Pregnancy after heterotopic heart transplant removal.

Heterotopic heart transplants were introduced in 1974. The technique allows the patient's native heart to be preserved in situ, alongside the transplanted heterotopic donor heart. We present the case of a nulliparous woman who underwent heterotopic heart transplant in infancy, and subsequent explantation of the donor heart eleven years later, when her native heart function recovered. In adulthood the patient attended pre-pregnancy counselling and was awaiting cardiac magnetic resonance imaging when she presented pregnant at 6 weeks-of-gestation. She attended the joint cardiac obstetric and anaesthetic clinic, where she was reviewed monthly and had bi-monthly echocardiograms. At 35 weeks-of-gestation she was admitted to hospital with preeclampsia. After blood pressure control and steroid administration, a category 3 caesarean delivery under spinal anaesthesia was performed. To our knowledge this is the first case report describing pregnancy in a patient with a removed heterotopic heart transplant.

Understanding biofilm in practice: a global survey of health professionals.

OBJECTIVE: The aim of this survey was to examine health professionals' views and practices relating to biofilm in chronic wounds. METHOD: A global online survey was conducted to assess the current understanding of biofilm and wound management practices. The survey consisted of 20 questions designed to evaluate health professional knowledge of biofilm, perception and understanding of biofilm behaviour, detection and diagnosis, and treatment. Respondents were classified as 'specialists' if wounds were their primary focus and they developed protocols and determined formularies. Respondents were classified as 'generalists' if wounds were part of multiple indications they treat and they were able to choose wound care products from a restricted list of products. The Pearson's chi-square or Fisher's exact test was used to assess whether the responses were independent of the clinician role, health-care setting and country. RESULTS: Overall, 3011 health professionals took part in the survey, of which 397 were excluded or disqualified. Of the remaining 2614 respondents, 1223 (46.8%) completed the entire survey. Although the majority of health professionals were aware of biofilm, knowledge gaps regarding its prevalence in chronic wounds were evident. In general, the majority indicated that they understood that biofilm is detrimental to wound healing. With the exception of wound stalling, there was a lack of consensus on other clinical signs in the detection and diagnosis of biofilm. Knowledge gaps were also evident over the treatment of biofilm and the efficacy of antimicrobial treatments, debridement and wound dressing. CONCLUSION: Our results show that though there is a broad recognition of biofilm and its possible role in chronic wounds, there is still a need to educate and increase knowledge on recognition and treatment of biofilm.

Mesendoderm cells induce oriented cell division and invagination in the marine shrimp Sicyonia ingentis.

The mesendoderm (ME) cells are the two most vegetal blastomeres in the early developing embryo of the marine shrimp Sicyonia ingentis. These two cells enter mitotic arrest for three cycles after the 5th cell cycle (32-cell stage) and ingress into the blastocoel at the 6th cycle (62-cell stage). Circumjacent to the ingressing ME cells are nine presumptive naupliar mesoderm (PNM) cells that exhibit a predictable pattern of spindle orientation into the blastopore, followed by invagination. We examined the role of ME cells and PNM cells in gastrulation using blastomere recombinations and confocal microscopy. Removal of ME progenitors prevented gastrulation. Removal of any other blastomeres, including PNM progenitors, did not interfere with normal invagination. Altered spindle orientations occurred in blastomeres that had direct contact with one of the ME cells; one spindle pole localized to the cytoplasmic region closest to ME cell contact. In recombined embryos, this resulted in an extension of the region of ME-embryo contact. Our results show that ME cells direct the spindle orientations of their adjacent cells and are consistent with a mechanism of oriented cell division being a responsible force for archenteron elongation.

[Measurement of the deposition of salbutamol in the bronchial lavage fluid of infants and young children]. / Bestimmung der Salbutamol-Deposition mittels Bronchiallavage bei Säuglingen und Kleinkindern.

BACKGROUND: Inhaled salbutamol is well known to be effective in obstructive lung disease in the paediatric age group. However, the individually necessary dose cannot be predicted in a single patient. Two inhalation techniques were compared measuring the concentrations of salbutamol in the bronchial lavage fluid. PATIENTS AND METHODS: Forty nine children, age 2 to 73 months, inhaled salbutamol immediately before bronchoscopy, either 200 microg using a metered dose inhaler (MDI) or 2000 microg using a nebulizer (NEB). The concentrations of salbutamol were measured in the bronchial lavage fluid. RESULTS: In spite of different inhaled doses the concentrations of salbutamol were similar in the nebulizer group (n = 29) and the metered dose inhaler group (n = 20) (medians 160 vs. 163 ng/ml; p = 0.27) with a wide range in both groups (12 to 1139 and 5.2 to 641 ng/ml, respectively). In a single patient the concentrations differed by the factor 159 between the right and the left lung. The concentrations of salbutamol were not age dependent (p = 0.06 for NEB, p = 0.28 for MDI). CONCLUSIONS: In infants and young children the concentrations of salbutamol in the bronchial lavage fluid vary widely after a single inhalation using either a metered dose inhaler or a nebulizer.

Sperm Penetration of the Vitelline Envelope of Sicyonia ingentis Eggs is Mediated by a Trypsin-like Lysin of Acrosomal Vesicle Origin: (sperm/fertilization/shrimp/vitelline envelope/protease).

The sperm of the decapod crustacean Sicyonia ingentis are nonmotile, unistellate cells. At spawning, mated females release both stored sperm and eggs. The sperm bind, via the tip of their anterior appendage, to the egg's vitelline envelope (VE), rapidly undergo acrosomal exocytosis, and penetrate the VE. In the present study we used protease inhibitors to show that sperm penetration of the VE is due to the activity of a sperm trypsin-like protease(s). Sperm extracts contained several proteases when examined using gelatin-substrate SDS-PAGE, with two major bands of relative molecular weight 46 kD and 30 kD. Using fluorescent peptidyl-MCA substrates, sperm extract showed trypsin-like and aminopeptidase-like activities, but no chymotrypsin-like activity. Sperm extracts were found to degrade isolated VEs. Using soybean trypsin inhibitor and anti-inhibitor antibodies, protease was localized at the light and electron microscope levels to the acrosome remnants of reacted sperm.

Two Extracellular Matrices From Oocytes of the Marine Shrimp Sicyonia ingentis that Independently Mediate Only Primary or Secondary Sperm Binding: (sperm/fertilization/shrimp/gamete binding/extracellular matrix).

During spawning, female Sicyonia ingentis simultaneously release ova and stored nonmotile sperm and mix them externally to initiate gamete interaction. Sperm bind to a thin vitelline envelope (VE) via their anterior appendage and within seconds are induced to undergo acrosomal exocytosis. The sperm penetrate the VE and become secondarily bound to the surface coat (SC), a glycocalyx on the oocyte surface. In this study, both extracellular matrices were isolated from S. ingentis oocytes. Isolated VEs mediated only primary sperm binding (i.e., before the acrosome reaction), while the isolated SCs mediated only secondary sperm binding (i.e., after acrosomal exocytosis). Isolated S. ingentis VEs were used to characterize primary sperm binding activity. The two extracellular matrices differ morphologically and possess different polypeptide profiles. Soluble fractions of isolated VEs inhibited primary sperm binding in a concentration dependent manner, and immunolocalization of VE components demonstrated highly localized VE binding sites at the tip of the sperm anterior appendage by which sperm bind eggs. Extensive Pronase digestion of VE components did not affect sperm binding activity of solubilized VE components, while complete deglycosylation with trifluoromethanesulfonic acid destroyed sperm binding activity. However, neither alkaline treatment nor enzyme digestion using glycosidases specific for asparagine and serine/ threonine linked oligosaccharides affected sperm binding activity.

Protease-Induced Formation of the Sperm Acrosomal Filament.

Filament extension during the sperm acrosome reaction in Sicyonia ingentis is triggered by an egg trypsin-like protease whose action can be mimicked using trypsin. Using biotinylated trypsin and either a fluorescently-labeled or colloidal gold-labeled antibody to biotin, trypsin binding was localized to the anterior granule of the sperm which is exposed upon acrosomal exocytosis. The binding was to proteinaceous material at the base of the granule juxtaposed to the inner acrosomal membrane. Other labeled proteins also bound in the same pattern but only in the presence of unlabeled trypsin; non-proteolytic proteins did not induce filament formation. Binding of all proteins tested occurred slowly over a period of about 30 min. A minimum of 30 min of trypsin exposure was required in order to trigger filament formation, and increasing trypsin concentration did not reduce this time requirement. These results indicate that the protease slowly uncovers a binding site for itself (or other proteins), and then its proteolytic activity is again required to induce filament formation. The protease kallikrein appeared to be a more potent inducer than trypsin, while thrombin and clostripain had no apparent inducing activity.

Sperm of the Shrimp Sicyonia ingentis Undergo a Bi-Phasic Capacitation Accompanied by Morphological Changes.

Sperm removed from seminal receptacles of female Sicyonia ingentis can be induced to undergo a bi-phasic acrosome reaction (AR), acrosomal exocytosis followed by filament formation, using egg water (EW). Sperm removed from males will not undergo any phase of the AR when incubated with EW, indicating that these sperm undergo a capacitation process after insemination. Freshly molted females (functional virgins) were placed in aquaria with males and monitored for copulation. Mated females were isolated and allowed to carry sperm for specific periods of time. At these time points, sperm were removed and assayed for the ability to undergo the AR using EW. The results indicate that sperm are competent to undergo acrosomal exocytosis after approximately 25 hr, while competency to form acrosomal filaments is not achieved until around 145 hr post-insemination. Morphological examination of sperm removed from males and sperm removed from females revealed dramatic differences. Microscopic evidence indicates that some of the morphological changes seen during capacitation are necessary for the successful completion of the AR.

Congenital adrenal hyperplasia. I: Gender-related behavior and attitudes in female patients and sisters.

Thirty-five female patients with congenital adrenal hyperplasia (CAH) were compared to a group of 16 healthy sisters in regard to gender-related behavioral patterns, present attitudes, and plans for the future. A semi-structured interview with the subjects, ages 11 to 41 yr, and their mothers concentrated on four to five age stages. Results of retrospective data from single items as well as from several related composite scales ("interests and behavior," "appearance," "overall scores") revealed significant group differences: Both in mother-assessment and self-assessment, CAH patients showed a "more masculine" orientation than their sisters, but this was far from consistent across all age stages, especially for single items. Unexpectedly, the gender-behavior differences between CAH patients and sisters did not hold for certain items and scales of "social behavior" (e.g., assertiveness, dominance, acceptance in peer groups) and, in contrast to some of the existing literature, also not for "high-energy expenditure." With regard to expectations for the future, CAH patients had less of a "wish to have their own children" and a higher preference for "having a career versus staying at home." Age, socioeconomic status, intelligence, and presence or absence of a sister as possibly intervening psychosocial/demographic factors could not explain the group differences in behavior. Degree of genital masculinization (Prader stages) or "onset and quality" of therapy as measures of pre- and postnatal androgenization, respectively, could also not account for the degree of the "more masculine" orientation in the CAH group. Nevertheless, the overall results are compatible with earlier findings on the masculinizing effects of prenatal androgens on behavior in humans and point to a time period after sexual differentiation of the genitalia and before birth as the most likely one for the effects of prenatal hormones on behavioral masculinization in humans.

Congenital adrenal hyperplasia. II: Gender-related behavior and attitudes in female salt-wasting and simple-virilizing patients.

The salt-wasting (SW) and simple-virilizing (SV) forms of congenital adrenal hyperplasia (CAH) are characterized by distinct prenatal hormonal milieus. To test whether these hormonal milieus differentially influence the development of a "more masculine" behavioral pattern in female CAH patients (Dittmann et al., 1990), SW patients (N = 13) were compared both to SV patients (N = 20) and healthy sisters of both groups (N = 16). The data are based on semi-structured interviews in which subjects (11-41 yr) and mothers were asked about aspects of "Gender-related interests and behavior," "Level of activity," "Social behavior," (reflecting e.g., assertiveness, dominance, and acceptance by peer groups) and "Appearance"; these areas of interest were represented by composite scales. On most scales, and by both mother-assessment and self-assessment, SW patients differed significantly from both SV patients and sisters in having a "more masculine" orientation. SW patients also showed a higher "Level of activity." These SW group results probably account for much of the CAH/sister differences reported in the companion article (Dittmann et al., 1990). In contrast, SV patients differed from the sister sample on only a few scales. There were no significant differences between SV and SW subjects in the degree of virilization of the external genitalia (indicating no group difference in prenatal androgenization). SW patients were treated "earlier" and "better" after birth (indicating less postnatal androgenization). However, these medical conditions, as well as several psychosocial/demographic variables, could not explain the group behavioral differences. These results do not support a primarily psychosocial explanation of behavioral development in CAH patients, especially those with the SW condition; they rather suggest differential organizational effects of two different hormonal environments (SV vs. SW) during critical periods of prenatal CNS development.

PHYSIOLOGICAL AND BIOCHEMICAL INVESTIGATIONS OF THE EGG JELLY RELEASE IN PENAEUS AZTECUS.

Following contact with seawater, Penaeus aztecus ova undergo a massive release of extracortical jelly precursor material which is transformed into a layer of jelly-like material surrounding the ova. Release and dissipation of the precursors can be irreversibly inhibited by the protease inhibitors N-a-p-tosyl-L-lysine chloromethyl ketone and soybean trypsin inhibitor, implicating trypsin-like proteases in the process. Treatment with the less-specific enzyme inhibitor phenylmethyl sulfonyl fluoride also irreversibly inhibits the release of the cortical material. Jelly precursor in whole mature ovaries stain positive with PAS. Staining with alcian blue reveals acid mucopolysaccharides in the investment coat of the ova but not in the jelly precursors. Precursors isolated from whole mature ovaries are approximately 25-30% carbohydrate (anthrone sulfuric acid reaction) and 70-75% protein (Lowry's and Bradford's protein determinations). No sialic acids are detected in the isolates (thiobarbituric acid assay). Trypsin is effective in dissipating the precursor isolates. Amino acid analysis reveals high ratios of cysteic acid. Significant biochemical differences between P. aztecus egg jelly material and sea urchin egg jelly are discussed.

Excystation and culturing of human and animal Giardia spp. by using gerbils and TYI-S-33 medium.

Mongolian gerbils were used as an animal model to excyst and host Giardia spp. isolated from meadow voles, dogs, beavers, and humans. Both cysts and trophozoites were used to establish infections. Gerbils were infected with Giardia duodenalis from beaver, dog, and human sources, and the trophozoites were extracted and cultured in Diamond TYI-S-33 medium. The use of gentamicin and ampicillin in the medium, coupled with treatment of gerbils with gentamicin before they were sacrificed, permitted the elimination of trophozoite purification techniques before culturing. An extract of whole bovine calf blood, CLEX, was substituted for fetal bovine serum in TYI-S-33 medium and was found to be both adequate and less expensive.

Spermiogenesis in the Marine Shrimp, Sicyonia ingentis: (penaeidae/sperm/spermiogenesis).

Spermiogenesis in the marine prawn Sicyonia ingentis was examined using transmission electron microscopy. The acrosomal vesicle, derived from the fusion of pro-acrosomal vesicles blebbed from the nuclear envelope, contains the membrane pouches, anterior granule and a spike. The anterior granule is formed from the coalescence of granular aggregates within the proacrosomal vesicles. Primordia underlying the apical acrosomal vesicle membrane polymerize to form a spike approximately 6 µm long. The convoluted pouch membranes arise from the posterior acrosomal vesicle membrane. Lateral and apical portions of the acrosomal vesicle are surrounded by a pentalaminar membrane comprised of the spermatid plasma membrane and the acrosomal vesicle membrane. Subacrosomal structures include the dense saucer plate, granular core and crystalline lattice. These components condense just posterior to the acrosomal vesicle and are separated from the chromatin by a nuclear plate. The spermatid nucleus becomes surrounded by rough endoplasmic reticulum (RER) and membranous lamellar bodies. RER gives rise to smooth endoplasmic reticulum. These membrane systems degenerate, forming a band of reticular elements around the lateral and posterior portions of the nucleus. The nucleus undergoes condensation followed by decondensation with concomitant breakdown of the nuclear envelope. The resultant chromatin is fibrillar in appearance.

[Areas of psychosocial care of children with cancer and their families. Evaluation of a model of comprehensive care]. / Bereiche psychosozialer Betreuung krebskranker Kinder und ihrer Familien. Auswertung eines ganzheitlichen Versorgungsmodells.

98 children with cancer and 7 children with other serious hematologic or oncologic illnesses and their families are counselled by psychologists in close teamwork with pediatric oncologists within the framework of a clinical pilot study in 1982. The problems and questions of interest posed on the psychologists in 646 such contacts are classified and presented according to frequency of problems and contacts. The resulting large number and variety of concerns demonstrates that in addition to the direct illness- and treatment-related coping strategies (52 %) and related social/economic questions (30 %), a considerable portion of the problems can be classified within the realm of the clinical psychologist: behavioral disorders (37 %), family problems (33 %), late effects (26 %).