Regulation of STAT3 signaling in IFNγ and IL10 pathways and in their cross-talk.

The pro-inflammatory IFNγ-STAT1 pathway and anti-inflammatory IL10-STAT3 pathway elicit cellular responses primarily utilizing their canonical STATs. However IL10 mediated STAT1 and IFNγ mediated STAT3 activation is also observed, suggesting crosstalk of these functionally opposing signaling pathways can potentially reshape the canonical dynamics both STATs and alter the expression of their target genes. Herein, we measured the dynamics of STATs in response to different doses of IL10 or IFNγ and in their co-stimulation and employed quantitative modeling to understand the regulatory mechanisms controlling signal responses in individual and co-simulation scenarios. Our experiments show, STAT3 in particular, exhibits a bell-shaped dose-response while treated with IFNγ or IL10 and our model quantiatively captured the dose-dependent dynamics of both the STATs in both pathways. The model next predicted and subsequent experiments validated that STAT3 dynamics would robustly remain IL10 specific when subjected to a co-stimulation of both IFNγ and IL10. Genes common to both pathways also exhibited IL10 specific expression during the co-stimulation. The findings thus uncover anovel feature of the IL10-STAT3 signaling axis during pathway crosstalk. Finally, parameter sampling coupled to information theory based analysis showed that bell-shaped signal-response of STAT3 in both pathways is primarily dependent on receptor concentration whereas robustness of IL10-STAT3 signaling axis in co-stimulation results from the negative regulation of the IFNγ pathway.

Esterified coir pith as an adsorbent for the removal of Co(II) from aqueous solution.

Coir pith was chemically modified for the adsorption of cobalt(II) ions from aqueous solution. Chemical modification was done by esterification using succinic anhydride followed by activation with NaHCO(3) in order to improve the adsorption of Co(II). Adsorptive removal of Co(II) from aqueous solution onto modified coir pith was evaluated in batch studies under varying conditions of agitation time and metal ion concentration to assess the kinetic and equilibrium parameters. A pseudo-second-order kinetic model fitted well for the sorption of Co(II) onto modified coir pith. Sorption kinetics showed that the loading of Co(II) by this material was quite fast under ambient conditions. The Langmuir and Freundlich equilibrium isotherm models provided excellent fits for the adsorption data, with R(2) of 0.99 and 0.98, respectively. After esterification, the maximum Co(II) sorption loading Q(0); was greatly improved. It is evident that chemically modified adsorbent exhibits better Co(II) removal capability than raw adsorbent suggesting that surface modification of the adsorbent generates more adsorption sites on its solid surface for metal adsorption. A complete recovery of the adsorbed metal ions from the spent adsorbent was achieved by using 1.0N HCl.

Prevalence of anemia among pregnant women and adolescent girls in 16 districts of India.

BACKGROUND: Nutritional anemia is one of India's major public health problems. The prevalence of anemia ranges from 33% to 89% among pregnant women and is more than 60% among adolescent girls. Under the anemia prevention and control program of the Government of India, iron and folic acid tablets are distributed to pregnant women, but no such program exists for adolescent girls. OBJECTIVE: To assess the status of anemia among pregnant women and adolescent girls from 16 districts of 11 states of India. METHODS: A two-stage random sampling method was used to select 30 clusters on the basis of probability proportional to size. Anemia was diagnosed by estimating the hemoglobin concentration in the blood with the use of the indirect cyanmethemoglobin method. RESULTS: The survey data showed that 84.9% of pregnant women (n = 6,923) were anemic (hemoglobin < 110 g/L); 13.1% had severe anemia (hemoglobin < 70 g/L), and 60.1% had moderate anemia (hemoglobin > or = 70 to 100 g/L). Among adolescent girls (n = 4,337)from 16 districts, the overall prevalence of anemia (defined as hemoglobin < 120 g/L) was 90.1%, with 7.1% having severe anemia (hemoglobin < 70 g/L). CONCLUSIONS: Any intervention strategy for this population must address not only the problem of iron deficiency, but also deficiencies of other micronutrients, such as B12 and folic acid and other possible causal factors.

Investigation of ethnic neutropenia by assessment of bone marrow colony-forming cells.

Healthy individuals with African ancestry have lower neutrophil counts than Caucasians. It has previously been postulated that this was consequent on either a diminished bone marrow granulocyte reserve or an altered distribution of neutrophils between the circulating and marginated granulocyte pools. Recent indirect evidence supports the former hypothesis. In this study we have compared the number of granulocyte plus granulocyte-macrophage colony-forming units (CFUs) in the bone marrow of healthy African and Afro-Caribbean subjects with the number of CFUs in the bone marrow of healthy age and sex-matched Caucasians. We found the group with African ancestry to have significantly fewer CFUs than the Caucasian group. There was no evidence of any qualitative difference between the CFUs of the two ethnic groups: they showed similar sensitivity to granulocyte-monocyte colony stimulating factor and similar enhancement of growth when cultured with a larger range of cytokines. These observations suggest that ethnic neutropenia observed in those with African ancestry is likely to result from reduced numbers of bone marrow progenitor cells in comparison with numbers present in Caucasians.

Immobilization of hormones for drug targeting.

Biological active compounds such as insulin, heparin, progesterone and labeled-LH were entrapped in glutaraldehyde cross-linked bovine serum albumin (BSA) and human serum albumin (HAS) microspheres. Studies were carried out for their binding capacity and biodegradability using new proteolytic enzymes. Effects of proteolytic enzymes such as trypsin, chymotrypsin, papain and pronase-E on microspheres were studied in order to understand the biodegradability of the cross-linked proteins. It has been observed that labeled-LG was entrapped 60% in BSA and HAS microspheres. Labelled-LH-BSA, Labelled-LH-HAS and insulin microspheres were injected into mice and rabbits. It was observed that these cross-linked microspheres were biodegradable and the process appeared to be slow one, useful for sustained release of hormones. It was also observed that these albumin microspheres exhibit fluorescence at 495 nm.

Study of the nonresorptive phenotype of osteoclast-like cells from patients with malignant osteopetrosis: a new approach to investigating pathogenesis.

Osteopetrosis manifests as failure of osteoclastic bone resorption. The cause of the disease lies either in the hematopoietic lineage or in the bone marrow stromal microenvironment. It has not been possible to define the cell type involved in the various forms of the human disease because of the inability to form human osteoclasts in vitro. Using the recently described method for generating human osteoclasts from peripheral blood in coculture with rat osteoblastic UMR 106 cells, we demonstrate that a defect lies in the mature osteoclast-like cells in four cases of this disease. Control and osteopetrotic cocultures generated large numbers of osteoclast-like cells (calcitonin and vitronectin receptor positive, and F-actin ring-positive cells) with similar morphology. Bone resorption did not occur in three of the four osteopetrotic cultures. In case 1, in which bone resorption was identified, the area of resorption was negligible compared with the number of osteoclast-like cells in the culture and was detected only by scanning electron microscopy. In contrast, up to 20% of the bone surface in controls was resorbed. The normal and osteopetrotic osteoclast-like cells had a similar phenotype except that two of the osteopetrotic cases did not express CD44 and two expressed CD44 weakly, whereas CD44 was strongly expressed in the controls. This study shows that it is possible to reproduce in vitro the pathological features of human osteopetrosis, and the assay provides a means of acquiring a greater understanding of the pathogenesis of human osteopetrosis.

Macrophage colony stimulating-factor transcripts are differentially regulated in rat bone-marrow by gender hormones.

There are at least three forms of macrophage colony-stimulating factor (M-CSF), a cytokine that is critical for osteoclast formation; and evidence exists that the membrane-bound form is involved in this process. We wished to test the hypothesis that the expression of the membrane form of M-CSF is modulated by the presence of gender steroids. This was achieved by analyzing M-CSF messenger RNA and protein in the bone-marrow of estrogen- and androgen-replete, and -deficient female rats. We found that the 1.4-kb M-CSF transcript was not detected in sham-operated rats but that the 4.6-kb transcript was expressed in abundance. In contrast, these transcripts were differentially expressed in ovariectomized rats, and this effect was reversed by 17beta-estradiol treatment. Administration of androstenedione to ovariectomized rats, so that androstenedione plasma levels were restored to just below that in sham-operated rats, also suppressed the expression of the 1.4-kb M-CSF transcript. This effect was abrogated by antiandrogen treatment, indicating that this was an androgen-mediated effect. The membrane-bound protein was detected in the bone-marrow of sham-operated rats and was elevated post ovariectomy, whereas ovariectomy had no effect on the soluble isoform. Our data support the hypothesis that the membrane form of M-CSF is modulated by gender hormones and that this isoform is involved in the estrogen- and androgen-mediated effects on the skeleton.

Macrophage colony-stimulating factor increases bone resorption by osteoclasts disaggregated from human fetal long bones.

Macrophage colony-stimulating factor (M-CSF) is known to play an important role in human and murine osteoclast formation. Although M-CSF has been shown to inhibit isolated neonatal rat osteoclasts from resorbing bone, its action on the mature human osteoclast has not been described. We now report that M-CSF increases osteoclastic bone resorption in a dose-responsive manner. Bone resorption by mature human fetal osteoclasts, including pit area, depth, and volume, was increased in the presence of M-CSF compared with vehicle. The number of osteoclasts in the cultures was similar after 2 and 18 h in the presence of M-CSF, whereas there was a significant reduction in osteoclast number, whether assessed as the number of tartrate-resistant acid phosphatase (TRAP)-positive or vitronectin receptor-positive cells after 18 h in M-CSF-free cultures. The number of nuclei per osteoclast after 2 or 18 h in M-CSF was also similar and there was no difference in the number of vitronectin receptor-positive mononucleate cells at 2 and 18 h. This suggests that the increased bone resorption is likely to be accounted for by enhanced osteoclast survival in M-CSF compared with controls rather than by formation of new osteoclasts.

Inhibition of bone resorption by 17beta-estradiol in human bone marrow cultures.

Estrogen deficiency puts individuals at risk of developing osteoporosis because it causes increased bone resorption. However, the mechanism by which this occurs is not known. We have shown, using a recently described two-phase human bone marrow culture system, that estradiol (17beta-E2) added to phase I results in inhibition of bone resorption by reducing the number of osteoclasts (identified as vitronectin receptor and/or calcitonin receptor-positive cells) formed in the cultures. The addition of 17beta-E2 in phase II was without effect, indicating that it does not interfere with the bone resorptive process. 17Beta-E2 down-regulated mRNA expression and protein synthesis of the membrane form of macrophage colony-stimulating factor (M-CSF). 17Beta-E2 did not the alter the expression of the 4.0 kb M-CSF transcript. However, it increased protein synthesis of the proteoglycan form of M-CSF, but not the 85 kDa soluble form in the same cultures. Finally, addition of M-CSF to the cultures reversed the 17beta-E2-induced inhibitory effect. These observations suggest that regulation of the synthesis of membrane-bound M-CSF plays a role in 17beta-E2-induced inhibition of bone resorption.

Study of the cell biology and biochemistry of cherubism.

AIMS: To establish whether the multinucleate cells in lesions of patients with cherubism are also osteoclasts and if this is the case whether they were responsive to calcitonin; to carry out cytogenetic studies on two members of the same family affected by cherubism in an attempt to identify any major chromosomal defects; and to perform an in-depth modern biochemical study of four children in the same family. SUBJECTS AND METHODS: Four related children with cherubism were studied. Tissue taken from one of the children at elective decompression of an optic nerve was submitted to in vitro bone resorption studies. Cytogenetic studies were done on two of the children and biochemical studies on all four. RESULTS: The multinucleate cells in the cherubic lesions were shown to be osteoclasts since they synthesised tartrate resistant acid phosphatase, expressed the vitronectin receptor, and resorbed bone. Bone resorption by the cultured multinucleate cells was significantly inhibited by calcitonin. High resolution cytogenetic studies failed to detect any chromosomal abnormalities in two children with cherubism. The biochemistry profile of all four children with cherubism showed that serum calcium, parathyroid hormone, parathyroid related hormone, calcitonin, and alkaline phosphatase were within normal levels. Urine analysis of pyridinium and deoxypyridinium cross links, hydroxyproline, and calcium in relation to urine creatinine were measured to assess bone resorption in these children, and the values were at the upper end of the normal range in all four. CONCLUSIONS: Further studies are required to determine whether calcitonin treatment will control this grossly deforming disease until the time when the physiological changes that occur at puberty rectify the pathology. It is not recommended that biochemical markers of bone resorption are used in isolation to monitor the activity of cherubism in individuals because the results are based on a small number of children and because of reports of marked interindividual variation in the levels of these markers, particularly in children.

Macrophage colony-stimulating factor induces substantial osteoclast generation and bone resorption in human bone marrow cultures.

Macrophage colony-stimulating factor (M-CSF) is essential for murine osteoclast formation and its role in human hematopoiesis in vitro is not fully defined. Therefore, we have investigated the effect of M-CSF on the formation of human osteoclasts in vitro. M-CSF was found to induce substantial bone resorption and osteoclast formation in a dose-responsive and time-dependent manner above that induced by 1,25 dihydroxyvitamin D3 (1,25 vitamin D3) in cultures of human bone marrow (BM) stromal cells sedimented onto devitalized bone. By day 14 there was a mean of approximately 50% of the surfaces of the bone slices resorbed compared with only 6% in cultures treated with 1,25 vitamin D3 alone. Osteoclasts were identified as 23c6+ cells (an antibody that recognizes the vitronectin receptor), 87.5% of which coexpressed the calcitonin receptor. The number of 23c6+ cells correlated strongly with bone resorption spatially, and in a dose-responsive and time-dependent manner; the correlation coefficient in cultures treated with 1,25 vitamin D3 alone was 0.856 and those treated with both M-CSF and 1,25 vitamin D3 was 0.880. Granulocyte-macrophage colony-stimulating factor, IL-1 beta, IL-3, IL-6, tumor necrosis factor-alpha, transforming growth factor-beta, leukemia inhibitory factor, and IL-11 did not increase bone resorption above that in 1,25 vitamin D3-treated cultures. We also found that 1,25 vitamin D3 increased, to a minor but significant degree, both bone resorption and the concentration of M-CSF in the culture supernatants above that in vehicle-treated cultures, indicating that M-CSF is present in our BM cultures, but that there is insufficient to induce substantial osteoclast formation. These results define a critical role for M-CSF in the formation of human osteoclasts.

Temporomandibular joint ankylosis: an Indian experience.

Seventy-five condylectomy and coronoidectomy specimens of temporomandibular joint ankylosis in 61 patients were studied. Fourteen patients had bilateral ankylosis, six of whom had fibrous ankylosis on one side. There were two types of ankyloses: intra-articular and juxta-articular. Intra-articular ankylosis was seen only in reankylosis or in postinfective cases. Sixty-six cases were posttraumatic juxta-articular ankylosis. A rudimentary temporomandibular joint with an atrophic condylar articular surface was found in all juxta-articular ankyloses. The size of new bone in the specimens varied from 0.5 to 3 cm. Fusion of the extra-articular bone mass with tympanic plate was also observed. Contracture of temporalis muscle was noted in all the cases, which made excision of the coronoid processes mandatory in all the arthroplasties. Arthroplasty early in childhood did not hamper growth; instead, facial remodeling was enhanced.

Effect of high tibial osteotomy on upper tibial venous drainage: study by intraosseous phlebography in primary osteoarthritis of knee joint.

The effect of high tibial osteotomy on the upper tibial venous pattern in primary painful osteoarthritis of the knee joint was studied by preoperative and three to six months postoperative intraosseous phlebograms. The normal phlebographic pattern was established by phlebograms in five patients with normal knees. The preoperative engorgement, tortuosity of the medullary sinusoids, and slow dye clearance showed a remarkable conversion to a near normal appearance after the operation. Rest pain disappeared in all patients after the osteotomy, suggesting that venous congestion is the cause of 'rest pain'.

Training folk practitioners as PHWs in rural India.

PIP: 4 pilot programs for training folk practitioners to provide specific types of primary health care (PHC) services in villages in the Varanasi area of India were described and evaluated. The programs were developed by staff members of Benaras Hindu University and by members of several other universities. The training programs included 1) 2 courses on the prevention and cure of diarrhea, including oral rehydration therapy; 2) a course providing birth attendants with training in delivery technics, including the use of delivery packs; and 3) a course in the diagnosis and treatment of 11 skin diseases. All programs were developed and taught by physicians, except for 1 of the diarrhea prevention courses which was developed and taught by a social anthropologist. The courses were conducted over a 3-5 day period. Methodology, not course content, was the focus of the discussion. An effort was made to make the practitioners feel accepted by the formal medical system. Courses were conducted in places and at times convenient to the trainees. The general mode of presentation was to give a short formal talk about a specific topic followed by a discussion period and then a review session. Each program was evaluated separately in regard to 1) the degree to which specific health messages were translated into behavioral changes, and 2) the amount of staff time which was required to motivate participation and to prepare and teach the course. In regard to the diarrhea prevention course taught by the physician, 14 practitioners were trained and of the 8 health messages conveyed in the course, 3 were being followed by the practitioners 0-4 weeks following the course. The diffusion rate was, therefore, 38%. Time inputs were 130 hours of medical staff time, including 45 hours of direct teaching time and 80 hours of nonmedical staff time. The message diffusion rate for the other diarrhea prevention course was 20% at 8 weeks following course presentation. 14 practitioners were trained. Staff time amounted to only 21 hours of nonmedical staff time, including 3 hours of direct teaching time. For the skin disease course, diffusion rate was 69% for drug related messages and 18% for health education messages at 0-4 weeks following the course. The course took 114 hours of physician time, including 45 hours of direct teaching time, and 55 hours of nonmedical staff time. 20 practitioners were trained in the course. In reference to the birth attendant course, of the 37 births which occurred in the area during the year following the course, 7 births were delivered with technique taught in the course. The diffusion rate was, therefore, 22%. A total of 19 birth attendants were trained. Time inputs were 46 hours of medical staff time and 88 hours of nonmedical staff time. Recommendations were that some of the approaches and technique developed in the pilot projects would be useful in developing a large scale training program in the Gangetic plain and that simplified courses should be taught by paramedical personnel rather than by physician. However, a physician should be present on the opening day of the course to demonstrate that the formal medical system appreciates and welcomes the assistance of the traditional practitioners. The characteristics of folk practitioners, the impact of the training on the status and income of the practitioner, and program acceptability by the villagers and by the practitioners were also discussed.^ieng