Regeneration of pleated filters used to concentrate enteroviruses from large volumes of tap water.

Pleated cartridge filters are capable of concentrating enteroviruses from large volumes (well over 2,000 liters) of tap water. These epoxy-fiberglass filters can be regenerated if they are treated with 0.1 N NaOH or autoclaved to inactivate any contaminating virus. The regenerated filters regained their ability to concentrate viruses from water at high flow rates.     

Ehlers Danlos syndrome type VIIB. Incomplete cleavage of abnormal type I procollagen by N-proteinase in vitro results in the formation of copolymers of collagen and partially cleaved pNcollagen that are near circular in cross-section.

We have shown that a child with Ehlers Danlos syndrome (EDS) type VII has a G to A transition at the first nucleotide of intron 6 in one of her COL1A2 alleles. Half of the cDNA clones prepared from the proband's pro alpha 2(I) mRNA lacked exon 6. The type I procollagen secreted by the proband's dermal fibroblasts in culture was purified, and collagen fibrils were generated in vitro by cleavage of the procollagen with the procollagen N- and C-proteinases. Incubation of the procollagen with N-proteinase resulted in a 1:1 mixture of pCcollagen and uncleaved procollagen. Incubation of this mixture with C-proteinase generated collagen and abnormal pNcollagen (pNcollagen-ex6) that readily copolymerized into fibrils. By electron microscopy these fibrils resembled the hieroglyphic fibrils seen in the N-proteinase-deficient skin of dermatosparactic animals and humans and were distinct from the near circular cross-section fibrils seen in the tissues of individuals with EDS type VII. Further incubation of the hieroglyphic fibrils with N-proteinase resulted in partial cleavage of the pNcollagen-ex6 in which the abnormal pN alpha 2(I) chains remained intact. These fibrils were not hieroglyphic but were near circular in cross-section. Fibrils formed from collagen and pNcollagen-ex6 that had been partially cleaved with elevated amounts of N-proteinase prior to fibril formation were also near circular in cross-section. The results are consistent with a model of collagen fibril formation in which the intact N-propeptides are located exclusively at the surface of the hieroglyphic fibrils. Partial cleavage of the pNcollagen-ex6 by N-proteinase allows the N-propeptides to be incorporated within the body of the fibrils. The model provides an explanation for the morphology and molecular composition of collagen fibrils in the tissues of patients with EDS type VII.     

Identification of CpG islands in a physical map encompassing the Friedreich's ataxia locus

The Friedreich's ataxia locus has been previously assigned to chromosome 9q 13-21.1 by the demonstration of tight linkage to two anonymous DNA markers. MCT112 (Z greater than 80, theta = 0) and DR47 (Z greater than 50, theta = 0). The absence of recombination between these three loci has prevented the resolution of gene/probe order in this region, impeding strategies for gene isolation. We report physical mapping over a 4-Mb genomic interval, linking the markers MCT112 and DR47 on a common 460-kb NotI fragment and identifying 11 CpG islands in the 1.7-Mb interval most likely to contain the Friedreich's ataxia locus. Four of these islands were detected only by analysis of three YAC clones spanning a 700-kb interval including the MCT112/DR47 cluster. Without clear evidence of the precise location of the disease locus from recombination events, each of these regions must be considered as specifying a potential "candidate" sequence for the mutated gene.     

Bacterial Streak Disease of Maize (Zea mays L.) in South Africa

Bacterial streak disease of maize is currently causing some concern among breeders in South Africa. The causal organism of this previously undescribed disease was successfully isolated and its pathogenicity established using KoCH's postulates. Standard physiological and biochemical tests used to identify phytopathogenic bacteria indicated that the bacterium is a Xanthomonas campestris pathovar. Comparisons between this organism and other recognized X. campestris pathovars of the Poaceae indicated that apart from some minor differences the maize streak pathogen is physiologically similar to X. campestris pv. holcicola. However, in repeated reciprocal inoculation experiments all attempts to induce disease symptoms in sorghum with the maize streak pathogen were unsuccessful. Conversely, X. campestris pv. holcicola did produce symptoms in maize leaves. In all the maize cultivars tested the symptoms produced by the maize streak pathogen were, however, always considerably more severe than those caused by X. campestris pv. holcicola. Notwithstanding its physiological similarity to X. campestris pv. holicola it would appear that on the grounds of host specificity the maize streak pathogen warrants new pathovar status. The name X. campestris pv. zeae is proposed.     

Influence of Estrogen and Progesterone on Glutamic Acid Decarboxylase Activity in Discrete Regions of Rat Brain

Abstract: Female Charles River rats were ovariectomized and treated for three days with 17/8-estradiol benzoate (E) (1.0 /μg/day), progesterone (P) (500 μg/day), vehicle, or a combined treatment (2 days E, one day P). Animals were killed on day 3 and the brains were dissected by the micropunch technique. Glutamic acid decarboxylase (GAD) activity was measured by collection of ¹⁴CO². Estradiol benzoate and progesterone were potent inhibitors of GAD activity in regions such as the arcuate nucleus, ventromedial hypothalamus and corticomedial amygdala. Estrogen reduced the V_max of GAD for glutamate as a substrate without changing the K_m. Estrogen also failed to change the K_m for pyridoxal phosphate. Combined treatment with estrogen and progesterone did not reduce GAD from ovariectomized levels except in the septum, indicating an interaction of the two hormones at the level of GAD. The suggestion is made that under conditions that inhibit LH secretion GAD activity is low, but when LH secretion is stimulated GAD activity may be comparatively high.     

Phosphatidylinositol-inositol exchange in a rabbit lung

A microsomal fraction prepared from rabbit lung tissue was found to catalyze CDPdiacylglycerol-independent incorporation of [3H]inositol into phosphatidylinositol. This incorporation resulted from CMP-dependent phosphatidylinositol-inositol exchange and did not constitute a net synthesis of phosphatidylinositol. The phosphatidylinositol-inositol exchange activity was distinct from the phospholipid-base exchange enzymes and was specific for inositol. Optimal in vitro phosphatidylinositol-inositol exchange activity was observed at pH 8.5--8.8 and either Mn2+ or Mg2+ was essential for activity. Mercaptoethanol stimulated phosphatidylinositol-inositol exchange and Hg2+ inhibited this activity. In the absence of CMP, no phosphatidylinositol-inositol exchange was observed. CDP (and to a smaller extent CTP) also supported phosphatidylinositol-inositol exchange and this appeared to occur via the generation of CMP during incubations. The apparent Km values of the phosphatidylinositol-inositol exchange enzyme for CMP and inositol were 0.4 mM and 11 microM, respectively. When CDPdiacylglycerol was present at a concentration optimal for CDPdiacylglycerol : inositol transferase activity, CMP-dependent phosphatidylinositol-inositol exchange activity was still observed. However, in the presence of Hg2+ CDPdiacylglycerol inhibited phosphatidylinositol-inositol exchange activity. Several properties of the phosphatidylinositol-inositol exchange enzyme resemble those of CDPdiacylglycerol : inositol transferase, but the two enzymes appear distinct on the basis of different degrees of inhibition by either Ca2+, Hg/+ or heat, and on the basis of different changes in activity during lung development.     

Purine nucleoside phosphorylase variation in the brook lamprey,Lampetra planeri (Bloch) (Petromyzone,Agnatha): Evidence for a trimeric enzyme structure

Genetic evidence for a trimeric structure for purine nucleoside phosphorylase in the brook lamprey is presented. This enzyme is encoded by a single locus with two alleles segregating at frequencies of 0.98 and 0.02 in a Welsh population. It is suggested that this enzyme is likely to be a trimer in all classes of vertebrates.B. J. McAndrew and G. P. Wallis were supported by NERC Research Studentships.     

Evaluation of the Epidemic Potential of Western Equine Encephalitis Virus in the Northeastern United States

The problem of evaluating the epidemic potential of western equine encephalitis in the northeastern United States is presented and possible reasons are discussed for the present lack of human and horse cases of this disease even though increased numbers of isolations of the virus have been obtained in the East during recent years. Epidemiologic factors of vector bionomics and virus strain variations are considered. It is concluded that while this virus strain can no longer be regarded as uncommon in the Northeast, the evidence indicates there is little potential for epidemic expression of this agent in the human and horse population. This appears to be due to differences in the bionomics of the mosquito Culiseta melanura, which serves as the primary enzootic vector in the northeastern United States and in the bionomics of Culex tarsalis that is the vector in the western region of the United States. Other limiting factors in the epidemic potential may be variations between virus strains located in the East and West.     

Concentration of viruses from large volumes of tap water using pleated membrane filters.

A method is described for the efficient concentration of viruses from large volumes of tap water in relatively short time periods. Virus in acidified tap water in the presence of aluminum chloride is adsorbed to a 10-inch (ca. 25.4 cm) fiberglass depth cartridge and a 10-inch pleated epoxy-fiberglass filter in series at flow rates of up to 37.8 liters/min (10 gallons/min). This filter series is capable of efficiently adsorbing virus from greater than 19,000 liters (5,000 gallons) of treated tap water. Adsorbed viruses are eluted from the filters with glycine buffer (pH 10.5) and the eluate is reconcentrated using an aluminum flocculation process. Viruses are eluted from the aluminum floc with glycine buffer (pH 11.5). Using this procedure, viruses in 1,900 liters (500 gallons) of tap water can be concentrated 100,000-fold in 3 h with an average recovery of 40 to 50%.     

Studies on the Survival and Fate of Enteroviruses in an Experimental Model of a Municipal Solid Waste Landfill and Leachate

In laboratory scale municipal solid waste lysimeters containing simulated refuse, and seeded with either laboratory or field strains of poliovirus type 1 and echovirus type 7, viruses were not detected in the lysimeter leachate produced over a 4-month period. In addition, viruses were detected in the lysimeter refuse contents after termination of lysimeter operation. These results appeared to be due to virus retention in the lysimeter caused by virus adsorption and virus inactivation. Evidence for virus inactivation was provided by the results of experiments on virus inactivation in composite leachate samples. Evidence for virus adsorption was supported by the rapid adsorption of viruses to various municipal solid waste components in the presence of a salt similar in composition to the major inorganic salts of leachates.