Light birefringence and molecular orientation in erythrocyte membranes and surface layers

The light birefringence in the surface layer of erythrocytes has been determined. A mathematical expression for the phase difference caused by birefringence in the spherical layer has been derived. Thickness of the anisotropic layer of erythrocytes has been evaluated. Birefringence in surface layers of erythrocytes has been shown to be caused not only by the membrane, but also by close to it structured proteins.     

Fertility factors in Pseudomonas putida: selection and properties of high-frequency transfer and chromosome donors.

The octane plasmid (OCT) in Pseudomonas putida strains has been shown to be transferred at low frequency. However, bacteria which had newly received this plasmid showed a transient increase in donor ability. Using Octane+ P. putida as the donor, the transfer of most chromosomal markers was shown to be independent of OCT transfer, whereas the mobilization of the octanoate catabolism genes (octanoic and acetate) was dependent on OCT plasmid transfer. The presence of a fertility factor termed FPo has been postulated to explain these results. Strains carrying only this fertility factor have been obtained from strains carrying both OCT and FPo plasmids. Strains in which the OCT plasmid was transferred at high frequencies have also been isolated, and chromosome mobilization by OCT and FPo has been compared. A different gradient of transmission by OCT and FPo has been observed. It has also been shown that chromosome transfer by OCT was dependent on the bacterial recombination system, whereas the chromosome transfer by FPo was unaffected by the presence of a rec mutation in the donor strain.     

Single-strand and double-strand cleavage at half-modified and fully modified recognition sites for the restriction nucleases Sau3a and Taqi

The influence of cytosine methylation on the cleavage of DNA by the restriction nucleases Sau3A and TaqI has been investigated. Bovine satellite DNA fragments containing a GATCGA sequence, i.e. a Sau3A site overlapping with a TaqI site have been used in this study. The methylation of these fragments has been determined by sequence analysis. It has been found that a TaqI site (TCGA) methylated at cytosine in both DNA strands is still sensitive to double-strand cleavage. A Sau3A site (GATC), however, is rendered resistant to double-strand cleavage by methylation of a single cytosine. Fragments containing the "half-modified" Sau3A site are nicked in the unmethylated DNA strand. It has been shown by sequence analysis of nicked DNA that the single-strand break occurs at the same position which is cleaved in unmodified DNA.     

Conformation and proteolysis of glucagon and insulin in surfactant and lipid solutions

The effects of micelles of nonionic, zwitterionic, anionic and cationic surfactants and lipids on the conformation of glucagon and insulin have been investigated by circular dichroism and intrinsic protein fluorescence. The influence of these amphipathic compounds on the hydrolysis, monitored by HPLC, of glucagon and insulin by trypsin and chymotrypsin has also been studied. The alpha-helix content of glucagon was increased to a similar extent by all the micelles, irrespective of their charge and of whether they were synthetic surfactants or phospholipids. The amphipathic compounds always induced a blue-shift in the wavelength of maximum emission of fluorescence of glucagon of about 9 nm, whereas the fluorescence intensity was increased in some cases and decreased in others. The circular dichroism of insulin was also modified in some cases. Some amphipathic compounds protected glucagon against proteolysis by trypsin and chymotrypsin very markedly, whereas others did not protect at all or only slightly protected the hormone. Two hypotheses have been formulated to explain the different results. Hydrolysis of insulin was generally not influenced by surfactants and lipids.     

Synthesis and partial characterization of hydrogels obtained via glutaraldehyde crosslinking of acetylated chitosan and of hyaluronan derivatives

Hydrophilic networks based on functionalized hyaluronic acid and on partially acetylated chitosan, respectively, have been obtained. In the case of hyaluronic acid (HA), primary amino functionalities have been introduced along the polysaccharide chains. The ensuing derivatives, i.e., HA-lysine (HA-K), HA-diamino pentane (HA-DAP), and HA-glycine-lysine (HA-GK), have been characterized by high field NMR spectroscopy. NMR 2D-DOSY experiments have allowed us to optimize the purification procedure. Chitosan was made soluble in water by partial acetylation. Cross-linking reactions have been performed using glutaraldehyde. The obtained networks have been qualitatively characterized by means of (13)C CP-MAS NMR technique. The hydrogels have been characterized also in terms of water uptake.     

Cerebrospinal Fluid Proteins Studied by Two-Dimensional Gel Electrophoresis and Immunoblotting Technique

The proteins of lumbar CSF have been investigated by two-dimensional gel electrophoresis, and their patterns have been compared with the corresponding serum protein patterns. Serum proteins in CSF have been identified by electroblotting and immunoreaction with antiserum against total human serum proteins. Proteins derived from brain have been identified with antiserum against human brain proteins. The most prominent CSF protein group has been identified as a multiple form of apolipoprotein E. The correct position of the glial fibrillary acidic protein has also been determined. The prefractionation of CSF proteins by size exclusion chromatography or by affinity chromatography followed by two-dimensional electrophoresis has facilitated the detection of trace components in CSF and the corresponding serum.     

Cloning of the Streptococcus thermophilus beta-galactosidase gene and its expression in Escherichia coli and Bacillus subtilis cells

The beta-galactosidase gene from the chromosome of Streptococcus thermophilus, strain 6 kb, has been cloned on a vector plasmid pBR322. The corresponding gene has been found to be located on the Pst1 DNA fragment. The restriction map of this 6 kb fragment has been constructed. The shortening of the DNA fragment carrying the beta-galactosidase gene has been achieved by digestion of the recombinant derivative of pBR322 by the restriction endonuclease Sau3A under the conditions of incomplete hydrolysis. The obtained fragments have been cloned into the BamHI site in the berepliconed shuttle vector pCB20 for grampositive and gramnegative bacteria. The obtained recombinant plasmids contained the beta-galactosidase gene in the inserted fragments of different length. Expression of the cloned beta-galactosidase gene in Escherichia coli and Bacillus subtilis cells has been studied.     

Degradation of polysaccharides and lignin in wood ozonation

Ozonation has been considered as a method for the pretreatment of plant biomass to obtain cellulose and monosaccharides. Ozone consumption by aspen wood with various moisture contents has been investigated. We have considered the gradual transformation of the substrate: wood to ozonated wood to cellulose-containing product (CP) to holocelluloze (HC) and to cellulose. Yields of ozonated wood (OW), the (CP), water-soluble ozonation products, HC, and cellulose have been determined. The lignin content in the CP has been estimated. Both HC and cellulose samples have been studied by IR spectroscopy. The degree of polymerization and molecular mass distribution of cellulose obtained from ozonated wood have been determined. It has been shown that wood destruction by ozone is accompanied by degradation of lignin, hemicelluloses, and cellulose.It has been found that physicochemical properties of cellulose obtained from ozonated wood can be regulated by the variation of the initial moisture content in the substrate. Both molecular ozone and radical species, which are generated in the course of ozone reactions with water present in the substrate structure, participate in wood destruction.     

The mangrove-based coastal and nearshore fisheries of Bangladesh: ecology,exploitation and management

The mangrove forest of Bangladesh, the largest continuous mangrove forest of the world, is one of the most important coastal features of the country. The existence of the mangrove has increased the values of other coastal and marine resources such as the coastal and marine fisheries by increasing productivity and supporting a wide biological diversity. The artisanal fishery, which is highly influenced by mangroves, has been contributing 85–95% of the total coastal and marine catch of Bangladesh. The mangrove also supports offshore and deep sea fisheries by playing a significant role as nursery ground for many deep sea fishes and shrimps including the giant tiger shrimp (Penaeus monodon) which is the major species of the industrial bottom trawl fishery of Bangladesh. The mangrove also contributes significantly in shrimp farming which has been the most significant export-oriented industry since the 1970s. However, the mangrove fisheries have been under intensive pressure from deleterious fishing activities and deliberate aquaculture development by destructing mangrove habitats. The impacts of mangrove have been reflected in the contribution of artisanal fishery catch that has been in a continuous decline since the 1980s. Shrimp farming has been the most destructive contributor to mangrove destruction with a corresponding loss of biological resources particularly the wild shrimp fishery. This paper reviews different aspects of the mangrove fisheries of Bangladesh and discusses the impacts of different fisheries. The paper identifies the importance of reviewing, amending and/or replacing the traditional management approaches by the new management techniques such as habitat restoration and stock enhancement in the natural environment; the paper also identifies the need for research findings in formulating and implementing new management approaches.     

Molecular-sieve chromatography and electrophoresis in polyacrylamide gels

1. The absolute electrophoretic mobilities of eight proteins have been measured at pH8.76, I 0.05, in polyacrylamide gels of 20 different compositions at 10 degrees C. 2. The partition coefficients of these proteins have been determined chromatographically under the same conditions by using columns of granulated polyacrylamide gel prepared simultaneously. 3. The electrophoretic mobilities are an exponential function of the gel concentrations when the latter are corrected for water uptake. The constants of this function have been determined by curvefitting methods. They have been shown to be related to the free solution mobility and to the mean molecular radius respectively. 4. The reduced mobilities have been shown to be a linear function of the partition coefficients by statistical analyses. 5. The physical significance of the relation between electrophoretic mobility and chromatographic phase distribution in gel media is discussed in the context of these results.     

Determination of amylase activity in bovine serum and its association with the Ami and Amll loci

Amylase activity has been estimated in the sera of 213 cattle by an amyloclastic method with potato amylose as the substrate with the addition of CaCl₂. The method has been developed from that of Street and Close to overcome the problems of the high colour of bovine serum which would otherwise interfere with the assay. A relationship has been found between the serum amylase genotype detected on starch-gel and the total enzyme activity. The enzymes AmI-B and AmII-B in homozygous combinations have been found to have the highest activities. Age of animal has no significant effect on the total enzyme activity. Activity has been investigated in sera from three separate cattle populations and it has been found to be a highly heritable character. Heritability estimates of 49 % and 39 % by sib analysis and 75 % by the twin method have been obtained.     

The isolation and identification of 2,4-diketones present in human and animal tissues and in urine.

The chemical isolation of 2,3-diketone fraction from hydrolysates of various mammalian tissues has been accomplished by the use of a modified Girard T procedure. This fraction, which constitutes a new lipid class, has been resolved by gas chromatography into a number of homologous 2,4-diketones, ranging in chain length from C13 to C25. After separation by preparative gas chromatography, the following compounds have been unequivocally identified: 2,4-heptadecanedione, 2,4-nonadecanedione, 2,4-heneicosanedione, 2,4-docosanedione, and delta12-2,4-heneicosenedione. The 2,4-diketones appear to exist in tissues in the free state or in labile combination. They are present also in human urine.     

Technology and medicine

Technology, which is older than science, has been of vital importance in the development of modern medicine. Even so, there are voices of dissent to be heard. The disenchantment with technology expressed by Aldous Huxley in Brave new world has been echoed by contemporary writers on the technology of modern medicine. Medicine is seen by some to have been dehumanized by technology, and techniques that are expensive are thought to be consuming a greater proportion of health resources than they deserve. The practice of medicine has, nevertheless, been transformed by modern technology and diagnostic techniques and therapeutic measures undreamed of a few short decades ago are now commonplace. There is no reason why these developments should be any more dehumanizing than the use of similar techniques in modern transportation or communication, nor is their expense out of proportion when compared with other demands on the nation's purse. British workers have been at the forefront of many recent advances. Yet, even though the National Health Service provides a ready market for the products of British medical technology, the nation depends to an inordinate degree on imported products. In the development of appropriate medical technology there is an urgent need for better communication between inventors, scientists, industrialists and the National Health Service. At the same time there is an equal need for improved evaluation of untried techniques. The pressure for a central integrating body to coordinate resources could well be supported by the establishment of evaluation units in the different health authorities in this country.     

Purification and substrate specificity of BcmI restriction endonuclease

A strain producing the new specific restriction endonuclease BcmI has been found in the Bacillus generum. The enzyme has been purified by chromatography on the blue sepharose, phosphocellulose PII, heparin sepharose. The analogous purification has been obtained when the blue sepharose has been substituted for the orange sepharose, the home produced sorbent. The BcmI enzyme has been shown by the substrate specificity definition to be an isoschizomer of the restriction endonuclease ClaI.     

Calmodulin-binding proteins from brain and other tissues.

The calmodulin contents of rabbit brain, lung, kidney and liver, of bovine aorta and uterus, and of chicken gizzard have been determined. 2. The calmodulin in all of these tissues has been shown to be present in the form of very stable complexes with several other proteins. 3. A calmodulin-binding protein of mol.wt. 22 000 has been purified in high yield from bovine brain. It has been shown to interact with calmodulin and rabbit skeletal-muscle troponin C in a Ca2+-dependent manner. 4. The 22 000-mol.wt. protein inhibits the activation of bovine brain phosphodiesterase by calmodulin, but has very little affect on the activation of myosin light-chain kinase. 5. Calmodulin-binding proteins of mol.wts. 140000, 77000 and 61000 have also been partially purified from rabbit brain by affinity chromatography and have been shown to interact in a Ca2+-dependent manner with calmodulin. 6. The apparent molecular weights of the calmodulin-calmodulin-binding protein complexes, determined by gel filtration in the presence of 6M-urea, have been shown to be similar for most of the mammalian tissues examined. 7. By using 125I-labelled calmodulin, similar complexes have been demonstrated in rabbit skeletal muscle, although they are present at much lower concentrations.     

The effect of urea, gamma- and UV-irradiation on physico-chemical characteristics of native and immobilized inulinase

The immobilization of inulinase by ionexchange AB-26 and AB-17-2P has been made by adsorbtion and glutaraldehyde methods. The effect of UV-radiation, carbamide and gamma-rays on the stability of native and immobilized enzyme has been investigated. The stability of inulinase in relation to denaturation agents has been shown to increase with the immobilization of ionexchange. The character of binding with the matrix affects greatly the stability of immobilized enzyme to physical factors.     

Starch phosphorylase enzymes in developing and germinating pea seeds

Phosphorylase has been fractionated during development and germination of seeds of smooth and wrinkled-seeded peas. The total phosphorylase levels have been compared. In addition, a number of other pea tissues and other legumes have been examined. Some kinetic properties of the two enzymes present have been measured. Both enzymes have been further purified by affinity chromatography on Sepharose 4B-starch columns and by sequential gel filtration in the absence and presence of amylopectin. The MW and sub-unit structures of the two enzymes have been examined and their possible roles discussed.     

General organization of the conjugal transfer genes of the IncW plasmid R388 and interactions between R388 and IncN and IncP plasmids.

The complete conjugal transfer gene region of the IncW plasmid R388 has been cloned in multicopy vector plasmids and mapped to a contiguous 14.9-kilobase segment by insertion mutagenesis. The fertility of the cloned region could still be inhibited by a coresident IncP plasmid. The transfer region has been dissected into two regions, one involved in pilus synthesis and assembly (PILW), and the other involved in conjugal DNA metabolism (MOBW). They have been separately cloned. PILW also contains the genes involved in entry exclusion. MOBW contains oriT and the gene products required for efficient mobilization by PILW. MOBW plasmids could also be mobilized efficiently by PILN, the specific pilus of the IncN plasmid pCU1, but not by PILP, the specific pilus of the IncP plasmid RP1.     

The growth stimulating effect of 5-bromouracil and uracil on chlorococcal algae

The growth stimulating effect of 5-bromouracil and uracil on two strains of chlorococcal algae has been found in cell colonies grown from single cells, which were inoculated onto an agar medium. Analyses of the effect recorded in the cell cycles after treatment have revealed that the growth stimulating effect required four, or more, cell cycles to become evident. This has been proved by the number of autospores released from the treated cells and by the length of the lag phase after inoculation. The differences between the control and the treated population in some experiments with 5-bromouracil inChlorella and with uracil inScenedesmus obliquus have been visible by the unaided eye, whereas in some other experiments, they have been proved statistically. Growth stimulation has not been found only in a small number of experiments. The inhibition of growth induced by 5-bromouracil has been recorded in one experiment withScenedesmus quadricauda.