The salivary flow rate and composition of whole and parotid resting and stimulated saliva in young and old healthy subjects

Resting and stimulated whole and parotid salivary composition and flow rate were examined in 63 healthy volunteers. No significant differences were found between the young and old in secretion rates and salivary concentrations of sodium, potassium, calcium, magnesium, and total protein. The activity of amylase in the resting and stimulated parotid saliva was significantly lower in the old.     

Milk progesterone measurement in dairy cows: Correlation with estrus and pregnancy determination

Progesterone levels in fore milk, determined by a highly specific radioimmunoassay, were compared for the assessment of estrus by a veteran herdsman and an experienced inseminator, in cows presented for insemination. In addition, an examination was made of the relative accuracy of using milk progesterone levels for the determination of pregnancy at 24, 40 and 44 days after insemination, as compared with rectal palpation at 45–50 days post-breeding.Fat-free fore milk progesterone levels were similar to jugular plasma levels at 24 days post-insemination and reached roughly 60% of the level of unextracted fore milk at this time. Accuracy of estrus diagnosis by herdsman, inseminator and milk progesterone level was 84%, 93% and 96%, respectively. For pregnancy diagnosis, milk progesterone determination in 85 cows showed 78% accuracy in predicting pregnancy and 100% accuracy in predicting non-pregnancy. At 40 days post-insemination false positives dropped to 10% and at 44 days only 7% of the cows were incorrectly diagnosed as pregnant. The false positives in this study were largely due to embryonic mortality as reflected by abnormal intervals of return to estrus. Two milk progesterone determinations, at 24 and either 40 or 44 days post-insemination ensure maximum reliability for early pregnancy diagnosis.     

The segregation pattern of a translocation quadrivalent

The segregation pattern of a translocation quadrivalent was studied in three hybrid families of the tetraploid (2n=28) oat, of the Avena strigosa polyploid complex. The rate of IV formation in F₁ was high and the fertility was normal. The adjacent-alternate orientation of this quadrivalent was very variable. Conspicuous variation in this frequency was found between anthers of the same floret and between florets. The alternate type was usually more common toward the end of MI. The adjacent-alternate ratio was found to be an unreliable measure for calculating the type of gametes produced by the F₁ and the F₂ plants derived from them. An attempt was made to determine the type of viable gametes produced on the F₁ and their frequency by examining the cytology of F₂ plants. The various combinations of the chromosomes of the translocation complex expected in the F₂ plants were derived and their expected frequencies calculated by taking into account chiasma formation at the chromosome ends and various restrictions of gamete viability. In none of the three hybrid families F₂ individuals were found to produce trivalents as the most complex chromosome configuration, indicating that one type of gamete derived from adjacent separation was not formed. The 11 ratio between F₂ plants having only bivalents (2II) and those with F ₁-like quadrivalent configuration R(c), expected when gametes resulting from alternate separation are fully functional, with the exclusion of other types, was not found. That ratio 2II/R(c), was 2-1/3 in the various families. The cytology of selected F₃ individuals basically followed the predictions based on chromosome association in the F₂.     

Congenital entropion in a litter of rabbits

Conjunctivitis and blepharospasm were observed in a litter of four, 2-week-old New Zealand white rabbits. Corneal opacity and neovascularization of the cornea and entropion of the upper eyelids were observed when the rabbits were examined 10 weeks later. Conjunctival cultures of these rabbits yielded normal bacterial flora. Treatment with a steroid-antibiotic ophthalmic ointment did not alter the ocular abnormalities. Surgical correction of the entropion resulted in a complete regression of all clinical signs.     

Stable germ line transformation of a leafy vegetable crop amaranth (Amaranthus tricolor L.) mediated by Agrobacterium tumefaciens

We have optimized a procedure for genetic transformation of a major leafy vegetable crop, Amaranthus tricolor L., using epicotyl explant co-cultivation with Agrobacterium tumefaciens. Two disarmed A. tumefaciens strains EHA 105 and LBA 4404, both carrying the binary plasmid p35SGUSINT harboring the neomycin phosphotransferase II gene (nptII) and the β-glucuronidase gene (gus), were evaluated as vector systems. The former displayed a higher transforming efficiency. Several key factors influencing the transformation events were optimized. The highest percentage of transformed shoots (24.24%) was achieved using hand-pricked epicotyl explants, a 10-min infection period, with 100 μM acetosyringone-pretreated Agrobacterium culture corresponding to OD₆₀₀???0.6 and diluted to 10⁹ cells ml^?1, followed by 4 d co-cultivation in the regeneration medium. Putative transformed explants capable of forming shoots were selected on medium supplemented with 75 μg?ml^?1 kanamycin, and transient as well as stable glucuronidase expression was determined by histochemical analysis. From a total of 48 selected shoot lines derived from independent transformation events with epicotyl explants co-cultivated with EHA 105, 32 showed positive PCR amplification for both the nptII and gus genes. Germ line transformation and transgene stability were evident in progeny of primary transformed plants (T₀). Among T₁ seedlings of 12 selected transgenic plant lines, kanamycin-resistant and kanamycin-sensitive seedlings segregated in a ratio typical of the Mendelian monohybrid pattern (3:1) as verified by the chi-square (χ ²) test. Southern hybridization of genomic DNA from kanamycin-resistant T₁ transgenic segregants to an nptII probe substantiated stable integration of the transgene. Neomycin phosphotransferase (NPTII) activity was detected in leaf protein extracts of selected T₁ transgenic plants, thereby confirming stable expression of the nptII gene.     

Structure and Coordination Determination of Peptide-metal Complexes Using 1D and 2D 1H NMR

Copper (I) binding by metallochaperone transport proteins prevents copper oxidation and release of the toxic ions that may participate in harmful redox reactions. The Cu (I) complex of the peptide model of a Cu (I) binding metallochaperone protein, which includes the sequence MTCSGCSRPG (underlined is conserved), was determined in solution under inert conditions by NMR spectroscopy.NMR is a widely accepted technique for the determination of solution structures of proteins and peptides. Due to difficulty in crystallization to provide single crystals suitable for X-ray crystallography, the NMR technique is extremely valuable, especially as it provides information on the solution state rather than the solid state. Herein we describe all steps that are required for full three-dimensional structure determinations by NMR. The protocol includes sample preparation in an NMR tube, 1D and 2D data collection and processing, peak assignment and integration, molecular mechanics calculations, and structure analysis. Importantly, the analysis was first conducted without any preset metal-ligand bonds, to assure a reliable structure determination in an unbiased manner.     

Beyond reified categories: multidimensional identifications among ‘black’ and ‘Indian’ groups in Columbia and Mexico

Analyses of multicultural state-dictated social categories are often governed by those same categories, even while they deconstruct them. Nonetheless, these categories are often used in public spheres such as national imaginary or ethno-political activism. Taking a different point of departure, that of representations rather than the categories themselves, the aim of our paper is to understand the modes of classification that are relevant among four populations in Colombia and Mexico who would, a priori, be categorized as ‘black’ or ‘Indian’. The daily reality of these groups indicates other possible internal, sometimes even intersecting, kinds of categorizations, which, far from naturalizing the ‘Indian’ and ‘black’ categories, in fact reveal place-based social identifications. These identifications seem closer to the everyday lives and practices of the people in question, and underscore the local conceptions of their presence and agency in a given spot.     

ALFY-Controlled DVL3 Autophagy Regulates Wnt Signaling,Determining Human Brain Size

Primary microcephaly is a congenital neurodevelopmental disorder of reduced head circumference and brain volume, with fewer neurons in the cortex of the developing brain due to premature transition between symmetrical and asymmetrical cellular division of the neuronal stem cell layer during neurogenesis. We now show through linkage analysis and whole exome sequencing, that a dominant mutation in ALFY, encoding an autophagy scaffold protein, causes human primary microcephaly. We demonstrate the dominant effect of the mutation in drosophila: transgenic flies harboring the human mutant allele display small brain volume, recapitulating the disease phenotype. Moreover, eye-specific expression of human mutant ALFY causes rough eye phenotype. In molecular terms, we demonstrate that normally ALFY attenuates the canonical Wnt signaling pathway via autophagy-dependent removal specifically of aggregates of DVL3 and not of Dvl1 or Dvl2. Thus, autophagic attenuation of Wnt signaling through removal of Dvl3 aggregates by ALFY acts in determining human brain size.