The assessment of human sperm morphology in surface replica preparations for transmission electron microscopy

Human spermatozoa were treated with the detergents Hyamine 2389, Triton X-100, or sodium dodecyl sulphate (SDS) and surface replica preparations and Papanicolaou-stained smears made. From the ultrastructural studies it was found that Hyamine caused a dissolution of the plasmalemma to reveal the outer acrosomal membrane that was often absent over the anterior cap region; whereas Triton, in addition, caused severe damage to the entire acrosome. Treatment with SDS rapidly cleaved the tails from the majority of the spermatozoa, stripped the plasmalemma from both the head and tail regions of the cells, and also removed the whole of the outer acrosomal membrane plus most of the acrosomal contents. However, when viewed in the Papanicolaou-stained preparations, the spermatozoa were all of apparently normal morphology. These studies make it clear that some caution must be used when interpreting human sperm morphology assessments made under the light microscope. While it is not suggested that the more involved technique of surface replication should be used in routine semen analyses, it does represent a valuable technique for use with research material.     

Metallic-Oxide encrustations of the nonprosthecate stalks of naturally occurring populations ofPlanctomyces bekefii

Naturally occurring metallic-oxide encrustations of the multifibrillar stalks of morphotype Ia (Planctomyces bekefii) of theBlastocaulis-Planctomyces group of budding and nonprosthecately appendaged prokaryotes from two sources-a pond in Budapest, Hungary (containing topotypical material) and Gebharts Teich, a fishpond in Austria-have been examined by energy-dispersive X-ray spectroscopy. Some rosettes of morphotype Ib (Planctomyces crassus) also occurred in the Budapest pond; only morphotype Ia was found in Gebharts Teich. Manganese was the major metallic element in the morphotype Ia stalk encrustations from the Budapest pond; iron was not detected. However, morphotype Ib rosettes occurring in the same Budapest pond sampes contained both manganese and iron in their stalk encrustations. Both iron and manganese were present in the morphotype Ia stalk encrustation of rosettes from Gebharts Teich.     

U-series and ESR analyses of bones and teeth relating to the human burials from Skhul

In order to resolve long-standing issues surrounding the age of the Skhul early modern humans, new analyses have been conducted, including the dating of four well-provenanced fossils by ESR and U-series. If the Skhul burials took place within a relatively short time span, then the best age estimate lies between 100 and 135 ka. This result agrees very well with TL ages obtained from burnt flint of 119+/-18 ka (Mercier et al., 1993). However, we cannot exclude the possibility that the material associated with the Skhul IX burial is older than those of Skhul II and Skhul V. These and other recent age estimates suggest that the three burial sites, Skhul, Qafzeh and Tabun are broadly contemporaneous, falling within the time range of 100 to 130 ka. The presence of early representatives of both early modern humans and Neanderthals in the Levant during Marine Isotope Stage 5 inevitably complicates attempts at segregating these populations by date or archaeological association. Nevertheless, it does appear that the oldest known symbolic burials are those of early modern humans at Skhul and Qafzeh. This supports the view that, despite the associated Middle Palaeolithic technology, elements of modern human behaviour were represented at Skhul and Qafzeh prior to 100 ka.     

Development of a functional genomics platform for Sinorhizobium meliloti: construction of an ORFeome

The nitrogen-fixing, symbiotic bacterium Sinorhizobium meliloti reduces molecular dinitrogen to ammonia in a specific symbiotic context, supporting the nitrogen requirements of various forage legumes, including alfalfa. Determining the DNA sequence of the S. meliloti genome was an important step in plant-microbe interaction research, adding to the considerable information already available about this bacterium by suggesting possible functions for many of the >6,200 annotated open reading frames (ORFs). However, the predictive power of bioinformatic analysis is limited, and putting the role of these genes into a biological context will require more definitive functional approaches. We present here a strategy for genetic analysis of S. meliloti on a genomic scale and report the successful implementation of the first step of this strategy by constructing a set of plasmids representing 100% of the 6,317 annotated ORFs cloned into a mobilizable plasmid by using efficient PCR and recombination protocols. By using integrase recombination to insert these ORFs into other plasmids in vitro or in vivo (B. L. House et al., Appl. Environ. Microbiol. 70:2806-2815, 2004), this ORFeome can be used to generate various specialized genetic materials for functional analysis of S. meliloti, such as operon fusions, mutants, and protein expression plasmids. The strategy can be generalized to many other genome projects, and the S. meliloti clones should be useful for investigators wanting an accessible source of cloned genes encoding specific enzymes.     

Ampicillin Levels in Human Bile in the Presence of Biliary Tract Disease

Ampicillin levels were measured in the serum and in the bile from both the gall bladder and the common bile duct in patients undergoing surgery for biliary tract diseases. In patients with radiologically non-functioning gall bladders ampicillin was either not present or its concentration was lower than normal. Therapeutic levels were present in the common bile duct of all patients except those with obstruction of the common bile duct. Hence ampicillin fails appreciably to penetrate the obstructed viscus in obstructive biliary tract disease, and it is unlikely to be effective in treating infection associated with this.