Geographically diverse lineage of mycobacterium tuberculosis in Sub-Himalayan region of Kangra, Himachal Pradesh.

We present the lineages of Mycobacterium tuberculosis (MTB) found using spoligotyping in the Sub-Himalayan Region of Kangra District in Himachal Pradesh. The DNA from clinical isolates was used for spoligotyping using a PCR based method to simultaneously detect and type Mycobacterium tuberculosis complex, performed by an outsource agency namely Mapmygenome, Hyderabad. Spoligotyping and database comparison was done. We found the following families: Indo Oceanic (51.2%) Central Asian (37.8%), West African (4.8%), East Asian (3.6%), Euro American and M.africanum (1.2%) each. There were genetically diverse strains of MTB causing pulmonary tuberculosis in Kangra region, indicating cosmopolitan nature of its population.

Per a 10 protease activity modulates CD40 expression on dendritic cell surface by nuclear factor-kappaB pathway.

Serine protease activity of Per a 10 from Periplaneta americana modulates dendritic cell (DC) functions by a mechanism(s) that remains unclear. In the present study, Per a 10 protease activity on CD40 expression and downstream signalling was evaluated in DCs. Monocyte-derived DCs from cockroach-allergic patients were treated with proteolytically active/heat-inactivated Per a 10. Stimulation with active Per a 10 demonstrated low CD40 expression on DCs surface (P < 0·05), while enhanced soluble CD40 level in the culture supernatant (P < 0·05) compared to the heat-inactivated Per a 10, suggesting cleavage of CD40. Per a 10 activity reduced the interleukin (IL)-12 and interferon (IFN)-γ secretion by DCs (P < 0·05) compared to heat-inactivated Per a 10, indicating that low CD40 expression is associated with low levels of IL-12 secretion. Active Per a 10 stimulation caused low nuclear factor-kappa B (NF-κB) activation in DCs compared to heat-inactivated Per a 10. Inhibition of the NF-κB pathway suppressed the CD40 expression and IL-12 secretion by DCs, further indicating that NF-κB is required for CD40 up-regulation. CD40 expression activated the tumour necrosis factor (TNF) receptor-associated factor 6 (TRAF6), thereby suggesting its involvement in NF-κB activation. Protease activity of Per a 10 induced p38 mitogen-activated protein kinase (MAPK) activation that showed no significant effect on CD40 expression by DCs. However, inhibiting p38 MAPK or NF-κB suppressed the secretion of IL-12, IFN-γ, IL-6 and TNF-α by DCs. Such DCs further reduced the secretion of IL-4, IL-6, IL-12 and TNF-α by CD4(+) T cells. In conclusion, protease activity of Per a 10 reduces CD40 expression on DCs. CD40 down-regulation leads to low NF-κB levels, thereby modulating DC-mediated immune responses.

Serine protease Per a 10 from Periplaneta americana bias dendritic cells towards type 2 by upregulating CD86 and low IL-12 secretions.

BACKGROUND: Serine protease activity of Per a 10 from Periplaneta americana induces airway inflammation and systemic Th2 response towards self and bystander allergen. OBJECTIVE: In the present study the effect of proteolytic activity of Per a 10 allergen on dendritic cells (DCs) polarization and consequent T cell response was investigated. METHODS: Non-atopic subjects with no family history of asthma/allergy were recruited for the study. CD14(+) peripheral blood monocytes were purified, differentiated to immature DCs and stimulated with proteolytically active/inactivated native or recombinant Per a 10. DCs phenotype was analysed with flow cytometry and antigen presenting function assessed by co-culturing with autologous CD4(+) T cells. Cytokine levels were determined using ELISA. RESULTS: Immature DCs differentiated into mature CD14(-)CD83(+)HLA-DR(+) cells after incubating with proteolytically active/inactivated or recombinant Per a 10. Proteolytically active Per a 10 induced significant CD86 up-regulation on DCs compared to inactivated or recombinant Per a 10 lacking enzymatic activity. Proteolytic activity of Per a 10 showed dose-dependent effect on expression of CD80, CD86, CD83, CD1a and HLA-DR. However, no significant differences were observed phenotypically in active or inactive forms except for CD86. Active Per a 10 stimulated DCs secreted significantly low IL-12 (P < 0.01) and high IL-6, compared to inactive forms of Per a 10. Naive CD4(+) T cells primed with active Per a 10 pulsed DCs also secreted significantly less IL-12 (P < 0.01) and high IL-4, IL-5 plus IL-6 (P < 0.01); in contrast to DCs pulsed with inactivated or recombinant Per a 10. CONCLUSION AND CLINICAL RELEVANCE: Proteolytic activity of Per a 10 modulates DCs towards type 2 by CD86 up-regulation, high IL-6 and reduced IL-12 secretions. Proteolytically inactive Per a 10 can be further explored for immunotherapy.

Multiple neural inductions in area opaca by grafts of the Hensen's node do not retard host chick embryo development.

Four hundred and forty four full primitive streak stage chick embryos were cultured in vitro and 261 were transplanted with 1, 3 or 5 Hensen's nodes in the area opaca. Irrespective of the number of grafts, neural induction was observed in 90% cases. The development of control and grafted embryos and the size of blastoderm area were monitored at the time of grafting and after 20 hr. We find that the induced neural tissue and differentiated tissue of graft-origin neither fuse with the host embryonic axis, nor retard its development.

Transurethral resection syndrome. A prospective study.

Significant hyponatraemia has been reported following transurethral prostatectomy (TURP) in 11-41% of cases. The majority of previous studies have been performed retrospectively. A prospective study was undertaken of 100 patients undergoing TURP. In all, a 24-Charr sheath with non-irrigating, resectoscope and 1.5% glycine as irrigant was used. Volume of irrigant used, weight of prostate and length of procedure were recorded. Serum electrolytes were measured at anaesthetic induction and immediately on transfer to the recovery room. In none of the 100 patients was there a statistically significant fall in serum sodium following resection. No clinical changes of transurethral resection (TUR) syndrome occurred. This study confirms that TUR syndrome and a significant fall in serum sodium can be virtually prevented in TURP and the use of an irrigating resectoscope or a trocar in the average case is not necessary.