Semen cryopreservation in golden‐headed lion tamarin,Leontopithecus chrysomelas

Wild animal genetic resource banking (GRB) represents a valuable tool in conservation breeding programs, particularly in cases involving endangered species such as the golden‐headed lion tamarin (Leontopithecus chrysomelas). Thus, we aimed to assess a sperm freezing protocol for golden‐headed lion tamarins using two different exenders: BotuBOV® (BB) and Test Yolk Buffer® (TYB). Ejaculates were collected by penile vibrostimulation from animals housed at São Paulo Zoological Park Foundation, São Paulo, Brazil, and after immediate analysis, two aliquots were diluted in BB and TYB. Postthawing samples were evaluated for total and progressive motility, plasma membrane and acrosome integrities, mitochondrial activity, susceptibility to oxidative stress, and sperm–egg‐binding. No differences between BB and TYB were found for most seminal parameters, except for acrosome integrity and susceptibility to oxidative stress (in both cases BB showed higher values). However, in spite of these differences and regardless of the extender used, postthaw sperm motility and viability with the described protocol were encouraging (on average >50% and >80%, respectively), indicating that sperm cryopreservation may be a short‐term measure for the conservation of golden‐headed lion tamarins.     

Lung T cells in hypersensitivity pneumonitis: phenotypic and functional analyses

Cells recovered from bronchoalveolar lavage (BAL) and tissue sections from transbronchial lung biopsies were studied in 16 patients with symptomatic hypersensitivity pneumonitis (HP) and in six subjects with a similar history of exposure but without features of disease by using a series of monoclonal antibodies (MoAb) detecting different lymphocyte subpopulations, including T and T subsets, B lymphocytes, and natural killer (NK) cells. Their functional activities in cytotoxic and suppressor assays and the microenvironment in the lung by using immunohistological techniques were also evaluated. It has been demonstrated that the majority of cells recovered from BAL of HP patients are represented by T8 lymphocytes, with a relevant imbalance of the T4/T8 ratio (p less than 0.001). HNK-1+ cells were markedly increased (p less than 0.001), whereas the frequency of cells bearing other NK-related markers (NK-15, VEP 13, Ab8.28, T10, M1, and Fc gamma R) were not significantly increased with respect to controls. Immunohistological study confirmed that the majority of cells infiltrating lung parenchyma are T8+ lymphocytes. The number of HNK-1+ cells detected on lung biopsies was very low in all cases, even in patients with the highest values on BAL suspensions. The evidence of cells bearing the proliferation-associated markers (Tac and T9 antigens) seems to support the hypothesis of a local proliferation in the lung. In terms of phenotypic analysis, the results observed in the group of asymptomatic individuals are qualitatively superimposable on those observed in the HP group, but the magnitude of the phenomenon is less prominent and therefore the data are not as statistically significant as that produced by the comparison between HP patients and the same controls. Functional analysis of BAL T cells from both HP patients and asymptomatic individuals showed suppressor activity in vitro, as determined by the ability to influence a pokeweed mitogen (PWM)-driven B cell differentiation assay. BAL cells from HP patients were also able to display a definite cytotoxic function in vitro, whereas BAL lymphocytes from asymptomatic subjects did not. Taken together, these data demonstrated that cells responsible for the alveolitis in patients with HP are characterized by the expansion of T cells with the phenotype and functions of both suppressor and/or cytotoxic lymphocytes. This expansion is likely to be related to a local immunologic response to the antigenic stimulus and may provide new insights into the pathogenetic mechanisms of this disease, its pathological pattern, and its management.     

The effect of post-heparin lipolytic activity on plasma lipoproteins

Summary Electron microscopy of negatively stained plasma shows that intravenous administration of heparin to healthy humans results in a fast degradation of very low density lipoproteins (VLDL), which are very rich in triglycerides (50% w/w). VLDL are also rapidly digested by post-heparin lipolytic activity when serum from non-heparinized subjects is incubated at 27°C in vitro with post-heparin serum of healthy controls.Abbreviations VLDL very low density lipoproteins - LDL low density lipoproteins - HDL high density lipoproteins - LP-X lipoprotein-X, abnormal lipoprotein characterizing obstructive jaundice     

Die Beziehung zwischen Mitochondrienmorphologie und Aktivitätsdauer verschiedener Flugmuskelfasern von Locusta migratoria L.

Zusammenfassung Die indirekten und direkten Flugmuskelfasern der Wanderheuschrecke enthalten parallel und voneinander getrennt verlaufende Myofibrillen. In den interfibrillären Räumen liegen zahlreiche, säulenförmig aneinandergereihte Mitochondrion, ein dichtes endoplasmatisches Retikulum und Tracheolen. In Höhe der A-I-Band-Grenzen befindet sich ein charakteristisches retikuläres System, dessen Queranordnung als Diade zu bezeichnen ist. Die chromatinreichen Kerne liegen peripher dicht unter dem Sarkolemm.Sowohl bei den indirekten als auch bei den direkten Flugmuskelgruppen kommt es im Verlaufe des Dauerfluges zu Transformationen der Mitochondrien, die sich in einer erheblichen Schwellung und in einer Umwandlung der Cristae zu Tubuli äußern. Diese Umwandlung wird als intramitochondriale Kompensation aufgefaßt, um das Verhältnis von Oberfläche der inneren membranösen Phase zum Volumen der inneren wäßrigen Phase — d. h. die spezifische Oberfläche — möglichst konstant zu halten. Eine Vesikulation der Tubuli könnte der morphologische Ausdruck für eine Dekompensation sein, da dann der Zusammenhang der äußeren wäßrigen Phase verlorengeht.Die Mitochondrien verschiedener Muskeln reagieren nicht synchron auf die Dauerbelastung, sondern ausgesprochen belastungs- und funktionsspezifisch: Zunächst transformieren sich die Mitochondrien der indirekten, dann erst die der direkten Flugmuskelgruppen, und der anfängliche Schwirrflug geht während fortschreitenden Dauerfluges in einen Flatterflug über. Die unterschiedlichen Erschöpfungszeitpunkte der indirekten Flugmuskeln werden durch die jeweils spezifische Funktion und die ontogenetische Entwicklung erklärt.     

Use of sequence characterised amplified region and RAPD markers in the identification of the white truffle Tuber magnatum Pico

Isolates of white truffles were identified as Tuber magnatum Pico species using a pair of primers selected from a sequence characterised amplified region (SCAR) and a specific random amplified polymorphic DNA (RAPD) marker. The present study reveals that PCR-fragment-pattern polymorphisms, the construction of probes and couples of primers from one or more of these polymorphic fragments may provide a useful and rapid tool for identifying species of ectomycorrhizal fungi in addition to conventional methods (morphological parameters).     

Evaluation by electron microscopy of the integrity of iodinated plasmalipoproteins

Summary Iodination of proteins and lipoproteins is a widely used in vitro labelling procedure in metabolic, autoradiographic and various other studies. However, all available iodination techniques have involved the possible damage to the proteins by self-irradiation, oxidizing agents, the alkaline milieu or by the introduction of iodine into the molecular structure itself. To evaluate the integrity of iodinated lipoproteins, we observed the electron microscopic appearance of normal and iodinated rabbit very low density lipoproteins (VLDL) by negative staining with phosphotungstic acid. Iodination up to a molar iodine/protein ratio of 2.89 did not result in any change of shape, size or aggregating tendency of the particles. No stacks or disk-like particles like those of various hyperlipoproteinemic states were found. We conclude that electron microscopy is a valuable tool in assessing the morphological appearance of lipoprotein iodination, but it should be complemented by other techniques.