Saksenaea vasiformis infections: A case of an immunocompetent adult after mild injury and a literature review.

Saksenaea vasiformis is an emerging human pathogen, belonging to the order Mucorales of the subphylum Mucormycotina, most often associated with rhino-cerebral, cutaneous and subcutaneous infections following trauma. A review of the published literature was attempted on the occasion of a cutaneous leg infection with favorable outcome in a young immunocompetent man after mild injury. The overall aim was the facilitation of the study and the integrated understanding of this kind of fungal infections.

Alternative approaches to the therapeutic drug monitoring of prophylactic posaconazole in haematology patients.

OBJECTIVE: The intention of the study was to assess whether a unique daily specimen is adequate for prophylactic posaconazole TDM in haematology patients and if bioassay and HPLC produce similar results and could be equally used in clinical setting. METHOD: Serum specimens from thirty haematology patients were collected at the end of the first and second week of treatment, just before the morning dose, 2 and 6 to 8hours afterwards. Levels were measured by bioassay in 157 specimens and additionally by HPLC in 51 of them. RESULTS: Bioassay levels were correlated inter and intra daily, with no statistical difference between them, irrespective of the timing. The same was true for HPLC measurements. There was no statistical difference between bioassay (median: 1.60mg/L, interquartile range: 0.60-2.30) and HPLC levels (median: 1.16mg/L, interquartile range: 0.56-1.72), while they were significantly correlated. CONCLUSION: In clinically stable haematology patients, a random specimen on any day after steady state serum concentrations have been achieved is probably adequate in order to monitor posaconazole levels. In the case of monotherapy, a bioassay is an acceptable alternative to HPLC.

Non-physician health-care workers and voluntary blood donation: an ambiguous relationship.

OBJECTIVES: This study aims to assess the knowledge, attitude and practice (KAP) of voluntary blood donation (BD) among the non-physician health-care workers of our institution, to investigate the relationship between these parameters and to determine the predictors of BD practice. BACKGROUND: KAP surveys provide a popular context-specific evidence base for the development of blood transfusion services' strategies and interventions to promote voluntary, non-remunerated BD. However, there are just few studies worldwide assessing KAP of BD among health-care workers. METHODS: This is a cross-sectional exploratory study, which took place at the 'Ippokrateio' General Hospital of Thessaloniki, Greece, a tertiary health-care institution. A specially designed, pre-tested questionnaire was distributed to all non-physician health-care workers of the hospital. A total of 1140 questionnaires were distributed. Participation in the study was optional and anonymous. Regarding data analysis, a binary logistic regression analysis was performed to describe the relationship between different elements of KAP of BD and to determine possible predictors of BD practice. RESULTS: The response rate was 25·5% (291 respondents). Logistic regression analysis revealed a positive attitude towards family replacement as an important predictor of both regular and voluntary BD, with male gender also a predictor of the latter. An inconsistency between knowledge, attitude and practice of BD was revealed. CONCLUSIONS: The present study adds valuable data on KAP of BD among health-care workers. This will help blood transfusion services to improve management practices among this group as they have the potential to provide a reliable, stable and safe source of blood products.

A comparative study of three extraction protocols of DNA from nails: Potential use in the diagnosis of onychomycoses.

Molecular techniques can be useful adjuncts to the diagnosis of onychomycoses. However, the nail presents difficulties in the extraction of its DNA. The comparison of three extraction protocols of DNA from nails and their ranking for possible use in the molecular diagnosis of onychomycoses are described. Extraction was performed on weighed nail clippings of equal size from positive (31) or negative (14) samples, according to the culture result. At Prot1, the extraction was performed according to Tahir and Watson, with an additional step implementing silica columns. At Prot2, the methodology proposed by the Statens Serum Institute of Copenhagen was used. At Prot3, DNA was extracted by the use of magnetic separation after homogenisation with glass beading. The evaluation parameters were DNA purity, DNA concentration, total DNA yield/g of tissue, cost and duration. The multiples of the means of medians of the first three parameters, for each protocol, were calculated. Prot3 showed the highest DNA purity. Prot2 presented the highest DNA concentration and DNA yield/g of tissue, while it was the cheapest and shortest. In total, the three protocols were graded as Prot2>Prot1>Prot3. The second method, although had a lower DNA purity, presented the higher DNA concentration and DNA yield, while its duration and cost were also favourable.

Epidemiology of superficial mycoses in Northern Greece: a 4-year study.

BACKGROUND: Superficial mycoses are defined as the fungal infections of skin, hair or nails that are caused by dermatophytes, yeasts and non-dermatophytic moulds. Dermatophytes are the most frequently isolated fungi from specimens of patients with superficial mycoses. OBJECTIVE: Studying the possible alteration of the epidemiology of superficial mycoses in Northern Greece during the last two to three decades. MATERIALS AND METHODS: Data concerning the superficial mycoses from patients coming mainly from the region of Macedonia, Northern Greece, between January 2010 and January 2014 were recorded and analysed. They included specimens from 438 patients (146 M/292 F), within an age range of 2-85 years old. 503 samples were collected from skin (81, 16.1%), hair (18, 3.6%) and nails (fingernails 84, 16.7%, toenails 320, 63.6%) lesions. RESULTS: Of a total of 222 positive cultures, 50 were considered as yielding clinically non-significant isolates (saprophytes). Among the rest (172), dermatophytes were the most prevalent isolates (102, 59.3%), followed by yeasts (51, 29.7%) and non-dermatophytic moulds (19, 11%). Trichophyton rubrum (55, 53.9%), Trichophyton mentagrophytes (18, 17.6%) and Microsporum canis (23, 22.5%) were the most common isolates among dermatophytes (total = 102). Candida parapsilosis (26, 51%), and Candida albicans (10, 19.6%) among yeasts (total = 51) whereas Fusarium (6, 31.6%) and Acremonium species (3, 15.8%) among the non-dermatophytic moulds (total=19). CONCLUSION: Compared to previous studies from Northern Greece, the epidemiology of superficial mycoses in the specific geographic region seems not to have been altered the last two to three decades.

Laboratory investigation of platelet function in patients with thalassaemia.

The aim of this study was to investigate platelet function in patients with thalassaemia and to detect any relation to chelation treatment (deferasirox or deferiprone/deferiprone plus desferioxamine). Thirty-three transfusion-dependent patients with thalassaemia were included. The investigation consisted of aggregation testing of platelet-rich plasma by light transmission aggregometry (LTA) with the use of 5 agonists as well as the global test of haemostasis by means of the PFA-100 platelet function analyser. In 66.67% of the patients, there was reduced LTA to at least one agonist and in 18.18% there was reduced LTA to two or more agonists. The PFA-100 test was prolonged in 60.6% of the cases. An abnormal LTA and a prolonged PFA-100 time were recorded in 33.3% of the patients and 27.4% had a normal aggregation and PFA-100 test. No correlation between chelation regimen and either LTA or PFA-100 test was found. The abnormal LTA can be explained either by the release of ADP from the haemolysed red blood cells, which leads to defective platelet aggregation, or by the presence of two platelet populations. An in vitro effect without an in vivo impact could be an alternative explanation. In patients with thalassaemia, the reduced LTA and the prolonged PFA-100 closure time could be an in vitro effect and has a close correlation to the bleeding phenotype of each patient.

The clinical utility of bone turnover markers in the evaluation of bone disease in patients with haemophilia A and B.

Haemophilia A and B have been associated with increased prevalence of low bone mineral density (BMD). However, the utility of bone turnover markers (BTM) remains unknown. The aim of this study was to evaluate bone metabolism in men with haemophilia and to investigate associations between BTM and bone disease. Serum N- (NTX-I), C-terminal telopeptide of type I collagen (CTX-I) and tartrate-resistant acid phosphatase band-5b (TRAP-5b), as bone resorption markers, and osteocalcin (OC) and bone-specific alkaline phosphatase (b-ALP), as bone formation markers, were assessed. Seventy men with haemophilia A (n = 59) or B (n = 11) were studied. Patients with low BMD had significantly higher b-ALP concentrations compared with those with normal BMD (12.8 ± 1.60 vs. 9.72 ± 0.58 µg/L, P = 0.009), without any differences in the other BTM. NTX-I and CTX-I concentrations were negatively associated with oestradiol levels and hip BMD and positively with human immunodeficiency virus infection, number of affected joints and arthropathy scores. B-ALP and OC concentrations were negatively associated with hip BMD, severity of haemophilia and fracture history, and positively with the number of affected joints and testosterone concentrations. After multivariate analysis, NTX-I levels remained negatively associated with oestradiol levels, whereas b-ALP concentrations negatively correlated with the level of physical activity and positively with the number of affected joints. Increased bone metabolism exists in men with haemophilia and low BMD. Increased b-ALP levels may identify patients at high risk for fracture. Increased number of target joints, low physical activity and low oestradiol concentrations are independently associated with increased bone metabolism.

Detection of anti-BP180NC16a and anti-BP230 autoantibodies in blister fluid of patients with bullous pemphigoid: the first survey in Greece.

Bullous pemphigoid (BP) is an acquired bullous disease with an increasing prevalence among elderly people worldwide, including in Greece. Blister formation in most patients with BP is caused by autoantibodies against structural components of the basement membrane zone of the skin, predominantly BP180NC16a and BP230 antigens on the hemidesmosome adhesion complex. Routine diagnostic methods such as histological examination and direct and indirect immunofluorescence are combined to determine diagnosis. In this study, an ELISA was used to measure levels of both anti-BP180NC16A and anti-BP230 autoantibodies in the blister fluid of 13 patients with newly diagnosed BP, before starting treatment. The aim of the study was to evaluate this method as a diagnostic tool in BP. Our results indicate that blister-fluid examination by ELISA can be a useful tool to diagnose bullous pemphigoid, especially in elderly patients who refuse biopsy or have poor venous access.

Cyclosporine and its metabolites before and 2 h post-dose: comparative measurements of a monoclonal and a polyclonal immunoassay.

The aim of the study was to investigate the better accuracy of the 2-h post-dose (C2) levels of cyclosporine (CyA), compared with the pre-dose (C0) levels and to evaluate the results measured by a monoclonal or a polyclonal immunoassay. The parent compound of CyA in C2 (monoclonal2) was measured in 53 kidney transplant patients by the monoclonal fluorescence polarization method, as well as the parent compound plus metabolites (polyclonal2) by the polyclonal fluorescence polarization method. Also, the parent compound was measured in 21 of the patients for the C0 (monoclonal0), whereas the parent compound plus metabolites in 36, for the C0 (polyclonal0). As level of metabolites was considered the difference between polyclonal and monoclonal values (polyclonal-monoclonal), either in C0 (metabolites0) or in C2 (metabolites2). The ratio polyclonal2/monoclonal2 gave a mean value of 1.7+/-0.2 (mean+/-SD), whereas the mean value of the ratio polyclonal0/monoclonal0 was 2.3+/-0.6, with almost double variation. The mean value of the ratio metabolites2/monoclonal2 was 0.7+/-0.2 and of the ratio metabolites0/monoclonal0 was 1.3+/-0.6. The difference between the two ratios is very significant (p = 0.000001) and they are not correlated with each other (r = 0.18, p = 0.44). The measurements of monoclonal0 and polyclonal0 or monoclonal2 and polyclonal2 are very significantly correlated (r = 0.94, p = 0.000001 and r = 0.97, p = 0.000001, respectively). In C0 the proportion of metabolites is higher than in C2, with a double variation, as the degree of metabolism is diverse. Consecutively, in monoclonal methods, as cross-reactions occur with metabolites, it is more accurate to use the C2 measurement for the evaluation of CyA. The application of both methods, the polyclonal and the monoclonal, could be a useful tool as it gives an estimation of metabolites whose degree of contribution to the immunosuppressive result is difficult to ascertain. Finally, if for reasons of clinical experience, the polyclonal method is used, then the mean therapeutic levels of polyclonal2 are 1.5-1.7 compared with monoclonal2.

Serum concentrations of insulin-like growth factor-I (IGF-I) in patients with liver cirrhosis.

BACKGROUND/AIMS: Insulin-like growth factor-I is an important anabolic polypeptide with various effects. The circulating insulin-like growth factor-I is mainly liver derived. The aim of this study was to determine insulin-like growth factor-I serum levels in patients with cirrhosis and to clarify their association with patients' clinical condition and the etiology of cirrhosis. METHODOLOGY: Forty patients with liver cirrhosis were enrolled. Cirrhosis was in 22 cases induced by virus, in 10 due to primary biliary cause and in the rest 8 of alcoholic origin. The Child score index was found as A (n = 26), B (n = 9), C (n = 5). Twenty, age-matched healthy subjects, were used as a control group. Serum insulin-like growth factor-I was measured by an immunoradiometric assay in all subjects. RESULTS: Serum insulin-like growth factor-I levels in liver cirrhosis were found very significantly lower than in healthy individuals (57.4 +/- 7.0 ng/mL vs. 198.8 +/- 16.3 ng/mL, p = 0.0000001). In liver cirrhosis insulin-like growth factor-I was negatively correlated with spleen enlargement (r = -0.46, p = 0.0031). Child B and C patients showed significantly reduced insulin-like growth factor-I levels in comparison to patients staged as Child A (28.9 +/- 3.0 ng/mL vs. 72.8 +/- 9.3 ng/mL, p = 0.0016). The comparison of 12 patients with viral induced cirrhosis (Child A) to 14 patients with non-viral cirrhosis, of the same clinical stage, showed non-significant difference (84.2 +/- 16 ng/mL vs. 63.1 +/- 10.3 ng/mL, p = 0.27). CONCLUSIONS: Insulin-like growth factor-I synthesis is disturbed in liver cirrhosis and reflects the severity of the clinical stage. It represents a good marker of hepatic function. The etiology of cirrhosis does not seem to influence its levels.