Sperm-preparation techniques for men with normal and abnormal semen analysis. A comparison

OBJECTIVE: To compare two commonly used sperm-preparation techniques, density gradient centrifugation and swim-up procedures, with respect to their effects on acrosome reaction (AR), hypoosmotic swelling (HOS) and nuclear maturity in men with abnormal and normal semen analyses. STUDY DESIGN: In accordance with World Health Organization criteria, 23 men with abnormal (group I) and 20 men with normal (group II) semen analyses were included in a prospective, controlled study. Each semen specimen was divided into aliquots in order to assess AR, HOS and nuclear maturity, determined with acridine orange staining, in both raw and processed semen samples using the density gradient centrifugation and swim-up techniques. RESULTS: Initial semen samples in group I revealed diminished AR, HOS and nuclear maturity rates in comparison to those in group II. In group I, density gradient centrifugation improved AR, HOS and nuclear maturity rates more than did swim-up. However, in group II it improved only the AR; HOS rates were better than with swim-up. There was a significant positive correlation between sperm concentration and HOS rate in raw semen samples from group I. In the same group, motility and morphology correlated with the nuclear maturity rate but not with AR and HOS rates. Semen samples with better motility (> 20%) or morphology (> 25%) showed better nuclear maturity rates (> 50%) in men with abnormal semen analyses. Motility had a sensitivity of 77% and specificity of 90% in predicting nuclear maturity. Morphology had similar sensitivity but lower specificity (70%). CONCLUSION: Density gradient centrifugation is superior to the swim-up technique in improving AR, HOS and nuclear maturity rates in men with abnormal semen analyses. However, when only nuclear maturity rate is taken into account, the swim-up technique seems to be sufficient for selecting spermatozoa in men with normal semen analyses. The nuclear maturity rate also correlates with sperm morphology and motility.     

The contribution of nitric oxide and endothelins to angiotensin: II. Evoked responses in the rat isolated uterus smooth muscle.

The present study was designed to determine the role of endogenous endothelin peptides and nitric oxide on angiotensin II (All) responses in the isolated nonpregnant rat uterine smooth muscle. AII (10, 20, or 50 ng/ml) increases rhythmic oscillations dose dependently (32.7 +/- 8.9, 55.96 +/- 10.3, and 62.78 +/- 17.7% increase, respectively). L-arginine methyl ester (L-NAME; 10(-5) M) did not affect the increase in rhythmic oscillations induced by All (10, 20, or 50 ng/ml) (17.5 +/- 12.1, 31.5 +/- 18.3, and 52.5 +/-11.8% increase, respectively, n = 6, p > 0.05). It reduced the contractile responses to AII (10 ng/ml: from 4.63 +/- 0.6 to 1.8 +/-0.7 cm2, p < 0.05: and 20 ng/ml: from 5.59 +/- 0.8 to 2.11 +/- 0.4 cm2, p < 0.05, n = 6). L-arginine (10 mM) decreased the contractile response obtained by AII (10 or 20 ng/ml) (1.93 +/- 1.05, p < 0.05 and 2.14 +/- 0.7 cm2, p < 0.05, respectively, n = 6). BQ 485 (50 ng/ml) decreased both the number of rhythmic oscillations and the contractility increased by AII. Bosentan (10(-5) M) induced an increase in the number of rhythmic oscillations but decreased the contractile responses to the higher concentrations of All. These data show that endogenous NO and endothelin peptides contribute to the motility changes induced by AII and may play an important role in the pathophysiological events of the uterine function.     

In vitro and in vivo evaluation of diclofenac sodium loaded albumin microspheres

The use of polymeric carriers in formulations of therapeutic drug delivery systems has gained widespread application, due to their advantage of being biodegradable and biocompatible. Among the microparticulate systems, microspheres have a special importance since it is possible to target drugs and provide controlled release. Diclofenac sodium (DS), is a potent drug in the NSAID group having non-steroidal, anti-inflammatory properties, and is widely used in the treatment of rheumatoid arthritis, osteoarthritis and ankylosing spondylitis. In this present study, it was aimed to prepare microsphere formulations of DS using a natural biodegradable polymer as a carrier for intraarticular administration to extend the duration period of the dosage form in the knee joint. Microsphere formulations of DS which were prepared were evaluated in vitro for particle size, yield value, encapsulation efficiency, surface morphology, and in vitro drug release. Two appropriate formulations were selected for in vivo trials. For the in vivo studies, Technetium-99m labelled polyclonal human immunogammaglobulin (99mTc-HIG) was used as the radiopharmaceutical to demonstrate arthritic lesions by gamma scintigraphy. After the induction of arthritis in knee joints of rabbits, the radio-labelled microspheres loaded with DS were injected directly into the articular cavity and at specific time points gamma scintigrams were obtained to find the residence time of the microspheres in knee joints in order to determine the most suitable formulation.     

Scintigraphic imaging of infections with 99m-Tc-labelled glutathione liposomes

In this study, liposomes containing glutathione were evaluated to detect infection in mice. Glutathione liposomes were labelled by using 99mTc-labelled-HMPAO (hexamethyl propylamine oxime). Mice were infected in the thigh by intramuscular injection with turbentine. Labelled liposomes were applied to the tail vein of the mice intravenously. At fixed time intervals they were sacrificed. The animals were imaged under a gamma camera. Then, tissue samples were excised and radioactivity of all organs was counted. Abscess-to-muscle, abscess-to-liver, and abscess-to-spleen ratios were calculated. The ratios of abscess-to-muscle were found to be 1.6 and 11.6 at 1 h and 24 h, respectively. According to these data, the abscess can be defined at 1 h and it became more clear with time.     

Assessment of mothers’ knowledge and perceptions of electroencephalography and determination of the short-term effect of an informational leaflet

ObjectiveThe goal of the study described here was to determine mothers’ knowledge and perceptions of electroencephalogram (EEG), to assess mothers’ understanding of the main aspects of electroencephalography (EEG), and to determine the effect of an informational leaflet on increasing knowledge and perception.MethodsA 20-item questionnaire was developed to assess mothers’ knowledge and perceptions of EEG. The questionnaire comprised 20 simple statements on aspects of the procedure, to which the mothers answered “yes” or “no.” Mothers were interviewed in person by an EEG technician at the beginning of the study. On completion of the questionnaire, the same technician provided the mothers with an informational leaflet. One month later, the mothers were telephoned and administered the same questionnaire over the phone.ResultsThe response rate was 86%. Before reading the informational leaflet, 89.5% of the mothers stated that they knew why their child was undergoing electroencephalography, and 67.6% knew what electroencephalography was. Furthermore, 78.1% of them believed that their child’s brain was mapped by electroencephalography. In addition, nearly 1 in 10 believed that EEG is a hazardous procedure and 6% believed it was addictive. Knowledge and perceptions changed after distribution of the informational leaflet. Comparison of mothers with different income levels, educational status, and numbers of electroencephalograms their child underwent revealed statistically significant differences with respect to knowledge and perceptions of electroencephalography.ConclusionWritten information is a simple, inexpensive, easy-to-implement, yet effective method of improving parental understanding of EEG. The present study has significant implications for informing individuals regarding medical procedures.     

Detection of bladder tumors with 3-dimensional sonography and virtual sonographic cystoscopy.

OBJECTIVE: Bladder tumors are among the most common types of malignant neoplasms of the urinary tract. The purpose of this study was to evaluate the potential value of 3-dimensional (3D) sonography and sonographic cystoscopy in detection of bladder tumors. METHODS: Thirty-one patients with suspected or known bladder tumors were included this study. All patients underwent 3D sonography and conventional cystoscopy within 15 days. The number, size, location, and morphologic features of the lesions were evaluated on gray scale, 3D virtual, and multiplanar reconstruction images obtained from the patients. The results of 3D sonographic cystoscopy were compared with the findings from conventional cystoscopy, which was considered the reference standard. RESULTS: Twenty-eight (90.3%) of 31 3D virtual sonographic cystoscopic studies had good or excellent image quality. Conventional cystoscopy revealed 47 lesions in 22 of 28 patients; 3D sonographic virtual cystoscopy showed 41 (87.2%) of 47 lesions. Three-dimensional virtual sonography alone had sensitivity of 96.2%, specificity of 70.6%, a positive predictive value of 93.9%, and a negative predictive value of 80% for tumor detection. The combination of gray scale sonography, multiplanar reconstruction, and 3D virtual sonography had sensitivity of 96.4%, specificity of 88.8%, a positive predictive value of 97.6%, and a negative predictive value of 84.2% for tumor detection. CONCLUSIONS: Three-dimensional sonography is a promising alternative noninvasive technique for use in detection of bladder tumors, their localization, and perivesical spreading. The location, size, and morphologic features of the tumors shown on 3D sonography agreed well with the findings of conventional cystoscopy.     

Glutathione S-transferase gene polymorphisms in presbycusis.

HYPOTHESIS: Glutathione and glutathione-related antioxidant enzymes are involved in the metabolism and detoxification of cytotoxic and carcinogenic compounds as well as reactive oxygen species. Reactive oxygen species generation occurs in prolonged relative hypoperfusion conditions such as in aging. The etiology of presbycusis is much less certain; however, a complex genetic cause is most likely. The effect of aging shows a wide interindividual range; we aimed to investigate whether profiles of (glutathione S-transferase (GST) M1, T1 and P1 genotypes may be associated with the risk of age-related hearing loss. PATIENTS AND METHODS: We examined 68 adults with presbycusis and 69 healthy controls. DNA was extracted from whole blood, and the GSTM1, GSTT1 and GSTP1 polymorphisms were determined using a real-time polymerase chain reaction and fluorescence resonance energy transfer with a Light-Cycler Instrument. Associations between specific genotypes and the development of presbycusis were examined by use of logistic regression analyses to calculate odds ratios and 95% confidence intervals. RESULTS: Gene polymorphisms at GSTM1, GSTT1, and GSTP1 in subjects with presbycusis were not significantly different than in the controls (p > 0.05). Also, the combinations of different GSTM1, GSTT1, and GSTP1 genotypes were not an increased risk of presbycusis (p > 0.05). CONCLUSION: We could not demonstrate any significant association between the GSTM1, GSTT1, and GSTP1 polymorphism and age-related hearing loss in this population. This may be because of our sample size, and further studies need to investigate the exact role of GST gene polymorphisms in the etiopathogenesis of the presbycusis.     

Autoimmune thyroid diseases in patients with chronic fasciolosis

BACKGROUND: The prevalence of autoimmune thyroid diseases and autoantibodies in patients with chronic fasciolosis (CF) was investigated. METHODS: Thyroid function of 32 patients with CF and 72 patients without fasciolosis infection was evaluated biochemically. Thyroid hormones, antithyroglobulin antibodies (TgAb) and antithyroid peroxidase antibodies (TPOAb) were measured. RESULTS: In patients with CF, the prevalences of autoantibodies against thyroid peroxidase and thyroglobulin were 21.9% (7/32) and 15.6% (5/32) respectively. However, thyroid function disorder was identified in only 3 (9.4%) cases. In the control group, TPOAb and TgAb were determined in 2 (2.7%) cases, but no one had thyroid function disorder. The prevalences of thyroid autoantibodies (chi 2: 7.948, p < 0.01) and autoimmune thyroid diseases (chi 2: 4.007, p < 0.05) were significantly higher in the CF group. CONCLUSIONS: The high prevalence of TPOAb and TgAb in CF suggests that patients with CF should be evaluated for autoimmune thyroid disorders at the time of diagnosis and during the follow-up period.     

PON1 L55M polymorphism is not a predictor of coronary atherosclerosis either alone or in combination with Q192R polymorphism in an Italian population

BACKGROUND: The present study evaluated the role of the PON1 L55M polymorphism independently and in conjunction with the Q192R polymorphism on the risk of coronary atherosclerosis in an Italian population. MATERIALS AND METHODS: Three hundred and ninety-one subjects with significant coronary stenosis (> 50%) (coronary artery disease-positive; CAD+), 196 subjects with normal coronary arteries (< 10% stenosis) (CAD-) and 178 healthy controls were screened using a combination of polymerase chain reaction and restriction enzyme digestion. RESULTS: In the pooled population, the frequencies of L and M alleles were 0.63 and 0.37, respectively; the most common haplotypes were QQ/LM (24.2%) and QR/LL (21.8%) and a strong linkage disequilibrium between L/55 and R/192 alleles was observed (D' = -0.91; P < 0.0001). CAD+ subjects did not show any significant differences in the distribution of PON1-55 genotypes as compared to CAD- subjects and population controls (chi2 = 1.5, P = 0.8). After controlling for other risk factors, the low-concentration M allele was not associated with a significant change of CAD risk (OR 1.02; 95% CI 0.80-1.29; P = 0.87). Moreover, the L55M polymorphism did not show any interaction with other risk factors such as smoking, diabetes, hypertension, low levels of high-density lipoprotein (HDL) or high ratios of low-density to high-density lipoproteins. The combination of L55M with the Q192R polymorphism did not show any effect on CAD risk. However, a marginal decrease in myocardial infarction risk was detected when QQ/MM carriers (OR 0.51; 95% CI 0.26-0.99; P = 0.048), but not LL/RR carriers, were compared with subjects not homozygous for an L or R allele. CONCLUSIONS: These findings did not indicate a major effect of the PON1 L55M polymorphism, either alone or in combination with the Q192R polymorphism, on CAD risk. Additional studies are needed for a better evaluation of the role of the 55/192 PON1 genotypes in combination on myocardial infarction risk.     

Evaluation of 99mTc-citrate, 67Gacitrate and 99mTc(V) dimercaptosuccinic acid for the scintigraphic visualization of acute appendicitis

An experimental study was planned to evaluate ^99mTc-citrate, ⁶⁷Gacitrate and ^99mTc(V) dimercaptosuccinic acid (DMSA) as agents for the visualization of acute appendicitis. Appendiceal ligation was performed through a midline incision in 24 rabbits. Twenty-four hours later the animals were divided into three equal groups. The rabbits were injected through the aurical vein with 1 mCi (37 MBq) ^99mTc-citrate in group I, 0.5 mCi (18.5 MBq) ⁶⁷Ga-citrate in group II and 1 mCi (37 MBq) ^99mTc(V) DMSA in group III. After 3 h, static images of the rabbits were obtained with a gamma camera. There were positive images in seven, six and five rabbits in groups I, II and III respectively. The image quality was better in group I than in the other groups. Also, the mean uptake in group I was significantly higher than those of other two groups (P < 0.05). There was no significant difference between groups II and III (P > 0.05). All rabbits had appendicitis confirmed histologically. In conclusion, these results show that 99mTc-citrate is preferable to 67Ga-citrate and ^99mTc(V) DMSA for the differential diagnosis of acute abdominal inflammations such as appendicitis, because of higher concentration ratios, simple and rapid preparation, low cost, excretion mainly through the kidneys and fast blood clearance.