Cytological evidence of a new male-sterile mutant in soybean (Glycine max (L.) Merr.)

Summary From one plant of soybean (Glycine max (L.) Merr.) with only two one-seeded pods, found in an F₄ population maintained by single-seed descent procedure, two fully fertile plants were obtained which, in turn, produced two progeny segregating for male sterility. Segregation ratios, observed on progeny from fertile plants in three successive generations, indicated that the male-sterility trait was under the control of a single recessive gene. Cytological observations made on malesterile, female-fertile plants showed the occurrence of a complete and properly timed cytokinesis with the formation of tetrad cells whose size was very variable, one of which sometimes had two nuclei. During pollen maturation binucleate microspores and grains with reduced size (micropollens) were frequently observed. Massive pollen degeneration occurred at a rather later stage. Structural evidence points to a normally functioning tapetum.On the basis of these cytological observations we conclude that the abnormalities observed in the mutant we studied have to be considered to be different from those caused by any other known ms allele. Tests of allelism with other sources of male sterility are in progress.     

Induction of micronuclei in polychromatic erythrocytes from mouse bone marrow following intraperitoneal administration of halogenated benzenes

The mutagenicity of halogenated benzenes, including three isomers of tri- and tetrachlorobenzenes (TCB, TeCB) was studied on male Swiss CD1 mice by MN test. The data presented show that all the halogenated benzenes tested were found to be clastogenic apart from 1,2,3,5-TeCB. No significant differences were observed in the clastogenic activities of TCB-isomers and TeCB-isomers.     

Nuove ricerche sull'orientamento e il senso del tempo di Arctosa perita (Latr.) (Araneae lycosidae)

Zusammenfassung Die Uferspinne Arctosa perita (Latr.) verfügt über einen astronomischen Orientierungsmechanismus, durch den die Tiere imstande sind, wenn sie auf dem Wasser ausgesetzt werden, in der Richtung nach dem Ufer zu fliehen. Die Spinnen orientieren sich auf Grund des Sonnenstandes und des polarisierten Himmelslichtes und haben die Fähigkeit, die Tageszeit einzukalkulieren (Papi 1955b und c).Wenn eine Gruppe von Tieren gefangengehalten wird, dann nimmt bei den Fluchtversuchen die Streuung der Fluchtrichtungen zu. Dabei ist die Streuung der gesamten Fluchtversuche — wenigstens während der ersten 16 Tage — statistisch nicht verschieden, ob die Tiere nun a) in den natürlichen Belichtungs- und Temperaturschwankungen, b) im Dauerdunkel und in den Temperaturschwankungen oder c) im Finstern unter konstanter Temperatur gehalten werden. Was die Genauigkeit der Richtungsorientierung betrifft, so bleibt, trotz der stärkeren Streuung, die durchschnittliche Fluchtrichtung bei Gruppe a) bis zu 21 Tage lang korrekt, während sie bei den Gruppen b) und c) von der theoretischen Richtung immer mehr abweicht.Tiere, die im Finstern unter konstanter Temperatur gefangengehalten werden, orientieren sich bezüglich einer unbeweglichen Lampe bei verschiedenen Tageszeiten ungefähr so, wie wenn sie die Sonne wäre.Exemplare, die 3 Tage lang einem gegen den natürlichen Tag um 6 Std verschobenen Belichtungsrhythmus ausgesetzt werden, nehmen Orientierungswinkel an, die zur Zeit ihres künstlichen Tages korrekt wären.Ein innerer Tagesrhythmus (innere Uhr) regelt die Abweichung des Orientierungswinkels der Tiere. Im Laufe des Tages ändert sich der Orientierungswinkel nicht mit einer konstanten Geschwindigkeit, sondern mit einer solchen, die die Azimutgeschwindigkeit der Sonne auszugleichen sucht.Wenn die Tiere einige Stunden bei einer Temperatur von 4–5°C oder in 2°C gehalten werden, dann orientieren sie sich so, wie es einige Stunden vorher korrekt wäre. Der Gang der inneren Uhr kann also durch sehr niedrige Temperaturen verzögert oder gestoppt werden.Unter experimentellen Bedingungen können die Tiere in 8–10 Tagen neue Fluchtrichtungen erlernen.In der Besprechung werden die Resultate mit jenen verglichen, die bei anderen, einer astronomischen Orientierung fähigen Tieren erhalten wurden.

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Dedicato al Prof. Karl von Frisch in occasione del suo 70 compleanno.     

The folding process of acylphosphatase from Escherichia coli is remarkably accelerated by the presence of a disulfide bond

The acylphosphatase from Escherichia coli (EcoAcP) is the first AcP so far studied with a disulfide bond. A mutational variant of the enzyme lacking the disulfide bond has been produced by substituting the two cysteine residues with alanine (EcoAcP mutational variant C5A/C49A, mutEcoAcP). The native states of the two protein variants are similar, as shown by far-UV and near-UV circular dichroism and dynamic light-scattering measurements. From unfolding experiments at equilibrium using intrinsic fluorescence and far-UV circular dichroism as probes, EcoAcP shows an increased conformational stability as compared with mutEcoAcP. The wild-type protein folds according to a two-state model with a very fast rate constant (k_F^H2O = 72,600 s^− 1), while mutEcoAcP folds ca 1500-fold slower, via the accumulation of a partially folded species. The correlation between the hydrophobicity of the polypeptide chain and the folding rate, found previously in the AcP-like structural family, is maintained only when considering the mutant but not the wild-type protein, which folds much faster than expected from this correlation. Similarly, the correlation between the relative contact order and the folding rate holds only for mutEcoAcP. The correlation also holds for EcoAcP, provided the relative contact order value is recalculated by considering the disulfide bridge as an alternate path for the backbone to determine the shortest sequence separation between contacting residues. These results indicate that the presence of a disulfide bond in a protein is an important determinant of the folding rate and allows its contribution to be determined in quantitative terms.     

Structure of PAK1 in an autoinhibited conformation reveals a multistage activation switch

The p21-activated kinases (PAKs), stimulated by binding with GTP-liganded forms of Cdc42 or Rac, modulate cytoskeletal actin assembly and activate MAP-kinase pathways. The 2.3 A resolution crystal structure of a complex between the N-terminal autoregulatory fragment and the C-terminal kinase domain of PAK1 shows that GTPase binding will trigger a series of conformational changes, beginning with disruption of a PAK1 dimer and ending with rearrangement of the kinase active site into a catalytically competent state. An inhibitory switch (IS) domain, which overlaps the GTPase binding region of PAK1, positions a polypeptide segment across the kinase cleft. GTPase binding will refold part of the IS domain and unfold the rest. A related switch has been seen in the Wiskott-Aldrich syndrome protein (WASP).     

Distributional niche of relatively rare sable antelope in a South African savanna: habitat versus biotic relationships

The geographic distribution of a species is governed by climatic conditions, topography, resources and habitat structure determining the fundamental niche, while the local distribution expressed via home range occupation may be compressed by biotic interactions with competitors and predators, restricting the realised niche. Biotic influences could be especially important for relatively rare species. We investigated how rainfall, geology, land type and abundance of other ungulate species serving as competitors or prey for predators contributed to the patchy distribution of sable antelope herds within Kruger National Park. Data were provided by annual aerial surveys of ungulate populations conducted between 1978 and 1988. Sable herds were more commonly present on granitic and sandstone substrates than on more fertile basalt. They occurred both in the moist south‐west and dry north of the park. They were most abundant in sour bushveld and mopane savanna woodland, and mostly absent from knob thorn‐marula parkland. The presence of sable was negatively associated with high concentrations of impala and wildebeest, less consistently related to the abundance of zebra, and positively associated with the occurrence of buffalo herds. Best supported models included the separate effects of the most abundant grazers along with land type. Interspecific relationships seemed more consistent with vulnerability to predation as the underlying mechanism restricting the distribution of sable herds than with competitive displacement. Sable favoured land types distinct from those where wildebeest, the most preferred prey of lions, and impala, numerically the most important resident prey species, were most abundant. Hence the risk of predation, associated with habitat conditions where abundant prey species are most concentrated, can exert an overriding influence on the distribution of rarer species in terms of their home range occupation.     

The intrachain disulfide bridge is responsible of the unusual stability properties of novel acylphosphatase from Escherichia coli

Acylphosphatase (AcP) activity in prokaryotes was classically attributed to some aspecific acid phosphatases. We identified an open reading frame for a putative AcP in the b0968 Escherichia coli gene and purified the recombinant enzyme after checking by RT-PCR that it was indeed expressed. EcoAcP has a predicted typical fold of the AcP family but displays a very low specific activity and a high structural stability differently from its mesophilic and similarly to its hyperthermophilic counterparts. Site directed mutagenesis suggests that, together with other structural features, the intrachain S–S bridge in EcoAcP is involved in a remarkable thermal and chemical stabilization of the protein without affecting its catalytic activity.     

Increased sensitivity of the neuronal nicotinic receptor alpha 2 subunit causes familial epilepsy with nocturnal wandering and ictal fear

Sleep has traditionally been recognized as a precipitating factor for some forms of epilepsy, although differential diagnosis between some seizure types and parasomnias may be difficult. Autosomal dominant frontal lobe epilepsy is characterized by nocturnal seizures with hyperkinetic automatisms and poorly organized stereotyped movements and has been associated with mutations of the alpha 4 and beta 2 subunits of the neuronal nicotinic acetylcholine receptor. We performed a clinical and molecular genetic study of a large pedigree segregating sleep-related epilepsy in which seizures are associated with fear sensation, tongue movements, and nocturnal wandering, closely resembling nightmares and sleep walking. We identified a new genetic locus for familial sleep-related focal epilepsy on chromosome 8p12.3-8q12.3. By sequencing the positional candidate neuronal cholinergic receptor alpha 2 subunit gene (CHRNA2), we detected a heterozygous missense mutation, I279N, in the first transmembrane domain that is crucial for receptor function. Whole-cell recordings of transiently transfected HEK293 cells expressing either the mutant or the wild-type receptor showed that the new CHRNA2 mutation markedly increases the receptor sensitivity to acetylcholine, therefore indicating that the nicotinic alpha 2 subunit alteration is the underlying cause. CHRNA2 is the third neuronal cholinergic receptor gene to be associated with familial sleep-related epilepsies. Compared with the CHRNA4 and CHRNB2 mutations reported elsewhere, CHRNA2 mutations cause a more complex and finalized ictal behavior.     

Home range occupation and habitat use of sable antelope in the Okavango Delta region of northern Botswana

Animals selectively utilize their environments within a hierarchical framework. Our study addressed how the home ranges of sable antelope selectively incorporated the landscape and habitat types available to them. It was conducted in a region of northern Botswana where the sable population was expected to be thriving, in contrast to their threatened status in the wild in South Africa. The movements and habitat use of three neighbouring sable herds were recorded by global positioning system (GPS) telemetry during parts of the seasonal cycle in a region adjoining the seasonally flooded Okavango Delta. Total home range extents covered by these herds were larger than those found for sable in other areas, and local population densities, taking into account the herd sizes, were accordingly lower than in these other areas. Access to surface water appeared to be the main limitation on seasonal home range occupation. Almost all of the local vegetation types were utilized, but sable herds generally favoured dryland grassland during the dry season and floodplain grassland during the wet season, contrary to what we had expected. Hence, it appeared that local home range occupation and habitat use by the three sable antelope herds could be influenced more by interactions with potential competitors and predators than by intrinsic habitat suitability.     

Apoptosis mapping in space and time of 3D tumor ecosystems reveals transmissibility of cytotoxic cancer death

The emerging tumor-on-chip (ToC) approaches allow to address biomedical questions out of reach with classical cell culture techniques: in biomimetic 3D hydrogels they partially reconstitute ex vivo the complexity of the tumor microenvironment and the cellular dynamics involving multiple cell types (cancer cells, immune cells, fibroblasts, etc.). However, a clear bottleneck is the extraction and interpretation of the rich biological information contained, sometime hidden, in the cell co-culture videos. In this work, we develop and apply novel video analysis algorithms to automatically measure the cytotoxic effects on human cancer cells (lung and breast) induced either by doxorubicin chemotherapy drug or by autologous tumor-infiltrating cytotoxic T lymphocytes (CTL). A live fluorescent dye (red) is used to selectively pre-stain the cancer cells before co-cultures and a live fluorescent reporter for caspase activity (green) is used to monitor apoptotic cell death. The here described open-source computational method, named STAMP (spatiotemporal apoptosis mapper), extracts the temporal kinetics and the spatial maps of cancer death, by localizing and tracking cancer cells in the red channel, and by counting the red to green transition signals, over 2–3 days. The robustness and versatility of the method is demonstrated by its application to different cell models and co-culture combinations. Noteworthy, this approach reveals the strong contribution of primary cancer-associated fibroblasts (CAFs) to breast cancer chemo-resistance, proving to be a powerful strategy to investigate intercellular cross-talks and drug resistance mechanisms. Moreover, we defined a new parameter, the ‘potential of death induction’, which is computed in time and in space to quantify the impact of dying cells on neighbor cells. We found that, contrary to natural death, cancer death induced by chemotherapy or by CTL is transmissible, in that it promotes the death of nearby cancer cells, suggesting the release of diffusible factors which amplify the initial cytotoxic stimulus.