Expression of PHT1 family transporter genes contributes for low phosphate stress tolerance in foxtail millet (Setaria italica) genotypes.

MAIN CONCLUSION: This is a first comprehensive study to analyze the 12 PHT1 family phosphate transporter genes in 20 foxtail millet genotypes for the improvement of millets and other crops for phosphate use efficiency. Phosphorus (P), absorbed from soil solutions as inorganic phosphate (Pi), is a limiting nutrient for plant growth and yield. Twenty genotypes of foxtail millet (Setaria italica) with contrasting degree of growth and Pi uptake responses under low Pi (LP) and high Pi (HP) supply were chosen based on a previous study. To gain molecular insights, expression dynamics of 12 PHosphate Transporter 1 (PHT1) family (SiPHT1;1 to 1;12) genes were analyzed in these 20 genotypes and compared with their Pi and total P (TP) contents. SiPHT1;1, 1;2, 1;3 and 1;8 genes were expressed in shoot tissues of three (ISe 1209, ISe 1305 and Co-6) of the LP best performing genotypes (LPBG); however, they were expressed in only one of the LP worst performing genotype (LPWG) (ISe 748). More importantly, this is correlating with higher shoot Pi and TP contents of the LPBG compared to LPWG. Apart from this condition, expression of SiPHT1 genes and their Pi and TP contents do not correlate directly for many genotypes in other conditions; genotypes with low Pi and TP contents induced more SiPHT1 genes and vice versa. Promoter analysis revealed that genotype ISe 1888 with a high level of SiPHT1;8 expression possesses two additional root box motifs compared to other genotypes. The PHT1 family genes seem to play a key role for LP stress tolerance in foxtail millet and further studies will help to improve the P-use efficiency in foxtail millet and other cereals.

Structure, Function, Regulation and Phylogenetic Relationship of ZIP Family Transporters of Plants.

Zinc (Zn) is an essential micronutrient for plants and humans. Nearly 50% of the agriculture soils of world are Zn-deficient. The low availability of Zn reduces the yield and quality of the crops. The zinc-regulated, iron-regulated transporter-like proteins (ZIP) family and iron-regulated transporters (IRTs) are involved in cellular uptake of Zn, its intracellular trafficking and detoxification in plants. In addition to Zn, ZIP family transporters also transport other divalent metal cations (such as Cd2+, Fe2+, and Cu2+). ZIP transporters play a crucial role in biofortification of grains with Zn. Only a very limited information is available on structural features and mechanism of Zn transport of plant ZIP family transporters. In this article, we present a detailed account on structure, function, regulations and phylogenetic relationships of plant ZIP transporters. We give an insight to structure of plant ZIPs through homology modeling and multiple sequence alignment with Bordetella bronchiseptica ZIP (BbZIP) protein whose crystal structure has been solved recently. We also provide details on ZIP transporter genes identified and characterized in rice and other plants till date. Functional characterization of plant ZIP transporters will help for the better crop yield and human health in future.

Phenotypic responses of foxtail millet (Setaria italica) genotypes to phosphate supply under greenhouse and natural field conditions.

Food insecurity is a looming threat for the burgeoning world population. Phosphorus (P), which is absorbed from soil as inorganic phosphate (Pi), is an essential macronutrient for the growth of all agricultural crops. This study reports phenotype analysis for P responses in natural field and greenhouse conditions, using 54 genotypes of foxtail millet (Setaria italica) representing wide geographic origins. The genotype responses were assessed in natural field conditions in two different seasons (monsoon and summer) under Pi-fertilized (P+) and unfertilized (P-) soil for eight above-ground traits. Enormous variations were seen among the genotypes in phenotypic responses for all the measured parameters under low P stress conditions. Variations were significant for plant height, leaf number and length, tillering ability and seed yield traits. Genotypes ISe 1234 and ISe 1541 were P+ responders, and the genotypes ISe 1181, ISe 1655, ISe 783 and ISe 1892 showed tolerance to low P for total seed yield. Genotypes that performed well under P- conditions were almost as productive as genotypes that performed well under P+ conditions suggesting some genotypes are well adapted to nutrient-poor soils. In the greenhouse, most of the genotypes produced changes in root architecture that are characteristic of P- stress, but to differing degrees. Significant variation was seen in root hair density and root hair number and in fresh and dry weight of shoot and root under P- stress. However, there was not much difference in the shoot and root total P and Pi levels of five selected high and low responding genotypes. We noticed contrasting responses in the greenhouse and natural field experiments for most of these genotypes. The leads from the study form the basis for breeding and improvement of foxtail millet for better Pi-use efficiency.

Finger Millet [Eleusine coracana (L.) Gaertn.] Improvement: Current Status and Future Interventions of Whole Genome Sequence.

The whole genome sequence (WGS) of the much awaited, nutrient rich and climate resilient crop, finger millet (Eleusine coracana (L.) Gaertn.) has been released recently. While possessing superior mineral nutrients and excellent shelf life as compared to other major cereals, multiploidy nature of the genome and relatively small plantation acreage in less developed countries hampered the genome sequencing of finger millet, disposing it as one of the lastly sequenced genomes in cereals. The genomic information available for this crop is very little when compared to other major cereals like rice, maize and barley. As a result, only a limited number of genetic and genomic studies has been undertaken for the improvement of this crop. Finger millet is known especially for its superior calcium content, but the high-throughput studies are yet to be performed to understand the mechanisms behind calcium transport and grain filling. The WGS of finger millet is expected to help to understand this and other important molecular mechanisms in finger millet, which may be harnessed for the nutrient fortification of other cereals. In this review, we discuss various efforts made so far on the improvement of finger millet including genetic improvement, transcriptome analysis, mapping of quantitative trait loci (QTLs) for traits, etc. We also discuss the pitfalls of modern genetic studies and provide insights for accelerating the finger millet improvement with the interventions of WGS in near future. Advanced genetic and genomic studies aided by WGS may help to improve the finger millet, which will be helpful to strengthen the nutritional security in addition to food security in the developing countries of Asia and Africa.

[Early aseptic loosening of cemented cup in metal-on-metal total hip arthroplasties]. / Echec par descellement aseptique précoce de la cupule acétabulaire cimentée de prothèses totales de hanche à couple de frottement métal-métal.

PURPOSE OF THE STUDY: The aim of this study was to report cases of acetabular failure of metal-on-metal total hip arthroplasties using cemented cups with a metal-metal bearing. MATERIAL AND METHODS: From 1997 to 1999, 137 patients underwent total hip arthroplasty with a 28-mm head and a metal-metal (Metasul) bearing. At mean follow-up of three years eight months (range 2-6 years), three patients had died and thirteen were lost to follow-up or had less than 2 years follow-up. The analysis thus included 118 patients (122 hips) 67 women and 51 men (mean age 61 years). The same transgluteal approach with two cemented implants (Palacos gentamycine) was used in all patients. The Postel-Merle-d'Aubigné score was used for clinical assessment with search for lucent lines in the Delee and Charnley zones and cup migration on the standard x-rays. Actuarial survival curves were drawn using revision and failure as the end events. RESULTS: In this series, there were no cases of dislocation or infection but there were five revisions for aseptic cup loosening: one at two years, two at four years, and two at five years. Among the remaining 117 implants, outcome was clinically very good in 101, good in 10 and fair in 6. Radiologically, there were no cases of femoral lucent lines, migration or osteolysis. For the cup, a lucent line measuring less than 1 mm and stable was observed in 81 hips. A progressive lucent line was observed in 18 hips with 11 showing signs of probable loosening. The five-year survival using surgical revision as the endpoint was 94%. Using cup failure (revision and progressive line), it was 80%. Revisions and radiographically probable loosenings were statistically more significant in small cups measuring 44 and 46 mm. DISCUSSION: In this series, cemented metal-on-metal bearings showed a rate of failure greater than observed in metal-polyethylene implants and metal-on-metal implants using a press fit cup. Other series using a metal-on-metal bearing with a cemented cup have found the same trend. CONCLUSION: We have discontinued this type of implant and recommend avoiding direct cementing of the polyethylene-metasul cup on the bone for total hip arthroplasty. The metal-on-metal bearing can still be used, but without cementing the cup.

In situ hybridization of GnRH mRNA in the rat and the mink hypothalamus using biotinylated synthetic oligonucleotide probes.

The cellular localization of GnRH messenger RNA (mRNA) in the rat and the mink hypothalamus has been examined using a newly developed highly sensitive non-radioactive in situ hybridization procedure. Synthetic oligonucleotides labeled by addition of a biotin-21-dUTP tail at their 3' end can be used to detect GnRH mRNA in both species. Streptavidin-alkaline phosphatase revealed with nitroblue tetra-zolium-bromo-chloro-indolyl-phosphate as substrate makes possible detection of the biotinylated oligonucleotides. In the rat, our findings confirm results previously obtained using synthetic radioactive probes, and demonstrate the potency of and interest in using biotinylated oligonucleotides to identify related sequences of bases in tissues. The principle advantages include rapid signal detection, excellent spatial resolution, and low background. In the mink, the in situ hybridization method clearly confirms the characterization of GnRH-producing cells and also allows detection of GnRH cell bodies in conditions in which they are not detected by immunohistochemistry. Adaptation of the in situ hybridization to the detection of GnRH mRNA in species like the mink which shows seasonal reproductive activity is a crucial step. This method offers a new approach to problems as fundamental as changes in gene expression depending on photoperiod or under a variety of experimental conditions.

Photic regulation of c-fos gene expression in the suprachiasmatic nucleus and the circadian rhythm of photosensitivity in the mink.

The relationship between the photic stimulation of the c-fos gene product in cells of the suprachiasmatic nuclei and the photoperiodic control of testicular activity were examined in mink. Mink were kept in a short photoperiod (control, LD4:20), or in 'asymmetric skeleton photoperiods' (groups A and B). The light period for groups A and B was divided into two fractions (3 h 30 min and 30 min); the shorter fraction occurred in the night, 4 h (group A) or 8 h (group B) after the end of the main light period. There was no photic activation of the proto-oncogene c-fos on the control or group A, and 4 weeks on this photoperiodic treatment produced marked testicular development. In contrast, in group B, c-fos mRNA was induced 30 min after the end of the secondary photofraction, was maximal 30 min later and then decreased. Fos-like immunoreactivity was detected 2 h after the end of the secondary photofraction, with activity peaking 1 h later. The animals of this group remained sexually quiescent. These results suggest that photo-induction of the proto-oncogene c-fos is implicated in the gonadal inhibition induced in this species when the light period, extends into the photosensitive phase of the circadian rhythm of photosensitivity.

Short-day stimulation of testicular activity and immunoreactivity of the hypothalamic GnRH system in mink following deafferentation of the pineal body by bilateral superior cervical ganglionectomy and melatonin replacement.

The effects of superior cervical ganglionectomy on testicular function (testis volume and plasma testosterone levels) and the immunocytochemical activity of the GnRH hypothalamic system were studied in the mink, a short-day breeder. Animals reared in a natural photoperiod were (i) ganglionectomized at four different times during the period extending from the end of summer to the end of autumn (September 15, October 20, October 28, and December 1), and (ii) reared for 50 days in a short gonadostimulatory photoperiod (4L:20D). Lastly, an attempt was made to overcome the effects of superior cervical ganglion removal by administering melatonin to mink reared in a natural photoperiod. In mink reared in a natural photoperiod, deafferentation of the pineal on September 15 (L = 12.5 h) or October 20 (L = 10.5 h) resulted in consistently low values of testicular volume and plasma testosterone until the end of the experiment (February). When the operation was performed on October 28 (L = 10 h) testicular activity was initiated but only lasted a short time and did not allow maximal gonadal development. When superior cervical ganglionectomy was carried out on December 1 (L = 8.5 h), during the phase of renewed testicular activity, the increases in testicular volume and testosterone levels were not affected by the operation and the subsequent variation of these parameters was identical to that observed in intact animals. Similarly, in mink reared for 50 days in a photoperiod of 4L:20D before superior cervical ganglionectomy, deafferentation of the pineal did not prevent gonadostimulation induced by short days.(ABSTRACT TRUNCATED AT 250 WORDS)

Localization of melatonin binding sites in the pars tuberalis of the mink at three times during the seasonal testicular cycle.

The strict photoperiodic dependence of gonadotropic function observed in mink provides an excellent physiological model for studying the activity of GnRH hypothalamic neurons. In mink, exposure to more than 10 h light inhibits the activity of this neurohormonal system. Melatonin plays an essential role in this type of regulation of gonadotropic function. In mink, contrary to results in many other photosensitive species, melatonin was observed to mediate the gonadostimulating effect of short days. However, the binding sites and action of this substance have not yet been defined. We thus attempted to identify melatonin binding sites in the mink brain. The study was carried out at three times during the seasonal testicular cycle in male minks maintained under natural environmental conditions. Coronal sections were taken from the whole brain and pituitary gland. Using 2-[125I]iodomelatonin, we observed a strong concentration of binding sites in the pars tuberalis and not in the part of the median eminence surrounded by the pars tuberalis. This concentrated localization of melatonin binding sites brings up the problem of defining the action of this substance in photoregulating the activity of the GnRH neurohormonal system.

[Bodian process. Ultrastructural study]. / Le procédé de Bodian. Etude ultrastructurale.

The Bodian method was used for an ultrastructural study on the optic nerve of the adult cat in order to compare the effects of two fixation techniques on the normal nerve and on the nerve during Wallerian degeneration after unilateral enucleation. Immersion fixation causes a predominately metallic deposit in the myelin sheath; the perfusion has no effect. During the Wallerian degeneration the two fixation methods lead to a metallic deposit located in the axon which is in large grains after immersion and extremely dense after perfusion. Various diffraction methods were used to identify the nature of the metallic deposit. These results emphasize the positive elements in systemically associating the electron microscope with the optic microscope for studies concerning Wallerian degeneration after metallic impregnation in the central nervous system

pS2 and PAI-1 in ovarian cancer: correlation to pathohistological parameters.

The estrogen regulated pS2 protein and the Plasminogen Activator Inhibitor type 1 (PAI-1) have been reported as important tumor parameters both in breast and in ovarian cancer. We analysed the cytosolic concentrations of pS2 in 111 ovarian carcinoma and the cytosolic concentrations of PAI-1 in 104 ovarian cancers by RIA and ELISA. Using a cut-off level of 2 ng/mg protein we found 27% pS2+ tumors. We observed 42% PAI-1+ tumors using a out-off level of 1 ng/mg. We found a statistically significant decline in the pS2 status corresponding with an increase in the PAI-1 status from well to poor differentiation grade. The highest levels of pS2 and the lowest levels of PAI-1 were measured in borderline carcinoma. Significantly higher concentrations of pS2 were measured in mucinous over serous carcinoma. We found no significant correlation between PAI-1 and histologic subtypes, or between pS2 or PAI-1 and estrogen receptor status, progesterone receptor status, age and tumor stage. To conclude, we found pS2 and PAI-1 concentrations to be correlated with the grade of differentiation. A correlation between protein status and histologic subtypes could be observed for pS2 but not for PAI-1.

The onset of synaptogenesis in rat temporal cortex.

The onset of synaptogenesis was studied in the temporal cortex of rat fetuses whose age ranged between 15 and 19 days of gestation. First synapses were found at a surprisingly early stage of cortical development: on day 16. These contacts showed relatively few vesicles and very inconspicuous membrane-thickenings. They were located in the marginal layer, above as well as below the narrow band formed by the newly arrived first neuroblasts of the prospective corticle plate. The postsynaptic structures were probably dendrites of the horizontally or obliquely orientated neurons scattered throughout the marginal layer (such neurons were seen even within the cell-dense band). On day 17, the cortical plate separated the differentiated cells definitely into a superficial and a deep population. As on the following days, synapses were found above and below the cortical plate but not within it. In addition to contacts showing the same features as those described on day 16, there were already synapses with numerous vesicles and clearly asymmetric membrane thickenings. On days 18 and 19 the borders of the cortical plate became more clear-cut. The well-differentiated neurons situated above and below this plate could now be identified as Retzius-Cajal cells of the prospective molecular layer and as polymorphous cells of the layer VI b respectively. The presence of axo-somatic contacts on these neurons provided direct evidence that both cell types are targets for synapses. Desmosome-like junctions were found even in the youngest fetuses studied. Their roughly symmetric membrane thickenings were clearly more conspicious than those of earliest synapses. Desmosome-like junctions occurred very frequently between structures which subsequently were never seen to become synaptically linked. During the entire period studied, numerous coated vesicles fused with cell membranes were noted. Such "open" vesicles were seen on neurons (sometimes in the immediate vicinity of synapses) but also on non-nervous, extracortical as well as intracortical structures. Thus there does not seem to be a specific relationship between desmosome-like junctions and coated vesicles on the one hand and synapse formation on the other.

Much ado about mentors.

An international management firm conducted a study of executives that included questions regarding compensations, personal data, and current position. It was of special interest to determine the effect of mentor and protege relationships on business careers. Of the individuals surveyed, two-thirds reported a relationship with a senior person who took a special interest in their career, during the first five years of their professional development. In general, executives who had a mentor are better educated, earn more money at an earlier age, more apt to follow a career plan and report high job satisfaction. The majority of sponsors are older businessmen holding positions of authority and who feel that personnel development and management succession are key responsibilities. The following characteristics of a mentor are judged most important: willingness to share experiences, knowledge of the organization, organizational power and respect from peers. Mentor-protege relationships frequently develop into lengthy friendships which in turn encourage young executives to eventually sponsor their own proteges.

[The optic nerve in the cat. I. Quantitative aspects in the adult cat]. / Le nerf optique du chat. I. Aspects quantitatifs chez lànimal adulte.

A quantitative study was carried out on the adult cat optic nerve near the eyeball by systematically measuring the perimeters of all the axons seen through the optic microscope. The main purpose of this study was to define the topographical distribution of these axons in function of their size. Statistical studies show the existence of an area of maximal concentration of large axons in crescent form situated in the temporal zone of the nerve. The neurophysiological implications are discussed.

[The optic nerve in the cat. II. Qualitative aspects of maturation in the adult cat]. / Le nerf optique du chat. II. Aspects qualitatifs de la maturation à l'âge adulte.

54 healthy cats were studied. In almost all cases, two types of degenerative signs could be seen: in the primary optic pathways, but also in the posterior funiculi and the spinal-cerebellar tracts, lesions of wide-diameter axons seen by their initial demyelination, accompanied by intense neuroglial reactions, and, on the other hand, a Wallerian degeneration of smaller axons situated in the axial part of the nerve. The percentage of these abnormalities must still be evaluated. Their eventual consequences and nature are discussed.

[Astrocytic response in alcoholic cirrhotic encephalopathy (immunohistochemical study)]. / La réponse astrocytaire au cours des encéphalopathies cirrhotiques alcooliques (étude immunohistochimique).

In human alcoholic cirrhotic encephalopathy, most of astrocytes of the cerebral cortex appear as Alzheimer type II cells. Thus, they cannot be visualized by the immunocytochemical detection of Glial Fibrillary Acidic protein (GFAP). However, fibrous astrocytes present in layer I of the cortex and in the vicinity of the white matter are GFAP positive. This special reactivity of layer I astrocytes could correspond to the persistance of the characters of primitive neuropil.

The cornu ammonis and alcoholic cirrhotic encephalopathy. A neuropathological and immunohistological study.

Two distinct categories of facts are deduced from a neuropathological study of the Ammon's horn, in particular the Sommer Sector, afflicted with alcoholic cirrhotic encephalopathy. In the pyramidal neurons of the CA1 layer, there are unquestionably paired filaments in a helical form, which correspond to the now classic "paired helical filaments". On the other hand, whereas the astrocyte population on the whole displays the characteristics of what can be called Alzheimer type II cells, a certain number of highly reactive areas persist, which can be detected by a GFAP-specific immunohistochemical method.

Peptidergic neurohormonal systems in the basal hypothalamus of the ferret and the mink: immunocytochemical study of variations during the annual reproductive cycle.

The hypothalamic systems secreting corticotropin-releasing hormone (CRF), somatostatin, oxytocin, vasopressin and luteinizing hormone-releasing hormone (LHRH) were characterized using immunochemistry, and variations were studied in relation to the recrudescence of testicular activity in the ferret and the mink, two species with opposite photoregulation of their annual reproductive cycles. Under the present conditions of study, the immunoreactivity of the CRF, somatostatin, and oxytocin systems showed no significant variation in either species. In contrast, in these two species, the immunoreactivity of the LHRH system varied considerably depending on the date of observation. The increase in the number and immunoreactivity of the LHRH-secreting neurons that occurred in November in the mink and in January in the ferret, is in agreement with previous results showing that the photoperiod plays an essential role in regulating the annual activity of the testis and that the photoperiodic environmental conditions required for the activation of the LHRH system differ between the species. Similarly, correlations could be found between an increase in immunoreactivity of the vasopressinergic axons projecting to the external median eminence and the recrudescence of testicular activity.