Usefulness of spoligotyping in molecular epidemiology of Mycobacterium bovis-related infections in South America

We studied the role of Ca++ and protein kinase C (PKC) in alpha-1A adrenergic receptor (AR)-mediated activation of mitogen-activated protein kinase pathways in PC12 cells. In PC12 cells stably transfected with the human alpha-1A AR, norepinephrine (NE) strongly activated both extracellular signal regulated kinases (ERKs) and c-jun-NH2-terminal kinases (JNK). Ten nanomolar thapsigargin (TG) increased cytoplasmic Ca++ at least as much as NE but did not activate ERKs or JNK. Higher concentrations of TG caused a small activation of ERKs but not JNK. Emptying [Ca++]i stores by pretreatment with TG prevented the NE-stimulated increase in [Ca++]i but not ERK or JNK activation. The Ca++ chelator bis(2-aminophenoxy)ethane-N-N-N'-N'-tetraacetate (BAPTA) dose dependently abolished NE-stimulated Ca++ responses but not ERK or JNK activation. NE increased tyrosine phosphorylation of Pyk2, and this response was neither blocked by BAPTA nor mimicked by TG. The phorbol ester tumor promoting agent (TPA) caused a dose-dependent activation of ERKs that was potentiated by 10 nM TG. TPA caused only a small activation of JNK relative to that caused by NE, which was not affected by TG. The potent PKC inhibitor bisindolylmaleimide I dose dependently inhibited ERK and JNK activation by TPA, but not NE. ATP and UTP activated similar mitogen-activated protein kinase responses through endogenous P2Y2 receptors, and these responses were not blocked by BAPTA or bisindolylmaleimide I, suggesting that these results may be generalizable to other Gq/11-coupled receptors. The results suggest that Ca++ release and PKC activation are neither necessary nor sufficient for alpha-1A AR-mediated activation of mitogenic responses in PC12 cells.     

Characterization and molecular cloning of the lectin from Helianthus tuberosus

A lectin called Helianthus tuberosus agglutinin or Heltuba has been isolated from tubers of the Jerusalem artichoke, a typical representative of the Asteraceae family. Heltuba is a tetrameric protein composed of four identical subunits of 15.5 kDa and exhibits a preferential specificity towards oligomannosides. Cloning of the corresponding cDNAs revealed that the mature lectin polypeptide comprises the entire open reading frame of the cDNA suggesting that the primary translation product is not processed and that the lectin is a cytosolic protein. Searches in the databases revealed sequence similarity with lectins from the taxonomically unrelated Convolvulaceae and Moraceae species. Therefore, the discovery of Heltuba is of great importance in view of the occurrence and molecular evolution of the jacalin-related lectins.     

Immunoglobulin levels in populations with low hemoglobin levels: study of five tribes of central India

Average hemoglobin levels for 424 individuals belonging to 5 tribes of central India vary between 10.64 g/dl and 11.30 g/dl among males and between 9.62 g/dl and 9.97 g/dl among females. These ranges indicate the poor state of health and nutrition in these tribes. Three immunoglobulin levels (IgG, IgA, and IgM) measured in a subset of the 424 individuals reveal that immunoglobulin and hemoglobin levels are inversely related to some extent. Levels of all three classes of immunoglobulins were found to be significantly correlated. This suggests that humoral immunity is not impaired even under acute nutritional stress.     

Alterations of low molecular weight acid phosphatase protein level in Alzheimer''s disease

We have previously reported that the activity of low molecular weight (LMW) acid phosphatase, which can remove tyrosine-linked phosphates of epidermal growth factor receptor, was significantly decreased in Alzheimer brains. In the present study, a specific antibody was prepared to analyze the protein level of this enzyme. Western blot analysis indicated that the level of LMW acid phosphatase protein was significantly reduced, whereas the activity of LMW acid phosphatase per enzyme molecule was not changed in Alzheimer brains. These results suggest that the reduction of LMW acid phosphatase activity in Alzheimer brains is due to its decreased protein level in Alzheimer's disease.     

Competing discourses of vital registration and personhood: perspectives from rural South Africa

Whilst birth and death data derived from civil registration systems are regarded as essential indicators of health status and important for population planning, in developing countries they are usually perceived by civil servants and researchers to be very incomplete. In South Africa in 1994 only 50% of deaths were registered and 18% of births in the first year of life. A rapid qualitative study was undertaken in a rural district of South Africa to ascertain why registration levels of births, still-births and infant deaths are so low. Fifty-five semi-structured interviews were conducted with 22 local Xhosa women and 33 "professional" key informants, ranging from local civil servants to hospital staff and grave diggers. The study found that local people had complex notions of personhood, before birth and in the years following. Personhood was viewed as a process rather than a stage which is achieved through live birth, as is implied in discourses of vital registration. The women interviewed knew about birth registration although most had registered some or none of their children; they did not know of death registration. There was little knowledge of why registration was necessary and perceptions of this among all informant groups mostly related to the need for a certificate to achieve something else, such as an identity document or welfare payment. Confusion about the procedures to be followed was found among both women and professionals, who advised them. In circumstances in which certificates were officially required, for school entry and burial, other documentation were reported to be accepted. This suggests that the dominance of vital registration as a means of establishing official identity was not recognized. Registration was regarded as a means of achieving something else rather than and end in itself, which discourses of statistical and juridical importance imply. In the light of this we suggest that the present system be replaced by one based on "passive" registration in health care settings if substantially greater levels of completeness are to be achieved.     

Five novel missense mutations of the Lewis gene (FUT3) in African (Xhosa) and Caucasian populations in South Africa

A new bivalent ligand of formula 3 which results from the covalent coupling of two sumatriptan molecules with a p-xylyl spacer at the level of the sulfonamide nitrogen has been prepared and evaluated as a 5-HT1B/1D receptors agonist. In vitro experiments at 5-HT1B human cloned receptors (Ki = 0.64 nM; EC50 = 0.58 nM) and at the level of the contraction of the New Zealand white rabbit saphenous vein (pD2 = 6.6) demonstrate the superior potency of dimer 3 as a 5-HT1B receptor agonist when compared to sumatriptan or zolmitriptan. Interestingly enough, the new bivalent agonist 3 was found to induce hypothermia in the guineapig upon oral administration suggesting good oral activity and access to the brain.     

Alterations in fibrinolytic and protein C pathways in gynaecological surgery: low molecular weight heparin prophylaxis

Several parameters of fibrinolytic and protein C pathways were evaluated in three groups of patients with high (HR), moderate (MR) and low (LR) postoperative thrombotic risk undergoing major gynaecological surgery. The HR and MR groups were subjected to low molecular weight heparin (LMW) prophylaxis. A significant increase in plasminogen activator inhibitor type 1 (PAI-1) antigen and activity levels was observed in the HR patient group in comparison with the MR and LR groups in the preoperative and early postoperative period. In all the groups studied, the maximum increase in the levels of PAI-1 was seen on day 1 after surgery. However, the D-dimeric levels reached the highest level on day 7. A significant increase in activated protein C:alpha 1 antitrypsin (APC:alpha 1AT) complex levels was observed in the HR group in comparison with the LR group, and a strong decrease in protein C inhibitor in the early postoperative period was detected in all the groups. In spite of heparin prophylaxis, 2 HR patients were diagnosed as deep vein thrombosis (DVT) during the postoperative period. Both patients showed pre-operative levels of PAI-1 antigen or activity and APC:alpha 1AT complexes above the mean + 1 SD of the pre-operative levels in the HR group. In conclusion, in HR patients a hypofibrinolytic and hypercoagulable state was detected in the pre-operative and early postoperative periods. The prophylactic LMW heparin dose used in the present report (20 mg/day x 7) was insufficient to prevent DVT in the HR group. At present our HR patients are given higher doses of LMW heparin (40 mg/day x 7).     

Capillary electrophoretic analyses of beta-trace protein and other low molecular weight proteins in cerebrospinal fluid from patients with central nervous system diseases

Ordinary capillary-zone electrophoresis (CZE), as well as CZE in a sodium dodecylsulfate-containing polymer solution (SDS-CZE) and capillary isoelectrofocusing (CIEF), was applied to the analysis of low molecular weight proteins in cerebrospinal fluid (CSF) from patients with various neuropsychiatric disorders. Under the CZE conditions employed, a peaks of beta-trace protein (beta TP), which is the most abundant low MW protein in CSF, was clearly detected on the electropherograms of all the samples examined, and the CSF beta TP level could be tentatively determined using allylamine added at a constant concentration as the internal standard. The results revealed that beta TP in CSF was non-specifically increased in organic disease in the central nervous system (CNS), especially in ones giving severe physical damage to the brain tissues. On the other hand, SDS-CZE allowed us to determine simultaneously the CSF minor low MW proteins other than beta TP, such as beta 2-microglobulin, gamma-trace protein, myelin basic protein, etc., while the CIEF electropherograms suggested that beta TP were separated into several fractions with the different PI values. These capillary electrophoresis systems seem to be powerful as aids in the biochemical examinations of beta TP and other low molecular weight proteins in CSF from patients with CNS diseases.     

Cloning of a lectin cDNA and seasonal changes in levels of the lectin and its mRNA in the inner bark of Robinia pseudoacacia

E. coli cells harboring pCG169 containing the secD secF locus possessed SecA protein almost entirely in an integral membrane form in which it displayed normal protein translocation activity. These results imply that integral membrane SecA is the catalytically active form of this enzyme and that products of the secD secF locus regulate SecA association with the inner membrane. Protease and biotinylation accessibility studies of right side-out and inside-out membrane vesicles derived from this strain revealed that SecA was exposed to the periplasmic surface of the inner membrane. These studies suggest a model of bacterial protein secretion, whereby insertion of SecA into the inner membrane and its association with SecY/E/G promotes assembly of active protein-conducting channels comprised in part of integral membrane SecA protein, and products of the secD secF locus regulate the channel assembly-disassembly reaction by modulating the SecA insertion-deinsertion step.     

Relationship between acute diarrhoea and low plasma levels of vitamin A and retinol binding protein

OBJECTIVE: We studied possible sex differences of the effect of fenofibrate on serum lipoproteins. Twenty-three patients with primary hypercholesterolaemia (10 postmenopausal women and 13 aged-matched men) were treated with slow-release fenofibrate for 96 weeks. RESULTS: Steady state lipoprotein concentrations were reached after 12 and 24 weeks of treatment in women and men, respectively. During the subsequent follow-up the lipoprotein concentrations remained constant. In women total and low-density lipoprotein (LDL) cholesterol decreased from 299 to 234 mg.dl-1 and from 210 to 151 mg.dl-1, respectively, and in men from 265 to 233 mg.dl-1 and from 192 to 160 mg.dl-1. The decrease in triglycerides was also more pronounced in women (-42%) than in men (-18%). High-density lipoprotein (HDL) cholesterol increased significantly in women from 53 to 63 mg.dl-1 but not in men (45 to 50 mg.dl-1). Since the changes in LDL and HDL cholesterol occurred in opposite directions, the decrease in LDL/HDL cholesterol ratio was accentuated in both groups. However, this ratio was decreased almost twofold in women (-41%) compared to men (-23%). Although the serum concentrations of fenofibric acid were 1.3-fold higher in women than in men, which was probably due to the higher body weight in men (1.2-fold), this difference can hardly explain the favorable effect on lipoproteins in women. CONCLUSION: The present study indicates that fenofibrate might be very effective by reducing the concentrations of atherogenic lipoproteins in postmenopausal women.     

Salmonella isolated from crocodiles and other reptiles during the period 1985-1994 in South Africa

Esophagorespiratory fistulas were frequently caused by malignant tumors, bougienage, laser therapy or radiochemotherapy. We here report the case of a patient with inoperable bronchial cancer, who developed a symptomatic esophagorespiratory fistula during combined radiochemotherapy with Cisplatin. A sufficient occlusion of the fistula could not be achieved with conventional plastic tubes or novel self-expanding silicone-coated Gianturco Song stents. After extraction of two Gianturco Song stents we inserted a Montgomery Salivary Bypass Stent into the esophagus and Dynamic stent into the trachea. This resulted in a total occlusion of the fistula. This present case suggests that the Montgomery stent may have little tendency to migrate due to its characteristic configuration and fixation and further demonstrates that the novel self-expanding silicone-coated Gianturco Song stents can be removed, if necessary.     

Release of low molecular weight silicones and platinum from silicone breast implants

We have conducted a series of studies addressing the chemical composition of silicone gels from breast implants as well as the diffusion of low molecular weight silicones (LM-silicones) and heavy metals from intact implants into various surrounding media, namely, lipid-rich medium (soy oil), aqueous tissue culture medium (modified Dulbecco's medium, DMEM), or an emulsion consisting of DMEM plus 10% soy oil. LM-silicones in both implants and surrounding media were detected and quantitated using gas chromatography (GC) coupled with atomic emission (GC-AED) as well as mass spectrometric (GC/MS) detectors, which can detect silicones in the nanogram range. Platinum, a catalyst used in the preparation of silicone gels, was detected and quantitated using inductive argon-coupled plasma/mass spectrometry (ICP-MS), which can detect platinum in the parts per trillion range. Our results indicate that GC-detectable low molecular weight silicones contribute approximately 1-2% to the total gel mass and consist predominantly of cyclic and linear poly-(dimethylsiloxanes) ranging from 3 to 20 siloxane [(CH3)2-Si-O] units (molecular weight 200-1500). Platinum can be detected in implant gels at levels of approximately 700 micrograms/kg by ICP-MS. The major component of implant gels appears to be high molecular weight silicone polymers (HM-silicones) too large to be detected by GC. However, these HM-silicones can be converted almost quantitatively (80% by mass) to LM-silicones by heating implant gels at 150-180 degrees C for several hours. We also studied the rates at which LM-silicones and platinum leak through the intact implant outer shell into the surrounding media under a variety of conditions. Leakage of silicones was greatest when the surrounding medium was lipid-rich, and up to 10 mg/day LM-silicones was observed to diffuse into a lipid-rich medium per 250 g of implant at 37 degrees C. This rate of leakage was maintained over a 7-day experimental period. Similarly, platinum was also observed to leak through intact implants into lipid-containing media at rates of approximately 20-25 micrograms/day/250 g of implant at 37 degrees C. The rates at which both LM-silicones and platinum have been observed to leak from intact implants could lead to significant accumulation within lipid-rich tissues and should be investigated more fully in vivo.     

Asp129 of low molecular weight protein tyrosine phosphatase is involved in leaving group protonation

Site-directed mutagenesis was used to explore the functions of a number of acidic residues of bovine low molecular weight protein tyrosine phosphatase. Residues Asp-129, Asp-56, and Asp-92 were mutated to Ala or Asn. The mutant enzymes D56A, D56N, and D92A showed no significant changes in Vmax values, although they did exhibit significantly altered Km values. In contrast, the D129A mutant enzyme exhibited a greater than 2000-fold reduction in Vmax, using p-nitrophenyl phosphate as a substrate. The Vmax values of D129A also exhibited a leaving group dependence, an altered solvent isotope effect of VmaxH/VmaxD of 0.78, and a lack of dependence on the presence of alternative phosphate acceptor alcohols, all properties that distinguish this mutant from wild type enzyme. The differences are due to a change of the rate-limiting step of the catalytic reaction. Asp-129 is concluded to be the proton donor to the leaving group in the phosphorylation step, and its mutation to alanine results in a reduced Vmax value and a change in the rate-limiting step of the catalysis from dephosphorylation to phosphorylation. Mechanistic considerations suggest that other phosphotyrosyl phosphatases having cysteine at the active site may be expected to have a similar requirement for a proton donor.     

Phylogenetic analysis of GB virus C: comparison of isolates from Africa, North America, and Taiwan

A group of flavi-like GB viruses (GBV-A, -B, and -C) has been discovered and partially characterized. Only GBV-C is found in humans. Nucleotide sequences of the putative helicase gene of the GBV-C genome were determined in 21 Taiwanese patients and compared with isolates from Africa and North America by phylogenetic analysis. The average similarities of Taiwanese isolates to those from West Africa, East Africa, Canada, and United States were 83%, 85%, 78%, and 82%, respectively. Phylogenetic analysis of the NS3 region showed that 81% of the Taiwanese isolates were more closely related to the East African isolate. GBV-C isolates from Taiwan could be classified into at least 3 groups. These data suggest that GBV-C isolates cloned from different geographic areas have genetic heterogeneity.     

A cholera outbreak and control in a rural region of South Africa

Dance injuries associated with cervical radiculopathy have not been described in the literature. This report describes the case of an international-style ballroom dancer who developed a cervical radiculopathy as a result of frequent lateral rotation and hyperextension of the cervical spine during dancing. The patient's symptoms and signs suggestive of a left C7 radiculopathy were confirmed and documented by both magnetic resonance imaging and electrodiagnostic testing. The patient was treated conservatively with activity modification, nonsteroidal anti-inflammatory drugs, and alternative medicine approaches, including herbs and acupuncture. Her neck pain and cervical radicular symptoms declined in severity, but continued even 4 1/2 months after the onset of her symptoms. She did not wish to try steroids either through an oral or epidural route and refused surgical intervention. This case report illustrates an unconventional manner in which a left cervical radiculopathy was clinically produced. The neck motions and positions of frequent hyperextension and lateral rotation demonstrated by this ballroom dancer simulated a pattern and sequence of movements that promoted the development of signs and symptoms of a left cervical radiculopathy.     

A field study of own-race bias in South Africa and England.

In shopping centers in South Africa and England, Black and White people were approached by either a Black or a White confederate. Later they were asked to identify the confederate from both a sequential lineup, where photographs are seen one at a time, and a forced-choice recognition test, where photographs are seen simultaneously. In both countries and with both identification tasks the confederates were better recognized by people of their own race. By observing this own-race bias in two countries with different demographics, using methods similar to those used in some criminal investigations of eyewitness memory, the authors demonstrated the robust nature of the own-race bias. In addition, witness confidence and accuracy were found to be correlated but only when the participant was the same race as the confederate. Implications for legal procedure are discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Molten globule-like state of peanut lectin monomer retains its carbohydrate specificity. Implications in protein folding and legume lectin oligomerization

A central question in biological chemistry is the minimal structural requirement of a protein that would determine its specificity and activity, the underlying basis being the importance of the entire structural element of a protein with regards to its activity vis à vis the overall integrity and stability of the protein. Although there are many reports on the characterization of protein folding/unfolding intermediates, with considerable secondary structural elements but substantial loss of tertiary structure, none of them have been reported to show any activity toward their respective ligands. This may be a result of the conditions under which such intermediates have been isolated or due to the importance of specific structural elements for the activity. In this paper we report such an intermediate in the unfolding of peanut agglutinin that seems to retain, to a considerable degree, its carbohydrate binding specificity and activity. This result has significant implications on the molten globule state during the folding pathway(s) of proteins in general and the quaternary association in legume lectins in particular, where precise subunit topology is required for their biologic activities.