Cloning of regions required for contact hemolysis and entry into LLC-MK2 cells from Shigella sonnei form I plasmid: virF is a positive regulator gene for these phenotypes.

Two distinct regions required for both contact hemolysis and entry into LLC-MK2 cells were cloned into Escherichia coli from the Shigella sonnei form I plasmid, pSS120. The first region was cloned into an E. coli HB101 strain containing noninvasive Tn1 insertion mutants of the form I plasmid, and expression of ipa (invasion plasmid antigen) gene products was restored. The plasmid carrying the first region was then transformed into E. coli lacking the form I plasmid, and additional DNA fragments from the form I plasmid were cloned into the same recipient on compatible vectors. Five of these double transformants were found to be positive for contact hemolysis activity. Restriction analysis of these five clones indicated that the previously reported ipa locus and the invA locus were present on the second plasmid region. Only the strains carrying both of these regions were active in contact hemolysis and cell invasion assays. Several proteins, including the a, b, c, and d proteins encoded by the ipa genes, were detected in the double transformants by Western blot (immunoblot) analysis with serum of a monkey convalescing from shigellosis. A positive regulator was suggested to exist in the first region, since the amounts of most of these proteins were simultaneously increased in the presence of this region. Subcloning and nucleotide sequencing indicated that this positive regulator gene was virF. Product analysis of the virF gene with minicells showed that two peptides (30 and 21 kilodaltons) were synthesized and that at least the 30-kilodalton protein was essential for regulation of the ipa genes.     

Relatedness of structures of a major immunogen in Trichomonas vaginalis isolates.

Solubilization of live Trichomonas vaginalis organisms with detergent caused the release of cysteine proteinases in the detergent extract which were inhibitable with N-alpha-p-tosyl-L-lysine chloromethyl ketone. The detergent extracts of all isolates tested possessed similar cysteine proteinase activities. These parasite proteinases rapidly degraded a prominent immunogen whose surface disposition undergoes phenotypic variation in some isolates. The relatedness of the forms of this immunogen among all isolates tested was confirmed by identical immunoblot patterns of autolysed immunogen, and data suggest the presence of repeating units or at least equidistant sites for proteinase cleavage within the immunogen molecule.     

Effect of extracellular fluid volume contraction and expansion on plasma atrial natriuretic peptide (ANP) concentration in patients with acromegaly

Atrial natriuretic peptide (ANP) is a hormone release into the circulation by atrial cardiocytes (Gutkowska et al. 1984). Extracellular fluid volume expansion acts as a powerful stimulus for ANP secretion and results in the augmentation of its plasma concentration (Lang et al. 1985). Patients with active acromegaly demonstrate the increased extracellular fluid volume (Falkheden et al. 1964), while a successful treatment of the disease results in the disappearance of hypervolemia (Strauch et al. 1977). We have recently demonstrated that in patients with active acromegaly the increased total body plasma volumes are accompanied by the elevated plasma ANP concentrations, whereas, in the successfully treated patients, both: total plasma volumes and plasma ANP levels do not differ significantly from these in healthy subjects (Czekalski et al. 1988b).     

Involvement of D1 dopamine receptors in the control of TSH secretion in the male rat

The effects of SCH 23390 (D1 dopamine receptor antagonist), SK&F 38393 (D1 dopamine receptor agonist), raclopride and remoxipride (D2 dopamine receptor antagonists) and ketanserin (5-hydroxytryptamine 2 receptor antagonist) on TSH serum levels (radioimmunoassay) and on brain catecholamine levels (Falck-Hillarp methodology in combination with quantitative histofluorimetry) were studied. SCH 23390 produced a dose-dependent increase in serum TSH levels in the lower dose range (0.01-0.03 mg kg-1, i.p.) administered 30 min before decapitation and in the higher dose range (1.0-3.0 mg kg-1) when given 2 h before decapitation. Following 30 min of treatment with the high doses of SCH 23390, reductions in serum TSH levels were found. The changes observed following SCH 23390 treatment occurred without affecting catecholamine levels in the median eminence and the peri- and paraventricular hypothalamic regions. Raclopride (0.1-10 mg kg-1, i.p.), remoxipride (1.0 mg kg-1, i.p.) or ketanserin (0.3 mg kg-1, i.p.) changed neither serum TSH levels nor brain catecholamine levels, SK&F 38393 (1.0-10 mg kg-1, i.p.) produced an increase in serum TSH levels. The results suggest the existence of inhibitory and facilitatory mechanisms regulating TSH secretion mediated via D1 dopamine receptors.     

Saponins from Thinouia coriacea]

The investigation of the stems of Thinouia coriacea Britton (Sapindaceae), an ichthyotoxic plant from South Brazil, afforded eight glycosides of oleanolic acid. Structures were assigned based on data from partial hydrolysis. 13C-NMR and mass spectral procedures as 3-O-alpha-L-arabinopyranoside (1), 3-O-alpha-L-rhamnopyranosyl-(1----2)-alpha-L-arabinopyranoside+ ++ (2), 3-O-beta-D-glucopyranosyl-(1----4)-alpha-L-arabinopyranoside (3), 3-O-beta-D-glucopyranosyl-(1----3)-alpha-L-rhamnopyranosyl-(1----2 )-alpha-L-arabinopyranoside (4), 3-O-alpha-L-rhamnopyranosyl-(1----2)[beta-D-glucopyranosyl-(1----4 )]-alpha-L-arabinopyranoside (5), 3-O-beta-D-xylopyranosyl-(1----3)-alpha-L-rhamnopyranosyl-(1----2) [beta-D-glucopyranosyl-(1----4)[alpha-L-arabinopyranoside (6), 3-O-beta-D-glucopyranosyl-(1----3)-alpha-L-rhamnopyranosyl-(1----2 )[beta-D-glucopyranosyl-(1----4)]-alpha-L-arabinopyranoside (8). Saponin 7 showed the same sugars as 8, but the attachment between the sugars could not be elucidated. The same saponins were present in the roots, but not in the leaves.     

Study of peripheral blood thrombocytes in smokers

Total circulating platelet count and thrombocytogram were examined in 60 tobacco smokers and in 30 nonsmoking normal subjects aged 23-32. The smokers were divided into two subgroups, 30 subjects in each: Group 1 included those with a history of smoking under 5 years and Group 2 those smoking for 5 years and Group 2 those smoking for 6-10 years. The platelet count was found increased in Group 1 smokers at the expense of old, atypical, and involutional forms. In Group 2 smokers the total platelet count and the counts of their differentiation forms were reduced. This evidences the effect of tobacco smoking on circulating platelets, this effect depending on the length of tobacco smoke inhalation.     

Liability for infectious diseases in day care: legal and practical considerations

A lawsuit directed by parents at a day care program following an outbreak of infectious disease would most likely be based on the legal theory of negligence--the failure to exercise due care as required by circumstances. In the determination of liability for infectious diseases in child day care, the element of causation is most critical. Did the day care facility's actions or failure to act cause the harm? Another important question is whether an infectious disease can be occupational and under what circumstances. Although worker's compensation laws initially had no provisions for occupational disease, now all states recognize responsibility for them. Awareness of potential problems with legal liability and establishment of procedures to minimize risk are important for day care providers, public health officials, clinicians, and parents.