The region Ser333-Arg356 of the alpha-chain of human C4b-binding protein is involved in the binding of complement C4b

Human C4b-binding protein (C4BP) functions as a cofactor to factor I in the degradation of C4b and accelerates the decay rate of the C4b2a complex. In this study we describe a monoclonal antibody directed against the alpha-chain of C4BP that inhibits the binding of C4b to C4BP. In order to identify the structural domain of the alpha-chain of C4BP that interacts with C4b, tryptic fragments of C4BP were generated. Amino acid sequence analysis of the fragments revealed that the residues Ser333-Arg356 of the alpha-chain of C4BP contain the epitope of this antibody, and as a consequence, that this part of the alpha-chain of C4BP is likely to be involved in the interaction with C4b.     

Collection of biological materials in biodiversity prospecting in India: problems and solutions

Forests are the chief resource for the collection and exploration of biological materials. The past few decades have witnessed a large scale deforestation in India due to substantial pressures generated by population growth, leading to demand for more land for agriculture, urbanization and industrial activities, in addition to increased demand for fuel wood and timber. This has resulted in the loss of soil cover, habitat destruction, environmental degradation and ecological imbalance. This scenario has created a progressive awareness for the conservation and restoration of habitats and, thus, the declaration of many forest areas into protected zones, such as national parks, biosphere reserves, etc., including the protection of some marine areas, by both the National and State Governments. Normally, permission for biological collecting is not granted in these protected areas. In India, forests are a State subject and grant for collection permission is vested with the State Forest Departments. In the absence of any rules, regulations and guidelines, either from National or State Governments, forest authorities impose their terms and conditions, which are arbitrary and even contradictory at times, in the process of granting collecting permits. A set of new rules to be applied throughout the country is needed.     

Antibody- and cell-mediated immune responses of Actinobacillus pleuropneumoniae-infected and bacterin-vaccinated pigs

The number involved in and the rate of migration of donor leucocytes into the following recipient organs (spleen, thymus, bone marrow, lung and mesenteric lymph nodes) were measured in two rat models of orthotopic liver transplantation (OLT) using donor-specific MHC class I antibodies. The first OLT model is one that does not require immunosuppression in order to achieve tolerance and involved the transplantation of DA (MHC haplotype, RT1a) livers into PVG (RT1c) recipients. The second model was one that required a 7-day (10 mg/kg) treatment with cyclosporin A (CsA) to achieve tolerance and used DA donors into Lewis (RT1(1)) recipients. Recipient organs were biopsied on days 3, 20 and 87 following OLT and donor leucocyte migration was quantified by immunohistochemistry and computer densitometry of immunoblots of detergent-solublized tissues in order to resolve both membrane-bound and soluble donor MHC class I antigen. In a separate experiment, spleen biopsies were taken following OLT on days 3 and 15 from the naturally tolerizing OLT model (DA into PVG), treated with and without CsA for 7 days and compared with the (DA into Lewis) model. The initial rate of leucocyte migration between days 3 and 21 following OLT was found to be the most rapid into the spleen, followed by the bone marrow and mesenteric lymph nodes in the naturally tolerant (DA into PVG) model when compared with the (DA into Lewis) model. The number of donor leucocytes in the spleen and mesenteric lymph nodes in both models was, however, approximately the same by 87 days. No real difference in the rate of leucocyte migration was seen in the thymus or the lung for both transplant models over the time course assayed. CsA was found to lower the rate of donor leucocyte migration only over the period it was administered. The rate of donor leucocyte migration into the spleen was still much lower 15 days after OLT in the (DA into Lewis) model compared with the (DA into PVG) model treated with and without CsA. Thus the differences in the rate of donor leucocyte migration into the spleen, bone marrow and mesenteric lymph nodes immediately following OLT may offer an explanation as to why the (DA into PVG) combination is able to accept a transplanted liver without immunosuppressive therapy.     

Quantitation of age-related mitochondrial DNA deletions in rat tissues shows that their pattern of accumulation differs from that of humans

We studied the postnatal development of the release of acetylcholine (ACh) and of presynaptic, release-inhibiting muscarinic autoreceptors in the rat hippocampus. To this end, hippocampal slices (350 microns thick) from rats of various postnatal ages (postnatal day 3 [P3] to P16) were preincubated with [3H]choline and stimulated twice (S1, S2: 360 pulses, 2 ms, 3 Hz, 60 mA) during superfusion with physiological buffer containing hemicholinium-3 (10 microM). In parallel, the activities of hemicholinium-sensitive high-affinity choline uptake (HACU, in synaptosomes) and of choline acetyltransferase (ChAT, in crude homogenates) were determined as markers for the cholinergic ingrowth. In hippocampal slices preincubated with [3H]choline, the electrically evoked overflow of 3H at S1 increased from 0.11 (P3) to 0.81% of tissue 3H (P16), the latter value being still much lower than that of hippocampal slices from adult rats (2.89% of tissue 3H). Already at P3 the evoked overflow of 3H was Ca(2+)-dependent and sensitive to tetrodotoxin, indicating an action potential-evoked exocytotic mechanism of ACh release. The muscarinic agonist oxotremorine (1 microM) significantly inhibited the evoked ACh release in hippocampal slices with increasing effectivity from P4 to P16; no significant effect was detectable at P3. The ACh esterase inhibitor physostigmine and the muscarinic antagonist atropine (1 microM, each) exhibited significant inhibitory and facilitatory effects, respectively, only at P15-16. The specific activities of both hippocampal HACU (pmoles/mg protein/min) and ChAT (nmoles/mg protein/min) continuously increased from P3 to P16. It is concluded (1) that cholinergic nerve terminals arriving at the hippocampal formation during postnatal ingrowth are already endowed with the apparatus for action potential-induced, Ca(2+)-sensitive (exocytotic) ACh release; (2) that, in contrast, the expression of presynaptic muscarinic autoreceptors on these cholinergic axon terminals is delayed; and (3) that autoinhibition due to endogenous ACh develops even later, probably when the density of presynaptic terminals in the hippocampus and hence, the concentration of released ACh has reached a suprathreshold value.     

Use of sky brightness measurements from ground for remote sensing of particulate polydispersions

The software code SKYEAD.pack for retrieval of aerosol size distribution and optical thickness from data of direct and diffuse solar radiation is described; measurements are carried out with sky radiometers in the wavelength range 0.369-1.048 μm. The treatment of the radiative transfer problem concerning the optical quantities is mainly based on the IMS (improved multiple and single scattering) method, which uses the delta-M approximation for the truncation of the aerosol phase function and corrects the solution for the first- and second-order scattering. Both linear and nonlinear inversion methods can be used for retrieving the size distribution. Improved calibration methods for both direct and diffuse radiation, the data-analysis procedure, the results from the proposed code, and several connected problems are discussed. The results can be summarized as follows: (a) the SKYRAD.pack code can retrieve the columnar aerosol features with accuracy and efficiency in several environmental situations, provided the input parameters are correctly given; (b) when data of both direct and diffuse solar radiation are used, the detectable radius interval for aerosol particles is approximately from 0.03 to 10 μm; (c) besides the retrieval of the aerosol features, the data-analysis procedure also permits the determination of average values for three input parameters (real and imaginary aerosol refractive index, ground albedo) from the optical data; (d) absolute calibrations for the sky radiometer are not needed, and calibrations for direct and diffuse radiation can be carried out with field data; (e) the nonlinear inversion gives satisfactory results in a larger radius interval, without the unrealistic humps that occur with the linear inversion, but the results strongly depend on the first-guess spectrum; (f) aerosol features retrieved from simulated data showed a better agreement with the given data for the linear inversion than for the nonlinear inversion.