Morphostructural investigation of the female reproductive system and molecular evidence for Wolbachia in Balclutha brevis Lindberg 1954 (Hemiptera, Cicadellidae).

Balclutha brevis Lindberg 1954 (Homoptera, Cicadellidae) is an allochthonous species that is rapidly spreading in Sicily and in mainland Europe due to the wide spread of its host plant and therefore could also compete with populations of native species. Considering these ecological implications, based on the lacking ultrastructural data about the reproductive systems of the Auchenorrhyncha and since previous investigations on the male reproductive system of B. brevis have shown some interesting features, we carried out morphostructural investigations on the female reproductive system of this alien leafhopper. Moreover, given the high interest in literature on Wolbachia entomoparasite and based on our previous studies, we provided a contribution to further investigations in applied sciences. For this aim we performed a molecular analysis on males and females of B. brevis to detect the possible presence of strains of the bacterium known to alter host reproductive biology. The female reproductive system has a morphological organization comparable to the general anatomical features of most of the Auchenorrhyncha species; however, comparing our data with the literature, some considerations are discussed. As for the histological and ultrastructural investigations, our results show a secretory activity of the various examined structures. In the spermatheca of B. brevis, in particular, the secretory activity is more marked in the sac-shaped tract, where histochemical investigations showed a lipid component of the secretion; possible origin of this component is discussed. Moreover, mainly free spermatozoa are found in the sac-shaped tract of the spermatheca and in the common oviduct. As for the latter, an interesting findings is the lack of cuticular intima on the epithelial surface of the common oviduct; furthermore, the observed features and the literature in this regards led us to review the significance of the structure called as spermatheca. The molecular screening study (PCR) with three Wolbachia specific genes (16SrRNA, ftsZ,wsp) strongly supports Wolbachia infections in both males and females of the host species. This is the first study demonstrating the presence of Wolbachia in Balclutha genus, though no morphostructural alteration commonly related to the presence of the bacterium has been found in all the examined specimens.

Morphostructural analysis of the male reproductive system and DNA barcoding in Balclutha brevis Lindberg 1954 (Homoptera, Cicadellidae).

Balclutha brevis Lindberg 1954 is an allochthonous leafhopper infesting an invasive grass, Pennisetum setaceum, in Sicily and in mainland Europe; therefore, this species could compete with populations of native species, thus contributing to the loss of biodiversity. Considering the ecological implications of B. brevis, investigations on all its biological aspects represent, therefore, a premise for further studies in applied sciences. Based on the lacking ultrastructural data about the reproductive systems of the Auchenorrhyncha, we carried out morphostructural investigations on the male reproductive system of B. brevis. Further, a first report of DNA barcoding analysis (amplification and sequencing of Cytochrome Oxidase I gene) has also been performed to characterize B. brevis compared to other congeneric species. From a morphological point of view, the male reproductive system of B. brevis has an organization comparable to the general anatomical features of most of the Auchenorrhyncha species; however, comparing our data with those concerning the different groups of Cicadomorpha, some considerations are discussed. As for the histological and ultrastructural investigations, our results show a secretory activity of the various examined structures, mainly in the lateral ejaculatory ducts and in the accessory glands. The latter, in particular, show morphostructural differences comparing the distal tract to the proximal one; moreover, the histochemical techniques showed the possible presence of a lipid component in the peculiar cytoplasmic granules found in the gland cells. The significance of these findings in the accessory glands is discussed. Finally, the ultrastructural features found in the seminal vesicles are different from those of the lateral ejaculatory ducts and are indicative of the different roles played by these structures in the organization of the spermatozoa bundles.

Genesis of spermatodesms in Tylopsis liliifolia (Orthoptera: Phaneropterinae) and their transit in the male genital tract.

The spermatodesms of Tylopsis liliifolia form in the most proximal follicular cysts and are composed of a large number of sperm held together by a cap located in the anterior region of the acrosome. The cap is formed by short thin fibrils, loosely arranged at random, probably derived from secretory activity of cells of the cyst wall. Compared to other Tettigoniidae, a peculiar feature is acrosomal wings that twist gradually around the anterior region of the nucleus; at the end of the twisting process, the region of the sperm acrosome, observed in cross section, shows a typical spiral form. Spermatodesms do not undergo any substantial changes in the spermiduct. The epithelial cells of the wall have secretory activity and many show marked spermiophagic activity, which is conducted by epithelial cell protrusions that envelop the gametes, taking them into the cytoplasm. When removed from seminal vesicles and observed in vivo, spermatodesms show accentuated corkscrew movement, and when observed by SEM, slight torsion. Thus organized, spermatodesms are transferred to the spermatophore during mating, where they are transformed before reaching the seminal receptacle.

Morphological and ultrastructural organization of the male genital apparatus of some Aphididae (Insecta, Homoptera).

Ultrastructural investigations on the amphigonic reproductive mechanisms in Aphidoidea (Homoptera, Sternorryncha) species, of which little is known in the literature, can provide useful information on their reproductive biology. Morphological and ultrastructural investigations were carried out on the reproductive tract and on spermatozoa from sexually mature males of five species belonging to three subfamilies of Aphidoidea. The organization of the reproductive tract and of spermatozoa appears simple and similar in the examined species. Each testis consists of three follicles containing many cysts arranged in a progressive order of maturation from the distal to the proximal tract; spermiogenesis only occurs in sexually mature males, ending with the organization of sperm bundles. Gametes are neatly arranged in each bundle and kept together by a cap showing a particular organization. The distal tract of each spermiduct is enlarged and full of gametes; close to the testis the two spermiducts merge together, except in Drepanosiphum platanoidis where the spermiducts run independently. The various tracts of each examined gland showed no peculiar differences either in the same species or among the species. Some interesting hypotheses are proposed in this work about the probable roles of the investigated structures in the reproductive mechanisms of these insects.

Formation and rearrangement of spermatodesms in males of some Orthoptera Tettigoniidae.

In the male genital tract of Tettigoniidae, the spermatodesms are composed of a limited number of spermatozoa whose nuclei and acrosomes are covered by a mucous cap. The formation of the cap begins in the testicular cyst during the lengthening of the apical prolongations of the spermatids and the spermatids' simultaneous division into small bundles or spermatodesms. The cap material is formed from a loosely arranged material in the lumen of the cyst, probably produced by the secretory activity of the delimiting cells. Another characteristic aspect of the Tettigoniidae is the rearrangement of the cap inside the spermiduct that seems to start when material from the lumen of the organ enters from the basal part of the cap. Except for the fibrils of the peripheral lamina, the fibrils of the cap undergo a marked degradation process. The final organization of the spermatodesm cap is reached inside the seminal vesicles; it consists of the fibrillar peripheral lamina delimiting an interior made up of loosely arranged fibrils immersed in a finely granular matrix.

Role of male accessory glands in spermatodesm reorganization in Orthoptera Tettigonioidea.

A preliminary examination of the spermatodesms of Orthoptera Tettigonioidea revealed a structure that is similar in individuals of the same sex but very different in specimens of opposite sex. This reorganization would seem to take place inside the spermatophore during transit from the male to female genital tracts. The results of incubating spermatodesms with the secretions of glandular extract (GE) obtained from male accessory glands, known to be involved in forming the spermatophore wall, revealed changes in the spermatodesm 'cap' that are comparable to those occurring in vivo. Moreover, incubation of spermatodesms with the extracts obtained separately from tubules of the 1st and 2nd orders (GE1, GE2) established that GE2 alone modifies the spermatodesms, thus excluding the possible implication of 1st order tubules in the rearrangement process. In conclusion, data from incubations of spermatodesms with the single fractions obtained by submitting GE2 to gel-filtration FPLC show that only the peak 4 maintains intact the biological activity of GE2, SDS-PAGE analysis of the fraction corresponding to peak 4 revealed a greater protein content of 29 kD, which also appears to a lesser degree in fraction 3. This material is responsible for a partial dismantling of the 'cap' in the incubated spermatodesms.

Potential interaction of troglitazone and cyclosporine.

BACKGROUND: Troglitazone (Rezulin) is a promising new oral hypoglycemic agent recently approved by the Federal Drug Administration for use in type II diabetes mellitus. Although troglitazone is not metabolized by the cytochrome p450 3A isozyme family, it is a potential inducer of this system. Other medications, e.g., rifampin and phenobarbital, which also induce p450 3A activity, have been reported to significantly decrease cyclosporine (CsA) concentrations. METHODS: We report a case of a stable renal transplant patient who had a decrease in CsA concentration after beginning troglitazone and who subsequently developed an acute rejection episode. We then reviewed all stable renal patients begun on troglitazone over the previous 6 months. RESULTS: The seven transplant patients who had been started on troglitazone therapy experienced a statistically and clinically significant decrease in CsA 12-hr trough levels immediately after the institution of troglitazone therapy. CONCLUSION: A potential interaction exists between troglitazone and CsA. Transplant patients on CsA who receive troglitazone therapy should be monitored closely.

Structure of spermatodesms of Orthoptera Tettigonioidea.

Examination of spermatodesms collected from the male and female genital tracts of numerous Orthoptera Tettigonioidea revealed an overall morphological and ultrastructural organization that is generally similar in individuals of the same sex but considerably different between males and females of even the same species. In the male genital tracts each spermatodesm is composed of a limited number of spermatozoa whose nuclei and acrosomes are covered by a mucous cap. The spermatozoa inside each bundle are mainly arranged in parallel rows and are always distinctly separate. The number of spermatozoa per spermatodesm may vary within the same individual although it does not seem to exceed a maximum value that we could only determine exactly in Tettigoniidae species. The most characteristic feature of spermatozoa of all the species examined is a conspicuous elongation of the plasma membrane in the acrosomal region that is not present in the female genital tracts. In addition, spermatodesms from females are composed of highly numerous tightly packed spermatozoa that are linked together via the acrosomal region. This characteristic of spermatodesms, never previously reported in other insect species, would involve their reorganization during transfer from the male to the female genital tracts and would seem to take place in the spermatophore. The probable role of spermatodesms in the reproductive physiology of Tettigonioidea might be related to the degree of maturity of the sex cells transferred to the female; the reorganization of the spermatozoa out of the male genital tracts seems to support this hypothesis.

Evaluation of voiding cystourethrography prior to renal transplantation.

In many centers, voiding cystourethrography is a routine part of pretransplantation assessment of the lower urinary tract. To assess the value of this investigation, a retrospective review of transplant candidates evaluated in our center over 2 years was undertaken. A total of 517 patients were fully evaluable. Only 13 voiding cystourethrograms (VCUGs) (2.5%) of 517 were found to be abnormal. Three patients with reflux alone did not require intervention before transplantation. Four patients with decreased bladder capacity underwent hydrodistention. Two patients increased their capacity to over 150 ml and two patients failed distention, one requiring an ileal conduit and the other requiring an augmentation cystoplasty. Three patients had increased postvoid residual (PVR). Two patients started clean intermittent catheterization. One required prostate resection for benign prostatic hypertrophy. One patient with reflux and decreased bladder capacity refused treatment. One patient with reflux combined with increased PVR started clean intermittent catheterization and was cleared for transplant surgery. One patient with decreased bladder capacity and increased PVR had a stroke and was excluded from transplantation. All 13 patients with abnormal VCUGs had a prior urologic history. In total, only 56 of 517 patients evaluated had a prior urological history. Each VCUG costs approximately $500. Limiting VCUG studies to those patients with a prior urological history would have resulted in a significant cost savings. Hence, we recommend that only patients with a prior urological history should undergo this costly and often distressing examination.

Ultrastructural and histochemical characteristics of the lateral oviducts of Baculum thaii Haus. (Phasm.).

The wall of the lateral oviducts of baculum thaii is composed of a monolayered epithelium, lacking a cuticular intima, which lies on a sheath of striated muscular fibres. Untreated oviductal specimens exhibit two distinct regions. The anterior region, into which the various ovarioles emerge, is opalescent. The posterior region is more dilated and darker in colour. On the basis of the ultrastructural characteristics of the epithelial cells, four successive zones are definable for each oviduct. The first two zones comprise the most anterior oviductal area while the third and fourth comprise the posterior region. In zone 1 only is the oviductal wall composed of a single cell type. In addition, differences in structural organization between mated and virgin females were observed in this zone alone. In the three remaining zones, two distinct cell types were distinguishable according to two models of fundamental organization: cells having numerous apical microvilli and dome-shaped cells. The results of histochemical analyses established that the abundant secretory product elaborated by the epithelial cells is rich in both proteins and predominantly acidic mucosubstances. The histochemical characteristics of the secretory product differentiate significantly not only between the four zones in the same individual but also when the corresponding zones are compared in the mated and virgin females.

Laparoscopic internal drainage of lymphocele in renal transplant.

Lymphoceles frequently develop following renal transplant surgery. The conventional methods used to drain lymphoceles are invasive and can be associated with complications. Laparoscopic drainage of posttransplant lymphoceles provides a safer alternative.

Lectin binding to the egg envelopes in Eyprepocnemis plorans (Charp.) (Orthoptera, Acrididae).

The distribution of glycoconjugates in the egg envelopes of Eyprepocnemis plorans was investigated using various FITC-conjugated lectins. In the epichorion, the lectins ConA, SBA and WGA each have particular binding patterns, while TPA binding is confined to its deepest regions only. The glycoconjugates of the micropylar wall present different characteristics from those of the surrounding chorion. The vitelline coat shows a marked binding for WGA and TPA only; below the inner micropylar openings, this binding pattern is uniform over the whole extent of the coat and therefore it is not possible to identify specific binding sites for these two lectins. Contrary to what has been observed in some other insect species, the vitelline coat does not seem to be involved in the structural organization of the mycropyles.

Mitomycin-induced hemolytic-uremic syndrome. Successful treatment with corticosteroids and intense plasma exchange.

Mitomycin has been associated with microangiopathic hemolytic anemia and renal failure. We describe a patient with the adult hemolytic-uremic syndrome due to mitomycin who was successfully treated with intense plasma exchange therapy and corticosteroid therapy. Patients receiving mitomycin should have their conditions monitored closely for acute renal failure, thrombocytopenia, and hemolytic anemia.

Cisplatin-induced hypocalcemia and hypomagnesemia.

Cisplatin (cis-diamminedichloroplatinum), a chemotherapeutic agent active against solid tumors, is a known cause of acute renal failure and renal tubular dysfunction. We saw a case of renal magnesium wasting and hypomagnesemia in a 58-year-old woman with genital pelvic malignancy. Her initial manifestation was that of chronic symptomatic hypocalcemia of four months' duration. The relatively reduced parathyroid hormone level in this patient and the possible sites of cisplatin-induced renal tubular dysfunction resulting in hypermagnesuria were evaluated. This case emphasizes the importance of serially observing cisplatin-treated patients for the possible development of hypomagnesemic hypocalcemia.

Anticipation of renal transplant failure by postanastomosis biopsy and immunofluorescence.

Ninety-six pre- and postanastomosis allograft biopsies from 48 transplant recipients were studied employing the direct fluorescent antibody procedure. Of the 48 transplants studied, 26 postanastomosis biopsies showed significant blood vessel wall deposits consisting predominantly of 2-4+ staining IgM and C3 deposits. The corresponding preanastomosis biopsies showed only patchy intimal surface staining. All but two of the grafts with positive biopsies had been preserved by the pulsatile perfusion method for various periods of time. Twenty-three of the twenty-six grafts with positive findings failed within 1 month while only 2 of the 19 negative postanastomosis biopsies failed within the 1-month period. Based on these findings actual graft failure occurred in 89% of the patients with postanastomosis transplant biopsies with positive findings. Although crossmatch studies were negative in all cases, further immunologic studies of eluates obtained from positive postanastomosis biopsies revealed the presence of lymphocytotoxic IgM antibodies in three of five cases studied. These findings suggest that rapid deposition of undetected preformed lymphocytotoxic antibodies accumulate in the vessel walls following anastomosis and contribute to transplant failure.