A New Ilarvirus Isolated from Grapevine in Greece.

In 1994, characteristic viruslike symptoms on grapevine were reported in the collection of the Grapevine Institute in Athens, Greece, on the hybrid Baresana × Baresana. The symptoms were sharp angular mosaic, leaf crinkle, and little leaf. The affected vines showed gradual decline and severe stunting or death. Such vines produced abortive flowers or very few berries with smaller, wrinkled, and nongerminating seeds. Serological testing, by enzyme-linked immunosorbent assay (ELISA), of the affected vines against the most common grapevine viruses Alfalfa mosaic, Arabis mosaic, Grapevine fanleaf, Grapevine fleck, Grapevine A, Rasberry ringspot, and grapevine leafroll-associated viruses gave negative results. A virus was isolated from affected grapevine young leaves by mechanical inoculation of Gomphrena globosa and single lesioned. The virus host range included G. globosa (local and systemic dark red or necrotic lesions), Chenopodium quinoa (necrotic local lesions and systemic mottle), and three tobacco cultivars (sharp necrotic local lesions, 1 to 3 mm in diameter). Pollination of C. quinoa with pollen from infected plant gave about 30% infected seedlings. The virus was purified from C. quinoa by differential centrifugation using 0.02 M phosphate buffer pH 8.0, containing 0.01 M DIECA and 0.01 M sodium thioglycolate as extraction buffers. In a purified preparation, quasisphaerical virus particles of about 29 nm were observed. Electrophoretic mobility of the viral coat protein showed a molecular weight of 30 kDa. Using purified preparations, an antiserum was obtained with a titer of 1:1024 in microprecipitin test and an optimum IgG dilution in ELISA of 1:10,000 for maximum absorption at OD405 nm Using degenerate primers designed from homologous regions in RNA-2 corresponding to a fragment of the polymerase gene of Ilarviruses, the expected 381-bp polymerase chain reaction product was obtained. This product was cloned and sequenced. Comparisons with sequence data from the homologous regions of RNA-2 of other known Ilarviruses, showed that the sequence of the above 381-bp amplicon shared 72% sequence similarity with Tobacco streak virus, 67% of Citrus variegation virus and Spinach latent virus, 66% of Asparagus virus 2 and Elm mottle virus, and 65% of Citrus leaf rugose virus. Based on the above data, it is concluded that the isolated virus is an Ilarvirus with closest similarity to Tobacco streak virus. From the relative bibliography (1-3) it appears that the virus reported here is different from Grapevine line pattern virus, a possible Ilarvirus, previously reported from Hungary. References: (1) J. Lehoczky et al. Kertgazdasag 19:61, 1987. (2) J. Lehoczky et al. Phytoparasitica 17:59, 1989. (3) J. Lehoczky et al. Phytopathol. Medit. 31:115, 1992.

Immunoglobulin E in serum and semen of infertile men.

Determination of IgE level in serum and semen of 92 men by an enzymatic immuno-assay method verified its presence in semen of most of them (92.3%) in much smaller amount than that present in serum (2.6%). IgE levels in serum and semen were significantly correlated. Serum levels were significantly higher in men with obstructive azoospermia, specially when associated with infection. Serum levels of IgE were higher in the fertile men, and in cases without infection as compared with those with prostatitis though this difference was not significant.

Diagnostic value of vaso-seminal-vesiculography.

The site of obstruction was evaluated by seminal vesiculography in 24 cases of azoospermia with normal testicular biopsies. In 21 cases, obstruction was located at the level of the tail of the epididymis; in 2 cases ejaculatory duct was obstructed; and in 1 case the injected dye was arrested at the level of internal inguinal ring, at the site of previous hernia operation. Vesiculogram in 24 obstructed cases and 4 cases of chronic seminal vesiculities without obstruction showed one normal picture, four with catarrhal inflammation, and the rest with chronic interstitial vesiculities. Radiological study of a patient complaining of aspermia showed multiple congenital anomalies. The high incidence of chronic vesiculities and postinflammatory obstruction is attributed to underlying bilharziasis.

Value of intraprostatic injection of zinc and vitamin C and of ultrasound application in infertile men with chronic prostatitis.

Seventy infertile men with chronic prostatitis were treated by prostatic massage and wide-spectrum chemotherapy as basic treatment to which intraprostatic injection of zinc or vitamin C with or without ultrasound application was added as a new line of treatment. Comparison showed no significant improvement of the additive treatment over the conventional treatment used alone. Pus cells in the expressed prostatic smear diminished significantly after treatment, which was associated with significant increase of percentage of motile spermatozoa and significant decrease of abnormal forms. Bacterial flora was studied in comparison with findings in 20 cases of infertile males without prostatitis; staphylococci predominated in both patient and control groups.

Zona-free hamster egg-sperm penetration assay: I. Testing of human semen fertilizability.

The zona-free hamster egg penetration test was performed on semen samples from 21 fertile donors, 57 infertile patients with normal semen parameters, and 63 infertile patients with subnormal spermiogram, as well as on 19 frozen semen samples. In the donor group 100% of the samples gave a positive range of penetration (PRP) of 20-96% with a mean penetration rate (MPR) of 53%, while 84% of the frozen samples showed a PRP of 18-44% and an MPR of 31%. Although 74% of the samples from infertile patients with normal spermiograms gave a PRP of 11-92% and an MPR of 34%, only 46% of the semen samples from the infertile patients with subnormal spermiogram showed a PRP of 21-100% but an MRP of 43%.

Zona-free hamster egg-sperm penetration assay: II. Correlative study with routine semen analysis.

This work studied the effect of variations in the different parameters of routine semen analysis on the penetration rate of the spermatozoa into the zona-free hamster eggs, using semen samples from 21 fertile donors, 57 infertile patients with normal spermiograms, 63 infertile patients with subnormal spermiograms, and 19 frozen semen samples. The results of the hamster test do not correlate with the percentage of motile spermatozoa, the percentage of live spermatozoa, or the percentage of spermatozoa with normal morphology in the semen samples. A positive correlation was found between the results of the hamster test and three other parameters of the semen analysis: the sperm concentration, the progressive type of motility, and a pH range of 7.2-7.7, but these positive correlations were statistically insignificant.

Testicular biopsy in azoospermia and severe oligozoospermia.

The continued experience of testicular biopsy application in 861 cases of azoospermia and 152 cases of severe oligospermia is reported adding to the previously published cases of 1075 patients with azoospermia. The most common finding in the whole series was that of normal testis denoting obstruction (48%), while among cases of functional azoospermia, Sertoli cell only and spermatogenic arrest were the most frequent (66%).

Treatment of asthenozoospermia by intratesticular testosterone therapy.

A single intratesticular injection of 0.5 cc of 50 mg/ml of an aqueous testosterone suspension was administered to 14 patients with asthenozoospermia. In 7 cases, injection was combined with ultrasound of 0.5 w/cm2 for 2 min weekly for 4 weeks. In another 4 cases, only ultrasound was applied. Intratesticular injection with or without ultrasound caused a significant increase of motility percentage. Sperm density improved in cases that were associated with oligozoospermia (less than 20 million/ml). Ultrasound therapy (0.5 w/cm2 for 2 min) used alone had no effect on semen parameters. Associated hormonal changes in blood plasma were evaluated.

FSH, LH, E2, progesterone, and testosterone in peripheral blood, spermatic vein, and semen of subfertile men with varicocele.

Peripheral blood (PB), spermatic vein (Sp. V.), and semen samples were collected from 31 subfertile men with varicocele. Levels of FSH, LH, estradiol, progesterone, and testosterone were determined in the three biological fluid compartments using RIA. Remarkable increases in both testosterone and estradiol in the spermatic vein samples, as compared to either peripheral blood or semen, were evident. Correlation coefficient indicated significant positive correlation between PB FSH and each of PB LH, Sp. V. FSH, and Sp. V. LH. Significant positive correlation existed between PB LH and each of Sp. V. FSH, LH, and testosterone. Significant positive correlations were found between PB testosterone and each of Sp. V. testosterone and semen testosterone, as well as between PB estradiol and each of Sp. V. FSH and progesterone. Significant positive correlations between Sp. V. FSH and Sp. V. testicular progesterone was significantly correlated with seminal estradiol.

C-reactive protein in semen and serum of men with chronic prostatitis.

Qualitative determination of C-RP in semen and serum of men with and without prostatitis showed that, the protein is present mostly in semen only in cases with prostatitis and with a significantly higher percentage incidence in the infertile than the fertile.

A double-blind trial of clomipramine in premature ejaculation.

A double-blind clinical trial was designed to study the effect of clomipramine in 50 patients complaining of rapid ejaculation. Patients were divided into two groups and each group received 2 periods of 6 consecutive weeks' treatment with drug and placebo, so that comparison can be made between drug and placebo within each group and between the two groups. Analysis of collected data showed a significantly higher percentage of satisfactory performances with the drug than with placebo and maintenance of the beneficial effect after the drug was replaced by placebo.

Sperm antibodies in serum and semen in men with bilateral congenital absence of the vas deferens.

Sperm antibodies were evaluated in serum and semen of 14 cases with bilateral congenital absence of the vas deferens, by agglutination and complement dependent tests. The level of the various immunoglobulin fractions were determined by the single radial immuno diffusion method. Sperm agglutinins were present in serum in 71% of cases and in 29% in semen. Immunoglobin (Ig) M was not detected in semen of both patient and control groups, while IgG and IgA were absent in the semen of patients.

Unconventional protein sources: "date seeds".

Different varieties of Egyptian date seed were evaluated biochemically. The protein content of which was in the range of 7.13-10.36%, while the fat content was from 6.32-9.28%. Phosphorus, calcium and iron were determined in all experimental samples. One variety of date seed (Samany) were used for extensive determination of the amino acids constituents by acid hydrolysis, quantitative determination of some amino acids, and evaluation of their biological value on attempt to use the seeds as new protein sources. Weanling albino rats fed on a diet of protein level 5% lost weight, but no toxic signs were observed. Analysis of blood serum of rats, for total proteins, albumin globulin ratio and free nonessential/essential amino acid protein, showed the date seed protein to be of low biological value.

Unconventional protein sources: apricot seed kernels.

Hamawy apricot seed kernels (sweet), Amar apricot seed kernels (bitter) and treated Amar apricot kernels (bitterness removed) were evaluated biochemically. All kernels were found to be high in fat (42.2--50.91%), protein (23.74--25.70%) and fiber (15.08--18.02%). Phosphorus, calcium, and iron were determined in all experimental samples. The three different apricot seed kernels were used for extensive study including the qualitative determination of the amino acid constituents by acid hydrolysis, quantitative determination of some amino acids, and biological evaluation of the kernel proteins in order to use them as new protein sources. Weanling albino rats failed to grow on diets containing the Amar apricot seed kernels due to low food consumption because of its bitterness. There was no loss in weight in that case. The Protein Efficiency Ratio data and blood analysis results showed the Hamawy apricot seed kernels to be higher in biological value than treated apricot seed kernels. The Net Protein Ratio data which accounts for both weight, maintenance and growth showed the treated apricot seed kernels to be higher in biological value than both Hamawy and Amar kernels. The Net Protein Ratio for the last two kernels were nearly equal.

Diagnostic value of determination of acid and alkaline phosphatase levels in the seminal plasma of infertile males.

To investigate the diagnostic value of phosphatases in seminal plasma, the levels of acid phosphatase and alkaline phosphatase were determined in 15 fertile subjects as well as in 26 cases of oligoasthenozoospermia. Statistical analysis of obtained data showed that acid phosphatase is a reliable parameter of prostatic function in cases of infection, while alkaline phosphatase may prove to be a non-specific parameter of subfertile semen. Alkaline phosphatase was significantly diminished in both oligozoospermia and azoospermia with and without infection or varicocele.

Clinical and hormonal studies of subfertile males with varicocele.

To evaluate the hormonal factor in subfertile males with varicocele, FSH, LH, estradiol (E2), and testosterone (T) were assayed both from cubital and spermatic veins. The hormonal profile of patients showed no significant differences from the normal control group. Blood hormonal levels of E2 and T in the testicular veins were much higher than those in the cubital veins while FSH and LH levels showed no significant difference. FSH, LH, and testosterone blood levels in cubital and spermatic veins were statistically correlated. A preoperative hormonal study would be helpful in excluding cases in which varicocele is not the underlying cause of infertility.

Endocrine profile of semen in subfertile males with varicocele.

The semen levels of FSH, LH, estradiol, progesterone, and testosterone were estimated in normal fertile males and in patients with oligozoospermia and varicocele. The only significant difference between the mean values of the two groups was noted for semen LH. Patients with oligozoospermia and varicocele had lower level than the normal fertile males. As a function of prostatic infection, no significant difference was obtained.

Lactate and pyruvate levels in the testicular vein of subfertile males with varicocele as a test for the theory of underlying hypoxia.

To test for the theory of testicular hypoxia in subfertile males with varicocele, the testicular mean blood level of lactate and pyruvate was compared with that of a control group. Statistical analysis of data showed a lower mean of lactate and of pyruvate in the testicular vein of varicocele patients. It is concluded that in varicocele there is no accumulation of lactic acid, thus excluding testicular hypoxia. Since lactate and pyruvate mean concentrations in testicular vein were correlated, it is suggested that in cases of varicocele there is defective glycolysis preceding the stage of pyruvate formation.

Bilharziasis and azoospermia.

Two patients in their twenties were investigated for a complaint of primary sterility. Semen analysis, showed azoospermia and testicular biopsy showed inhibited spermatogenesis in association with bilharzia ova in the interstitium in both cases. No similar cases were reported previously in the literature.