Surface pretreatment for prolonged survival of cemented tibial prosthesis components: full- vs. surface-cementation technique]

Aseptic loosening of tibial components due to degradation of the interface between bone cement and metallic tibial shaft component is still a persistent problem, particularly for surface-cemented tibial components. The surface cementation technique has important clinical meaning in case of revision and for avoidance of stress shielding. This study was done to prove crack formation in the bone cement near the metallic surface when this is not coated. We propose a newly developed coating process by SiOx-PVD layering to avoid crack formation. A biomechanical model for a vibration fatigue test was done to prove that crack formation can be significantly reduced in the case of coated surfaces. It was found that coated tibial components showed a highly significant reduction of cement cracking near the metal/bone cement interface (p < 0.01) and a significant reduction of gap formation in the metal-to-bone cement interface (p < 0.05). Coating dramatically reduces hydrolytic- and stress-related crack formation at the prosthesis metal/bone cement interface. This leads to a more homogenous load transfer into the cement mantle which should reduce the frequency of loosening in the metal/bone cement/bone interfaces. With surface coating of the tibial component it should become possible that surface-cemented TKAs reveal similar loosening rates as TKAs both surface- and stem-cemented. This would be an important clinical advantage since it is believed that surface cementing reduces metaphyseal bone loss in the case of revision and stress shielding for a better bone health.     

Estimating elephant densities from wells and droppings in dried out riverbeds

In this paper, we present a new method for estimating elephant densities by counting elephant wells and dung boli within dry seasonal flooding rivers. A combination of aerial and ground counts of elephant wells and dung boli in the Ewaso Ngiro River were related to elephant numbers, obtained from an on‐going monitoring program of individually identified elephants in Samburu and Buffalo Spring National Reserves, Kenya. The total number of elephant observations was highly correlated with both densities of wells and dung boli at a spatial resolution of 4‐km river‐section. This indicates that both wells and droppings can be used for estimating relative densities at such spatial resolution. The method can be used as a quick and reliable way of estimating relative elephant densities in semiarid regions but is sensitive to differences in the time when different parts of the river dry out and will be unreliable in areas with secondary un‐censused water sources. A short 4‐week period between the river dry out and the count is recommended, because of an error induced by a level of well reuse and the difficulties in counting areas of high well densities from the air.     

Heterologous Production of Antimicrobial Peptides in Propionibacterium freudenreichii

Heterologous bacteriocin production in Propionibacterium freudenreichii is described. We developed an efficient system for DNA shuttling between Escherichia coli and P. freudenreichii using vector pAMT1. It is based on the P. freudenreichii rolling-circle replicating plasmid pLME108 and carries the cml(A)/cmx(A) chloramphenicol resistance marker. Introduction of the propionicin T1 structural gene (pctA) into pAMT1 under the control of the constitutive promoter (P₄) yielded bacteriocin in amounts equal to those of the wild-type producer Propionibacterium thoenii 419. The P. freudenreichii clone showed propionicin T1 activity in coculture, killing 90% of sensitive bacteria within 48 h. The pamA gene from P. thoenii 419 encoding the protease-activated antimicrobial peptide (PAMP) was cloned and expressed in P. freudenreichii, resulting in secretion of the pro-PAMP protein. Like in the wild type, PAMP activation was dependent on externally added protease. Secretion of the antimicrobial peptide was obtained from a clone in which the pamA signal peptide and PAMP were fused in frame. The promoter region of pamA was identified by fusion of putative promoter fragments to the coding sequence of the pctA gene. The P₄ and P_pamp promoters directed constitutive gene expression, and activity of both promoters was enhanced by elements upstream of the promoter core region.     

Construction, characterization, and preliminary BAC-end sequencing analysis of a bacterial artificial chromosome library of white clover (Trifolium repens L.).

White clover (Trifolium repens L.) is a forage legume widely used in combination with grass in pastures because of its ability to fix nitrogen. We have constructed a bacterial artificial chromosome (BAC) library of an advanced breeding line of white clover. The library contains 37 248 clones with an average insert size of approximately 85 kb, representing an approximate 3-fold coverage of the white clover genome based on an estimated genome size of 960 Mb. The BAC library was pooled and screened by polymerase chain reaction (PCR) amplification using both white clover microsatellites and PCR-based markers derived from Medicago truncatula, resulting in an average of 6 hits per marker; this supports the estimated 3-fold genome coverage in this allotetraploid species. PCR-based screening of 766 clones with a multiplex set of chloroplast primers showed that only 0.5% of BAC clones contained chloroplast-derived inserts. The library was further evaluated by sequencing both ends of 724 of the clover BACs. These were analysed with respect to their sequence content and their homology to the contents of a range of plant gene, expressed sequence tag, and repeat element databases. Forty-three microsatellites were discovered in the BAC-end sequences (BESs) and investigated as potential genetic markers in white clover. The BESs were also compared with the partially sequenced genome of the model legume M. truncatula with the specific intention of identifying putative comparative-tile BACs, which represent potential regions of microsynteny between the 2 species; 14 such BACs were discovered. The results suggest that a large-scale BAC-end sequencing strategy has the potential to anchor a significant proportion of the genome of white clover onto the gene-space sequence of M. truncatula.     

Stunting at 2 Years in Relation to Body Composition at 9 Years in African Urban Children**

Objective: To determine whether African urban children who were stunted at 2 years of age demonstrated an altered body composition by the end of childhood, before entering puberty, at 9 years of age. Research Methods and Procedures: This was a mixed‐longitudinal study of 330 prepubertal African children (182 boys) from Soweto‐Johannesburg, South Africa. Anthropometric data at 2 years of age were compared with anthropometric, DXA‐determined body composition and fat patterning in late childhood (7 to 9 years). Results: Children who had been stunted at 2 years were significantly shorter and lighter than non‐stunted children at 7 to 9 years, but there were no differences in their BMI or centralization of body fat. Previously stunted status significantly predicted reduced weight and height at 7 to 9 years but did not predict BMI, body composition, or fat patterning after controlling for potential confounding factors. The odds ratio for stunting at 2 years as a predictor of overweight at 7 to 9 years was not significant at 1.09 (95% confidence limits: 0.30, 3.98). Discussion: Greater BMI in stunted infants does not demonstrate a tendency toward overweight or obesity but is a reflection of the greater reduction in height rather than weight in stunted children. Stunted children may be programmed to accumulate greater body fat at central sites during adolescence, but we have been unable to show that these changes are evident before the initiation of pubertal development.     

PREPARATION OF PLASMA MEMBRANE FROM ISOLATED NEURONS

A bulk fraction enriched with respect to neuronal cell bodies was used as starting material for the isolation of neuronal plasma membrane The cells were gently homogenized in isotonic sucrose and a crude membrane containing fraction sedimented at 3000 g. Subsequently, the membrane fraction was purified on a discontinuous sucrose density gradient between 35% and 25 5% sucrose (w/w). Enzymatic analyses showed a 4–5-fold enrichment in plasma membrane markers, and a 10–15% contamination of mitochondrial and microsomal material. Electron micrographs of the membrane fraction confirmed the enzymatic data Fragmented membranes were found, mainly in vesicular form No ribosomes, but a few mitochondria and some multilamellar membranes were seen     

The effect of gonadotrophins on the 3',5'-AMP levels of seminiferous tubules

The addition of follicle-stimulating hormone (FSH) to isolated tubules from hypophysectomized rats was shown to increase the level of adenosine 3′,5′-monophosphate (3′,5′-AMP). In contrast, luteinizing hormone (LH) exerted no effect in this system. The results presented are consistent with the concept that FSH exerts a direct effect upon cells within the seminiferous tubule, possibly on Sertoli cells, whereas the effects of LH on spermatogenesis are primarily due to the stimulation of androgen production by the interstitial cells of the testis.