The O-specific polysaccharide chain of Campylobacter fetus serotype A lipopolysaccharide is a partially O-acetylated 1,3-linked alpha-D-mannan

A polysaccharide fraction liberated from Campylobacter fetus subsp. fetus serotype A lipopolysaccharide by mild acid hydrolysis followed by gel-permeation chromatography contained a partially O-acetylated D-mannan chain, as an O-specific polysaccharide, with a core oligosaccharide attached. The structure of the polysaccharide was studied by O-deacetylation, methylation, and 1H- and 13C-NMR spectroscopy, including computer-assisted analysis of the 13C-NMR spectrum. A structure of -->3)-alpha-D-Manp2Ac-(1--> was established as the structure of the O-specific polysaccharide, the degree of O-acetylation of the mannose residues at position 2 being estimated as 80-90%. As judged by the ratio of mannose to core constituents, the D-mannan chain consists on average of 10-12 monosaccharide units.     

Kinetic evidence for peptide-induced oligomerization of the molecular chaperone DnaK at heat shock temperatures

The pre-steady-state kinetics of the binding of a fluorescent peptide (dansyl-KLIGVLSSLFRPK, fVSV13) to the Escherichia coli molecular chaperone DnaK were investigated over a range of temperatures (25-42 degrees C). At 42 degrees C, over a wide range of peptide concentrations, the fVSV13 peptide bound to DnaK with biphasic kinetics: a rapid burst in the DnaK-fVSV13 signal in the first 5 s was followed by a gradual reduction in the signal over the next 100 s. The descending portion of each biphasic trace followed the equation F(t) = DeltaF exp(-kdt) + Finfinity, where DeltaF, kd, and Finfinity are the amplitude, the apparent first-order rate constant, and the fluorescence end point, respectively. Both DeltaF and kd increased with increasing concentrations of DnaK, which suggests that the loss of the DnaK-fVSV13 signal is caused by a bimolecular reaction. We propose that (i) the fVSV13 peptide binds to and induces a conformational change in the DnaK monomer [E + P right harpoon over left harpoon (EP)*]; and (ii) the conformational change promotes the formation of oligomeric DnaK-peptide complexes [En + (EP)* right harpoon over left harpoon En-EP]. The term (EP)* denotes a monomeric DnaK-peptide complex in which the bound peptide is fluorescent; En-EP denotes an oligomeric DnaK-peptide complex in which the fluorescence of the bound peptide is quenched. Numerical fitting of the stopped-flow data to reactions (i) and (ii) yielded values for the four rate constants. When the proposed kinetic model was tested by conducting experiments in the presence of excess peptide or excess ATP&sbd;conditions which inhibit oligomerization&sbd;DnaK-fVSV13 complex formation proceeded to stable asymptotes, with no reduction in the DnaK-fVSV13 signal at long times.     

Endosulfan poisoning in Northern India: a report of 18 cases

Eighteen cases of endosulfan poisoning by accidental overexposure during spray, admitted between October 1995 to September 1997, were observed and analyzed. These accounted for approximately one third of the total number of poisoning cases admitted in our unit during this period. Nausea, vomiting abdominal discomfort, tonic and clonic convulsions, confusion, disorientation, and muscular twitchings were cardinal manifestations. None of the patients succumbed to their illness. Analysis of various incriminating factors revealed that accidental overexposure was due to failure to adhere to the instructions for spray either due to ignorance or due to illiteracy. All the patients avoided preventive measures and developed toxicity both due to inhalation and absorption through skin. Endosulfan (a chlordiene derivative) poisoning is gaining up momentum in this part of world and has become an important matter for public health in India.     

Hierarchy of lens proteins requiring protection against heat-induced precipitation by the alpha crystallin chaperone

Tooth to denture base discrepancy (the discrepancy) is the difference between the dental arch length and the sum of crown diameters of teeth in the jaw, a concept which was originally developed in orthodontics. Since the cause-effect relationship between a soft diet and the discrepancy has been demonstrated, the size of the discrepancy should indicate the amount of load on the masticatory system from chewing foods in jaws from archaeological periods. The dietary condition of 71 citizens compared to that of 186 slaves from the Yin-Shang period of China was reconstructed through a study of the discrepancy. The prevalence of the discrepancy in the Yin-Shang period was around 15%, almost the same as it was during the later Jomon to Yayoi (3000-2000 BP) periods, when rice agriculture was introduced into Japan, and also the same as for present-day pastoralists around Lake Turkana, Kenya. Although the frequency of the discrepancy was slightly higher in male citizens, there were no significant differences in the frequencies between male citizens and female citizens or slaves. The differences in diet may not have been fundamental since the Yin-Shang period would be at the very beginning of the age in which differences of diet according social class began to appear, with implications for the load on the masticatory system. At that time agriculture may not been sufficiently intensified in variety or quantity to have produced a differentiation of the diet between social classes.     

CD14-dependent and CD14-independent signaling pathways in murine macrophages from normal and CD14 knockout mice stimulated with lipopolysaccharide or taxol

The antitumor agent, Taxol, shares with bacterial LPS the ability to activate murine macrophages, and its LPS-mimetic effects are blocked by LPS analogue antagonists. Since CD14 is central to the recognition of LPS by macrophages, we sought to examine a role for CD14 in the response to Taxol vs LPS. A comparison of responses of macrophages from wild-type mice with those from mice lacking CD14 due to a targeted disruption of the CD14 gene (CD14-deficient knockout (CD14KO)) revealed that like LPS, Taxol induces both CD14-dependent and -independent pathways of gene activation, although the CD14 dependency of Taxol stimulation is much less striking than that observed with LPS. The macrophage interaction with low concentrations of LPS (< or = 10 ng/ml) is largely CD14 dependent, as evidenced by the lack of induction of TNF-alpha, IL-1beta, and interferon-inducible protein-10 (IP-10) genes by CD14KO macrophages cultured in the absence of soluble CD14 (i.e., in autologous CD14KO -/- mouse serum). However, at high concentrations of LPS or Taxol, a CD14-independent pathway of activation is observed: this pathway leads to minimal IP-10 gene induction, even though induction of TNF-alpha and IL-1beta occurs. Measurements of TNF secretion followed a similar pattern to that observed at the level of steady state mRNA. These data suggest the existence of two pathways of activation by both LPS and Taxol: one that is CD14 dependent and leads to induction of TNF-alpha, IL-1beta, and IP-10 gene induction, and a CD14-independent pathway that results in the induction of TNF-alpha and IL-1beta, with minimal induction of IP-10.     

Overproduction of Campylobacter ferritin in Escherichia coli and induction of paracrystalline inclusion by ferrous compound

Levels of the proteoglycans hyaluronic acid, heparan sulfate, dermatan sulfate and chondroitin sulfate in brains were measured. The samples were from 25 brains obtained at autopsy from patients who had senile dementia of Alzheimer's type (SDAT) (60 to 97 years old) and 10 brains from non-demented elderly subjects (75 to 101 years old). Proteoglycan levels in the brains of patients with SDAT were twice as high in the hippocampus and four times higher in the superior frontal gyrus than the levels in brains of non-demented elderly subjects. The content of heparan sulfate proteoglycan was greatest nine times higher in the hippocampus and seven times higher in the superior frontal gyrus. By immuno-electronmicroscopic examination, heparan sulfate proteoglycan was found in the basement membranes of capillaries, and in a portion of the cores of the senile plaques.     

Effectiveness of bacillus Calmette-Guerin (BCG) vaccination in the prevention of leprosy; a case-finding control study in Nagpur, India

Among numerous available materials for osseous repair and reconstruction, those presenting osteoinductive characteristics and promoting bone regeneration are preferable. Fresh autologous bone is one of the most effective, but it has some disadvantages and risks. Demineralized bone matrix (DBM) is considered to be a valid alternative, because it seems to show osteogenic potential, ascribed to the presence of bone morphogenetic proteins. In addition it can be prepared without difficulty and preserved without losing osteoinductive properties. The aim of the study was to evaluate the osteoinductive ability of xenogenic DBM, by testing DBM powder obtained from rabbit long bones, in cell culture of murine fibroblasts, alone or associated with electromagnetic field (EMF), that are known to exhibit biologic effects on cells: in particular they are used in orthopedics to improve bone formation. At the end of experiment, alkaline phosphatase (ALP) activity, calcium levels and cell proliferation and morphology were evaluated. A statistically significant stimulation of ALP activity and cell proliferation and a morphological change of fibroblasts were found. The results obtained show how DBM and EMF have different effects on cells, and that together they have synergic action toward bone induction.