Red blood cell derived extracellular vesicles during the process of autologous blood doping.

The purpose of this pilot study was to investigate the effects of the transfusion of one erythrocyte concentrate on the number of circulating red blood cell extracellular vesicles (RBC-EVs) and their clearance time. Six, healthy volunteers donated their blood and were transfused with their RBC concentrate after 35-36 days of storage. One K2 EDTA and one serum sample were collected before donation, at four timepoints after donation and at another six timepoints after transfusion. RBC-EVs were analyzed on a Cytek Aurora flow cytometer. A highly significant increase (p < 0.001) of RBC-EVs from an average of 60.1 ± 19.8 (103 /µL) at baseline to 179.3 ± 84.7 (103 /µL) in the first 1-3 h after transfusion could be observed. Individual differences in the response to transfusion became apparent with one volunteer showing no increase and another an increased concentration at one timepoint after donation due to an influenza infection. We concluded that in an individualized passport approach, increased RBC-EVs might be considered as additional evidence when interpreting suspicious Athletes Biological Passport (ABPs) but for this additional research related to sample collection and transport processes as well as method development and harmonization would be necessary.

Prevalence of Toxoplasma gondii infection in diabetic patients in Makkah AL Mukarramah, Saudi Arabia.

Toxoplasmosis is a parasitic and zoonotic disease caused by Toxoplasma gondii. The disease has worldwide distribution and all people maybe under the risk of getting infected by the parasite. The overall aim of this research was to detect the prevalence rate of antiToxoplasma gondii IgM and IgG among diabetic patients in Makkah, Saudi Arabia. Blood samples were collected from diabetic patients. Toxo IgM and IgG combo rapid test cassette were used to screen the samples, and the results were confirmed by using Enzyme-Linked Immunosorbent Assay (ELISA) to detect anti-IgM and anti- IgG antibodies on the plasma of 90 diabetic patients who attended the mobile clinic or AL Noor hospital in Makkah area. The subject was asked to complete a structured questionnaire. The questionnaire data and serological results were analyzed by using SPSS 20. Chi-square was used to compare different variables. Out of 90 samples, 39 (43.3%) were positive to anti-Toxoplasma gondii IgG wherein 33 (36.6%) of them were male and 6 (6.7%) were female. The age ranged between 13-85 years with the mean of 49.9 years. The study found that there were statistical differences between the age groups with higher prevalence in the 50-65 years age group. Detection of IgM against T. gondii gave negative results. The results of the study indicate that latent T. gondii in diabetic patient are relatively high especially among the 50-65 age group. There were significant associated between direct contact with a cat and infection by T. gondii (p<0.05).

Testicular sperm aspiration (TESA) for infertile couples with severe or complete asthenozoospermia.

The aim of the study was to evaluate reproductive outcomes in a cohort of infertile couples with severe and complete asthenozoospermia undergoing TESA (testicular sperm aspiration) with ICSI. We conducted a retrospective study of 28 couples with complete or severe asthenozoospermia who underwent TESA between January 2010 and December 2015. We compared TESA-ICSI outcomes of these couples to ejaculate ICSI outcomes of 40 couples with severe asthenozoospermia treated during the same time period at our institution. Couples with female factor infertility and/or female aged ≥39 were excluded. Sperm retrieval rates and ICSI outcomes [(MII oocytes, fertilization rate, good embryo rate (transferred and frozen), couples with embryo transfer (per cycle started), clinical pregnancy (per embryo transfer)] were recorded. Patients were grouped based on whether they had ejaculated (Ej-group) or testicular (TESA-group) spermatozoa used. Testicular sperm patients were further classified based on whether they had complete asthenozoospermia (0% total motility) (Tc-group) or severe asthenozoospermia (≤1% progressive motility) (Ts-group). Mean (±SD) male and female ages were 36 ± 6 and 32 ± 4, respectively. Sperm recovery by testicular sperm aspiration (TESA) was successful in 100% (28/28) of the men. The overall clinical pregnancy rate (CPR) per cycle started was 34% (23/68) with a mean of 1.1 ± 0.4 embryos transferred per transfer. Fertilization rates were significantly lower in TESA-group compared to Ej-group (52% vs. 67%, respectively; p = 0.001), while male age was significantly higher in TESA-group compared to Ej-group (34 ± 6 vs. 37 ± 6, respectively; p = 0.03). Moreover, female age was significantly higher in Tc-group compared to Ts-group (30 ± 4 vs. 33 ± 3, respectively; p = 0.0285). However, there were no significant difference in clinical pregnancy rate per embryo transfer in the Tc-group, Ts-group, and Ej-group (50% vs. 45% vs. 57%, respectively; p = 0.8219). The data suggest that testicular sperm-ICSI is no better than ejaculated sperm-ICSI in couples with severe or complete asthenozoospermia. Randomized, controlled trials comparing ejaculated vs. testicular spermatozoa are needed to assess the true benefit of TESA-ICSI in these couples.

Biochemical modulation of cell energy by 2-deoxyglucose and malonate in 7,12-dimethylbenz(a)anthracene-induced carcinogenesis in rats.

The goal of this study was to explore the impact of 2-deoxglucose or malonate individually or in combination on the level of cell energy (adenosine-5'-triphosphate) and oxidative stress in 7,12-dimethylbenz(a)anthracene (DMBA)-induced mammary proliferation in rats. A total of 60 adult female Sprague Dawley rats were randomly divided into five groups (12 rats each): group I serves as the control group. Rats in groups (II-V) were administrated intragastrically a single dose of 50 mg/kg body weight (bw) of DMBA. A day after DMBA administration, rats in group III were injected intraperitoneally (ip) with 100 mg 2-deoxyglucose (2-DG)/kg bw daily. Rats in group IV were injected ip with 10 mg sodium malonate/kg bw daily. Rats in group V were injected ip with 100 mg 2-DG/kg bw and 10 mg sodium malonate/kg bw (treatment for 90 days). The results obtained showed that DMBA induced oxidative stress by decreasing the activities of glutathione reductase (GRase) and superoxide dismutase (SOD), glutathione peroxidase (GPx) and elevating the levels of malondialdehyde (MDA) and nitric oxide (NO) in mammary tissues when compared with control. The combined treatment protected against the previous deleterious changes by a significant elevation in the activities of GRase and SOD, GPx and lowering the levels of MDA and NO more potentially when compared with individual treatment. Apoptosis, as indicated by a significant release of cytochrome c from mitochondria into the cytosol, observed in DMBA-injected rats was positively significantly correlated with the elevation of the level of NO. These data explained the possible additive effect of 2-DG and malonate by depleting the cell energy by their protective effects against the earlier stages of carcinogenesis.

Effects of a water-soluble curcumin protein conjugate vs. pure curcumin in a diabetic model of erectile dysfunction.

INTRODUCTION: Curcumin is involved in erectile signaling via elevation of cyclic guanosine monophosphate (cGMP). AIM: Assessment of the effects of water-soluble curcumin in erectile dysfunction (ED). METHODS: One hundred twenty male white albino rats were divided into: 1st and 2nd control groups with or without administration of Zinc protoporphyrin (ZnPP), 3rd and 4th diabetic groups with or without ZnPP, 5th diabetic group on single oral dose of pure curcumin, 6th diabetic group on pure curcumin administered daily for 12 weeks, 7th and 8th diabetic groups on single dose of water-soluble curcumin administered with or without ZnPP, 9th and 10th diabetic groups on water-soluble curcumin administered daily for 12 weeks with or without ZnPP. All curcumin dosage schedules were administered after induction of diabetes. MAIN OUTCOME MEASURES: Quantitative gene expression of endothelial nitric oxide synthase (eNOS), neuronal NOS (nNOS), inducible NOS (iNOS), heme oxygenase-1 (HO-1), nuclear transcription factor-erythroid2 (Nrf2), NF-Кß, and p38. Cavernous tissue levels of HO and NOS enzyme activities, cGMP and intracavernosal pressure (ICP). RESULTS: Twelve weeks after induction of diabetes, ED was confirmed by the significant decrease in ICP. There was a significant decrease in cGMP, NOS, HO enzymes, a significant decrease in eNOS, nNOS, HO-1 genes and a significant elevation of NF-Кß, p38, iNOS genes. Administration of pure curcumin or its water-soluble conjugate led to a significant elevation in ICP, cGMP levels, a significant increase in HO-1 and NOS enzymes, a significant increase in eNOS, nNOS, HO-1, and Nrf2 genes, and a significant decrease in NF-Кß, p38, and iNOS genes. Water-soluble curcumin showed significant superiority and more prolonged duration of action. Repeated doses regimens were superior to single dose regimen. Administration of ZnPP significantly reduced HO enzyme, cGMP, ICP/ mean arterial pressure (MAP), HO-1 genes in diabetic groups. CONCLUSION: Water-soluble curcumin could enhance erectile function with more effectiveness and with more prolonged duration of action.

Pea choline kinase: purification, properties and isolation of a cDNA.

Choline kinase has been partially purified from pea seedlings and its properties studied. Using sequence information from soya bean and other choline kinases, we have also isolated a cDNA encoding the enzyme. It encodes a protein of 343 amino acids (calculated molecular mass of 39785 Da), which shows 82% homology with the soya bean choline kinase. The protein has been expressed in Escherichia coli with very good activity and high expression levels.

Nasolacrimal tuberculosis in a patient with conjunctivodacryocystorhinostomy.

PURPOSE: To present an unusual case of nasolacrimal tuberculosis that was diagnosed with polymerase chain reaction (PCR) and to discuss the role of PCR as a diagnostic aid. METHODS: Case presentation with diagnostic modalities including PCR, cytopathologic, and histopathologic examinations with special stains. RESULTS: A 40-year-old woman with bilateral lacrimal drainage system obstruction underwent a right dacryocystorhinostomy with stents and left conjunctivodacryocystorhinostomy with Jones bypass tube. Although the findings of direct and endoscopic examinations of the nasal cavity were within normal limits, the tissue removed during surgery from the middle turbinate revealed caseating granulomatous lesions on histopathologic examination. Acid-fast organisms were demonstrated in postoperative nasal washings, and Mycobacterium tuberculosis infection was confirmed with PCR. CONCLUSION: Primary tuberculosis of the nasolacrimal mucosa is rare, and its occurrence without any symptoms, as in this patient, is even less frequent. The most interesting aspect of this case was the identification of the M. tuberculosis DNA in the mucosa of anterior turbinate and fibrous tissue within the previous surgical site. Histopathologically, the presence of caseating granulomas was seen only in the nasal mucosa, indicating that PCR may be of use in identifying the causative organism even in the absence of typical histopathologic features.