Herpes simplex virus type 1 immediate-early protein Vmw110 reactivates latent herpes simplex virus type 2 in an in vitro latency system.

Reactivation of latent herpes simplex virus type 2 (HSV-2) by the immediate-early protein Vmw110 was studied by using an in vitro latency system. Adenovirus recombinants that express Vmw110 reactivated latent HSV-2. An HSV-1 mutant possessing a deletion in a carboxy-terminal region of Vmw110 reactivated latent HSV-2, whereas mutant FXE, which has a deletion in the second exon, did not. Therefore, Vmw110 alone is required to reactivate latent HSV-2 in vitro, and the region of Vmw110 defined by the deletion in FXE is important for this process.     

Hormonal stimulation of tyrosinase activity in human foreskin organ cultures

Summary A human foreskin organ culture system has been developed to study the response of human skin to hormonal stimulation. Foreskins are maintained in culture on floating plastic supports which allows the epidermal surface to be exposed to air while the dermis is bathed in nutrient medium. Both black and white human foreskins can be maintained in organ culture for at least 1 wk with no change in the tissue structure or cell viability as determined by histochemical staining and by dopa reaction staining. Tyrosinase activity in both black and white human foreskin cultures decays markedly during the first 2 d of culture to a new steady state level which remains stable throughout the culture period. Both black and white foreskin cultures consistently demonstrate 2- to 10-fold increases in tyrosinase activity when treated with theophylline (1 mM). Approximately 90% of all skin cultures examined showed an increase in enzyme activity when treated with this phosphodiesterase inhibitor. Dibutyryl cAMP (0.1 mM) and [Nle⁴, D-phe⁷]-alpha MSH (10⁻⁸ M), were also found to markedly stimulate tyrosinase activity in some skin cultures, whereas alpha-MSH and prostaglandin E₁ produced only an inconsistent and small increase in the activity of the enzyme. Histamine (1 μM), vitamin D₃ (1 μM), and retinoic acid (1μM) failed to stimulate tyrosinase activity in either white or black foreskin cultures. This hormone-responsive organ culture system can be utilized to characterize the molecular processes responsible for the regulation of tyrosinase and pigmentation in human skin. This work was supported by a research contract from the Oklahoma Center for the Advancement of Science and Technology (OCAST) and by a research grant from the Presbyterian Health Foundation.     

A Second Gene for Cerulean Cataracts Maps to the β Crystallin Region on Chromosome 22

Congenital cataracts are one of the most common major eye abnormalities and often lead to blindness in infants. At least a third of all cases are familial. Within this group, highly penetrant, autosomal dominant forms of congenital cataracts (ADCC) are most common. ADCC is a genetically heterogeneous group of disorders, in which at least eight different loci have been identified for nine clinically distinct forms. Among these, Armitageet al.(Nature Genet.9: 37–40, 1995) mapped a gene for cerulean blue cataracts to chromosome 17q24. Bodkeret al.(Am. J. Med. Genet.37: 54–59, 1990) described a large family with cerulean blue cataracts, in which the affected daughter of affected first cousins was presumed to be homozygous for the purported gene. We report linkage in this family to the region on chromosome 22q that includes two β crystallin genes (CRYBB2, CRYBB3) and one pseudogene (CRYBB2P1). The affected female in question is homozygous at all markers.     

A METHOD FOR ISOLATION OF CAMPYLOBACTER SPP. (JEJUNI, COLI AND LARI) FROM SHELLFISH AND THE MARINE ENVIRONMENT

A method has been developed to detect thermophilic species of Campylobacter in shellfish, marine and tributary waters, sediment and farm runoff by-products such as manure and silage. The method consists of a 48 h enrichment incubation and subcultured to selective agars. Presumptive colonies confirmed with a latex agglutination (antibodies) to common flagellar antigens of C. jejuni, C. coli and C. lardi. Over an 8 year period, West Coast estuaries (Washington, Oregon, and California) were sampled, resulting in analysis of a total of 512 samples. Results suggest that Campylobacter spp. are well distributed in the marine environment. Two enrichment broths were compared for the recovery of campylobacters from environmental samples. The method described in the Food and Drug Administration Bacteriological Analytical Manual (FDA/BAM) (1984), was compared to a modified method. Use of the modified method described here resulted in higher recovery rates of Campylobacter spp. Recoveries of campylobacters from sediment, shellfish, and water were 10,13, and 28% higher for the modified method, respectively.     

Fine mapping of the Autosomal Dominant Split Hand/Split Foot Locus on Chromosome 7, Band q21.3-q22.1

Split hand/split foot (SHFD) is a human developmental defect characterized by missing digits, fusion of remaining digits, and a deep median cleft in the hands and feet. Cytogenetic studies of deletions and translocations associated with this disorder have indicated that an autosomal dominant split hand/split foot locus (gene SHFD1) maps to 7q21-q22. To characterize the SHFD1 locus, somatic cell hybrid lines were constructed from cytogenetically abnormal individuals with SHFD. Molecular analysis resulted in the localization of 93 DNA markers to one of 10 intervals surrounding the SHFD1 locus. The translocation breakpoints in four SHFD patients were encompassed by the smallest region of overlap among the SHFD-associated deletions. The order of DNA markers in the SHFD1 critical region has been defined as PON–D7S812–SHFD1–D7S811–ASNS. One DNA marker, D7S811, detected altered restriction enzyme fragments in three patients with translocations when examined by pulsed-field gel electro-phoresis (PFGE). These data map SHFD1, a gene that is crucial for human limb differentiation, to a small interval in the q21.3-q22.1 region of human chromosome 7.     

Source of an egg kairomone for Trissolcus basalis, a parasitoid of Nezara viridula

Abstract. The eggs of the southern green stink bug, Nezara viridula (L.) (Hemiptera: Pentatomidae), are successfully attacked by Trissolcus basalis (Woll.) (Hymenoptera: Scelionidae) and are recognized as hosts by a secretion applied to the egg chorion. This secretion is produced by the follicular cells in the proximal region of the ovariole of the female pentatomid and functions as an adhesive for attaching the eggs to the oviposition substrate. The adhesive and kairomone activity could be partially removed with water. This water extract elicited host recognition behaviour in T. basalis when applied to glass beads which stuck together as the extract dried. The adhesive and kairomonal activity was removed completely with acetone since acetone-washed host eggs were not recognized by T. basalis. Application of the acetone extract to glass beads stimulated ovipositional probes by T. basalis. The adhesive appeared to be composed of a mucopolysaccharide–protein complex.     

Minimizing growth regulators in shoot culture of an endangered plant,Hackelia venusta (Boraginaceae)

Summary Hackelia venusta (Boraginaceae) is an endangered perennial herb endemic to the interior northwestern United States. Because of seed scarcity, micropropagation (anex situ conservation strategy) could produce true-to-type plantlets suitable for reintroduction. We hypothesized that clones of predetermined size could be rapidly produced by supplementing multiplication and rooting media with minimal levels of cytokinin and auxin. Microshoots derived from shoot expants were cultured on Murashige and Skoog (1962) media supplemented with 1% (wt/vol) agar and 0.0001 to 10 μM benzyladenine. Inverse regression estimates on 3 genotypes predicted that a target of 2.5 axillary microshoots per explant would require a minimal level of 0.04±0.02 μM benzyladenine. Culture of 25 genotypes with 0.04 μM benzyladenine resulted in an average of 2.3±0.1 axillary microshoots per explant. Elongated microshoots were transferred to media supplemented with 0.1 to 25 μM indoleacetic acid. Clones rooted from 36% to 100% success after 4 wk in 2.0 μM indoleacetic acid. Plantlets transplantedex vitro with three or more roots survived at 84% versus 46% of plantlets with fewer roots. Up to 84% of the plantlets survived in a planting trial. The data suggest that shoot culture ofHackelia venusta, with minimal growth regulators, can produce axillary microshoots for reintroduction.