bluuebunny/crossref_metadata_2025 huggingface dataset:
Influence of photoinhibition on GABA and glutamic acid levels, and on glutamate decarboxylase activity in the testis and epididymis of the golden hamster
Authors: FRUNGIERI M. B. , GONZALEZ‐CALVAR S. I. , CALANDRA R. S.
Published: May 2008

Gamma‐aminobutyric acid (GABA) is found in the gonads and accessory reproductive organs, and a direct effect on steroidogenesis and sperm viability and motility has been described. The golden (Syrian) hamster is a seasonal breeder, and a pattern of regression‐recrudescence in their reproductive organs is observed when adult animals are exposed to less than 12.5 h daylight for an extended period of time. The purpose of this study was to investigate: (1) the presence of GABA in the testis and epididymis of golden hamsters undergoing regression and spontaneous recrudescence; (2) glutamic acid levels and glutamate decarboxylase (GAD) activity in both tissues, and (3) testicular and epididymal testosterone, dihydrotestosterone and 5α‐androstane‐3α, 17β‐diol concentrations. Adult golden hamsters were exposed to long (LP 14L:10D) or short (SP 6L:18D) photoperiods for 9, 12, 14, 16, 18 or 22 weeks. When animals were exposed to SP for 14–16 weeks, the testis and epididymis reached maximal involution. Testicular and epididymal androgen levels showed a marked decrease (p<0.05) during the regression period, and after 18–22 weeks, values began to recover. Between 12 and 18 weeks in SP, the testicular and epididymal content of GABA and glutamic acid was reduced significantly. The concentration of GABA in both tissues showed a sharp rise (p<0.05), while the concentration of glutamic acid diminished during the period of maximal involution (p<0.05). In the testis, GAD activity was increased (p<0.001) after 14 weeks in SP, with no change in the epididymis. In conclusion, glutamic acid via GAD activity could be the main source of GABA in the testis, but not in the epididymis. Furthermore, the presence of GABA in testicular cells and its subsequent photoperiodic variations might act as an important autocrine and/or paracrine modulatory signal in gonadal processes.
Infection with Trypanosoma brucei potentiates the antifertility effect of gossypol, especially in the protein‐malnourished male rat
Authors: AKINGBEMI B. T. , AIRE T. A. , OKE B. O.
Published: May 2008

Reproductive parameters were investigated in gossypol‐treated male rats that had been infected experimentally with Trypanosoma brucei and then placed on one of two diets of differing (low and normal) protein content. The endpoints assessed were reproductive organ weights, semen epididymal sperm counts, serum testosterone and histological, stereological and histomorphometric evaluation of the testis and accessory reproductive organs. Most of the parameters studied were lowest in the protein‐malnourished, gossypol‐treated, trypanosome‐infected animals, when compared to those obtained from the corresponding animals that were only gossypol‐treated or trypanosome‐infected. Mean testicular size and the diameter and the total length of seminiferous tubules were especially reduced, indicating that the overall volume of the seminiferous epithelium in these animals was smaller. These findings suggest that reproductive capacity could be impaired in protein‐malnourished, trypanosome‐infected animals fed on gossypol‐containing products, even when there are no obvious clinical signs of disease. This could translate into increased production costs in a farm enterprise. Screening for haemoparasitism would also seem to be worthwhile prior to evaluation and/or use of gossypol (or perhaps other potential contraceptives) in human subjects, especially in communities that are confronted with severe food shortages.
Expression of protamine P2 in the testis of the common marmoset and man visualized using non‐radioactive in‐situ hybridization
Authors: SAUNDERS P. T. K. , GAUGHAN J. , SAXTY B. A. , KERR L. E. , MILLAR M. R.
Published: May 2008

Information on the organization of the spermatogenic cycle of the common marmoset (Callithrix jacchus), a small New World primate, is limited to a single histological report on the differentiation of spermatids. In the present study we have used non‐radioactive in‐situ hybridization with a cRNA probe directed against marmoset protamine 2, on fixed sections of marmoset and human testis to elucidate the organization of mature germ cells within the seminiferous epithelium. Specificity of the probe was checked on Northern blots; mP2 hybridized exclusively to mRNA in samples extracted from marmoset and human testis. In sections from human and marmoset testis, positive staining for mRNA was confined to round and elongating spermatids and in the human was reduced in samples from patients with incomplete spermatogenesis. In the human, P2 mRNA was present in groups of cells consistent with the presence of more than one stage of the spermatogenic cycle in transverse sections of individual tubules. In the marmoset, P2‐positive cells were detected as a continuous ring of staining in the majority of sections of tubules whilst in others only a group(s) of cells was positive. We conclude that the arrangement of the spermatogenic wave in this New World primate may be intermediate between that seen in rodents (segmental) and in the human (helical).
Acrosome reaction stimulated by the GTP non‐hydrolizable analogue GTPγS is blocked by phospholipase A2 inhibitors in human spermatozoa
Authors: DOMÍNGUEZ L. , YUNES R. M. F. , FORNÉS M. W. , MAYORGA L. S.
Published: May 2008

Phospholipase A2 (PLA2, EC 3.1.1.4) is involved in the cascade of signalling events leading to the acrosome reaction in human spermatozoa. In order to study the role of PLA2 in the acrosome reaction triggered by GTPγS, a non‐hydrolizable analogue of GTP, two well‐known PLA2 inhibitory reagents were used: dexamethasone (1 mM, a synthetic glucocorticoid), and 2‐(p‐amylcinnamoyl)amino‐4‐chlorobenzoic acid (ONO‐RS‐082, 320 μg/ml). Normal human spermatozoa were incubated for 3 h under capacitating conditions and treated with several reagents [GTPγS, dexamethasone, ONO‐RS‐082, arachidonic acid (AA) and lysophosphatidylcholine (LPC)], alone or in different combinations. In confirmation of earlier reports, GTPγS induced the acrosome reaction. On the other hand, dexamethasone and ONO‐RS‐082 were both able to inhibit the acrosome reaction induced by GTPγS. However, when AA or LPC was added after dexamethasone or ONO‐RS‐082, the acrosome reaction reached values close to those obtained using GTPγS alone. It is concluded that PLA2 probably plays an active role in the acrosome reaction triggered by GTP‐binding proteins.
Systemically administered gonadotrophin‐releasing hormone enhances copulatory behaviour in castrated, testosterone‐treated hyperprolactinaemic male rats
Authors: DENNISON E. , BAIN P. A. , BARTKE A. , MELISKA C. J.
Published: May 2008

Hyperprolactinaemic male rats exhibit deficits in copulatory behaviour which can be reversed by a single injection of GnRH. We tested whether systemically administered GnRH can stimulate copulatory behaviour independently of LH‐mediated increases in plasma testosterone levels. Gonadectomized, pituitary‐grafted adult male Fischer 344 rats bearing implants of 5, 10 or 20 mm capsules of testosterone were administered a single injection of 500 ng GnRH or saline s.c., 30 min prior to copulation tests. Pituitary‐grafted castrates displayed copulatory deficits, relative to sham‐operated castrates with identical levels of testosterone replacement. Administration of 500 ng GnRH to pituitary‐grafted castrates bearing 10 mm testosterone implants significantly increased the proportion of rats that mounted, intromitted and ejaculated during a 30 min test. This treatment also reduced significantly the latency of intromission and ejaculation, and increased significantly the frequency of intromission. The copulatory behaviour of the sexually unresponsive, pituitary‐grafted castrates bearing 5 mm testosterone implants, or of the more sexually responsive castrates bearing 20 mm testosterone implants, was not altered significantly by GnRH injections. These results support the hypothesis that copulatory deficits in moderately hyperprolactinaemic rats are due in part to reduced hypothalamic GnRH release, and suggest that GnRH can stimulate sexual behaviour in these animals via mechanisms that are independent of luteinizing hormone‐induced testosterone release. However, a threshold level of testosterone (achieved with 10 mm implants) appears to be required for GnRH to elicit this effect.
Influence of Escherichia coli on motility parameters of human spermatozoa in vitro
Authors: DIEMER T. , WEIDNER W. , MICHELMANN H. W. , SCHIEFER H.‐G. , ROVAN E. , MAYER F.
Published: May 2008

The influence of E. coli on human sperm motility was studied in vitro. Semen samples were prepared by a swim‐up technique and adjusted to 22 times106 spermatozoa/ml. Samples were then inoculated with different concentrations of a uropathogenic strain of E. coli, serotype 06, with initial sperm/bacteria ratios varying between 10:1 and 10000:1.

Motion parameters were analysed by computer‐aided motility analysis directly, and 2, 4 and 6 h after inoculation. In a second series of experiments, bacterial replication was inhibited by addition of chloramphenicol. In a third series, the effect of E. coli culture filtrates on sperm motility was investigated.

The direct inhibitory effect of E. coli on progressive motility of spermatozoa was found to depend upon the bacterial concentration. A distinct inhibitory effect was observed only at a sperm/bacteria ratio of approximately 1, achieved by growth of E. coli during the experiments. For modality of motion, no distinct changes were observed. When growth of bacteria was prevented by chloramphenicol, no inhibitory effect on sperm motility was detected. Sperm motility was not inhibited by E. coli culture filtrates. Analysis by electron microscopy revealed multiple adhesions of E. coli to spermatozoa, causing variable ultrastructural damage as probable morphological correlates of immobilization.
Epididymal oxytocin in the rat: its origin and regulation
Authors: HARRIS G. C. , FRAYNE J. , NICHOLSON H. D.
Published: May 2008

Oxytocin is localized to the Leydig cells of the testis in the rat and several other species where it is postulated to play a role in steroidogenesis and seminiferous tubule contractility. Oxytocin has also been detected in the epididymis of the ram where active uptake of the peptide from luminal fluid has been demonstrated. This study was performed to investigate whether oxytocin is present in the rat epididymis, and the origin of the peptide. Immunoactive oxytocin was detected in the epididymis of all control animals examined (147.7±41.7 pg/g). Total epididymal oxytocin was reduced significantly following castration (p<0.05). Testosterone treatment also reduced the epididymal concentration of the peptide in both intact and castrated rats. Efferent duct ligation (EDL) did not affect the presence of oxytocin in the epididymis. Immunoactive oxytocin was localized in discrete cells of the epithelium of the caput epididymis, with less staining apparent in the initial segment and cauda epididymis. Staining disappeared from the caput epididymis following castration, but reappeared following testosterone supplementation. No obvious alteration in staining was observed in the cauda epididymis after EDL.

These data demonstrate for the first time the presence of oxytocin in the epididymis of the rat and that the peptide may be regulated by androgens. They further suggest an epididymal source of the peptide.
Smoking and male reproduction: a review
Authors: VINE M. F.
Published: May 2008

Because of the large number of men worldwide who smoke and the fact that cigarette smoke contains known mutagens and carcinogens, there has been concern that smoking may have adverse effects on male reproduction. A review of the epidemiological literature indicates that cigarette smoking is associated with modest reductions in semen quality including sperm concentration, motility and morphology. The associations between male smoking and sperm concentration and motility are stronger among studies of ‘healthy’ men (e.g. volunteers and sperm donors) than among men from infertility clinic populations. Smoking has also been associated with alterations in hormone levels in males, for example increases in the levels of oestrone and oestradiol. Despite modest reductions in semen quality and altered hormone levels among smokers compared to non‐smokers, studies have not shown a reduction in male fertility in association with paternal smoking. There is some evidence to suggest that paternal smoking is associated with congenital anomalies and childhood cancer (with ORs in positive studies generally < 2.0). Smoking has not been shown to be mutagenic to human spermatozoa, although studies have been small and have had methodological shortcomings. It is concluded that, although smokers as a group may not experience reduced fertility, men with marginal semen quality who wish to have children may benefit from quitting smoking, since several small studies indicate the potential for improved semen quality after quitting smoking. More research on the benefits of quitting smoking among men with marginal semen quality is needed. Methodological improvements in studies aimed at clarifying the association between paternal smoking and reproductive and developmental outcomes in offspring should include obtaining accurate paternal smoking dose information, evaluating smoking exposure information in relation to various time windows (e.g. prior to conception, during gestation), and controlling for potential confounders and modifying factors such as age and maternal smoking habits. More sensitive and specific laboratory assays and increased sample sizes are required to establish whether smoking induces mutations in human spermatozoa.
Assessment of sperm motion characteristics in infertile renal transplant recipients using computerized analysis
Authors: EID M. M. , ABDEL‐HAMID I. A. , SOBH M. A. , EL‐SAIED M. A.
Published: May 2008

A total of 34 kidney transplant recipients (18 infertile and 16 fertile) and 31 non‐transplant persons (15 infertile and 16 fertile) were included in this study. All subjects were assessed clinically and by measurement of basal concentrations of total testosterone, FSH, cyclosporine whole blood trough levels, serum creatinine, haemoglobin and semen analysis using computer‐aided sperm analysis (CASA) as well as scrotal ultrasonography to evaluate testicular dimensions. Our results demonstrate a significant decrease (p<0.05) in sperm concentration, the percentage of motile spermatozoa, straight line velocity (VSL), linearity (LIN) and velocity of average path (VAP) among infertile transplant patients in comparison with the fertile transplant group. Serum testosterone, FSH levels and testicular dimensions did not differ significantly (p>0.05) between fertile and infertile transplant recipients. Both sperm concentration and VSL were inversely correlated to the cyclosporine whole blood trough levels (p<0.05). The time spent on haemodialysis was inversely correlated (p<0.05) with the percentage of motile spermatozoa and the amplitude of lateral head displacement (ALH). In conclusion, CASA is valuable in evaluation of sperm motility in infertile renal transplant patients. Stabilization of the cyclosporine whole blood trough level within the target therapeutic level and correction of anaemia (if any) could improve the fertility potential in kidney transplant recipients.
Characterization of human sperm antigens reacting with antisperm antibodies from autologous sera and seminal plasma: comparison among infertile subpopulations
Authors: PARADISI R. , BELLAVIA E. , PESSION A. L. , VENTUROLI S. , BACH V. , FLAMIGNI C.
Published: May 2008

Immunoblotting techniques were used to characterize the reactivity of human sperm antigens with sperm antibodies from different groups of infertile subjects. Sperm antigens of each subject were tested with autologous sperm antibodies present in serum and seminal plasma in order to assess whether there were qualitative differences in the antigenic structure of spermatozoa among the various infertile groups. Sixty‐one infertile males, divided into six groups according to changes in the two main semen parameters (sperm concentration and motility), entered the study. Immunoreactivity to several antigens, in both serum and seminal plasma, was usually demonstrated in infertile subjects. Using a complex statistical package no significant difference was found in the distribution of antigens between groups; this included previously termed ‘fertility‐related antigens’ (28, 35, 57 and 62kDa proteins). This suggests that the most relevant sperm antigens involved in reproduction have no particular relationship to sperm concentration and motility.
Sperm concentrations and quality in β‐thalassaemia major
Authors: JENSEN C. E. , ABDEL‐GADIR A. , COX C. , TUCK S. M. , WONKE B.
Published: May 2008

We report semen analyses in eight patients with β‐thalassaemia major, with poor results in all but the youngest. The causation is multi‐factorial with iron deposition in the pituitary gland resulting from life‐long dependence on blood transfusions being a major factor. We speculate on other contributing causes, but further research is required to elucidate these. Improving haematological care means that these men are increasingly surviving to adult life. Relevant techniques to enable them to achieve their desire for fatherhood are considered.
Detection and enrichment of carcinoma‐in‐situ cells in semen by an immunomagnetic method using monoclonal antibody M2A
Authors: MENG F. J. , ZHOU Y. , SKAKKEBÆK N. E. , MARKS A. , GIWERCMAN A.
Published: May 2008

Carcinoma‐in‐situ (CIS) is a precursor of germ cell cancers in the testis. Early detection and treatment of CIS can prevent the development of invasive cancer. We have developed an immunomagnetic approach for enrichment of CIS cells from semen. Immunomagnetic beads were coated with monoclonal antibody (mAb) M2A, which is specific for seminoma and CIS. In preliminary mixing experiments, cultured human HEY cells, which express the M2A surface antigen, were added to semen samples and recovered selectively by incubation with M2A‐coated immunomagnetic beads followed by magnetic activated cell sorting (MACS). Application of this procedure to ejaculates from five subjects with CIS and 22 control subjects with no evidence of testicular neoplasia resulted in the preferential recovery of a population of large round cells from a 0.5 ml semen sample obtained from CIS subjects (median 1 times 104, range 2 times 103 to 2 times 104) relative to controls (median 2 times 102; range 0 to 103), with no overlap between the two groups (p<0.001). These results suggest that a subpopulation of large round cells recovered from the semen of CIS patients by incubation with M2A‐coated immunomagnetic beads and MACS are CIS cells. Immunomagnetic enrichment of CIS cells in semen will facilitate the definitive identification of exfoliated CIS cells in ejaculates and could be an important first step in a non‐invasive screening procedure for testicular CIS.
Oral health in preschool children with asthma
Authors: STENSSON MALIN , WENDT LILL‐KARI , KOCH GÖRAN , OLDAEUS GÖRAN , BIRKHED DOWEN
Published: May 2008

Objective.  The aim of this study was to investigate oral health and its determinants in 3‐year‐old and 6‐year‐old children with asthma.

Methods and subjects.  Caries and gingivitis were examined in 127 asthmatic (all children with asthma in a selected area and born during a specific time period) and 117 matched, healthy control children.The parents were interviewed regarding various oral‐health‐related factors.

Results.  The mean dfs (± standard deviation) in the 3‐year‐old with asthma was 1.4 ± 3.2 compared with 0.5 ± 1.2 in the controls (P < 0.05). The corresponding figures for the 6‐year‐old were 2.5 ± 3.9 and 1.8 ± 2.8. The 3‐year‐old asthmatic children had more gingival bleeding than the healthy controls (P < 0.05). There were no significant differences in gingivitis in the 6‐year‐old children. Asthmatic children reported higher consumption of sugar‐containing drinks and were more frequently mouthbreathers than healthy children (P < 0.05). In 3‐year‐old children with asthma and immigrant background, the mean dfs was higher compared with immigrant children in the control group (P < 0.01).

Conclusion.  The results indicate that preschool children with asthma have higher caries prevalence than healthy children. The factors discriminating for caries in asthmatic children are higher intake of sugary drinks, mouth breathing, and immigrant background.
The clinically related predictors of dental fear in Taiwanese children
Authors: LEE CHEN‐YI , CHANG YONG‐YUAN , HUANG SHUN‐TE
Published: May 2008

Background.  Dental fear has been singled out as one of the most troublesome problems facing paediatric dentistry today. Children with dental fear may avoid visiting dentists; therefore, their oral health protection is often compromised. However, the aetiology of dental fear is still not entirely understood.

Objective.  This study investigated the dental visiting habit, the previous dental experiences, the conditioning pathway, and the clinically related predictors of dental fear in children.

Design.  The dental history of 247 children (2–10 years old) was obtained when they came to a dental clinic for treatment. The level of dental fear in these children was assessed using the Children's Fear Survey Schedule–Dental Subscale (CFSS‐DS). Observers rated the clinically anxious responses and uncooperative behaviour towards dental treatment in these children. Three stepwise regression analyses were performed to determine significant predictors of CFSS‐DS score, clinically anxious responses, and uncooperative behaviour of children, respectively.

Results.  We found that the CFSS‐DS score and clinical anxiety have different predictors, but age ≤ 3.99 years old and cooperativeness in the first dental visit were important predictors for both the CFSS‐DS score and the clinical anxiety. Furthermore, the other predictors of the CFSS‐DS score were maternal dental fear, unbearable pain during the first dental visit, and visiting dentists in a regular dental clinic; the other predictors of clinical anxiety were first‐born, regular dentist, and CFSS‐DS score. Finally, the only significant predictor for uncooperative behaviour was clinical anxiety.

Conclusion.  Children's dental fear and their anxious response during dental treatment were dynamic processes that consisted of many different factors. The direct conditioning of subjective experience of pain was more important than the objective pathway of child dental fear, and the indirect conditioning does not seem influential in this study sample.
The pharmacogenomics of P‐glycoprotein and its role in veterinary medicine
Authors: MARTINEZ M. , MODRIC S. , SHARKEY M. , TROUTMAN L. , WALKER L. , MEALEY K.
Published: May 2008

Despite advancements in pharmacogenetics in human medicine, the incorporation of pharmacogenetics into veterinary medicine is still in its early stages of development. To date, efforts to understand the pharmacologic impact of genetic variation in veterinary species have largely focused on genes encoding for the membrane transporter, P‐glycoprotein (P‐gp). The emphasis on the role of P‐gp is largely because of safety concerns associated with the use of some macrocyclic lactones in dogs. Because of the body of information available on this topic, we use P‐gp as a platform for understanding the importance of population diversity in veterinary medicine. The impact of P‐gp on drug pharmacokinetics and pharmacodynamics is considered, along with endogenous and exogenous factors that can modulate P‐gp activity. The review includes discussion of how population diversity in P‐gp activity can lead to susceptibility to certain diseases or alter patient response to environmental stress or pharmaceutical intervention. In addition, phenotypic diversity also needs to be considered, as demonstrated by the impact of P‐gp up‐regulation and drug resistance. The aim of this review was to set the stage for further exploration into the impact of genetic and phenotypic variability on drug pharmacokinetics, disease propensity, product formulation and drug response in both companion and food‐producing animals.
Early transient presence of implanted bone marrow stem cells reduces lesion size after cerebral ischaemia in adult rats
Authors: Keimpema E. , Fokkens M. R. , Nagy Z. , Agoston V. , Luiten P. G. M. , Nyakas C. , Boddeke H. W. G. M. , Copray J. C. V. M.
Published: May 2008

Aims: Previous studies on the therapeutic time window for intravascular administration of bone marrow stem cells (BMSCs) after stroke have shown that early intervention (from 3 h after onset) in the middle cerebral artery occlusion (MCAO) rat model is the most effective approach to reduce ischaemic lesion size. We have confirmed these observations but noticed that 2 weeks after transplantation, almost none of the grafted BMSCs could be detected in or around the lesion. The present experiments aimed to assess the fate and kinetics of intravascularly injected BMSCs shortly after administration in correlation to the development of the ischaemic lesion after MCAO. Methods: We administered a syngeneic suspension of complete (haematopoietic and mesenchymal) BMSCs via the carotid artery to rats at 2 h after MCAO onset. We examined the distribution and tissue location of BMSCs within the first 24 h after arterial administration by perfusion‐fixating rats and performing immunohistochemical analysis at different time points. Results: The vast majority (>95%) of BMSCs appeared to become trapped in the spleen shortly after injection. Six hours after implantation, together with the appearance of activated microglia, the first BMSCs could be detected in and around the lesion; their number gradually increased during the first 12 h after implantation but started to decrease at 24 h. The implanted BMSCs were surrounded by activated and phagocytotic microglia. Conclusion: Our results show that ischaemic lesion size reduction can already be achieved by the early transient presence at the lesion site of intravascularly implanted BMSCs, possibly mediated via activated microglia.
Generation and evaluation of movement protein‐specific single‐chain antibodies for delaying symptoms of Tomato spotted wilt virus infection in tobacco
Authors: Zhang M.‐Y. , Zimmermann S. , Fischer R. , Schillberg S.
Published: May 2008

This study investigated whether single‐chain antibodies (scFvs) specific for a viral movement protein could accumulate in the plant cell cytosol and restrict viral systemic infection in plants. Nine chicken scFv fragments against the Tomato spotted wilt virus (TSWV) movement protein (NSM) were isolated by phage display. Soluble scFvs were produced in bacteria and the NSM binding activity of purified scFvs was confirmed. The nine scFv genes were cloned into a plant expression vector enabling recombinant protein accumulation in the plant cell cytosol. Immunoblot analysis demonstrated that two of the nine chicken scFvs accumulated to high levels (5·9 and 8·0% of total soluble protein). Bioassays of viral infection using transgenic tobacco plants producing NSM‐specific chicken scFvs showed delayed symptom development when compared to non‐transgenic control plants, indicating that expression of antibodies recognizing the TSWV movement protein is a potential strategy for generating resistant plants.
Characterization of Tomato curly stunt virus: a new tomato‐infecting begomovirus from South Africa
Authors: Pietersen G. , Idris A. M. , Krüger K. , Brown J. K.
Published: May 2008

The biological and molecular characterization of a virus recognized as a distinct begomovirus species, Tomato curly stunt virus (ToCSV), first observed in South Africa in 1997, is reported here. Whitefly‐transmission and host‐range studies were carried out using a Bemisia tabaci colony identified as the B‐biotype. The experimental host range of ToCSV spanned primarily species in the Solanaceae and Fabaceae. The complete ToCSV genome (2·766 kb) was amplified by PCR, cloned, and the DNA sequence determined. Phylogenetic analysis revealed that ToCSV was most closely related to Tobacco leaf curl Zimbabwe virus (TbLCZV), at 84% nucleotide identity, indicating that ToCSV is a new species in the genus Begomovirus that is probably endemic to southern Africa. The ToCSV genome sequence contained all of the hallmark coding and non‐coding features characteristic of other previously recognized monopartite begomoviruses. ToCSV is only the second begomovirus described from southern Africa that infects solanaceous species. Neither a begomoviral DNA‐B component nor a satellite‐like DNA molecule was detected by PCR in extracts of ToCSV‐infected plants.
Genome‐wide identification of pathogenicity genes in Xanthomonas oryzae pv. oryzae by transposon mutagenesis
Authors: Wang J. C. , So B. H. , Kim J. H. , Park Y. J. , Lee B. M. , Kang H. W.
Published: May 2008

A transposon mutant library was constructed from the bacterial blight pathogen Xanthomonas oryzae pv. oryzae (Xoo) KACC10331 by Tn5 transposon mutagenesis. The susceptible rice cultivar Milyang 23 was inoculated with a total of 24 540 mutants resistant to kanamycin and 67 avirulent or reduced‐pathogenicity mutant strains were selected for study. Southern hybridization verified that 84 mutant strains had single‐copy insertions and their single‐transposon insertion sites were identified by sequencing analysis combined with thermal asymmetric interlaced (TAIL)‐PCR. The single‐transposon‐tagged sequences of 21 mutant strains belonged to pathogenicity‐related genes previously reported in Xanthomonas species, while the other 46 single‐transposon‐tagged sequences included diverse functional genes encoding, five cell‐wall‐degrading enzymes, three fimbrial and flagella assembly regulators, five regulatory proteins, 15 metabolic regulators and 18 hypothetical proteins, which were identified as novel pathogenicity genes of Xoo.
Unravelling the conundrum of river response to rising sea‐level from laboratory to field. Part II. The Fly–Strickland River system, Papua New Guinea
Authors: PARKER GARY , MUTO TETSUJI , AKAMATSU YOSHIHISA , DIETRICH WILLIAM E. , WESLEY LAUER J.
Published: May 2008

The most recent deglaciation resulted in a global sea‐level rise of some 120 m overca12 000 years. A moving boundary numerical model is developed to predict the response of rivers to this rise. The model was motivated by experiments at small scale, which have identified two modes describing the transgression of a river mouth: (i) autoretreat without abandonment of the river delta (no sediment starvation at the topset–foreset break); and (ii) sediment‐starved autoretreat with abandonment of the delta. In the latter case, transgression is far more rapid, and its effects are felt much further upstream of the river mouth. A moving boundary numerical model that captures these features in experimental deltas is adapted to describe the response of the Fly–Strickland River system, Papua New Guinea. In the absence of better information, the model is applied to the case of sea‐level rise without local climate change in New Guinea. The model suggests that: (i) sea‐level rise has forced the river mouth to transgress over 700 km since the last glacial maximum; (ii) sediment‐starved autoretreat has forced enough bed aggradation to block a tributary with a low sediment load and create the present‐day Lake Murray; (iii) the resulting aggradation was sufficient to move the gravel–sand transition on the Strickland River upstream; (iv) the present‐day Fly Estuary may be, in part, a relict river valley drowned by sea‐level rise and partially filled by tidal effects; and (v) the Fly River is presently reforming its bankfull geometry and prograding into the Fly Estuary. A parametric study with the model indicates that sediment concentration during floods plays a key role in determining whether or not, and to what extent, transgression is expressed in terms of sediment‐starved autoretreat. A sufficiently high sediment concentration can prevent sediment‐starved autoretreat during the entire sea‐level cycle. This observation may explain why some present‐day river mouths are expressed in terms of deltas protruding into the sea, and others are wholly contained within embayments or estuaries in which water has invaded landward.
Testing for IgG4 against foods is not recommended as a diagnostic tool: EAACI Task Force Report*
Authors: Stapel Steven O. , Asero R. , Ballmer‐Weber B. K. , Knol E. F. , Strobel S. , Vieths S. , Kleine‐Tebbe J.
Published: May 2008

Serological tests for immunoglobulin G4 (IgG4) against foods are persistently promoted for the diagnosis of food‐induced hypersensitivity. Since many patients believe that their symptoms are related to food ingestion without diagnostic confirmation of a causal relationship, tests for food‐specific IgG4 represent a growing market. Testing for blood IgG4 against different foods is performed with large‐scale screening for hundreds of food items by enzyme‐linked immunosorbent assay‐type and radioallergosorbent‐type assays in young children, adolescents and adults. However, many serum samples show positive IgG4 results without corresponding clinical symptoms. These findings, combined with the lack of convincing evidence for histamine‐releasing properties of IgG4 in humans, and lack of any controlled studies on the diagnostic value of IgG4 testing in food allergy, do not provide any basis for the hypothesis that food‐specific IgG4 should be attributed with an effector role in food hypersensitivity. In contrast to the disputed beliefs, IgG4 against foods indicates that the organism has been repeatedly exposed to food components, recognized as foreign proteins by the immune system. Its presence should not be considered as a factor which induces hypersensitivity, but rather as an indicator for immunological tolerance, linked to the activity of regulatory T cells. In conclusion, food‐specific IgG4 does not indicate (imminent) food allergy or intolerance, but rather a physiological response of the immune system after exposition to food components. Therefore, testing of IgG4 to foods is considered as irrelevant for the laboratory work‐up of food allergy or intolerance and should not be performed in case of food‐related complaints.
Meta‐analysis of nebulized amphotericin B to prevent or treat pulmonary aspergillosis in immunosuppressed animals
Authors: Ho K.M. , Duff O. , Chambers D. , Murray R.
Published: May 2008

Objectives: Pulmonary aspergillosis in immunosuppressed patients is associated with significant mortality and morbidity. We assessed the prophylactic and therapeutic effect of nebulized amphotericin B (AmB) on mortality of immunosuppressed animals with pulmonary aspergillosis in this meta‐analysis.

Methods: Literature search was based on MEDLINE (1966 to January 15, 2007), EMBASE, and Cochrane‐Controlled Trials Register (2006 issue 4) databases. Only randomized‐controlled studies comparing nebulized AmB with placebo on immunosuppressed animals were included. Two reviewers reviewed and extracted the data independently.

Results: Eight studies including 839 immunosuppressed animals were considered. The overall mortality of the immunosuppressed animals with pulmonary aspergillosis treated with nebulized AmB was reduced (69.4% versus 87.7%, odds ratio [OR] 0.12, 95% confidence interval [CI]: 0.08–0.19; P<0.0001). The effect of nebulized AmB on mortality was more significant when the drug was administered as prophylaxis before the inoculation of Aspergillus fumigatus (OR 0.07, 95% CI: 0.03–0.12, P=0.0001; I2=0) than as a therapeutic agent (OR 0.20, 95% CI: 0.11–0.36, P<0.0001; I2=0) (ratio of the 2 ORs=2.86, 95% CI: 1.03–6.11; P=0.04). The effectiveness of AmB desoxycholate (OR 0.16, 0.06–0.38, P<0.0001; I2=0) and lipid‐associated (liposomal, or lipid complex or colloidal dispersion) amphotericin (OR 0.11, 95% CI: 0.06–0.20, P<0.0001; I2=0) was not significantly different (ratio of the 2 ORs=1.46, 95% CI: 0.49–4.39; P=0.50). Three studies evaluated the potential side effects of nebulized amphotericin and showed that there was no significant renal toxicity but the amphotericin desoxycholate had some detrimental effects on pulmonary surfactant function.

Conclusions: The use of nebulized amphotericin, either as a prophylactic or therapeutic agent, is effective in reducing mortality of immunosuppressed animals with pulmonary aspergillosis. A large randomized controlled trial is needed to confirm whether combining nebulized amphotericin with systemic antifungal agents will reduce mortality in immunosuppressed patients with suspected or proven pulmonary aspergillosis.
Male sand crickets trade‐off flight capability for reproductive potential
Authors: SAGLAM I. K. , ROFF D. A. , FAIRBAIRN D. J.
Published: May 2008

In this paper, we test the hypothesis that male sand crickets, Gryllus firmus, experience a trade‐off between flight capability and reproductive potential expressed as reduced testis weight in flight‐capable morphs. We used a half‐sib design with 130 sires, three dams per sire and an average of 5.66 males per dam family, for a total of 2206 F1 offspring. Traits measured were head width, somatic dry weight, testis weight, wing morph (micropterous/macropterous), weight of the dorso‐longitudinal flight muscles (DLM) and the functional status of these muscles. Heritabilities of all traits were significant and ranged from 0.14 to 0.43. All traits were positively correlated with body size, but removal of this covariance revealed a highly significant trade‐off, both phenotypically and genetically, between testes size and flight capability as measured by wing morph, DLM size or DLM status. The possible implications of this for morph‐specific reproductive tactics are discussed.
Kaolin particle films suppress many apple pests, disrupt natural enemies and promote woolly apple aphid
Authors: Markó V. , Blommers L. H. M. , Bogya S. , Helsen H.
Published: May 2008

Multiple applications of hydrophobic kaolin particle film in apple orchards suppressed numbers of blossom weevil (Anthonomus pomorum), brown leaf weevil (Phyllobius oblongus), attelabid weevil (Caenorhinus pauxillus), leafhoppers (Empoasca vitis and Zygina flammigera) and green apple aphid (Aphis pomi) colonies. The kaolin treatments reduced the apple sawfly (Hoplocampa testudinea) fruit infestation on cultivar J. Grieve, and the fruit damage caused by oyster scale (Quadraspidiotus ostreaeformis), mussel scale (Lepidosaphes ulmi), early caterpillars, leaf rolling moths (Tortricidae), fruitlet‐mining tortrix moth (Pammene rhediella) and codling moth (Cydia pomonella). There was no effect on the number of colonies of rosy leaf curling aphid (Dysaphis devecta), nor on the fruit damage caused by common earwig (Forficula auricularia) and apple sawfly on cv. G. Delicious. The level of infestation of rosy apple aphid (Dysaphis plantaginea), leaf miner moths (Phyllonorycter blancardella, Lyonetia clerkella), and agromyzid flies (Phytomyza heringiana) increased in the kaolin‐treated plots. Kaolin treatments promoted woolly apple aphid (Eriosoma lanigerum) infestation, which became severe, while it reduced the abundance of polyphagous predators like F. auricularia, predaceous Heteroptera and Coleoptera, the red velvet mite (Allothrombium fuliginosum), spiders (Araneae) and the abundance of common black ant (Lasius niger). The treatments also reduced parasitism of the apple sawfly by the ichneumonid Lathrolestes ensator. Many weeks after ending the kaolin treatments, the number of predaceous Coleoptera and especially the number of spiders remained low in the kaolin‐treated plots.
Destruxin fromMetarhizium anisopliaeinduces oxidative stress effecting larval mortality of the polyphagous pestSpodoptera litura
Authors: Sree K. Sowjanya , Padmaja V.
Published: May 2008

Nine‐day‐oldSpodoptera litura(Fab.) larvae were treated with crude destruxin (dtx) extracted from a high‐virulent (M‐19) and a low‐virulent (M‐10) isolate of the entomopathogenic fungus,Metarhizium anisopliae(Metch.), at doses that caused 30%, 50% and 90% mortality in the treated groups after 48 h. Destruxins produced a dose‐dependent decrease in the body weight of the larvae after 1, 24 and 48 h of treatment. There was a considerable hike in the activity of lipoxygenase and lipid peroxidation levels in the treated larvae with increased time of exposure to mycotoxin. The activities of total superoxide dismutase, total catalase, total peroxidase and specific ascorbate peroxidase in the larval body also registered alterations in the dtx‐treated larvae, suggesting that exposure of larvae to crude dtx induces oxidative stress which is countered by the antioxidant enzymes to an extent governed by the concentration and time of treatment, beyond which the larvae succumb to the ecofriendly biotoxin.
Efficacy of cultural control measures against the banana weevil, Cosmopolites sordidus (Germar), in South Africa
Authors: De Graaf J. , Govender P. , Schoeman A. S. , Viljoen A.
Published: May 2008

The banana weevil, Cosmopolites sordidus, is the most important insect pest of banana and plantain in the world. Cultural control methods were investigated over 2 years in southern KwaZulu‐Natal, South Africa. Harvesting at ground level and dissection of remnants (treatment 1), and covering the base of the mat (entire plant consisting of several meristems) with soil and moving debris to the inter‐row (treatment 2), were compared to a positive control that involved treatment of plants with a registered pesticide (treatment 3), and a negative control that involved harvesting at 150 cm from the collar with no soil or sanitation amendments (treatment 4). Yield, weevil damage and pseudostem girth of plants were measured from August to November annually, while adult beetle densities were assessed over 4 weeks in October/November and April. Nematode samples were taken and analysed in October/November every year. Damage parameters included the coefficient of infestation, the percentage coefficient of infestation (PCI) at two intervals, the summed PCI value, the percentage cross‐sectional damage of the central cylinder and cortex, and the mean cross‐sectional damage percentage. A randomized block design with three replicates was used in the trial. The parameters were similar before the onset of the trial. Fruit yield and plant girth, corrected by nematode densities, were not significantly different in any treatment, nor were the nematodes controlled. Soil cover and recession of remnants was the only effective treatment, significantly reducing the CI, but not the adult density or the other damage parameters. Soil cover showed promise as a cultural control method because it only needs to be applied seasonally and reduced the percentage cross‐sectional damage of the central cylinder, the damage parameter most closely related to yield, by 14%.
Quantification of the DNA Difference, and Separation of X‐ and Y‐Bearing Sperm in Alpacas (Vicugna pacos)
Authors: Morton KM , Rückholdt M , Evans G , Maxwell WMC
Published: May 2008

Contents

Sperm sexing is an emerging reproductive technology which has been successfully used to produce offspring of a pre‐determined sex in domestic and wildlife species but has yet to be applied to New World camelids. The aims of the present study were to (i) optimize the Hoescht 33342 (H33342) staining concentration for the flow cytometric separation of X and Y chromosome‐bearing alpaca (Vicugna pacos) sperm nuclei, (ii) separate alpaca sperm nuclei into high purity (>90%) populations bearing the X‐ and Y‐chromosome and (iii) determine the DNA difference between X‐ and Y‐bearing sperm in alpacas. Semen was collected from alpacas and sperm nuclei stained with H33342, incubated and analysed using a high‐speed cell sorter (SX‐MoFlo®). H33342 staining concentrations of 36, 54, 72 or 90 μm did not affect the proportion of correctly oriented sperm nuclei (43.3 ± 3.9, 46.4 ± 3.7, 44.5 ± 4.0 and 51.1 ± 2.5% respectively) nor the speed of sorting (1381 ± 160, 1386 ± 123, 1371 ± 133 and 1379 ± 127 sperm nuclei/s). Sort reanalysis determined high levels of purity for X‐ and Y‐enriched populations (96.6 ± 0.7% and 96.1 ± 1.1% respectively). The DNA difference, based on fluorescence intensity (determined by the SX‐MoFlo®), was 3.8 ± 0.06%. These data demonstrate for the first time that alpaca sperm nuclei can be separated into high purity populations and the potential for applying sperm sexing technology to New World camelids.
Bovine Oocyte Quality in Relation to Ultrastructural Characteristics of Zona Pellucida, Polyspermic Penetration and Developmental Competence
Authors: Santos P , Chaveiro A , Simões N , Moreira da Silva F
Published: May 2008

Contents

In the present study, the effect of bovine oocyte quality related to ultrastructural characteristics of zona pellucida (ZP), polyspermic penetration and embryo developmental competence was evaluated. Cumulus–oocyte complexes were punctioned from 453 ovaries, classified as 1, 2, 3 and 4 according to their morphological aspect, matured for 24 h and then divided into two groups. In group A, oocytes were fixed in 2.5% glutaraldehyde and 0.1 m sodium cacodylate and examined under a scanning electron microscope. Photomicrographs were taken and ZP’s pores were evaluated in squares of 6.4‐μm width. In group B, oocytes were fertilized in vitro. After 48 h, non‐cleaved oocytes were fixed for polyspermy evaluation. On days 7, 9 and 10, embryos were classified as developed (blastocysts and hatched blastocysts). Results showed that quality 1 oocytes revealed a ZP pore diameter of 0.50 ± 0.07 μm, which was smaller than the observed on oocytes of quality 2 (0.83 ± 0.10 μm), quality 3 (1.02 ± 0.22 μm) and quality 4 (1.38 ± 0.59 μm) (p ≤ 0.05). For In Vitro Fertilization (IVF), results showed that embryos originating from oocytes classed as 3 and 4 had lower cleavage rate (68.4% and 43.8%) than those belonging to class 1 and 2 (79.5% and 69.3%) (p ≤ 0.05). None oocyte classified as 3 and 4 developed to hatch blastocysts, while for oocytes belonging to quality 1 and 2, these values were, respectively, 15.2% and 12.5%. Concerning polyspermy, oocytes class 1 and 2 had lower polyspermic penetration than those belonging to class 3 and 4 (respectively 4.1%, 4.5%, 11.1% and 9.8%, for class 1, 2, 3 and 4). In conclusion, the present study demonstrated that oocytes with low qualities result in lower developmental competence and with high percentage of polyspermy after IVF, which can be the result of the ZP structure such as the number and the pore’s diameter.
Oviductal Fluid Proteins Associated with the Bovine Zona Pellucida and the Effect on In Vitro Sperm–Egg Binding, Fertilization and Embryo Development
Authors: Gonçalves RF , Staros AL , Killian GJ
Published: May 2008

Contents

Studies have demonstrated that oviductal fluid (ODF) proteins associate with eggs of numerous species including the bovine. In this study, the association of three ODF proteins, the bovine oestrus‐associated protein, osteopontin (OPN), lipocalin‐type prostaglandin D synthase (L‐PGDS), with the bovine zona pellucida (ZP) was demonstrated by immunohistochemistry and western blot. The biological function of ODF derived egg‐associated OPN and L‐PGDS in sperm binding, fertilization and embryonic development was also explored. In vitro matured bovine oocytes were pre‐incubated with ODF collected by cannula from cows in oestrus, or ODF with antibodies to OPN, L‐PGDS and bovine serum albumin (BSA). Following incubation, oocytes were inseminated with 1 × 105 frozen‐thawed spermatozoa, and they were evaluated for sperm binding, fertilization and embryonic development in vitro. Pre‐treatment of ODF with antibodies to all of proteins reduced sperm binding to the ZP and fertilization in vitro. Cleavage rates were not significantly different among incubations, but rates of embryo development were significantly decreased. We conclude that antibodies to OPN, L‐PGDS and BSA react with oocytes incubated with ODF and inhibit sperm binding, fertilization and embryonic development in vitro, suggesting a potential role of these proteins in these events.