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W B Schill

Publications and source records attributed to W B Schill.

At least 19 recordsLinked to original sources

Classification of cutaneous malignant melanoma: a reassessment of histopathologic criteria for the distinction of different types.

BACKGROUND: Human cutaneous malignant melanoma currently is classified into four principle types: nodular, superficial spreading, lentigo maligna, and acral lentiginous. The criteria for the histopathologic diagnosis of these types are not applied consistently. Nevertheless, the classification has become the foundation of many clinical, histopathologic, epidemiologic, and molecular studies. The results of those studies can have validity only if the classification itself is valid. For this reason, the authors reassessed histopathologic criteria advocated for the distinction of the different types of melanoma and searched for other repeatable constellations of findings that may serve to define distinct subsets of the neoplasm. METHODS: Nine hundred fifteen melanomas were examined with regard to 72 parameters that are considered to be important for histopathologic diagnosis. The results were analyzed statistically with special attention to findings that have been reported to be characteristic of the four principle types of melanoma. RESULTS: The histopathologic criteria advocated for the distinction of different types of melanoma were found not to correlate with one another. A logistic regression analysis did not detect any other repeatable constellation of morphologic findings that may reflect a distinct biologic subgroup. CONCLUSIONS: The validity of the current classification of cutaneous malignant melanoma into four principle types could not be substantiated. Malignant melanoma may present with many different forms, but these forms appear to be part of a continuous spectrum rather than examples of distinct biologic entities.

Adult

Use of failed-fertilized oocytes for diagnostic zona binding purposes after sperm binding improvement with a modified medium.

PURPOSE: Because the availability of prophase oocytes for zona binding testing is limited, we compared sperm binding to the zona of failed-fertilized intracytoplasmic sperm injection (ICSI) and in vitro fertilization (IVF) oocytes after incubation in a standard IVF medium and a specially composed binding improvement medium. METHODS: Semen samples from nine patients and nine fertile donors were separated in parallel by the standard swim-up method in both media. Subsequently, hemizona assays were performed with prophase, failed-fertilized ICSI and IVF oocytes. RESULTS: Sperm separation resulted in a significantly higher sperm count (P < 0.01) and progressive motility (P = 0.018) in binding improvement medium. Moreover, spermatozoa coincubated with hemizonae (prophase, failed-fertilized ICSI and IVF oocytes) in binding improvement medium bound significantly more to hemizonae than in the controls (P < 0.01). However, the hemizona index did not differ. CONCLUSIONS: Thus, the limited number of human zonae can be increased by the use of oocytes that failed to fertilize during ICSI or IVF. This will lead to a qualitative improvement of the diagnostic spectrum in male-factor infertility.

Adolescent

Reactive oxygen species influence the acrosome reaction but not acrosin activity in human spermatozoa.

It is now widely accepted that the higher levels of reactive oxygen species (ROS) produced by damaged or deficient spermatozoa are associated with a loss of motility and a decreased capacity for sperm-oocyte fusion. Furthermore, earlier studies show, under physiological conditions, that some ROS may be involved in capacitation and hyperactivation of human spermatozoa. We measured ROS levels, acrosome reaction (AR) and acrosin activity (AA) in semen samples from suspected subfertile men to reveal the influence of ROS on AR and AA of human spermatozoa. Semen samples were obtained from 60 patients. Samples with > or = 1 x 10(6) leukocytes/mL were excluded from the study. ROS production was determined using a chemiluminescence technique. AR was determined using a triple stain technique. The percentage of acrosome-reacted spermatozoa after low temperature induction of the AR (test value), and the inducibility of AR (= the difference between the test value and the control), were calculated. The AA was analysed by determining the proteolytic potential of spermatozoa on gelatin plates. The mean halo diameter and percentage of halo formation in each sample were measured as AA parameters. Scatter plots of ROS levels and AR parameters showed that the percentage of acrosome reacted spermatozoa and AR inducibility were better in samples with low rather than high ROS levels. On the other hand, there were no apparent similarities between ROS and the AA parameters. Therefore, the percentage of acrosome-reacted spermatozoa and AR inducibility were significantly higher in the low than in the high ROS group (p = 0.028, p = 0.0001, respectively). In addition, there was no significant difference in AA parameters between groups. These findings suggest that lower ROS in semen may have a role in AR but excessive ROS may exert a negative influence on AR, while ROS in semen has no relationship to AA.

Acrosin

Species specificity of human and murine anti-ZP3 synthetic peptide antisera and use of the antibodies for localization and identification of ZP3 or ZPC domains of functional significance.

The mammalian zona pellucida has an important function in the fertilization process. The zona pellucida protein 3 (ZP3 or ZPC) is the ligand for primary sperm binding and induces the acrosome reaction. In various species, ZP3 primary structures are highly conserved as revealed by cDNA cloning. The objective of these studies was to localize ZP3 protein using antisera generated against defined synthetic peptides that are specific for mouse or for human ZP3. Immunohistochemistry and transmission electron microscopy were applied to murine and human ovary sections. Immunochemical studies were performed in hemizonae pellucidae from microbisected human oocytes. Using the competitive hemizona assay and various anti-ZP3 antibodies, we further intended to identify human ZP3 epitopes of functional significance. Our results showed that antiserum AS ZP3-9 (mouse specific) detected mouse ZP3 protein in mouse oocytes and in immunoblots, whereas AS ZP3-14 (human specific) detected human ZP3 protein in human ovary sections, native hemizonae pellucidae and in immunoblots. ZP3 material was also detected in cumulus cells by immunohistochemistry. Ultrastructural studies showed an equal distribution of ZP3 throughout the zona pellucida. The human competitive hemizona assay revealed that none of the anti-ZP3 synthetic peptide antisera affected sperm binding suggesting that those epitopes are not involved in primary sperm binding. Anti-porcine ZP3 beta protein antibodies (polyclonal) blocked human sperm-zona pellucida binding. In summary, these anti-ZP3 synthetic peptide antibodies specifically reacted with intact ZP3 protein (murine and human) but did not inhibit human sperm-zona pellucida binding; anti-ZP3 antibodies can therefore be used as biomarkers for ZP3 localization and function.

Animals

[From neuropeptides and cytokines to psychotherapy. Skin diseases between psychoneuroimmunology research and psychosomatic treatment].

Most therapists recognize that emotional factors influence the course of neurodermitis (ND). Most studies concerning the influence of experimental, acute stressors brought only inconsistent results in ND. Most likely, the experimental stressors chosen thus far were only able to poorly simulate the reactions to real stressors in neurodermitis patients. More recent articles, however, were able to demonstrate changes in psychoimmunological parameters under experimental stress conditions in ND in comparison with a control group of persons with healthy skin. Retrospective studies, serial analyses over time and psychotherapeutic evaluation studies were able to demonstrate even more clearly that emotional factors influence the course of ND.

Complementary Therapies

Different cumulative pregnancy rates in patients with repeated IVF- or ICSI cycles: possible influence of a male factor.

The low rate of ongoing pregnancies in IVF cycles leads to a high number of repeated cycles in couples with previously failed attempts. Therefore it would be helpful to have a prediction about the chance of becoming pregnant in a repeated cycle. In a retrospective study the data of about 4246 cycles were analysed. Because the pregnancy rates in IVF- and ICSI cycles are generally different, these two groups were distinguished between and the outcome in patients with one, two or more attempts was analysed. The rate of ongoing pregnancies per patient was lower after IVF (24.9%) than after ICSI (32.9%), but was similar or even slightly increased in patients with more than one attempt. On the other hand, there was a high pregnancy rate with ICSI in the first two cycles (35.9%), but patients with more than two ICSI cycles had a significantly lower chance of becoming pregnant (20.7%). Factors that are known to influence the pregnancy rate, such as stimulation protocol, oocyte quality or number of transferred embryos, were similar in all groups. However, significantly reduced embryo quality with successive cycles was only observed in ICSI patients. There might be a negative selection of patients with poor embryo quality and previously failed attempts after ICSI, possibly due to an andrological factor. The differences between IVF- and ICSI patients are based on treatment indications, and andrological diseases are the predominant indication for ICSI. Although no correlation was found between changes in conventional sperm parameters and number of treated cycles, there might be a subgroup of andrological patients selected by repeatedly failed ICSI cycles. Chromosomal or genetic disturbances in spermatozoa of this subgroup could be the reason for failure.

Adult

Performance of serum-free broth media for growth of Renibacterium salmoninarum.

Growth of Renibacterium salmoninarum was compared in 14 different broth media; 13 serum-free, and 1 that contained newborn calf serum, KDM2+M. Supplementation with 1% v/v R. salmoninarum MCO4M metabolite was evaluated for 6 of the media that do not utilize it as part of their ingredients. Viable cells were enumerated on Days 10, 20, and 30 post inoculation to evaluate performance. The experiment was repeated 3 times using high, low, and medium (trials 1 to 3, respectively) cell concentrations as inoculum. In general there was no optimal medium and all performed well. The choice of which to employ depends on the ease of preparation and presence of certain ingredients that might affect subsequent assays. In trials 2 and 3, the pH was estimated using test papers at the same time as cells were counted. Maximum pH increase occurred with KDM2+M and those media containing charcoal. For most media, a simple pH determination could be used as a means to check that growth has occurred in a culture, particularly if charcoal was added directly to the media and a visual inspection could not be made to detect growth.

Analysis of Variance

[Balneophotochemotherapy in disseminated granuloma annulare].

A 56-year-old female patient with generalized granuloma anulare was treated with bath-PUVA therapy with 8-methoxypsoralen. Her diabetes mellitus renal insufficiency excluded oral photochemotherapy. A series of 30 treatments resulted in complete disappearance of the lesions on the trunk and neck and significant improvement of those on the limbs. No side-effects were observed. Bath-PUVA therapy seems to have positive effects not only on epidermal, but also on dermal processes.

Balneology

Influence of gossypol on the secretory function of cultured rat sertoli cells.

Cottonseed gossypol is a potent male contraceptive in several mammalian species including man. Sertoli cells play a crucial role in spermatogenesis. Therefore, the antifertility competence of gossypol may reflect a change in Sertoli cell function. Rat primary cultures were used to examine the effect of gossypol on cell viability, mitochondrial dehydrogenase function, lactate production and secretion of the Sertoli cell-specific protein inhibin. Exposure for 24 h to gossypol (3-6 microM) significantly enhance secretion of lactate but reduce secretion of inhibin without affecting cell viability. At 9-15 microM, the observed decrease of both lactate and inhibin accumulation apparently resulted from Sertoli cell degeneration and death, because viability and mitochondrial function were also reduced. The results suggest that mitochondria of Sertoli cells are a possible target for gossypol-induced infertility.

Animals

Production and characterization of monoclonal antibodies to the major protein of boar outer dense fibers.

Outer dense fibers (ODF) are structural elements in the mammalian sperm tail which surround the axoneme in the midpiece and principal piece and probably may help to maintain the elastic structures and elastic recoil of the sperm tail. In the present study, we have generated and characterized and describe a series of monoclonal antibodies (mAbs) against the 30 kDa major protein from boar ODF. For antibody screening an ELISA was developed using a newly developed method to fix the ODF proteins to the solid phase. A total of seven mAbs were selected and characterized by ELISA, Western blot and immunofluorescence. The mAbs recognize the major protein component of boar ODF on preparative Western blot and mark the mid- and principal piece of demembranated flagella. These mAbs also recognize the mid- and principal piece of demembranated human spermatozoa from normozoospermic patients, but not from those with asthenozoospermia. For the first time, we succeeded in obtaining hybridoma cell lines that secrete mAbs of class IgM, which react with the 30 kDa protein of boar ODF.

Animals

Omega-3 fatty acid-based lipid infusion in patients with chronic plaque psoriasis: results of a double-blind, randomized, placebo-controlled, multicenter trial.

BACKGROUND: Profound changes in the metabolism of eicosanoids with increased concentrations of free arachidonic acid (AA) and its proinflammatory metabolites have been observed in psoriatic lesions. Free eicosapentaenoic acid (EPA) may compete with liberated AA and result in an antiinflammatory effect. OBJECTIVE: Our purpose was to assess the efficacy and safety of intravenously administered fish-oil-derived lipid emulsion on chronic plaque-type psoriasis. METHODS: A double-blind, randomized, parallel group study was performed in eight European centers. Eighty-three patients hospitalized for chronic plaque-type psoriasis with a severity score of at least 15 according to the Psoriasis Area and Severity Index (PASI) participated in a 14-day trial. They were randomly allocated to receive daily infusions with either a omega-3 fatty acid-based lipid emulsion (Omegavenous; 200 ml/day with 4.2 gm of both EPA and docosahexaenoic acid (DHA); 43 patients) or a conventional omega-6-lipid emulsion (Lipovenous; EPA+DHA < 0.1 gm/100 ml; 40 patients). The groups were well matched with respect to demographic data and psoriasis-specific medical history. Efficacy of therapy was evaluated by changes in PASI, in an overall assessment of psoriasis by the investigator, and a self-assessment by the patient. In one center neutrophil 4- versus 5-series leukotriene (LT) generation and platelet 2- versus 3- thromboxane generation were investigated and plasma-free fatty acids were determined. RESULTS: The total PASI score decreased by 11.2 +/- 9.8 in the omega-3 group and by 7.5 +/- 8.8 in the omega-6 group (p = 0.048). In addition, the omega-3 group was superior to the omega-6 group with respect to change in severity of psoriasis per body area, change in overall erythema, overall scaling and overall infiltration, as well as change in overall assessment by the investigator and self-assessment by the patient. Response (defined as decrease in total PASI of at least 50% between admission and last value) was seen in 16 of 43 patients (37%) receiving the omega-3 emulsion and 9 of 40 patients (23%) receiving omega-6 fatty acid-based lipid emulsion. No serious side effects were observed. Within the first few days of omega-3 lipid administration, but not in the omega-6 supplemented patients, a manifold increase in plasma-free EPA concentration, neutrophil leukotriene B5 and platelet thromboxane B3 generation occurred. CONCLUSION: Intravenous omega-3-fatty acid administration is effective in the treatment of chronic plaque-type psoriasis. This effect may be related to changes in inflammatory eicosanoid generation.

Adult

Ultrastructural localization of angiotensin-converting enzyme in ejaculated human spermatozoa.

Angiotensin-converting enzyme (ACE) is known to be released from human spermatozoa during capacitation. However, it has not yet been localized ultrastructurally in ejaculated sperm cells. Therefore, the purpose of the present study was to demonstrate the location of ACE by means of immunoelectron microscopy and direct immunofluorescence. In addition, ACE activity of spermatozoa was correlated with standard semen parameters. The activity of angiotensin-converting enzyme was measured in spermatozoa from 115 donors and patients attending the andrological outpatient department. Progressive motility was negatively correlated with sperm ACE activity (Spearman rank correlation r=-0.364, P < 0.0001), whereas no statistically significant correlations with sperm concentration, total motility and morphology were observed. Immunoelectron microscopy demonstrated that ACE is mainly located at the plasma membrane of the acrosomal region, equatorial segment, postacrosomal region and midpiece. In contrast, only weak ACE-like immunoreactivity was found at the flagellum. In cases of cells with missing plasma membranes ACE seems also to be located at the surface of the outer acrosomal membrane. By means of immunohistochemical methods, different patterns of ACE-like immunofluorescence were observed: (i) fluorescence of the acrosome or the entire sperm head, midpiece and flagellum; (ii) fluorescence of the postacrosomal region, midpiece and flagellum; (iii) bright fluorescence of the equatorial segment with less intensive labelling of the postacrosomal region and flagellum. Induction of the acrosome reaction by calcium ionophore A23187 resulted in an increase of spermatozoa with weak acrosomal fluorescence, indicating loss of the plasma membrane.

Acrosome

Influence of urogenital infections on sperm functions.

Many studies have examined the impact of genital tract infections on male fertility; however, the effect of bacteriospermia on sperm quality is still controversial. Bacterial infections are more frequently found in semen samples from asymptomatic infertile patients than in those from fertile men. Bacteriospermia is also a common problem of male partners from couples undergoing IVF. Therefore, the effects of microorganisms on human sperm acrosome reaction of oocytes have been studied in vitro and in vivo. Incubation of spermatozoa with Escherichia coli or Mycoplasma hominis in vitro resulted in reduced sperm motility and inducibility of acrosome reaction (delta AR) after exposure to calcium ionophore A23187. To show possible effects of E. coli and mycoplasma species on sperm functions in vivo, data from 488 patients were evaluated, in whose ejaculates microbiological examinations and determinations of acrosome reaction after exposure to low temperature had been performed. U. urealyticum and E. coli were found in semen samples from 52 and 31 men, respectively. M. hominis was only present in a minor number of samples and was not included in this study. Semen concentrations of E. coli and U. urealyticum ranged between 500-100,000 cfu x ml-1 and 100-80,000 cfu x ml-1. No correlation was found between delta AR and concentration of bacteria (Spearman rank correlation coefficient, E. coli: r-0.081, P = 0.6644; U. urealyticum: r = -0.081, P = 0.5698). In 69% of cases with U. urealyticum infection and reduced inducibility of acrosome reaction, this sperm function was normal after antibiotic therapy. However, improvement of acrosomal function may only be due to intra-individual variations of acrosome reaction. While E. coli and mycoplasma species affect sperm functions in vitro, the present data and a review of the literature fail to demonstrate similar effects in vivo.

Acrosome

Sperm separation in patients with urogenital infections.

A total of 196 patients attending the Center of Dermatology and Andrology, Giessen, Germany, were examined for fertility problems. Polymorphonuclear elastase, reactive oxygen species (ROS) and the number of round and peroxidase-positive cells were investigated in addition to routine semen analysis. The ejaculates were also analysed before and after sperm separation by means of swim-up or glass wool filtration. In 20 cases of leukocytospermia, sperm concentration, motility, viability, production of reactive oxygen species, and the number of peroxidase-positive cells were evaluated before and after glass wool filtration. The results show that ROS production by viable spermatozoa is highly correlated with the concentration of PMN elastase and the number of both peroxidase-positive and round cells. Multiple regression analysis with motility as dependent parameter showed the number of round cells (n = 91; r = -0.332; P = 0.0030) to be the most important parameter affecting motility, while ROS mainly affects the viability of spermatozoa (n = 69; r = 0.250; P = 0.0107). In the case of leukocytospermia, glass wool filtration significantly reduced the number of peroxidase-positive cells and ROS production (P = 0.0098 and P = 0.0005, respectively). Receiver operating characteristic (ROC) curve analysis for ROS production in the ejaculate using a concentration of 1.000 ng ml-1 PMN elastase as decisive parameter resulted in a cut-off value of 49,489.9 counts 10(-7) viable spermatozoa. The statistical parameters were: Sensitivity: 63.2%, specificity: 100%, positive predictive value: 100%, negative predictive value: 36.1%.

Cell Separation

Evaluation of ZP2 domains of functional importance with antisera against synthetic ZP2 peptides.

The mouse zona pellucida protein ZP2 plays an important role in the process of fertilization by mediating secondary sperm binding to mammalian oocytes. ZP2 primary structures are highly conserved as revealed by cDNA cloning. The aim of the study was to identify ZP2 domains of functional relevance. Antisera were raised against synthetic peptides that are either conserved in the structure of ZP2 from different mammalian species (AS ZP2-20) or present in the human ZP2 but not in the mouse ZP2 amino acid sequence (AS ZP2-26). Antibody binding to zona pellucida proteins was assessed by assaying the antisera with human hemizonae. Using human zonae pellucidae, we demonstrated that anti-ZP2 common antibodies and anti-ZP2 human peptide antibodies react with human zona pellucida antigens. For the first time, ZP2 domains of functional relevance for human sperm-oocyte interaction could be identified applying the competitive hemizona assay. Antiserum AS ZP2-20 significantly inhibited binding of spermatozoa to test hemizonae, whereas treatment of hemizonae with AS ZP2-26 did not influence sperm-oocyte interaction. These results show that antibodies against synthetic ZP2 peptides react with ZP2 protein and that AS ZP2-20 identifies a linear ZP2 epitope that is of possible functional importance for sperm-oocyte interaction.

Amino Acid Sequence