The β-Hemolysin and Intracellular Survival of Streptococcus agalactiae in Human Macrophages

S. agalactiae (group B streptococci, GBS) is a major microbial pathogen in human neonates and causes invasive infections in pregnant women and immunocompromised individuals. The S. agalactiae β-hemolysin is regarded as an important virulence factor for the development of invasive disease. To examine the role of β-hemolysin in the interaction with professional phagocytes, the THP-1 monocytic cell line and human granulocytes were infected with a serotype Ia S. agalactiae wild type strain and its isogenic nonhemolytic mutant. We could show that the nonhemolytic mutants were able to survive in significantly higher numbers than the hemolytic wild type strain, in THP-1 macrophage-like cells and in assays with human granulocytes. Intracellular bacterial multiplication, however, could not be observed. The hemolytic wild type strain stimulated a significantly higher release of Tumor Necrosis Factor-α than the nonhemolytic mutant in THP-1 cells, while similar levels of the chemokine Interleukin-8 were induced. In order to investigate bacterial mediators of IL-8 release in this setting, purified cell wall preparations from both strains were tested and found to exert a potent proinflammatory stimulus on THP-1 cells. In conclusion, our results indicate that the β-hemolysin has a strong influence on the intracellular survival of S. agalactiae and that a tightly controlled regulation of β-hemolysin expression is required for the successful establishment of S. agalactiae in different host niches.     

Molecular characterization of a patient with del(1)(q23–q25)

Summary We report a patient (S.T.) with multiple congenital anomalies and developmental delay associated with an interstitial deletion of 1q23–1q25. Molecular analysis of the deletion was performed using DNA markers that map to 1q. Five DNA markers, MLAJ-1 (D1S61), CRI-L1054 (D1S42), HBI40 (D1S66), OS-6 (D1S75), and BH516 (D1S110), were demonstrated to be deleted. Informative polymorphisms demonstrated this to be a de novo deletion of the maternally derived chromosome. Deletion status was determined using restriction fragment length polymorphism (RFLP) analysis supplemented with densitometry in the experiments where RFLP analysis was not fully informative. Deletions were confirmed by Southern analysis using genomic DNA from a somatic cell hybrid retaining the del(1)(q23–q25) chromosome that was constructed from patient S.T. Flow karyotyping confirmed the deletion and estimated that the deletion encompassed 11,000–16,000 kb. The clinical and cytogenetic characteristics of S.T. are compared with those of ten previously described patients with monosomy 1q21–1q25.     

The interactions with Ro60 and La differentially affect nuclear export of hY1 RNA.

Ro RNPs are evolutionarily conserved ribonucleoprotein particles that consist of a small RNA, known as Y RNA, associated with several proteins, such as La, Ro60, and Ro52. The Y RNAs (Y1-Y5), which are transcribed by RNA polymerase III, have been shown to reside almost exclusively in the cytoplasm as Ro RNPs. To obtain more insight into the nuclear export pathway of Y RNAs, hY1 RNA export was studied in Xenopus laevis oocytes. Injection of various hY1 RNA mutants showed that an intact Ro60 binding site is a prerequisite for nuclear export, whereas the presence of an intact La binding site resulted in strong nuclear retention of hY1 RNA. Competition studies with various classes of RNAs indicated that, in addition to Ro60, another titratable factor was necessary for nuclear export of hY1 RNA. This factor appears also to be involved in nuclear export of tRNA. Because export of hY1 RNA could not be blocked by a synthetic peptide containing the recently identified nuclear export signal of the HIV-1 Rev protein, nuclear export of hY1 RNA does not seem to be dependent on a Rev-like nuclear export signal.     

Hyperpolarization response of an identified neuron of Planobarius corneus mollusks to several cholinomimetic substances]

Two kinds of responses to cholinomimetics were found on the identified neuron (P-2) of Planorbarius corneus pedal ganglion using microelectrode technique. Nicotinomimetics caused depolization whereas some muscarinomimetics caused hyperpolarization of the neuron membrane. Acetylcholine usually depolarized the neuron membrane but after the blockade of nicotinic receptors with tubocurarine one can reveal a hyperpolarizing action of acetylcholine. These findings suggest that two kinds of receptors exist on the P-2 neuron membrane and these receptors differ both in pharmacological characteristics and in ionic permeability changes they control.     

Three novel components of the human exosome

The yeast exosome is a complex of 3' --> 5' exoribonucleases. Sequence analysis identified putative human homologues for exosome components, although several were found only as expressed sequence tags. Here we report the cloning of full-length cDNAs, which encode putative human homologues of the Rrp40p, Rrp41p, and Rrp46p components of the exosome. Recombinant proteins were expressed and used to raise rabbit antisera. In Western blotting experiments, these decorated HeLa cell proteins of the predicted sizes. All three human proteins were enriched in the HeLa cells nucleus and nucleolus, but were also clearly detected in the cytoplasm. Size exclusion chromatography revealed that hRrp40p, hRrp41p, and hRrp46p were present in a large complex. This cofractionated with the human homologues of other exosome components, hRrp4p and PM/Scl-100. Anti-PM/Scl-positive patient sera coimmunoprecipitated hRrp40p, hRrp41p, and hRrp46p demonstrating their physical association. The immunoprecipitated complex exhibited 3' --> 5' exoribonuclease activity in vitro. hRrp41p was expressed in yeast and shown to suppress the lethality of genetic depletion of yeast Rrp41p. We conclude that hRrp40p, hRrp41p, and hRrp46p represent novel components of the human exosome complex.     

The role of Brunner's glands in the mucosal protection of the proximal part of duodenum

In the course of a prospective study authors examined the role of the hyperplasia of Brunner's glands in the mucosal protection of proximal part of duodenum in patients with peptic ulcer, chronic pancreatitis and chronic renal insufficiency. Their method for this study was the histological examination of the endoscopic biopsy specimens. The hyperplasia of Brunner's glands occurs with significant frequency in all the three examined groups of patients, while we found it less frequently in the controls. Hyperplasia of Brunner's glands is one of the protective mechanisms of the organism, which serves the protection of the duodenal mucosa between the pyloric ring and the papilla of Vater, against the damaging effect of the hydrochloric acid.     

Proteus-typing by proticin production and susceptibility

A Proteus-typing method based on proticin production and proticin susceptibility (c.f. Senior, 1977) has been modified to increase its sensitivity. Proticins were prepared in fluid medium and applied to agar-plates shortly before seeding the plates with indicator bacteria. A given 10 proticin producer strains, which are responsible for the susceptibility patterns of the indicator-bacteria (S-types), form the foundation for this typing method. Using this producer-set an indicator-set (28 strains) was selected which was suitable for the typing of strains with different proticin activities (P-types). Standardization of the temperature for proticin production proved to be necessary. The degree of similarity between proticins was further elaborated by testing all indicator strains for susceptibility to proticin titrations. In the group of 148 clinical Proteus-isolates (four species) used for the development of the typing system 28 S-types and 34 P-types were observed. By combining the S- and P-type parameters 86 S-P-types were obtained for the 4 species combined. Seven strains were not typable. A separate group of 100 clinical Proteus-isolates was tested in order to prove the usefulness of the method. 39 new S-P-types were found. Repeated isolations from the same patients yielded the same patterns. Proteus S-P-typing is a useful method for the typing of Proteux vulgaris and Proteus mirabilis, but proves inadequate for the typing of Proteus rettgeri and Proteus morganii.     

Abiotic factors driving cyanobacterial biomass and composition under perennial bloom conditions in tropical latitudes

Hydrobiologia - While warming and eutrophication have increased the frequency and magnitude of harmful cyanobacterial blooms globally, the scenario for many eutrophic tropical freshwaters is a...