Chloroplast ultrastructure and fluorescence response of oilseed rape containing male-sterile radish cytoplasm

Summary The production of hybrid seed is facilitated if one parent possesses a male-sterile cytoplasm. Introduction of the cytoplasm of male-sterile radish (Raphanus sativus L.) into rapeseed (Brassica napus L.) results not only in transfer of the desirable male-sterile trait but induces a chlorophyll defect in the backcrossed male-sterile plants. In this study we show that the defect manifests itself in two different ways in the alloplasmic plants: a) smaller and fewer chloroplasts with an impaired ultrastructure and b) an increase in chlorophyll fluorescence. Defective chloroplasts were characterized by a reduction in both the number and size of grana, the latter due to poor stacking of thylakoids and with frequent discontinuity in the intergranal thylakoid systems. The changed chloroplast morphology and the increase in chlorophyll fluorescence are probably the cause of the lowered photosynthetic efficiency associated with the alloplasmic plants. We propose that the deficiency is the result of incompatibility between the genomes of the radish chloroplast and the rapeseed nucleus. Supporting this hypothesis are studies of male-sterile rapeseed plants in which, by protoplast fusion, the radish chloroplasts were substituted by those of normal male-fertile rapeseed. Such plants showed complete restoration of their photosynthetic potential and displayed both normal chloroplast ultrastructure and normal levels of chlorophyll fluorescence.     

Das Cellulassenzymsystem während Wachstum und Entwicklung von Acanthamoeba castellanii

Zusammenfassung In Extrakten wachsender Kulturen von Acanthamoeba castellanii konnte ein cellulose-abbauendes Enzymsystem nachgewiesen werden. Es besteht aus einer reduzierende Zucker abspaltenden Komponente mit einem pH-Optimum bei 4, einer viscositätsverändernden Komponente mit einem pH-Optimum bei 6 und einer -Glucosidase mit einem pH-Optimum von 3,5. Bei pH 4 sind die Celluloseabbauprodukte Cellobiose und Glucose, bei pH 6 höhermolekulare Oligosaccharide.Während der Entwicklung in einem nährstofffreien Salzmedium nehmen die Cellulaseaktivitäten ab: Vor dem Start der Cellulosesynthese sind noch etwa 30% der ursprünglich vorhandenen Celluloseaktivität nachzuweisen, fertige Cysten besitzen noch etwa 10% der Aktivität.Die Bedeutung des Cellulassenzymsystems wird ausgehend von der Tatsache diskutiert, daß die Excystierung ohne Abbau der Cystenwand, in die die Cellulose eingelagert ist, stattfindet.

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The cellulase enzymes system during growth and development of Acanthamoeba castellanii

It could be shown that extracts of growing cultures of Acanthamoeba castellanii contained a cellulose degrading system. Reducing sugars are split off by one component of this system at an optimum of pH 4, another enzyme changes the viscosity at an optimum of pH 6, and a third component is a -glucosidase with an optimum at pH 3.5. At pH 4 the cellulose degradation products are cellobiose and glucose; at pH 6 higher molecular weight oligosaccharides are produced.During the development from trophozoites to cysts in a nutrient-free medium, the activities of both cellulases decline: Prior to the start of cellulose synthesis only 30%, and in cysts only 10% of the original existing activities are detectable.The biological function of the cellulase enzyme system is discussed together with a consideration of the fact that excystment takes place without digestion of the cyst wall in which the cellulose is deposited.

     

Neuroprotection by Argon Ventilation after Perinatal Asphyxia: A Safety Study in Newborn Piglets

MethodsEight newborn piglets (weight 1.4–3.0 kg) were used. Heart rate, blood pressure, regional cerebral saturation, and electrocortical brain activity were measured continuously. All experiments had a 30 min. baseline period, followed by three 60 min. periods of argon ventilation alternated with 30 min argon washout periods. Two animals were ventilated with increasing concentrations of argon (1h 30%, 1 h 50%, and 1 h 80%), two were subjected to 60 min. hypoxia (FiO₂ 0.08) before commencing 50% argon ventilation, and two animals received hypothermia following hypoxia as well as 50% argon ventilation. Two animals served as home cage controls and were terminated immediately.ResultsArgon ventilation did not result in a significant change of heart rate (mean ± s.d. −3.5±3.6 bpm), blood pressure (−0.60±1.11 mmHg), cerebral oxygen saturation (0.3±0.9%), electrocortical brain activity (−0.4±0.7 µV), or blood gas values. Argon ventilation resulted in elevated argon concentrations compared to the home cage controls (34.5, 25.4, and 22.4 vs. 7.3 µl/ml).ConclusionVentilation with up to 80% argon during normoxia, and 50% argon after hypoxia did not affect heart rate, blood pressure, cerebral saturation and electrocortical brain activity. Clinical safety studies of argon ventilation in humans seem justified.     

Proof of concept: visual categorization of carotenoid pigmentation in Arctic charr (Salvelinus alpinus L) can predict stress response

Carotenoid pigmentation in Salvelinus alpinus has been connected to stress responsiveness in earlier studies. This has, however, only been tested with time-consuming image analysis from photos. Here, we used quick visual categorization of carotenoid pigmentation to investigate the stress responsiveness of the extreme groups. The visually selected charr were then exposed to a net restraint stressor. Arctic charr with few spots also had a lower stress responsiveness compared to charr with many spots. Thus, visual selection could be used as a simple method within aquaculture.     

Phenotypical variation within 22 families with Pompe disease

Background

Pompe disease has a broad clinical spectrum, in which the phenotype is partially explained by the genotype. The aim of this study was to describe phenotypical variation among siblings with non-classic Pompe disease. We hypothesized that siblings and families with the same genotype share more similar phenotypes than the total population of non-classic Pompe patients, and that this might reveal genotype-phenotype correlations.

Methods

We identified all Dutch families in which two or three siblings were diagnosed with Pompe disease and described genotype, acid α-glucosidase activity, age at symptom onset, presenting symptoms, specific clinical features, mobility and ventilator dependency.

Results

We identified 22 families comprising two or three siblings. All carried the most common mutation c.-32-13 T?>?G in combination with another pathogenic mutation. The median age at symptom onset was 33 years (range 1–62 years). Within sibships symptom onset was either in childhood or in adulthood. The median variation in symptom onset between siblings was nine years (range 0–31 years). Presenting symptoms were similar across siblings in 14 out of 22 families. Limb girdle weakness was most frequently reported. In some families ptosis or bulbar weakness were present in all siblings. A large variation in disease severity (based on wheelchair/ventilator dependency) was observed in 11 families. This variation did not always result from a difference in duration of the disease since a third of the less affected siblings had a longer course of the disease. Enzyme activity could not explain this variation either. In most families male patients were more severely affected. Finally, symptom onset varied substantially in twelve families despite the same GAA genotype.

Conclusion

In most families with non-classic Pompe disease siblings share a similar phenotype regarding symptom onset, presenting symptoms and specific clinical features. However, in some families the course and severity of disease varied substantially. This phenotypical variation was also observed in families with identical GAA genotypes. The commonalities and differences indicate that besides genotype, other factors such as epigenetic and environmental effects influence the clinical presentation and disease course.