (weakly) and vagal motoneuron (strongly) oscillations over a wide frequency range—one that extends well below the frequency of actual breaths. Second, breathing frequency broadly modulates vagal baroreflex gain, with peak gains registered in the low frequency range. Third, breathing frequency does not influence median levels of sympathetic or vagal activity over time. Fourth, phase relations between arterial pressure and sympathetic and vagal motoneurons are unaffected by breathing, and are therefore likely secondary to intrinsic responsiveness of these motoneurons to other synaptic inputs. Finally, breathing frequency does not affect phase coherence between diastolic pressure and muscle sympathetic oscillations, but it augments phase coherence between systolic pressure and R-R interval oscillations over a limited portion of the usual breathing frequency range. These results refine understanding of autonomic oscillatory processes and those physiological mechanisms known as the human respiratory gate.

We describe an analysis of cardiac and respiratory time series recorded from 189 subjects of both genders aged 16–90. By application of the synchrosqueezed wavelet transform, we extract the respiratory and cardiac frequencies and phases with better time resolution than is possible with the marked events procedure. By treating the heart and respiration as coupled oscillators, we then apply a method based on Bayesian inference to find the underlying coupling parameters and their time dependence, deriving from them measures such as synchronization, coupling directionality and the relative contributions of different mechanisms. We report a detailed analysis of the reconstructed cardiorespiratory coupling function, its time evolution and age dependence. We show that the direct and indirect respiratory modulations of the heart rate both decrease with age, and that the cardiorespiratory coupling becomes less stable and more time-variable.

The prevalence of hepatitis C virus (HCV) genotypes depends on geographical location. HCV genotyping is important for epidemiological investigations and treatment management. The aim of this study was to determine the HCV genotype prevalence in the most prominent risk groups in the Republic of Macedonia in the last 5 years and to evaluate its association with patient's age, gender, and mode of transmission. A total of 1,167 HCV positive patients, divided into three risk groups (intravenous drug use, chronic hemodialysis, and other risk factor), were genotyped using an in-house ASO hybridization method with genotype-specific oligonucleotide probes. The genotypes 1, 2, and 3 were present with 52.2%, 0.6%, and 47.0%, respectively. Genotype 1 was most prevalent in hemodialysis (89.0%) and other risk factor group (53.8%). It was found associated independently with hemodialysis, age40 and female gender. Genotype 3 predominated in intravenous drug users (64.0%) and was associated significantly also with age ≤40 and male gender. Multivariable logistic regression analysis pointed out hemodialysis (P 0.0001, Exp (B) = 12.0) as a positive predictor factor for genotype 1 and age ≤40 (P = 0.021, Exp (B) = 1.8) and intravenous drug use (P 0.0001, Exp (B) = 8.4) as a positive predictor factors for genotype 3. In conclusion, the main transmission route of HCV infection in the Republic of Macedonia is intravenous drug use, followed by hemodialysis. HCV genotypes 1 and 3 dominate in these two most prominent risk groups in the Republic of Macedonia.

Frequency-domain analyses of simultaneously recorded skin and muscle sympathetic nerve activities may yield unique information on otherwise obscure central processes governing human neural outflows. We used wavelet transform and wavelet phase coherence methods to analyse integrated skin and muscle sympathetic nerve activities and haemodynamic fluctuations, recorded from nine healthy supine young men. We tested two null hypotheses: (1) that human skin and muscle sympathetic nerve activities oscillate congruently; and (2) that whole-body heating affects these neural outflows and their haemodynamic consequences in similar ways. Measurements included peroneal nerve skin and tibial nerve muscle sympathetic activities; the electrocardiogram; finger photoplethysmographic arterial pressure; respiration (controlled at 0.25 Hz, and registered with a nasal thermistor); and skin temperature, sweating, and laser-Doppler skin blood flow. We made recordings at ∼27°C, for ∼20 min, and then during room temperature increases to ∼38°C, over 35 min. We analysed data with a wavelet transform, using the Morlet mother wavelet and wavelet phase coherence, to determine the frequencies and coherences of oscillations over time. At 27°C, skin and muscle nerve activities oscillated coherently, at ever-changing frequencies between 0.01 and the cardiac frequency (∼1 Hz). Heating significantly augmented oscillations of skin sympathetic nerve activity and skin blood flow, arterial pressure, and R-R intervals, over a wide range of low frequencies, and modestly reduced coordination between skin and muscle sympathetic oscillations. These results suggest that human skin and muscle sympathetic motoneurones are similarly entrained by external influences, including those of arterial baroreceptors, respiration, and other less well-defined brainstem oscillators. Our study provides strong support for the existence of multiple, time-varying central sympathetic neural oscillators in human subjects.

In order to assess outcome and possible predictors of early good results, a prospective study on 22 patients who were treated with save surgical hip dislocation for symptomatic isolated cam-type femoroacetabular impingement (FAI) was performed. After a follow-up of 6 and 12 months, standard clinical and radiographic parameters were recorded. A statistically significant improvement of the clinical status according to the Harris hip score could be assessed at six months (p-value = 0.003) and 12 months (p-value = 0.001) post-surgery. By comparing standard clinical and radiographic preoperative parameters with various follow-up outcomes, we revealed no specific parameter with predictive value. These findings are important for centers that have just started to use this surgical technique and are still identifying their learning curve.

Background The 1984/86 published neurogram results showing only rare sympathetic nerve activity (SNA) to the muscles and skin in tetraplegia are still accepted. The present study by a different method attempted to confirm or deny those findings. Methods and results The effect of basal SNA to the microcirculation of the feet and calf in 10 complete (AIS A) traumatic tetraplegic and 10 healthy age matched subjects were evaluated by wavelet transform of laser Doppler flowmetry (LDF) recordings. The results clearly indicated there is significant basal SNA from the decentralized spinal cord in tetraplegia. In addition, wavelet analysis allowed a study of other influences on the microcirculation besides SNA. Collectively, in tetraplegia compared with controls, the powers of the low frequency oscillations in blood flow were reduced; in that the endothelium caused less vasodilatation while the SNA and intrinsic vascular smooth muscles induced smaller degrees of vasoconstriction. However, the high frequency and especially the cardiac powers were greater. The latter presenting an obvious important factor for the preservation of blood flow in the microcirculation. Conclusions It is suggested that basal SNA to the cutaneous microcirculation occurs in complete tetraplegia, and the significant levels of circulating noradrenaline reported by others indicate this is also true in other parts of the body. This may explain the usual absence of severe, incapacitating, autonomic deficiency in this condition.

For realizing our study the supervised tooth-brushing program was carried out among 100 schoolchildren at the age of 9-12 and 13-16 years with low and moderate mental handicap in Skopje. To evaluate the results of six months intervention program, concentrated on encouragement of independent manual skills, OHI levels were detected by Green-Vermillion and CPITN index levels to characterize the gingival and periodontal health.For comparative analyzes of date-base OHI levels and after six months of intervention program, we detected that the mean date-base OHI index level for mentally handicaped children are 2.46, and at the end of the program (after six months) it was 0.73. CPITN index levels at the beginning and after six months of intervention programmed for mentally handicaped children in both age groups, also confirmed r statistical significance for this examined parameter, with evident reduction of CPITN mean levels from 2.11 to 0.95. Correlation among date-base OHI levels and levels at the end of our intervention program means high positive correlation between these index levels at the beginningand final examinations.This program gave promising results and was effective in reducing the plaque and gingivitis scores, so the key to long-term success of the program is to maintain the subjects' motivation to make oral hygiene a part of their daily routine and thus sustain this improvement (Tab. 1, Fig. 4, Ref. 12). Full Text (Free, PDF) www.bmj.sk.

The pulse transit time (PTT) of a wave over a specified distance along a blood vessel provides a simple non-invasive index that can be used for the evaluation of arterial distensibility. Current methods of measuring the PTT determine the propagation times of pulses only in the larger arteries. We have evaluated the pulse arrival time (PAT) to the capillary bed, through the microcirculation, and have investigated its relationship to the arterial PAT to a fingertip. To do so, we detected cardiac-induced pulse waves in skin microcirculation using laser Doppler flowmetry (LDF). Using the ECG as a reference, PATs to the microcirculation were measured on the four extremities of 108 healthy subjects. Simultaneously, PATs to the radial artery of the left index finger were obtained from blood pressure recordings using a piezoelectric sensor. Both PATs correlate in similar ways with heart rate and age. That to the microcirculation is shown to be sensitive to local changes in skin perfusion induced by cooling. We introduce a measure for the PTT through the microcirculation. We conclude that a combination of LDF and pressure measurements enables simultaneous characterization of the states of the macro and microvasculature. Information about the microcirculation, including an assessment of endothelial function, may be obtained from the responses to perturbations in skin perfusion, such as temperature stress or vasoactive substances.

Laser Doppler flowmetry (LDF) of forearm skin blood flow, combined with iontophoretically-administered acetylcholine and sodium nitroprusside and wavelet spectral analysis, was used for noninvasive evaluation of endothelial function in 17 patients newly diagnosed with New York Heart Association class II-III congestive heart failure (CHF). After 20+/-10 weeks' treatment with a beta(1)-blocker (Bisoprolol), the measurements were repeated. Measurements were also made on an age- and sex-matched group of healthy controls (HC). In each case data were recorded for 30 min. In HC, the difference in absolute spectral amplitude of LDF oscillations between the two vasodilators manifests in the frequency interval 0.005-0.0095 Hz (p0.01); this difference is initially absent in patients with CHF, but appears following the beta(1)-blocker treatment (p0.01). For HC, the difference between the two vasodilators also manifests in normalised spectral amplitude in 0.0095-0.021 Hz (p0.05). This latter difference is absent in CHF patients and is unchanged by treatment with beta(1)-blockers. It is concluded that there are two oscillatory skin blood flow components associated with endothelial function. Both are reduced in CHF. Activity in the lower frequency interval is restored by beta(1)-blocker treatment, confirming the association between CHF and endothelial dysfunction but suggesting the involvement of two distinct mechanisms.

In coupled nonlinear oscillators approach, the framework that has been used for the studies of cardiovascular and brain oscillations. As background, it describes the human CVS and present results of time-frequency analysis using wavelet transforms of several noninvasive measurements of cardiovascular signals. Studies of neuronal oscillations have been undertaken since the first human electroencephalographic (EEG) recording, and the recent resurgence of interest in neuronal oscillations. It is concluded that interactions occur between the oscillatory processes, both within and between the cardiovascular and the neuronal systems. The strengths and directions of these interactions may be used, in principle, for characterization of the state of the organism as demonstrated here for the case of deep anesthesia.

Bispectral analysis, introduced recently as a technique for revealing time-phase relationships among interacting noisy oscillators, has been used to study the nature of the coupling between cardiac and respiratory activity. Univariate blood flow signals recorded simultaneously by laser-Doppler flowmetry (LDF) on both legs and arms were analysed. Coupling between cardiac and respiratory activity was also checked by use of bivariate data and computation of the cross-bispectrum between simultaneously recorded ECG, blood pressure and respiratory signals. Recordings were taken during spontaneous breathing and paced respiration. It is shown that cardiorespiratory coupling exists during spontaneous as well as during paced respiration and that coupling information among cardiac and respiratory processes in young, healthy subjects is spatially invariant, but can be expected to change with cardiovascular diseases.

Background/Aims: Laser Doppler flowmetry (LDF) was used to determine the influence of acute myocardial infarction (AMI) and of successful reperfusion treatment on basal skin blood flow and its oscillatory components. Methods: Skin LDF was performed on all extremities in 58 patients 4–9 days after AMI (Killip class I), and in 71 healthy age- and sex-matched controls. Wavelet analysis was applied to evaluate oscillatory components within the interval 0.005–2 Hz. Results: AMI patients had reduced mean flow (p < 0.01) and oscillatory components (p < 0.04) in all extremities. Reperfused (n = 40), compared to nonreperfused (n = 18), patients had higher mean flow and total spectral amplitude at all recording points. The difference was statistically significant only in legs (group median LDF in the left leg was 9.68 AU for reperfused and 5.71 AU for nonreperfused patients, p < 0.04, and 11.47 and 4.24 AU in the right leg, p < 0.01). Reperfused patients had significantly higher total spectral amplitude in both legs (p < 0.04). Conclusions: In AMI patients, reduced skin blood flow and its oscillatory components may reflect ongoing neurohumoral activation despite absence of clinically apparent heart failure. The reduction of blood flow and its oscillatory components was larger in nonreperfused AMI patients, although they had a comparable left ventricular function.

We hypothesized that, associated with the state of anaesthesia, characteristic changes exist in both cardio-respiratory and cerebral oscillator parameters and couplings, perhaps varying with depth of anaesthesia. Electrocardiograms (ECGs), respiration and electroencephalograms (EEGs) were recorded from two groups of 10 rats during the entire course of anaesthesia following the administration of a single bolus of ketamine-xylazine (KX group) or pentobarbital (PB group). The phase dynamics approach was then used to extract the instantaneous frequencies of heart beat, respiration and slow delta-waves (within 0.5-3.5 Hz). The amplitudes of delta- and theta-waves were analysed by use of a time-frequency representation of the EEG signal within 0.5-7.5 Hz obtained by wavelet transformation, using the Morlet mother wavelet. For the KX group, where slow delta-waves constituted the dominant spectral component, the Hilbert transform was applied to obtain the instantaneous delta-frequency. The theta-activity was spread over too wide a spectral range for its phase to be meaningfully defined. For both agents, we observed two distinct phases of anaesthesia, with a marked increase in theta-wave activity occurring on passage from a deeper phase of anaesthesia to a shallower one. In other respects, the effects of the two anaesthetics were very different. For KX anaesthesia, the two phases were separated by a marked change in all three instantaneous frequencies: stable, deep, anaesthesia with small frequency variability was followed by a sharp transition to shallow anaesthesia with large frequency variability, lasting until the animal awoke. The transition occurred 16-76 min after injection of the anaesthetic, with simultaneous reduction in the delta-wave amplitude. For PB anaesthesia, the two epochs were separated by the return of a positive response to the pinch test at 53-94 min, following which it took a further period of 45-70 min for the animal to awaken. delta-Waves were not apparent at any stage of PB anaesthesia. We applied non-linear dynamics and information theory to seek evidence of causal relationships between the cardiac, respiratory and slow delta-oscillations. We demonstrate that, for both groups, respiration drives the cardiac oscillator during deep anaesthesia. During shallow KX anaesthesia the direction either reverses, or the cardio-respiratory interaction becomes insignificant; in the deep phase, there is a unidirectional deterministic interaction of respiration with slow delta-oscillations. For PB anaesthesia, the cardio-respiratory interaction weakens during the second phase but, otherwise, there is no observable change in the interactions. We conclude that non-linear dynamics and information theory can be used to identify different stages of anaesthesia and the effects of different anaesthetics.

Background There are only few assays available for the detection of Crimean-Congo Hemorrhagic Fever Virus (CCHFV)-specific antibodies in animals, and data about diagnostic sensitivity and specificity are incompletely documented for most of these tests. This is unfortunate since CCHFV antibodies in animals can be used as indicator for virus circulation in a geographic area and therewith potential risk of human exposure. This paper therefore reports on a novel ELISA for the detection of CCHFV-specific antibodies in cattle and on its application for testing ruminant sera from the Former Yugoslav Republic of Macedonia. Principal Findings A highly sensitive and specific ELISA was developed to detect CCHFV-specific IgG antibodies in cattle. The assay was validated by using 503 negative serum samples from a country where CCHFV has never been detected until now, and by using 54 positive serum samples. The positive sera were verified by using two commercially available assays (for testing human serum) which we have adapted for use in animals. The sensitivity of the novel ELISA was 98% and its specificity 99%. The presence of Hyalomma ticks was demonstrated in the Former Yugoslav Republic of Macedonia and depending on the region antibody prevalence rates up to 80% were detected in the cattle population. Conclusion This article describes a fully validated, highly sensitive and specific ELISA for the detection of CCHFV-specific IgG antibodies in cattle. Using this assay, CCHFV-specific antibodies were detected for the first time in cattle in the Former Yugoslav Republic of Macedonia, giving evidence for an active circulation of this virus in the country. Supporting this conclusion, the occurrence of the main vector of CCHFV was demonstrated in the present work for the first time in Former Yugoslav Republic of Macedonia., Author Summary Crimean-Congo hemorrhagic fever virus (CCHFV) infections in humans can cause a deadly hemorrhagic fever with high fatality rates. Most humans are infected by ticks or by contact with blood. No vaccine prophylaxis or therapeutic interventions are available. Ticks of the genus Hyalomma function as vector of CCHFV. Domestic ruminants play a crucial role in the transmission and amplification of the virus. Since infected ruminants do not develop clinical signs, CCHFV infections can in most cases only be detected by serology. There are only few assays available for the detection of CCHFV-specific antibodies in animals. In general, the real distribution of CCHFV is fairly unknown. Here we present a novel, highly sensitive and specific ELISA, and the adaptations for two commercially available assays for the detection of CCHFV-specific antibodies in cattle. With these assays, we detected CCHFV-specific antibodies in cattle in the Former Yugoslav Republic of Macedonia. This is the first indication of an active circulation of CCHFV in this country. This knowledge will assist decision makers and public health authorities in deciding on effective countermeasures and can help to prevent outbreaks.

Spectral analysis of the laser Doppler flow (LDF) signal in the frequency interval from 0.0095-2.0 Hz reveals blood flow oscillations with frequencies around 1.0, 0.3, 0.1, 0.04 and 0.01 Hz. The heartbeat, the respiration, the intrinsic myogenic activity of vascular smooth muscle, the neurogenic activity of the vessel wall and the vascular endothelium influence these oscillations, respectively. The first aim of this study was to investigate if a slow oscillatory component could be detected in the frequency area below 0.0095 Hz of the human cutaneous blood perfusion signal. Unstimulated basal blood skin perfusion and enhanced perfusion during iontophoresis with the endothelium-dependent vasodilator acetylcholine (ACh) and the endothelium-independent vasodilator sodium nitroprusside (SNP) were measured in healthy male volunteers and the wavelet transform was computed. A low-frequency oscillation between 0.005 and 0.0095 Hz was found both during basal conditions and during iontophoresis with ACh and SNP. Iontophoresis with ACh increased the normalized amplitude to a greater extent than SNP (P = 0.001) indicating modulation by the vascular endothelium. To gain further insight into the mechanisms for this endothelium dependency, we inhibited nitric oxide (NO) synthesis with N(G)-monomethyl-L-arginine (L-NMMA) and prostaglandin (PG) synthesis by aspirin. L-NMMA did not affect the increased response to ACh vs. SNP iontophoresis in the 0.005-0.0095-Hz interval (P = 0.006) but abolished the difference in the 0.0095-0.021-Hz interval (P = 0.97). Aspirin did not affect the difference in response to ACh and SNP in either of the two frequency intervals. Thus, other endothelial mechanisms, such as endothelium-derived hyperpolarizing factor (EDHF), might be involved in the regulation of this sixth frequency interval (0.005-0.0095 Hz).

Potential differences have been measured during transdermal iontophoresis in order to establish the effect of voltage, as opposed to current, on cutaneous blood flow. It is known that, even in the absence of drugs, the iontophoresis current can sometimes produce increased blood flow. The role of voltage in this process is studied through single-ended measurements (between electrode and body) of the potential difference during iontophoresis with 100-/spl mu/A, 20-s current pulses through deionized water, saturated 20.4% NaCl solution, 1 % acetylcholine, and 1 % sodium nitroprusside. It is found that the voltage needed to deliver the current varied by orders of magnitudes less than the differences in the conductance of these different electrolytes, and it is concluded that, at least for the present current protocol, the voltage as such is not an important factor in increasing the blood flow.

Iontophoresis currents are used in the transcutaneous delivery of vasoactive substances for noninvasive assessment of skin vascular properties. The blood flow rate can be recorded by laser Doppler flowmetry (LDF), its average value and the amplitudes of its oscillatory components being used to evaluate the effect of the drugs. Because non-drug-specific, current-induced, vasodilation could confound the interpretation of the response, we have investigated the effect of currents of both polarities on the spectral components of the LDF signal in the absence of vasoactive substances. It was recorded for healthy volunteers with both high conductance (5 mol/l NaCl) and low conductance (deionized water) electrolytes. The oscillatory components were analysed by wavelet transform within 0.0095–1.6 Hz, divided into five sub-intervals. Only cathodal iontophoresis with deionized water increased the oscillatory energy and amplitude. It did so at all frequencies, but none of the sub-intervals associated with vasodilation (0.0095–0.145 Hz) was selectively affected compared to the others.

Background: Microvascular blood flow in the human skin is subject to rhythmic variations reflecting the influence of heartbeat, respiration, intrinsic myogenic activity, neurogenic factors and endothelial activity. The aim of our study was to test the hypothesis that basal skin blood flow (BSBF) and its dynamic components differ (1) among diabetic patients without autonomic neuropathy and with it and healthy control subjects, and (2) among the upper and lower extremities. Patients and Methods: BSBF at four recording sites with predominantly nutritive capillary circulation (right and left caput ulnae, right and left medial malleolus) was measured by laser Doppler flowmetry in 25 diabetic patients without cardiovascular autonomic neuropathy (D), 18 neuropathic diabetic patients (DAN) and 36 healthy controls (C). Wavelet transform was applied to the laser Doppler signal. Results: In absolute terms, mean flow, mean amplitude of the total spectrum and mean amplitudes at all frequency intervals were highest in C, followed by DAN and lowest in D. However, these differences were statistically significant only in the left arm. Within all three groups, mean flow and spectral amplitudes were significantly higher in the arms than in the legs, besides there was a significant difference between the two arms in D. Conclusion: We have confirmed the differences in BSBF among D, DAN and C, and demonstrated differences among the four recording sites which have not been previously described. The latter indicates an uneven progression of autonomic neuropathy and allows for speculation that the left arm is the latest to be affected.

Nitric oxide (NO) and prostaglandines (PGs) are important in regulation of vascular tone and blood flow. Their contribution in human cutaneous circulation is still uncertain. We inhibited NO synthesis by infusing N(G)-monomethyl-L-arginine (L-NMMA) in the brachial artery (16 micromol/min for 5 min) and reversed it by intraarterial infusion of L-arginine (40 micromol/min for 7.5 min). PG synthesis was inhibited by the cyclooxygenase inhibitor aspirin (600 mg over 5 min intravenously). Basal cutaneous perfusion and perfusion responses during iontophoresis with the endothelium-dependent vasodilator acetylcholine (ACh) and the endothelium-independent vasodilator sodium nitroprusside (SNP) were recorded by laser Doppler flowmetry (LDF). We performed wavelet transforms of the measured signals. Mean spectral amplitude within the frequency interval from 0.0095 to 1.6 Hz and mean and normalized amplitudes of five intervals around 1, 0.3, 0.1, 0.04, and 0.01 Hz were analysed. The oscillations with frequencies around 1, 0.3, 0.1, and 0.04 Hz are influenced by the heartbeat, the respiration, the intrinsic myogenic activity of vascular smooth muscle, and the neurogenic activity of the vessel wall, respectively. We have previously shown that the oscillation with a frequency around 0.01 Hz is modulated by the vascular endothelium. L-NMMA reduced mean value of the LDF signal by approximately 20% (P = 0.0067). This reduction was reversed by L-arginine. Mean value of the LDF signals during ACh and SNP iontophoresis did not change after infusion of L-NMMA. Aspirin did not affect mean value of the LDF signal or the LDF signal during ACh or SNP iontophoresis. Before interventions the only significant difference between the effects of ACh and SNP was observed in the frequency around 0.01 Hz, where ACh increased normalized amplitude to a greater extent than SNP. L-NMMA abolished this difference, whereas it reappeared after infusion of L-arginine (P = 0.0084). Aspirin did not affect this difference (P = 0.006). We conclude that basal cutaneous blood flow and the endothelial dependency of the oscillation around 0.01 Hz are partly mediated by NO, but not by endogenous PGs. Other aspects of human cutaneous circulation studied are not regulated by NO or PGs.

Haemophilia A is an X-linked bleeding disorder caused by mutations in the factor VIII gene. In our efforts to elucidate molecular defects in the haemophilia A patients from the Republic of Macedonia, we employed nonradioactive single-strand conformation polymorphism analysis followed by direct sequencing, for identifying point mutations in the factor VIII gene. In the present study we report the detection of three novel missense mutations: Met19 --Arg; Ala78 --Pro and Cys2174 --Gly, all causing haemophilia A.