Missense mutation in exon 7 of the common gamma chain gene causes a moderate form of X-linked combined immunodeficiency.

Clinical and immunologic features of a recently recognized X-linked combined immunodeficiency disease (XCID) suggested that XCID and X-linked severe combined immunodeficiency (XSCID) might arise from different genetic defects. The recent discovery of mutations in the common gamma chain (gamma c) gene, a constituent of several cytokine receptors, in XSCID provided an opportunity to test directly whether a previously unrecognized mutation in this same gene was responsible for XCID. The status of X chromosome inactivation in blood leukocytes from obligate carriers of XCID was determined from the polymorphic, short tandem repeats (CAG), in the androgen receptor gene, which also contains a methylation-sensitive HpaII site. As in XSCID, X-chromosome inactivation in obligate carriers of XCID was nonrandom in T and B lymphocytes. In addition, X chromosome inactivation in PMNs was variable. Findings from this analysis prompted sequencing of the gamma c gene in this pedigree. A missense mutation in the region coding for the cytoplasmic portion of the gamma c gene was found in three affected males but not in a normal brother. Therefore, this point mutation in the gamma c gene leads to a less severe degree of deficiency in cellular and humoral immunity than that seen in XSCID.

Humoral immunological profile of workers exposed to asbestos in asbestos mines.

Humoral immunological profile including immunoglobulins IgG, IgA, IgM, C-reactive protein, rheumatoid factor, antinuclear antibodies and circulating immune complexes were studied in a representative sample of 36 workers suffering from asbestosis (group A), 35 workers who are exposed to asbestos but not having evidence of asbestosis (group B) and 28 control workers (group C). Mean IgG and IgA levels were found to be significantly higher in the two exposed groups than in the controls. Circulating immune complexes of IgG, IgA and IgM class were detected in a significant percentage of cases in exposed groups than in controls. In groups A and B, the percentage of positive ANF cases was much higher than in the controls. The results suggest that immunological changes are associated with exposure to asbestos and these may play an important role in the pathogenesis of the disease process.

Pollution and its control in asbestos milling processes in India.

An intervention study has been conducted in chrysotile asbestos milling processes with an interval of one year. The 'Membrane Filter Method' adopted in the former study for fiber concentration estimation has been followed in the present study, to estimate the environmental samples under the same experimental conditions. Though very high fiber levels compared to the prescribed permissible limit in India, have also been obtained in this study, a general reduction trend in the values of mills common to the both studies are observed. A further suitable control measure has been recommended in the paper to reduce fiber levels in the milling processes.

The correlation of chest radiograph and pulmonary function tests in asbestos miners and millers.

An environmental-cum-medical survey was carried out in asbestos mines and milling units at Pullivendalla, Cuddaph (A.P.) India. This was done in two mines and six milling units with 95% of the total work force being surveyed. Out of a total of 633 registered workers, 329 (52%, all males) were employed in mines while 135 (21.4%) workers of whom 114 (84%, all females) were employed in the milling units. All subjects underwent limited medical examination, spirometry and chest radiographs. The levels of asbestos fiber concentration was much below threshold limit value (TLV) in underground mines but several times higher than TLV in milling units. The percentage of workers with abnormal pulmonary function tests (PFT) and chest radiographs increased with duration of exposure in smokers as well as non-smokers. Restrictive pattern of lung functions (159 workers-16.27%) was more common than obstructive (33 workers-5.21%) and combined type (22 workers-3.4%). Similarly, the parenchymal changes (156 workers-24.6%) were more common than pleural (27 workers-4.3%). As most of the males were employed in mines, where the fiber levels were much below TLV, the number of male workers with normal PFT and chest radiographs were ten times (61.3%) more than male workers with both the parameters abnormal (6.3%). As most of the females (114 our of 120--95%) were employed in milling units, where the levels of fibers were several times higher than TLV, the number of females having both the parameters normal 29 (24.1%) or abnormal 35 (29.2%) were similar.

The relation of exposure to asbestos and smoking habit with pulmonary function tests and chest radiograph.

Environmental cum medical study was conducted in asbestos cement factory. The environment was evaluated for asbestos fiber by the methods recommended by BIS. Total 355 exposed and 312 suitably matched control workers were investigated by spirometer, Wright's peak flow meter and full sized postero-anterior chest radiograph. The levels of asbestos fiber were 2 to 3 times higher than TLV i.e. 2 f/ml in pipe cutting dept., crude fiber grinding inlet count was more than the ACGIH recommended limit i.e. 5 mpccf of air in pipe cutting dept. and silica mill. In the rest of the department, fiber level as well as dust particle count were below prescribed limit. The comparison of mean values of PFT parameters of workers with 16-20 years exposure history with control one was showing statistically significant decline in mean values of FVC only suggesting restrictive type of PFT impairment in this group of workers. But in workers with more than 20 years exposure, the mean values of all the parameters studied were reduced as compared to control one suggesting combined type of PFT impairment. When the mean values of PFT parameters of exposed smokers were compared with exposed non-smokers there was statistically no significant difference. This can be due to marginal contribution of smoking habit in impairment of PFT parameters of exposed smokers. The percentages of workers with parenchymal and pleural changes due to asbestos exposure were nearly two times more in more than 20 years exposure groups as compared to 11-20 years exposure groups. The parenchymal and pleural changes due to asbestos exposure were more common in exposed smokers as compared exposed non-smokers. However the detailed analysis revealed that if smoking contributes to the development of interstitial fibrosis, the contribution is a marginal one in comparison to the effect of asbestos dust exposure.

Role of achara rasayana and sadvrta in the prevention of AIDS.

Acquired Immuno Deficiency syndrome (AIDS) has emerged as a real threat for mankind since last decade. Lack of knowledge and awareness about AIDS is an important factor responsible for rte rapid spread of the disease. Ayurveda as a part of its holistic approach offers the concepts of Achara Rasayana and sadvrta i.e good conduct for the self and for society. Such concepts can prevent the disease which are basically caused by Prajnaparadha which also appears responsible for the spread of AIDS.

A monitoring study of workers handling pesticides in warehouses and godowns.

Monitoring observations made on 60 operators involved in pesticide application work in godowns and warehouses and 60 matched control workers are reported. Occupational exposure history and medical history are noted. Biochemical investigations, plasma and RBC cholinesterase estimations are included along with medical examination of the workers. Workers were found to be mostly exposed to Celphos, DDVP, Malathion, Pyrethrum, etc, and the use of protective devices were very limited. Cases of significant reduction in plasma and RBC cholinesterase activity were found. Frequency of symptoms like dizziness, headache, lachrymation, burning sensation in eyes, nausea and anorexia, etc, were much more in the exposed workers. No cases of clinical poisoning attributable to occupational exposure to pesticides were reported by the workers.

A new look at the promoter of the human monoamine oxidase A gene: mapping transcription initiation sites and capacity to drive luciferase expression.

Monoamine oxidase (MAO) A (EC 1.4.3.4) oxidizes norepinephrine and serotonin and is expressed in a cell type-specific manner. Recent evidence that MAO A-deficient males in a large Dutch kindred suffer from mild mental retardation and occasional episodes of impulsive aggressive behavior makes it important to understand how the human MAO A promoter is regulated. Conventional primer extension analyses of MAO A mRNA in earlier studies predicted incorrect transcription initiation sites for the human MAO A promoter. Reverse transcription and polymerase chain reaction (PCR) readily detected MAO A mRNA initiated 5' to -135 bp but not 5' to -226 bp (5' to the ATG initiation codon). PCR-assisted primer extension and RNase protection assays reveal that most MAO A mRNA is initiated between -30 and -40, which resembles a eukaryotic initiator element. Depending on the tissue source, a minor, variable proportion of MAO A mRNAs is initiated more distally at approximately -95 and -136, within the more proximal of two 90-bp GC-rich tandem repeats. Genomic DNA segments spanning -4 to -200 and -465 or -935, but not -4 to -82, drive robust luciferase expression in mammalian cells. We conclude that (a) the primary transcription initiation site occurs at a putative initiator (lnr) element located between -30 and -40, with a minor, tissue-specific proportion of additional initiation near -95 and -136; and (b) MAO A-luciferase reporter constructs that contained all the known transcription initiation sites exhibited no evidence for inhibitory cis elements between -200 and at least -935. The apparent inhibitory activity previously reported for sequences 5' to the most proximal PvuII site may have resulted from the use of partial promoter constructs that omitted the putative lnr element.

Post-transcriptional effects of extracellular pH on tumour necrosis factor-alpha production in RAW 246.7 and J774 A.1 cells.

The present studies determined the effects of extracellular pH (pH(o)) on the production of tumour necrosis factor-alpha (TNF-alpha) in the macrophage-like cell lines RAW 246.7 and J774 A.1. The cells were activated with lipopolysaccharide (LPS) at pH(o) 5.5, 6.5 or 7.4. TNF-alpha gene transcription was monitored by Northern blot analysis. Synthesis of the cytokine was monitored by ELISA measurements of the TNF-alpha content of cell-conditioned media (extracellularly released TNF-alpha) and cell lysates (cytosolic TNF-alpha). The magnitude of the TNF-alpha response differed markedly between the two cell lines. RAW cells were more responsive to LPS than were J774 cells. However, the effects of pH(o) on TNF-alpha production were similar in the two cell lines. TNF-alpha gene transcription was insensitive to experimental pH(o). The pH(o) had no effect on the abundance of TNF-alpha mRNA at 2, 4 or 18 h. Nonetheless, synthesis of TNF-alpha was affected significantly by pH(o). The TNF-alpha contents of cell-conditioned medium and cell lysate at 18 h were reduced progressively at lower pH(o) values. The data indicate that pH(o) alters TNF-alpha production in RAW and J774 cells at a post-transcriptional level. These findings suggest that pH(o) influences the phenotypic responses of macrophages to activating stimuli and modifies the role that macrophages play in inflammatory and immune actions.

An environmental survey in chrysotile asbestos milling processes in India.

Environmental monitoring to determine airborne asbestos fiber levels has been carried out in four different mills processing chrysotile asbestos in the Cuddapah District (Andhra Pradesh) of India. The "membrane filter method" comprising standard asbestos sampling techniques, acetone-triacetin method for sample preparation, fiber counting, and sizing using the phase contrast optical microscope were adopted in the study. Fiber concentrations both with respect to personal exposures and processing areas were found in most of the cases to be much higher than the prescribed Threshold Limit Value (TLVs) of the developed and developing countries for chrysotile asbestos. By optical microscopy, fiber length distribution showed 70% of fibers in the milling processes were in size range > 5-10 microns, whereas in > 10-20 and > 20 microns, 20% and 8%, respectively. Fiber identification for major elemental content, also done by using scanning electron microscope equipped with an energy dispersive X-ray analyzer, indicated the presence of tremolite along with chrysotile. The study stresses the urgent need to adopt suitable engineering controls at the dust generating sources to reduce the fiber level in the mill environment below the threshold limit.

Effects of extracellular pH on tumour necrosis factor-alpha production by resident alveolar macrophages.

Cellular acid-base status has been found to exert selective actions on the effector functions of activated macrophages (mphi). We examined the effects of extracellular pH (pH(o)) on the production of tumour necrosis factor-alpha (TNF-alpha) induced by lipopolysaccharide (LPS) in resident alveolar mphi. Cells were obtained by bronchoalveolar lavage of rabbits, activated in vitro with LPS, and cultured at pH(o) 5.5, 6.5 or 7.4 for up to 18 h. The relative abundance of TNF-alpha mRNA peaked at approximately 2 h. The peak transcript abundance was increased at lower pH(o) values. This finding probably reflected pre-transcription/transcription effects of pH, in as much as the stability of TNF-alpha mRNA induced with phorbol ester was unaffected by the experimental pH(o) values. TNF-alpha secretion by LPS-treated mphi decreased at lower pH(o) values. The TNF-alpha content of mphi-conditioned media decreased progressively with decrements in pH(o). The reduced TNF-alpha secretion at pH(o) 5.5 was accompanied by an increase in the cytosolic TNF-alpha content (compared with that at pH(o) 7.4), indicating that pH(o) altered TNF-alpha secretion due, in part, to the intracellular retention of synthesized cytokine (i.e. a post-translation effect). The data show that pH(o) has multiple effects (pre-transcription/transcription and post-translation) on TNF-alpha production induced by LPS in resident alveolar mphi. These results suggest that the role of alveolar mphi in inflammatory responses is modulated by pH(o), which may be important in tumours/abscesses and sites of infection where the external milieu is acidic.

The chitinase PfCHT1 from the human malaria parasite Plasmodium falciparum lacks proenzyme and chitin-binding domains and displays unique substrate preferences.

Within hours after the ingestion of a blood meal, the mosquito midgut epithelium synthesizes a chitinous sac, the peritrophic matrix. Plasmodium ookinetes traverse the peritrophic matrix while escaping the mosquito midgut. Chitinases (EC 3.2.1.14) are critical for parasite invasion of the midgut: the presence of the chitinase inhibitor, allosamidin, in an infectious blood meal prevents oocyst development. A chitinase gene, PgCHT1, recently has been identified in the avian malaria parasite P. gallinaceum. We used the sequence of PgCHT1 to identify a P. falciparum chitinase gene, PfCHT1, in the P. falciparum genome database. PfCHT1 differs from PgCHT1 in that the P. falciparum gene lacks proenzyme and chitin-binding domains. PfCHT1 was expressed as an active recombinant enzyme in Escherichia coli. PfCHT1 shares with PgCHT1 a substrate preference unique to Plasmodium chitinases: the enzymes cleave tri- and tetramers of GlcNAc from penta- and hexameric oligomers and are unable to cleave smaller native chitin oligosaccharides. The pH activity profile of PfCHT1 and its IC(50) (40 nM) to allosamidin are distinct from endochitinase activities secreted by P. gallinaceum ookinetes. Homology modeling predicts that PgCHT1 has a novel pocket in the catalytic active site that PfCHT1 lacks, which may explain the differential sensitivity of PfCHT1 and PgCHT1 to allosamidin. PfCHT1 may be the ortholog of a second, as yet unidentified, chitinase gene of P. gallinaceum. These results may allow us to develop novel strategies of blocking human malaria transmission based on interfering with P. falciparum chitinase.