bluuebunny/crossref_metadata_2025 huggingface dataset:
Investigation of epididymal sperm maturation in the golden hamster
Authors: WEISSENBERG R. , YOSSEFI S. , OSCHRY Y. , MADGAR I. , LEWIN L. M.
Published: May 2008

During passage of hamster spermatozoa through the epididymis their maturation is shown to involve changes in the sperm head, midpiece (mitochondria) and tail. The sum of these changes results in a dramatic increase in the fertilizing potential of the spermatozoa. When comparable numbers of spermatozoa from the caput or corpus epididymis were injected into one uterine horn of mature females, following ovulation induction, and spermatozoa from the cauda epididymis were injected into the contralateral horn, no fertilization was observed with caput epididymal spermatozoa, 1.7% of oocytes were fertilized by corpus epiddymal spermatozoa, whereas 79.5% fertilization was obtained with cauda epididymal spermatozoa. Total sperm numbers increased from caput to corpus to cauda [28.3 ± 12.2, 40.6 ±20.8, 1434 ±62 mihon, respectively]. The percentage of progressively motile spermatozoa increased from 27.9 ±6.4 to 33.8 ± 4.8 to 70 ± 10.7 during this passage. Viability, measured by exclusion of the dye, propidium iodide, was significantly less in spermatozoa from the cauda than from the proximal or mid‐caput epididymis. The percentage of the live cells that were stained intensely by rhodamine‐123 (a measure of mitochondrial membrane potential) increased during epididymal passage from 22.8 ±7.8% in the proximal caput epididymis to 57.2 ± 16.5% in the cauda epididymis. Staining with acridine orange (a measure of DNA packaging in the sperm head) indicated an increase in chromatin condensation in cauda epididymal spermatozoa, when compared to those obtained from the caput or corpus.
Development of sperm motility patterns in the murine epididymis
Authors: SOLER C. , YEUNG C. H. , COOPER T. G.
Published: May 2008

The maturation of sperm motility in the epididymis of the mouse was assessed using a computer‐assisted sperm analysis system. Spermatozoa were immotile in the most proximal regions of the epididymis but developed motility rapidly in the proximal caput epididymis; the percentage motility remained high thereafter. In the caput, flagellar beat was erratic with little progression, but in the proximal corpus region circular movement patterns were reflected in reduced linearity (LIN) and straightness (STR) of the sperm tracks, although velocities were little changed and wobble (WOB) increased. In the mid‐corpus region, however, all velocities, LIN, STR and WOB, increased markedly. In more distal regions there was little change in these parameters. Distribution curves of the kinematic parameters of spermatozoa obtained from each region indicated that the most heterogeneous population was that from the mid‐corpus epididymis; the most homogeneous was that from the mid‐cauda region. Individual sperm tracks revealed slowly progressing spermatozoa in the distal caput, transforming to motion in small circles, interrupted by more linear progression. More distally, linear progression was interrupted by looping movements and a generally progressive path was observed thereafter, with less deviation from the average path as the spermatozoa matured. Spermatozoa displaying motion compatible with passing the uterotubal junction were first found in the proximal corpus epididymis, in agreement with earlier in‐vivo fertilization studies on where fertilizing capacity is achieved with epididymal spermatozoa.
Androgen regulation of intra‐ and extra‐thymic T cells and its effect on sex differences in the immune system
Authors: KIMURA MOTOHIKO , TOMITA YOSHIHKO , WATANABE HISAMI , SATO SHOTARO , ABO TORU
Published: May 2008

Male predominance of malignant diseases and female predominance of autoimmune diseases is widely known in humans. To elucidate one of the underlying mechanisms, we examined whether sex differences exist at the level of extra‐thymic T cells in various organs of mice under physiological conditions. Effects of castration and testosterone administration were also examined. Intra‐ and extra‐thymic T cells with T cell receptors (TCR) of bright and intermediate intensities, respectively, and with nil and high levels of the interleukin‐2 receptor β‐chain, respectively, were identified by immunofluorescence. Of the three strains tested, it was demonstrated that intermediate TCR cells were predominant in the liver of female mice. In the liver of male mice after bilateral orchidectomy, mononuclear cells increased and intermediate TCR cells predominated as if they were female; testosterone treatment prevented this immunological consequence of orchidectomy. As extra‐thymic T cells comprise a considerable proportion of self‐reactive forbidden clones and possess cytotoxic activity against syngeneic tumour cells, the present results suggest a possible relationship between (a) physiologically low levels of extra‐thymic T cells in males and the male predominance of malignancy, and (b) the high levels of extra‐thymic T cells in females and female predominance of autoimmune diseases.
Tight underpants and trousers and risk of dyspermia
Authors: PARAZZINI F. , MARCHINI M. , LUCHINI L. , TOZZI L. , MEZZOPANE R. , FEDELE L.
Published: May 2008

The association between the wearing of tight underpants and trousers and risk of dyspermia has been analysed using data from a case–control study. Cases included infertile men with a diagnosis of unexplained dyspermia. Normospermic men of infertile couples were eligible as controls. In comparison with men usually wearing loose underpants, the odds ratio (OR) of dyspermia was 1.9 (95% confidence interval, CI, 0.9–4.1) in those wearing tight underwear. Likewise, the OR of dyspermia was 1.6 (95% CI 0.9–3.0) in men reporting usually wearing tight trousers (including jeans), in comparison with those wearing loose trousers.
Reagents that activate GTP‐binding proteins trigger the acrosome reaction in human spermatozoa
Authors: DOMÍNGUEZ L. , DÍZ A. , FORNÉS M. W. , MAYORGA L. S.
Published: May 2008

The acrosome reaction is a specialized exocytotic process. In the mouse there is compelling evidence that receptor‐mediated activation of GTP‐binding proteins by factors in the zona pellucida of oocytes is a central event in the acrosome reaction. Several reagents are able to affect GTP‐binding proteins directly, bypassing the receptor–ligand step for activation. We have assessed the effect of several of these compounds on human spermatozoa, monitoring cell vitality and the acrosome reaction simultaneously using the triple‐stain technique. GTPγS and aluminium fluoride complexes promote sperm activation very efficiently; amphiphilic peptides capable of activating Go and Gi, also elicit the acrosome reaction. The results indicate that activation of heterotrimeric GTP‐binding proteins is sufficient to trigger acrosome exocytosis in human spermatozoa.
A strategy for identifying candidate sperm antigens for immunocontraception: isolation of human testis cDNA clones using polyclonal antisera directed against hamster acrosomal membrane preparation
Authors: ADOYO P. A. , MOORE A. , MOORE H. D. M.
Published: May 2008

Components of the mammalian sperm acrosome that have been conserved during evolution are probably essential for fertilization and are therefore potential antigens for the development of an immunocontraceptive vaccine. In order to identify such protein components, a series of specific polyclonal antisera were generated by immunizing rabbits with purified acrosomal membrane fractions from hamster epididymal spermatozoa. Antisera were finally selected using immunological and in‐vitro fertilization assays, and used to then screen a human testis Λgt11 cDNA library. As a result of this screening over 70 clones were identified, selected and purified. The cDNAs were amplified by polymerase chain reaction (PCR) and the inserts characterized by restriction enzyme digestion and oligonucleotide probing techniques. The functional activity of β‐galactosidase fusion proteins expressed by these clones was tested by a hamster in‐vitro fertilization assay (IVF). Fusion protein from three clones (HA5‐2, HA6‐2 and HB4‐1) inhibited significantly fertilization and reduced spermzona binding compared to controls. To date, sequence data has been obtained from HB4‐1 (1.75 kb). The first 1132 nucleotides displayed > 96% homology to human testis‐specific lactate dehydrogenase (LDH‐C4) gene, the product of which is a known candidate antigen for a contraceptive vaccine. This finding suggests that a strategy involving the screening across species for conserved moieties of the mammalian acrosome may be useful for identifying candidate antigens for immunocontraception.
Subcapsular intratesticular assay: a preliminary screening method for putative male antifertility drugs
Authors: XU Y. E. , WANG Y. I. , LIN N. , ZHANG J.‐W. , QIAN S. Z.
Published: May 2008

The subcapsular intratesticular assay was developed for the preliminary screening of male antifertility agents. In the assay, small amounts of the test sample are injected twice, 1 week apart just beneath the tunica albuginea of the testes of rats. The rats were killed within 3 weeks of treatment to examine epididymal spermatozoa and the histology of the testis and epididymis.

There is an excellent agreement between this method and the modified MB‐50 assay developed by the World Health Organization. To date, more than 300 samples have been assayed, of which the results of 10 plant‐derived compounds are reported here. Not one active sample has been missed as confirmed by the modified MB‐50 method, although one compound which was positive in the subcapsular assay did not show antifertility action in the in‐vivo assay. Compared with the latter assay, the advantages of this method are: simplicity, time‐saving, rat‐saving and the need for far smaller amounts of test sample. The absence of false‐negative results makes the subcapsular intratesticular assay applicable as a preliminary screening method to test potential male fertility regulating agents.
Severe asthenozoospermia: a structural and functional study
Authors: GOPALKRISHNAN K. , PADWAL V. , D'SOUZA S. , SHAH R.
Published: May 2008

Ejaculates from 25 patients with severe asthenozoospermia (all spermatozoa immotile or only non‐progressively motile) were studied to identify individually the cause of impaired motility. Multiple tests were performed, viz. light and electron microscopic studies and sperm function tests. An objective scoring was applied to both the routine and the functional analyses. Three categories of samples were identified: (1) necrozoospermia (n=9), where sperm viability was very poor; (2) structural tail abnormality as seen by light microscopy (n=4); and (3) ultrastructural abnormality (n=12). In the last category, one case showed absence of dynein arms; this was associated with mitochondrial abnormalities. Mitochondrial abnormality with normal tail components was observed in the majority (n=7) and accessory fibre abnormality in four cases. The scoring system revealed that, functionally, all samples were abnormal whereas routine analysis showed 15 samples to be subnormal and 10 to be abnormal, which indicates the need for functional analysis. Because of the multiple defects seen in these samples, there is a need for a battery of sperm function tests. This study indicates that mitochondrial defects are one of the causes that may account for the loss of sperm motility in the patient population.
Effects of progesterone on human spermatozoa prepared for in‐vitro fertilization
Authors: EMILIOZZI C. , CORDONIER H. , GUÉRIN J. F. , CIAPA B. , BENCHAÏB M. , FÉNICHEL P.
Published: May 2008

Progesterone has been tested in vitro with human spermatozoa to verify its physiological effects and its possible therapeutic use in cases of male infertility. Progesterone induced a rapid, dose‐dependent influx of calcium in capacitated and non‐capacitated spermatozoa with a half‐maximally effective dose of 30 nM. The agonist, 19‐nortestosterone, was much less potent that progesterone itself. Progesterone‐induced calcium influx was not inhibited by a similar concentration (0.1 μg/ml) of RU 486, a classical progesterone antagonist. The increase in intracytoplasmic calcium levels was unable to induce the acrosome reaction (AR) even after incubation for 5 h, when this was evaluated by double staining, using a monoclonal antibody GB24 raised against the inner acrosome membrane and ethidium homodimer as a vital probe. However, after incubation for 5 h, progesterone was able to enhance the tyrosine phosphorylation of a 95 kD sperm protein, which is phosphorylated progressively during capacitation in well‐defined culture media. Incubation of spermatozoa with 1 or 10 μg/ml progesterone for 3 or 30 min did not induce major modifications of hyperactivated movement when analysed by computer‐assisted semen analysis. Progesterone secreted by cumulus cells may physiologically increase sperm intracytoplasmic free calcium during capacitation. This priming effect may facilitate the acrosome reaction, induced by binding to the zona pellucida, without enhancing spontaneous acrosome reaction prematurely. It therefore seems useful to propose progesterone as a means of accelerating capacitation during in vitro fertilization in cases of male infertility.
Prolactin and testosterone: their role in male sexual function
Authors: CARANI C. , GRANATA A. R. M. , FUSTINI M. FAUSTINI , MARRAMA P.
Published: May 2008

The role of androgens in the sexuality of men is still not completely clear. Men with severe hyperprolactinaemia frequently show mild hypogonadism, and many complain of loss of libido and penile erectile dysfunction (ED). We studied the night‐sleep related erections and the penile response to visual erotic stimuli (VES) in 44 men: 13 with severe hypogonadism (Group 1; serum testosterone <1.4 ng/ml), 10 with mild hypogonadism (Group 2; serum testosterone 2–3.5 ng/ml), nine with severe hyperprolactinaemia and mild hypogonadism (Group 3) and 12 control men (Group 4). All of the patients complained of loss of libido and ED. Group 1 showed significantly impaired night erections when compared with any of the other three groups, but no differences were detected between Groups 2, 3 and 4. The penile response to VES did not show any significant difference between the four groups, but was lower in Group 1 than in Group 4. These data confirm that night erections are androgen‐dependent, but also suggest that there are two thresholds for serum testosterone: one below which sexual behaviour is impaired with normal night erections, and a lower threshold below which night erections are also impaired. The penile response to VES was confirmed as being only partially androgen‐independent. Furthermore hyperprolactinaemia does not affect night erections or the penile response to VES, suggesting that its effect on libido and sexual behaviour is due mainly to modulation of the psychological pattern of the patient.
In‐vitro initiation of forward motility in testicular spermatozoa
Authors: JAISWAL B. S. , MAJUMDER G. C.
Published: May 2008

Initiation of forward motility in vitro was investigated in goat and ram spermatozoa obtained from the rete testis. No forward motility was generated in the immotile testicular spermatozoa when they were incubated in a modified Ringer's solution containing theophylline (30 mM) and epididymal plasma (2 mg protein/ml). However, these reagents induced non‐progressive flagellar movement in approximately 25% of spermatozoa. Bicarbonate (25 mM) induced forward motility in approximately 16% of the goat/ram testicular spermatozoa. Theophylline was essential for the bicarbonatemediated activation of sperm motility, but epididymal plasma had no significant effect on this activation process. Theophylline activated progressive motility in testicular spermatozoa in a dose‐dependent manner, the maximum effect occurring after incubation for 10 min with 30 mM theophylline. The initiation profile of in‐vitro motility of goat/ram spermatozoa from the caput epididymis closely resembled that of testicular spermatozoa except that induction of motility in the caput spermatozoa was dependent both on bicarbonate and epididymal plasma. The data indicate that, unlike caput epididymal spermatozoa, initiation of motility in testicular spermatozoa is not dependent on motility‐promoting protein(s) in epididymal plasma.
Detection of Chlamydia trachomatis in the prostate by in‐situ hybridization and by transmission electron microscopy
Authors: CORRADI Gy. , BUCSEK M. , PÁNOVICS J. , VEREBÉLYI A. , KARDOS M. , KÁDÁR A. , FRANG D.
Published: May 2008

Chlamydia trachomatis infection has been investigated in the prostate gland using in‐situ DNA hybridization and transmission electron microscopy. Sixty‐four samples of tissue removed by trans‐urethral resection or by open surgery from men with benign prostatic hypertrophy (BPH), were examined histologically and 20 of the cases were found to be positive for mononuclear cell infiltration. Some samples were then investigated using in‐situ hybridization and transmission electron microscopy. Out of 20 tissue samples with mononuclear cell infiltration, nine were found to be positive for C. trachomatis infection.
Immunohistochemical detection of immature Sertoli cell markers in testicular tissue of infertile adult men: a preliminary study
Authors: STEGER K. , REY R. , KLIESCH S. , LOUIS F. , SCHLEICHER G. , BERGMANN M.
Published: May 2008

Testicular biopsy specimens from oligozoospermic infertile patients are characterized by different types of spermatogenic impairment in adjacent seminiferous tubules, a phenomenon called mixed atrophy. In order to evaluate possible involvement of the state of Sertoli cell differentiation, the distribution pattern of anti‐Müllerian hormone (AMH), vimentin and cytokeratin intermediate filament proteins was investigated by means of immunohistochemistry. AMH immunoactivity occurs in Sertoli cells of the normal postnatal prepubertal testis, but it is absent in the adult testis with normal spermatogenesis. In the case of mixed atrophy, AMH immunoactivity was found in Sertoli cells of tubules showing spermatogenic arrest at the level of spermatogonia and in tubules showing Sertoli‐cell‐only (SCO) syndrome. Vimentin was expressed regularly in Sertoli cells independent of spermatogenic impairment or the state of Sertoli cell differentiation. Cytokeratin immunoactivity occurs in Sertoli cells of the normal postnatal prepubertal testis. Furthermore, cytokeratin expression was found in Sertoli cells of tubules showing spermatogenic arrest at the level of spermatogonia and in some SCO tubules. Co‐expression of AMH and cytokeratin suggests that spermatogenic impairment such as spermatogenic arrest and SCO syndrome in human seminiferous tubules is associated with a population of Sertoli cells showing a prepubertal stage of development. The different pattern of AMH and cytokeratin expression in SCO tubules indicates that Sertoli cells in SCO syndrome show a mosaic pattern of differentiation.
Testicular gamma‐aminobutyric acid and circulating androgens in Syrian and Djungarian hamsters during sexual development
Authors: FRUNGIERI M. B. , GONZALEZ‐CALVAR S. I. , CHANDRASHEKAR V. , RAO J. N. , BARTKE A. , CALANDRA R. S.
Published: May 2008

Several factors, besides luteinizing hormone (LH), participate in the modulation of testicular function. A number of neurotransmitters are reported to be involved in this process, including a stimulatory action of gamma‐aminobutyric acid (GABA) on steroidogenesis in the rat testis. The purpose of this study was to investigate the testicular pattern of GABA and glutamic acid, one of its main precursors, during sexual maturation in two seasonally breeding species: Syrian (golden) and Djungarian hamsters. Plasma androgen levels were also measured. The animals were maintained under long‐day photoperiod (16:8, L:D) and were killed at 23, 30, 36, 46, 60, and 90 days of age. A different pattern of developmental changes in body and testicular weight was observed in these two species. GABA was present in the testes at all ages studied. GABA concentration and content showed a sharp elevation in the prepubertal period in golden as well as Djungarian hamsters. However, glutamic acid concentrations remained nearly constant during development in both species. Glutamic acid content increased gradually with age in the golden hamster, while a marked peak at 36 days of age was detected in the Djungarian hamster. Plasma testosterone and dihydrotestosterone levels were maximal at pubertal age in both species. The plasma levels of 5α‐androstane‐3α,17β‐diol increased significantly at 30 days of age in the golden hamster while in Djungarian hamsters this steroid remained unchanged. These results suggest that glutamic acid may serve as a precursor for GABA biosynthesis in the testis. In addition, changes in testicular GABA and plasma androgen levels might reflect a modulatory effect of this neurotransmitter in the acquisition of steroidogenic capability during development.
Sertoli cell barrier dysfunction and spermatogenetic cycle breakdown in the human testis: a lanthanum tracer investigation
Authors: MEYER J. M. , MEZRAHID P. , VIGNON F. , CHABRIER G. , REISS D. , RUMPLER Y.
Published: May 2008

The introduction of nitrate of lanthanum in fixative solutions for testicular biopsies improves ultrastructural observations of the blood–testis barrier. In complete (normal) spermatogenesis, junctional specializations impede the diffusion of lanthanum into the adluminal compartment. They clearly exhibit a three‐storied disposition in orthogonal sections. In maturation arrest, lanthanum passes easily through the junctional specializations surrounding the germ cells and up to the lumen. In irregular hypospermatogenesis, the Sertoli cell barrier is permeable, but diffusion of the tracer is less significant and variable. Unexpectedly, in germ cell aplasia, the barrier remains efficient.
Leydig cells secrete factors which increase vascular permeability and endothelial cell proliferation
Authors: COLLIN O. , BERGH A.
Published: May 2008

Adult intact control rats, and animals treated with human chorionic gonadotrophin (hCG) or with ethane dimethane sulphonate (EDS) to deplete Leydig cells, were injected with bromodeoxyuridine (BrdU) to label proliferating cells. Apoptotic cells were visualized by in‐situ end labelling (ISEL) of fragmented DNA. Three per cent of testicular endothelial cells were labelled with BrdU and few were apoptotic in intact testes. The BrdU endothelial cell labelling index was increased by hCG‐treatment and decreased in Leydig cell‐depleted testes. Immunohistochemical staining showed that Leydig cells and testicular macrophages contain immunoreactive vascular endothelial growth factor (irVEGF). The ability of testicular cells to stimulate angiogenesis was studied further by transplanting interstitial cells or seminiferous tubule segments under the kidney capsule. A prominent vascular network was observed around interstitial cell grafts, but not around tubule grafts. Treatment of transplanted rats with human chorionic gonadotrophin (hCG, 50 i.u.) resulted in an accumulation of PMN‐leukocytes and an increase in vascular permeability in the remaining testis and in interstitial cell grafts. Interstitial cells from Leydig cell‐depleted (EDS‐treated) testes were also transplanted under the kidney capsule. This type of graft caused only a discrete stimulation of angiogenesis, and there was no increase in vascular permeability around the graft after hCG treatment.

It is suggested that Leydig cells secrete angiogenic factors and that they are the source of the inflammation mediator(s) produced in the testis after hCG treatment. The high proliferation rate in endothelial cells suggests continuous remodelling of the testicular microvasculature, but the functional significance of this remains unknown.
Induction of apoptotic cell death in the seminiferous tubule of the adult rat testis: assessment of the germ cell types that exhibit the ability to enter apoptosis after hormone suppression by oestradiol treatment
Authors: BLANCO‐RODRÍGUEZ J. , MARTÍNEZ‐GARCÍA C.
Published: May 2008

The involvement of apoptosis in germ cell degeneration in the adult rat testis after gonadotrophin suppression has recently been shown in several studies, which have focused on the localization of apoptotic cells to the stages of the spermatogenic cycle. However, the precise germ cell types and maturing steps at which apoptosis is elicited remain controversial.

The present study used oestradiol treatment to produce hormone suppression and to study induced germ cell degeneration. Adult male rats were administered a daily injection of 50 μg oestradiol benzoate for 5, 10 or 15 days. Characterization of the ultrastructural features of the dying cells and in‐situ 3′‐end labelling of DNA showed clearly that the deaths of all the germ cell types occurred by apoptosis. High‐resolution light microscopy, although time‐consuming, resulted in a precise method for analysis of the localization of the cells involved. Stages IV–X of the seminiferous epithelium were found to be the most sensitive to degeneration in response to oestradiol treatment. Our results are discussed in the light of current knowledge about the hormonal control of the spermatogenic cycle. Oestradiol treatment has proved to provide a suitable in‐vivo model to study germ cell death.
Treatment with human chorionic gonadotrophin for cryptorchidism: clinical and histological effects
Authors: KALEVA M. , ARSALO A. , LOUHIMO I. , RAPOLA J. , PERHEENTUPA J. , HENRIKSÉN K. , TOPPARI J.
Published: May 2008

The efficacy of hCG treatment was studied in 182 cryptorchid patients. The efficacy of the treatment correlated with the initial position of the testis. None of the abdominal testes reached a normal position, whereas 90% of high scrotal testes descended during the treatment. Fifty‐four boys who were treated unsuccessfully with hCG and 29 untreated boys were biopsied. The biopsies were performed on 32 scrotal and 87 maldescended testes to examine the state of the organ and the effects of hCG treatment. In biopsies, the volume densities of seminiferous tubules, interstitial tissue and blood vessels were counted, and the sections screened for interstitial bleeding. Significant differences between scrotal and maldescended testes were found in all of the volume densities measured. Interstitial bleeding occurred rarely in scrotal testes, whereas in maldescended testes it was frequently apparent. hCG treatment induced a significant increase in the volume density of both interstitial tissue and blood vessels. Even though the hCG treatment induced measurable, possibly harmful, changes both in scrotal and maldescended testes, our data do not prove that hCG treatment causes permanent damage to the testis.
Anxiety‐induced failure in erectile response to intracorporeal prostaglandin‐E1 in non‐organic male impotence: a new diagnostic approach*
Authors: AVERSA A. , ROCCHIETTI‐MARCH M. , CAPRIO M. , GIANNINI D. , ISIDORI A. , FABBRI A.
Published: May 2008

Intracavernous injection (ICI) of prostaglandin‐E1 (PGE1) is used widely as the first diagnostic test in the study of erectile dysfunction. However, a lack of full erection after a maximal dose is frequent. As well as vascular incompetence, this may be due to stress‐induced changes, related to the ICI procedure. The aim of this study was to investigate the influence of emotional disturbances on erectile response to ICI in impotent patients. Initially, 24 young men with non‐organic impotence (age 34.6 ± 1.5 years; mean ± SEM) were selected and randomized single‐blind to pharmacoerection with PGE1 alone (20 μg/mL) or a mixture (cocktail) containing 20 μg PGE1 plus an α‐adrenergic receptor blocker, phentolamine (Phe, 0.5 mg/mL). Additional studies were also performed double‐blind on 10 men with non‐organic impotence (age 37.6 ± 1.2 years) utilizing higher PGE1 dosages for ICI (25 μg/mL alone or in combination with Phe, 0.5 mg/mL). After a 7‐day interval, all subjects were crossed‐over to receive the alternative treatment. The presence of emotional disturbances was assessed in all patients by the administration of rapid tests (Stai‐X1 and Stai‐X1r for state‐anxiety before and after ICI, respectively; Stai‐X2 for trait‐anxiety; Zung‐test for depression) at the first and at the remaining (Stai‐X1 and Stai‐X1r) ICI sessions.

ICI with 20 and 25 μg/mL PGE1 led to a comparable percentage of patients who reported a valid‐for‐intromission (VFI) erection (63 and 60%, respectively). In contrast, use of the cocktails significantly increased the percentage of subjects with a VFI (87 and 90% of the total number of patients tested, respectively; p <0.05). Moreover, a strong inverse correlation between state‐anxiety scores (Stai‐X1) and the erectile response to ICI with 20 and 25 μg PGE1 was found (r= ‐0.69, p<0.001); such a correlation was not present in patients who underwent ICI with the cocktails. Two cases of prolonged erection occurred (one after 20 μg PGE1 and the other after 20 μg PGE1 plus Phe) which were reversed promptly by the intracavernous injection of metharaminol.

It is concluded that the lack of a full erectile response after ICI with PGE1 can be related to the presence of a high ‘state‐anxiety’ in the patients. In such patients, a VFI erectile response can be induced by the administration of a cocktail test‐dose.
Association of chlorhexidine and fluoride for plaque control and white spot lesion remineralization in primary dentition
Authors: DE AMORIM RODRIGO GUEDES , LEAL SORAYA COELHO , BEZERRA ANA CRISTINA BARRETO , DE AMORIM FERNANDA PENNA LIMA GUEDES , DE TOLEDO ORLANDO AYRTON
Published: May 2008

Background.  Plaque control and caries arrest still remain a challenge for dentists.

Objective.  This study was conducted to assess the effect of the combined use of chlorhexidine varnish and fluoride varnish on the visible plaque index (VPI) and white spot lesion (WS) remineralization in primary dentition.

Methods.  A total of 80 caries‐active preschool children (3–5 years) were randomly divided into four groups. Group 1 received a chlorhexidine varnish application every week during 4 weeks. Group 2 received a fluoride varnish application every week during 4 weeks. Group 3 received alternated applications of chlorhexidine and fluoride varnish during 4 weeks. Group 4 served as control (without any type of cariostatic agent).

Results.  There was no statistically significant difference in the VPI and WS remineralization among the groups after 1 month. However, 3 months follow‐up demonstrated that group 3 (chlorhexidine + fluoride) showed significantly better results for both VPI and WS remineralization.

Conclusion.  The combined application of chlorhexidine and fluoride varnishes is more effective on plaque and remineralization of incipient caries after 3 months than the same agents applied separately.
Executive functions and cognitive subprocesses in patients with obstructive sleep apnoea
Authors: LIS STEFANIE , KRIEGER STEPHAN , HENNIG DOROTHEE , RÖDER CHRISTIAN , KIRSCH PETER , SEEGER WERNER , GALLHOFER BERND , SCHULZ RICHARD
Published: May 2008

In recent years, special interest has been focused on impairments of executive functions in patients with obstructive sleep apnoea syndrome (OSAS). However, the majority of studies have not clearly separated deficits in executive functions from impairments in other cognitive processes involved in task solving. In the present study, working memory (WM) functions of 20 patients with OSAS were compared with those of 10 age‐, sex‐ and education‐matched healthy subjects. Cognitive functions were measured four times a day; each of these measurements was accompanied by an assessment of subjective and objective daytime sleepiness. To separate dysfunctions of WM from those of additionally involved processes, n‐back tasks were applied embedded in a reaction‐time‐decomposition approach. Deficits in n‐back tasks could be observed in OSAS patients in accuracy and reaction times. However, the slowing could already be observed in simple reaction time tasks. The drop in 1‐back accuracy in the morning was related to daytime sleepiness. During the afternoon, accuracy of OSAS patients dropped in 2‐back tasks, an effect which correlated neither with sleepiness nor with the extent of sleep apnoea or oxygen desaturation. In conclusion, our data reflect a complex perspective upon cognitive deficits in OSAS. Cross‐group differences in processing time on the higher level WM task appeared to be attributable to slowing at a more elementary cognitive processing level. In contrast, reduced accuracy during the WM task in the OSAS group could not be explained by deficits in more elementary cognitive processes.
Evidence from population genetics that the ectomycorrhizal basidiomycete Laccaria amethystina is an actual multihost symbiont
Authors: ROY MELANIE , DUBOIS MARIE‐PIERRE , PROFFIT MAGALI , VINCENOT LUCIE , DESMARAIS ERICK , SELOSSE MARC‐ANDRE
Published: May 2008

It is commonly assumed that ectomycorrhizal (ECM) fungi associated with temperate forest tree roots are not host‐specific. Because this assumption relies on species delineations based on fruitbodies morphology or ribosomal DNA sequences, host‐specific, cryptic biological species cannot be ruled out. To demonstrate that Laccaria amethystina has true generalist abilities, we sampled 510 fruitbodies on three French sites situated 150–450 km away from each other. At each site, populations from monospecific stands (Abies alba, Castanea europea and Fagus sylvatica) or mixed stands (F. sylvatica + Quercus robur or Q. robur +Carpinus betulus) were sampled. Three different sets of markers were used for genotyping: (i) five microsatellite loci plus the ribosomal DNA intergenic spacer, (ii) the mitochondrial large ribosomal DNA subunit, and (iii) direct amplification of length polymorphism (DALP), a new method for fungi providing dominant markers. Evidence for allogamous populations (with possible inbreeding at local scale) and possibly for biparental mitochondrial inheritance was found. All markers congruently demonstrated that L. amethystina populations show little structure at this geographical scale, indicating high gene flow (as many as 50% of founding spores in all populations being of external origin). Our results also showed that host species contributed even less to population differentiation, and there was no evidence for cryptic biological species. This first in situ demonstration of a true multihost ability in an ECM species is discussed in terms of ecology and evolutionary biology.
Molecular diagnosis of Puccinia psidii (guava rust) – a quarantine threat to Australian eucalypt and Myrtaceae biodiversity
Authors: Langrell S. R. H. , Glen M. , Alfenas A. C.
Published: May 2008

A species‐specific, nested polymerase chain reaction (PCR)‐based detection assay, using two primer sets designed from the rRNA ITS region, was developed for Puccinia psidii. Detection sensitivities of one urediniospore alone, or one or two urediniospores in the presence of pollen or leaf tissues, respectively, were observed. The assay reliably, accurately and sensitively detected the rust from naturally infected, geographically widespread eucalypt and fruit tree plantation and nursery species from diverse tissues types (e.g. leaves, flowers, fruits, pollen, seeds and woody material) including symptomless or cryptically contaminated plants or plant tissue. Independent testing in Brazil and Australia demonstrated the international inter‐laboratory transferability of the P. psidii assay required for germplasm screening, disease monitoring and quarantine and incursion management, towards which the assay has already been employed.
Apical leaf necrosis as a defence mechanism against pathogen attack: effects of high nutrient availability on onset and rate of necrosis
Authors: Van Den Berg F. , Van Den Bosch F. , Powers S. J. , Shaw M. W.
Published: May 2008

An outdoor experiment was conducted to increase understanding of apical leaf necrosis in the presence of pathogen infection. Holcus lanatus seeds and Puccinia coronata spores were collected from two adjacent and otherwise similar habitats with differing long‐term N fertilization levels. After inoculation, disease and necrosis dynamics were observed during the plant growing seasons of 2003 and 2006. In both years high nutrient availability resulted in earlier disease onset, a higher pathogen population growth rate, earlier physiological apical leaf necrosis onset and a reduced time between disease onset and apical leaf necrosis onset. Necrosis rate was shown to be independent of nutrient availability. The results showed that in these nutrient‐rich habitats H. lanatus plants adopted necrosis mechanisms which wasted more nutrients. There was some indication that these necrosis mechanisms were subject to local selection pressures, but these results were not conclusive. The findings of this study are consistent with apical leaf necrosis being an evolved defence mechanism.
Snow mould caused by a Pythium sp.: a potential vascular plant pathogen in the maritime Antarctic
Authors: Bridge P. D. , Newsham K. K. , Denton G. J.
Published: May 2008

The pathogenicity of an isolate of a Pythium species from Signy Island in the South Orkney Islands was tested against the Antarctic hairgrass Deschampsia antarctica. The isolate was found to infect plants at 8ºC and to cause foliar and root symptoms similar to those seen in other Pythium infections in grasses. Analysis of ribosomal RNA sequences placed it, together with another isolate from Antarctica, in a clade that included the known snow moulds caused by Pythium spp. Sporangia and oogonia were produced in culture, but the isolate differed from other Pythium spp. in producing chlamydospores in older cultures and plant tissue. This is the first report of a pathogen of an eukaryotic vascular plant in the maritime Antarctic region.
Genotypic diversity and migration of clonal lineages of Botrytis cinerea from chickpea fields of Bangladesh inferred by microsatellite markers
Authors: Isenegger D. A. , Macleod W. J. , Ford R. , Taylor P. W. J.
Published: May 2008

An analysis of allelic diversity at nine microsatellite loci provided an insight into the population structure of Botrytis cinerea from four fields (sampled in 2003 and 2004) that represented important regional locations for chickpea production in Bangladesh. Although three populations were limited by sample size after clone‐correction, a total of 51 alleles were amplified among 146 B. cinerea isolates from Bangladesh, which revealed a high amount of within‐population and overall genetic diversity (HS = 0·48 and HT = 0·54, respectively). The percentage of maximal genotypic diversity (G) ranged between populations (G = 23–40), with a total of 69 haplotypes detected (G = 25). Bayesian cluster analysis depicted two major clusters distributed among the four Bangladesh populations, indicating population admixture from two origins that have spread throughout these regions. Genotype flow between regions was detected and indicated the spread of clonal lineages, consistent with relatively low differentiation among the four populations (mean GST = 0·1, P < 0·05). These results highlighted the potential threat of host resistance breakdown as a result of considerable genetic diversity, genotype flow and the evolutionary potential of B. cinerea.
Maternal country of birth and previous pregnancies are associated with breast milk characteristics
Authors: Amoudruz Petra , Holmlund Ulrika , Schollin Jens , Sverremark‐Ekström Eva , Montgomery Scott M.
Published: May 2008

Populations in high infectious exposure countries are at low risk of some immune‐mediated diseases such as Crohn’s disease and allergy. This low risk is maintained upon immigration to an industrialized country, but the offspring of such immigrants have a higher immune‐mediated disease risk than the indigenous population. We hypothesize that early life exposures in a developing country shape the maternal immune system, which could have implications for the offspring born in a developed country with a low infectious load. The aim of this study was to investigate if exposures in childhood (indicated by country of origin) and subsequent exposures influence immunologic characteristics relevant to stimulation of offspring. Breast milk components among 64 mothers resident in Sweden, 32 of whom immigrated from a developing country, were examined using the ELISA and Cytometric Bead Array methods. Immigrants from a developing country had statistically significantly higher levels of breast milk interleukin‐6 (IL‐6), IL‐8 and transforming growth factor‐β1. A larger number of previous pregnancies were associated with down‐regulation of several substances, statistically significant for soluble CD14 and IL‐8. The results suggest that maternal country of birth may influence adult immune characteristics, potentially relevant to disease risk in offspring. Such a mechanism may explain the higher immune‐mediated disease risk among children of migrants from a developing to developed country. Older siblings may influence disease risk through the action of previous pregnancies on maternal immune characteristics.
Free IAA in stigmas and styles during pollen germination and pollen tube growth of Nicotiana tabacum
Authors: Chen Dan , Zhao Jie
Published: May 2008

Although many studies have emphasized the importance of auxin in plant growth and development, the thorough understanding of its effect on pollen–pistil interactions is largely unknown. In this study, we investigated the role of free IAA in pollen–pistil interactions during pollen germination and tube growth in Nicotiana tabacum L. through using histo and subcellular immunolocalization with auxin monoclonal antibodies, quantification by HPLC and ELISA together with GUS staining in DR5::GUS‐transformed plants. The results showed that free IAA in unpollinated styles was higher in the apical part and basal part than in the middle part, and it was more abundant in the transmitting tissue (TT). At the stage of pollen germination, IAA reached its highest content in the stigma and was mainly distributed in TT. After the pollen tubes entered the styles, the signal increased in the part where pollen tubes would enter and then rapidly declined in the part where pollen tubes had penetrated. Subcellular localization confirmed the presence of IAA in TT cells of stigmas and styles. Accordingly, a schematic diagram summarizes the changing pattern of free IAA level during flowering, pollination and pollen tube growth. Furthermore, we presented evidence that low concentration of exogenous IAA could, to a certain extent, facilitate in vitro pollen tube growth. These results suggest that IAA may be directly or indirectly involved in the pollen–pistil interactions. Additionally, some improvements of the IAA immunolocalization technique were made.
Parasitoids of Paratachardina lobata (Hem., Kerriidae): surveys for biological control of the invasive lobate lac scale
Authors: Schroer S. , Pemberton R. W. , Selvaraj P.
Published: May 2008

Paratachardina lobata is an invasive pest in southern Florida, threatening a great number of economically important and native plants. The lobate lac scale does not cause problems in its area of origin, India and Sri Lanka, presumably due to various parasitoid wasps. In an attempt to discover promising parasitoids for biological control against the invasive pest in Florida, P. lobata infesting Pongamia pinnata (Fabaceae) was collected bimonthly from August 2005 to June 2006 at 14 sites in southern India. Four parasitoids were demonstrably associated with P. lobata: a eulophid Aprostocetus bangaloricus Narendran, an encyrtid Ooencyrtus kerriae Hayat and two aphelinids, Marietta leopardina Motschulsky and an undescribed Coccophagus Westwood sp. These chalcidoid wasps were found regularly at all heavily infested sites with an average emergence number per collection period of 62.8, 31.7, 11.9 and 90.5 respectively. The mean number emerged 20–28 days after the collection date, excluding the Coccophagus sp., which occurred significantly later, on average 41 days after collection. Species emergence was examined and parasitized scales dissected. Parasitoid remains were interpreted to understand the mode of parasitism. The Coccophagus sp. was found to be a secondary parasitoid. Marietta leopardina occurred as a primary parasitoid, but only in low number and this species is also known to be hyperparasitic on chalcidoid wasps. Aprostocetus bangaloricus and O. kerriae are promising candidates for the lobate lac scale control in Florida. They are primary parasitoids of P. lobata and occurred at almost every collection site, especially where P. lobata was very abundant.
Noradrenergic Control of GnRH Release from the Ewe Hypothalamus In Vitro: Sensitivity to Oestradiol
Authors: Ghuman SPS , Jones DN , Prabhakar S , Smith RF , Dobson H
Published: May 2008

Contents

The present study investigates the influence of α1‐adrenoreceptors in GnRH release in vitro and determines whether oestradiol modulates α1‐adrenoreceptor‐GnRH interaction. Within 10 min after ewe sacrifice, saggital midline hypothalamic slices were dissected, placed in oxygenated Minimum Essential Media‐α (MEM‐α) at 4°C and within 2 h were singly perifused at 37°C with oxygenated MEM‐α (pH 7.4; flow rate 0.15 ml/min), either with or without oestradiol (24 pg/ml). After 4‐h equilibration, 10‐min fractions were collected for 4 h interposed with a 10‐min exposure at 60 min to specific α1‐adrenoreceptor agonist (methoxamine) or antagonist (thymoxamine) at various doses (0.1–10 mm). The α1‐adrenoreceptor agonist (10 mm) increased (p < 0.05) GnRH release at 90 min both in presence and absence of oestradiol. However, in presence of oestradiol, α1‐adrenoreceptor agonist (10 mm)‐induced GnRH release remained elevated (p < 0.05) for at least 60 min. The bioactivity of the released GnRH was studied using a hypothalamus–pituitary sequential double‐chamber perifusion. Only after exposure of hypothalamic slices to α1‐adrenoreceptor agonist (10 mm), did the hypothalamic eluate stimulate LH release from pituitary fragments (n = 9, 7.8 ± 12.3–36.2 ± 21.6 ng/ml) confirming that the α1‐adrenoreceptor agonist stimulated release of biologically active GnRH. In summary, GnRH release from the hypothalamus is under stimulatory noradrenergic control and this is potentiated in the presence of oestradiol.
Repeated administration of a dopamine D1 receptor agonist reverses the increased proportions of striatal dopamine D1High and D2High receptors in methamphetamine‐sensitized rats
Authors: Shuto Takahide , Seeman Philip , Kuroiwa Mahomi , Nishi Akinori
Published: May 2008

Repeated administration of psychostimulants produces a behavioural sensitization. Amphetamine‐sensitized animals are known to have a higher proportion of high‐affinity states of dopamine D2 receptors (D2High receptors) in the striatum. We recently reported that repeated administration of a dopamine D1 receptor agonist, R‐(+)‐SKF38393, reverses the established behavioural sensitization to methamphetamine (MAP). To investigate the mechanisms for reversal of behavioural sensitization, we examined the effect of repeated administration of the dopamine D1 receptor agonist on the proportions of D2High receptors and the high‐affinity states of dopamine D1 receptors (D1High receptors) in the striatum. In the striatum from the MAP‐sensitized rats, the proportions of D1High and D2High receptors (28.5 ± 1.96 and 57.5 ± 3.58%) were higher than those in the saline‐control rats (12.0 ± 1.01 and 21.9 ± 1.60%, respectively). Repeated administration of R‐(+)‐SKF38393 to the MAP‐sensitized rats reduced the increased proportions of D1High and D2High receptors to 12.4 ± 1.57 and 31.0 ± 2.14%, respectively, which were similar to the proportions in the saline‐control rats. The total densities of dopamine D1 and D2 receptors were not altered in each treatment condition. The results demonstrate that the proportions of D1High and D2High receptors in the striatum are elevated in MAP‐sensitized rats, and that repeated administration of the dopamine D1 receptor agonist to the MAP‐sensitized rats reverses the increased proportions of D1High and D2High receptors. The findings reveal postsynaptic mechanisms for the development of behavioural sensitization to MAP and the reversal of established sensitization by repeated administration of the dopamine D1 receptor agonist.