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Several antigens of Mycobacterium tuberculosis have been identified and specificity to one or multiple antigens could determine the distinction between protective and pathogenic host reaction. Therefore T cell immune response to combinations 38 kDa/CFP‐10, 38 kDa/MPT‐64, ESAT‐6/MPT‐64 and ESAT‐6/CFP‐10 (each related to a single protein of Mycobacterium tuberculosis) in individuals from tuberculosis endemic areas have been examined. ELISA was used to detect IFN‐γ production in PBMC priming with single proteins and combinations in a panel of 105 individuals: 38 tuberculosis patients (6 untreated and 32 treated) and 67 healthy controls with tuberculin skin test positive or negative (TST). Brazilian TB patients highly recognized ESAT‐6 (66%), but combinations improved response in the following order: ESAT‐6/MPT‐64 (89%) > ESAT‐6/CFP‐10 (73%) > 38 kDa/CFP‐10 (70%), the last combination showing the highest specificity (TST+=42% and TST–=83%). Average IFN‐γ production in TB patients was significantly higher for 38 kDa/CFP‐10 (P=0.012) and 38 kDa/MPT‐64 (P<0.035), when compared to single antigens. None of the combinations was able to discriminate TB patients from TST+ controls; however, 38 kDa/CFP‐10 displayed a borderline significance (P=0.053). Similar to the ESAT‐6/CFP‐10 combination, IFN‐γ response to 38 kDa/CFP‐10 showed an increased tendency in treated patients, although not significant (P=0.16). We demonstrated for the first time that 38 kDa/CFP‐10 had prediction sensitivity for TB patients similar to the ESAT‐6/CFP‐10 combination and also significant response improvement related to the single proteins with more selective reactivity among TST‐positive individuals, which could be of potential interest for diagnostic evaluation for tuberculosis infection.

The aim of this work is to correlate the histological gonad analysis of adult male Macrobrachium amazonicum of a wild population with morphotypes identified in populations reared in earthen ponds. A total of 500 adult male specimens were collected in the Mosqueiro Island, Municipality of Belém, State of Pará, Brazil. The animals were classified on the basis of body and cheliped colour, second right cheliped spination, total length, cheliped length and weight. The gonads of each morphotype were fixed, processed by histological techniques and stained with haematoxylin and eosin. All four morphotypes, translucent claw (TC), cinnamon claw (CC), green claw 1 (GC1) and green claw 2 (GC2), were identified in the wild population, indicating the same population structure described for cultured animals. The M. amazonicum testes are composed of multiple lobes. The TC gonads showed a population of germ cells in the eccentric portion of the seminiferous tubule and few spermatozoa in the central region. Cinnamon claw males exhibited a large cell population with intense meiotic activity and sparse spermatozoa. Regardless of the different external morphologies of GC1 and GC2, no histological differences were found in their gonads; both had a large number of gametes and similar spermatogenic cell population. Despite the external morphological evidences of the four morphotypes, the histological criteria indicate the existence of only three morphotypes: TC, CC and green claw (GC).

The objective of this study was to determine the number, morphology and ultrastructure of preantral ovarian follicles of buffalo (Bubalus bubalis) foetuses at different ages. Quantification revealed number of primordial, primary and secondary follicles of 48,857 ± 17,506, 26,000 ± 20,452, 18,428 ± 10,875 and 18,375 ± 19,690, 225 ± 349, 326 ± 288 at 12-34 cm and 35-60 cm crown rump length (CRL), respectively. Follicular diameter values were 28.9 (± 3.4), 34.7 (± 5.9) and 59.4 (± 12.6) μm; oocyte diameters were 21.7 (± 2.8), 24.3 (± 3.4) and 33.0 (± 7.7) μm, and the numbers of follicular cells in the follicle equatorial section were 7.1 (± 1.4), 12.0 (± 2.4) and 13.8 (± 2.4) for primordial, primary and secondary follicles, respectively. The primordial follicle consisted of an oocyte surrounded by a layer of flattened follicular cells with a normally eccentric oocyte nucleus. Dispersed Golgi complex, smooth endoplasmic reticulum, rounded mitochondria and several lipid vesicles were observed in the cytoplasm and cell junctions between the follicle cell membranes and the oocyte. This work describes the number, morphometry and ultrastructure of preantral follicles of buffalo foetuses, concluding that folliculogenesis is established between 8 and 34 cm CRL and that follicle number varies individually and according to age and that further studies are needed in this species.

Smooth muscle cells (SMC) of normal and varicose human saphenous intima were studied on cryostat sections by immunohistochemistry with alpha-smooth muscle actin (ASMA), type IV collagen, and laminin antibodies and also by transmission electron microscopy. The findings suggest two structurally distinct subtypes of smooth muscle cells with thin and thicker external lamina. Thin external lamina SMC were characterized by laminin, type IV collagen, weaker external lamina reactivity, and intense cytoplasmic alpha-smooth muscle actin immunoreactivity. Ultrastructurally, they exhibited abundant cytoplasmic microfilaments and thin external lamina. These cells were found isolated or, more frequently, clustered in fascicles close to the subendothelium in focal or zonal cushions, or in diffuse enlargement of the intima. In contrast, thicker external lamina smooth muscle cells were intensely immunolabeled for laminin and collagen IV, showing irregular cytoplasmic ASMA reaction. Single or clustered thicker external lamina SMC were seen predominantly in zonal cushions and in intima diffuse enlargement. It is very likely that these cells secrete these matrices in a nonpolarized fashion. The thicker external lamina of these SMCs showed a fine granular amorphous aspect sometimes intermingled with microfibrils. These external lamina were interposed between neighboring cells and exposed to collagen fibrils and elastic fibers. The cells also exhibited rarefaction of the cytoplasmic filaments. Intermediary cells exhibiting both features were rarely seen. Thicker external lamina SMC should be discussed in the context of an adaptive/proliferative response leading to dysfunction of the fibroelastic properties of the vein wall.

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