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Purpose: To determine whether two families diagnosed with X-linked retinoschisis contained mutations in the XLRS1 gene., Methods: DNA from the patients was obtained from blood lymphocytes using commercially available kits. Single-strand conformation assay was performed in an electrophoresis apparatus using 10% acrylamide TBE gels at 10 degrees C. The gels were stained with SYB green II and were scanned in a phosphoimager. DNA was sequenced using an automated fluorescence sequencer., Results: A deletion that eliminates exon 2 was found in one family. An abnormal sequence replacement in exon 4 was found in the other family. Both mutations have severe effects in the coding region by inserting premature stop codons., Conclusions: Both of the families have mutations in the XLRS1 gene. One of these mutations points to a novel mechanism. The mutation is caused by a replacement of 17 bp of a normal sequence with 20 bp of a sequence originating from two different places in the antisense strand. This suggests that early Okazaki fragments were incorporated into the sense strand of exon 4, replacing the normal sequence.

Activins, a subgroup of the transforming growth factor-beta (TGF-beta) superfamily, have been extensively studied in vertebrates for their roles in growth and development. However, activins are not thought to be expressed in invertebrates. The identification of the first invertebrate activin gene is reported here. A genomic clone representing 102 F region of the Drosophila chromosome 4 is found to encode a putative activin beta. The predicted protein sequence has a multibasic protease site that would generate a mature C-terminal peptide containing 113 amino acids showing > 60% similarity to the vertebrate activin beta B (inhibin beta B) sequences. A TGF-beta family signature as well as all 9 cysteine residues conserved in the vertebrate activins are also present in this mature peptide sequence. Northern blot and RT-PCR analyses indicated that the activin beta gene is expressed in embryo, larva and adult stages of Drosophila.

Purpose: [gamma]-Crystallins are major structural proteins of the eye lens. While other crystallins have revealed distinct non-lens functions and patterns of expression, [gamma]-crystallins have generally appeared to be the most lens-specific of the crystallins. Here we examine the mouse [gamma]S-crystallin ([gamma]S) gene and its expression., Methods: The cDNA and gene for mouse [gamma]S were cloned and sequenced. The Crygs gene was mapped using genetic crosses. Expression patterns in mouse and cow were examined by northern blot, PCR and western blot using a specific peptide antibody., Results: The Crygs gene was sequenced and mapped to mouse chromosome 16, at or near the locus for the genetic cataract Opj. Northern blots of tissues from new born mice, showed lens specific expression of [gamma]S. However, in the mature mouse eye there was, in addition, clear non-lens expression of [gamma]S. In the adult bovine eye RT-PCR shows that [gamma]S is expressed in lens, retina and cornea. A peptide antibody directed against [gamma]S detects bands of the expected size in western blots of mouse lens and in 33 day old mouse retina., Conclusions: These results suggest that [gamma]-crystallins have a non-crystallin role outside the lens, one which may predate the lens in evolutionary terms. Non-lens expression seems to increase with age in young mice, hinting that [gamma]S may have a role similar to that of a stress protein in tissues of the eye, perhaps related to accumulating insults resulting from light exposure.

Pax-6 is essential for eye development and for tissue-specific gene expression within the eye. Splicing of an alternative exon (5a) leads to variant DNA binding specificity in Pax6. Here we show that in the mature bovine eye the relative abundance of the alternative Pax6-5a mRNA varies markedly, particularly between the lens and iris. Additional alternative splicing products were identified which may lead to "domain swapping" or truncation of Pax6 proteins. Comparison of amphibian (Xenopus) and mammalian (bovine) Pax-6 genes revealed highly conserved intronic sequences flanking exon 5a, including potential lariat sites which may have a role in the mechanism of alternative splicing.

A 13 kDa protein from bovine lens was identified and characterized by protein microsequencing and by rapid amplification of cDNA ends (RACE) PCR. Its complete sequence shows that this protein belongs to a family of fatty acid-binding proteins (FABPs), including myelin and adipocyte P2, that are associated with cellular differentiation. The bovine lens protein, designated LP2, shows very close similarity to human epidermal FABP (eFABP) and human eFABP was detected in human lens, suggesting that the two proteins might be orthologous. Reverse transcriptase-PCR (RT-PCR) was used to compare expression patterns of LP2 with those for actin and for the differentiation markers gamma B-crystallin and gamma s-crystallin in lens. Actin was most abundant in the relatively undifferentiated epithelial cells and decreased with lens cell differentiation. In contrast gamma B-crystallin and gamma s-crystallin were detected only in fibres (nuclear and cortical respectively). LP2 transcripts were detected most abundantly in fibre cells and apparently increased with cellular differentiation. Molecular modelling confirms that the sequence of LP2 fits the tertiary template of adipocyte P2 but reveals the presence of two close pairs of cysteine residues that might be susceptible to intramolecular disulphide bond formation under appropriate oxidizing conditions. LP2 is thus another potential target for oxidative stress during cataract formation in lens.

Background: In this time of federal government cutbacks, federal agencies are looking at the cost effectiveness of their programs. This pilot study is a first attempt at quantifying the cost effectiveness of Indian Health Service (IHS) optometry., Methods: Using a super bill system and the Indian Health Service's own value figures per procedure, the cost of providing optometry services at one clinic was compared with the value of the care given in terms of dollars., Results: The Indian Health Service receives about $2.44 in care for each dollar spent on optometric services at the clinic studied., Conclusions: If the results found are typical of all the IHS clinics, then the optometry program saves U.S. taxpayers more than $14 million per year. Further studies at multiple IHS sites should be performed to confirm these figures.

alpha A-crystallin, a major structural protein in the ocular lenses of all vertebrates, has been a valuable tool for molecular phylogenetic studies. This paper presents the complete sequence for human alpha A-crystallin derived from cDNA and genomic clones. The deduced amino acid sequence differs at two phylogenetically informative positions from that previously inferred from peptide composition. This led us to examine the same region of the alpha A-crystallin gene in 12 other mammalian species using direct sequencing of PCR-amplified genomic DNA. New sequences were added to the database, and corrections were made to all anthropoid sequences, defining clear synapomorphies for anthropoids as a clade distinct from prosimians. Within the anthropoids there are further synapomorphies delineating hominoids, Old World monkeys, and New World monkeys. Significantly, sequence revisions and the addition of new sequence for a prosimian, the sifaka, eliminate the previous support for the proposed anthropoid affinities of the tarsier inferred from alpha A-crystallin protein sequences. In addition, DNA sequences provide greater resolution of certain relationships. For example, although they are identical in protein sequence, comparison of DNA sequences clearly separates mouse and the common tree shrew, grouping the tree shrew closer to prosimians. These results show that adding DNA sequences to the existing alpha A-crystallin database can enhance its value in resolving phylogenetic relationships.

Background: Mentally retarded patients are often found to have exposure corneal disease. No study could be found that documented the prevalence of this corneal disorder among these patients., Methods: A retrospective study was conducted on a population of 210 mentally retarded adults to determine the prevalence of clinically significant exposure keratopathy., Results: 17.6 percent of this population was found to have this disorder mostly due to incomplete blinking., Conclusions: Exposure keratopathy was found to be a significant ocular health problem among this population. Since many of these people cannot verbalize their discomfort it is recommended that the optometrist tailor his or her examination so this disorder is not overlooked and untreated.

Approximately 2 kb of 5'-flanking sequences of the lens-specific alpha A-crystallin genes from human and mouse are presented and compared with similar regions of the chicken gene. A repetitive element was found approximately 1 kb upstream from the coding sequences of the alpha A-crystallin gene in all three species (Alu in human, B2 in mouse, and CR1 in chicken), suggesting that they may have an important functional or structural role. Despite the ability of alpha A-crystallin promoters to function across species, dot matrix analyses show only limited similarity among the 600 bp 5' to the structural genes of these three species. The human 5'-flanking sequence is more similar to that of the mouse and chicken than the mouse and chicken are to each other. Numerous short sequences (8-13 bp) are common to all three genes but are distributed differently in each species. The locations and conservation of these sequence motifs suggest functional roles, possibly as cis-regulatory elements of transcription. One motif is similar to the alpha A-CRYBP1 binding site implicated earlier in the transcriptional regulation of the mouse alpha A-crystallin gene, and other motifs correspond to sites previously mapped by methylation interference studies in the mouse alpha A-crystallin promoter. The modular arrangement of conserved sequence motifs is consistent with evolutionary changes occurring at the level of gene regulation.

The crystallin genes encode the major soluble proteins of the lens. Some of the crystallin genes are expressed exclusively in the lens while others are also expressed in different tissues. The two alpha-crystallin genes, alpha A and alpha B, differ in their tissue specificity. Transcription of the alpha A-crystallin gene occurs only in the lens, while the alpha B-crystallin gene is also expressed in other tissues, including heart, skeletal muscle, kidney, lung and brain. MIP (also called MP26), the major intrinsic protein of the lens fiber membranes, is also expressed exclusively in the lens. Correct expression of both alpha-crystallin and MIP are required for normal lens function. Here we review our studies on the molecular basis of expression of the alpha-crystallin and MIP genes in the lens. The 5' flanking sequences containing the initiation site of transcription of the alpha A-crystallin, alpha B-crystallin and MIP genes were fused to the bacterial chloramphenicol acetyltransferase (CAT) gene, and the expression of this reporter gene was studied in transient assays and transgenic mice. DNA sequences flanking the 5' end of the alpha A-crystallin gene contain regulatory elements responsible for the lens-specific expression and developmental regulation of the CAT gene in transgenic mice. Interestingly, although some of the murine alpha A-crystallin regulatory sequences are conserved in the human and chicken genes, different functional regulatory elements appear to control the expression of the murine and chicken alpha A-crystallin genes. The 5' flanking sequence of the alpha B-crystallin gene preferentially directs expression of the CAT gene to the lens and to skeletal muscle. Different regulatory elements of the alpha B-crystallin gene appear to be responsible for its transcription in various tissues. The 5' flanking sequence of the MIP gene also contains regulatory elements that direct expression of the CAT gene to lens cells; these sequences are not functional in transfected non-lens cells and are different from the cis regulatory elements controlling alpha-crystallin gene expression. The multiplicity of cis-regulatory elements controlling the transcription of these three genes indicates the complexity of the mechanisms that regulate gene expression in the lens.

Presented is an unusually severe case of acute necrotizing ulcerative gingivitis (ANUG) in association with systemic lupus erythematosus (SLE), in which the patient was observed to slough a considerable amount of attached gingival tissues and eventually to lose a number of teeth and supporting bone. The usual limited and treatable nature of ANUG is reviewed, and attention is directed to the mechanisms by which SLE and therapeutic amounts of steroids can contribute to the increased severity of the oral disease.

Six general-purpose fumigants and one fungicide were applied by different methods and evaluated for control of nematode-fungus complexes on cabbage grown for transplant production. All chemicals reduced populations of nematodes and soil-borne fungi but varied greatly in effectiveness. Methyl bromide + chloropicrin (98% methyl bromide + 2% chloropicrin) (MBR-CP gas), DD + methyl isothiocyanate (DD-MENCS), methyl bromide + chloropicrin (67% methyl bromide + 31.75% cbloropicrin) (MBR-CP gel), and chloropicrin were more effective than sodium methyl dithiocarbamate (metham), pentachloronitrobenzene (PCNB), and potassium N-hydroxy-methyl-N-methyldithiocarbamate (Bunema) against Meloidogyne incognita. Populations of Pythium spp. and Fusarium spp. were reduced markedly by all treatments except PCNB. Plant growth, uniformity, and yield were greater when nematodes and fungi were controlled.

Three crops of cucumber were grown in succession in beds by use of trickle irrigation, plastic film mulch, and soil chemical treatments over a 17-month period, including a fallow winter season. Total yield for the three crops was highest (1208 quintals/ha) in film-mulched plots treated with MBR-CP, and next-highest in film-mulched plots treated with DD-MENCS (1094 quintals/ha); total yield was only 456 quintals/ha in film-mulched control (untreated) plots. Yield in untreated film-mulched plots was 256% of that in untreated unmulched plots (178 quintals/ha). Plant growth and yields were greatest when populations of nematodes and soil-borne fungi were suppressed to very low levels. The residual control by soil treatments lasted longest on Meloidogyne incognita and Fusarium solani.

Field plots in Tifton loamy sand were treated with various soil pesticides in 1973 and 1974 and either left exposed or covered with biodegradable flint mulch. Test crops were cantaloup, slicing and pickling cucumber, squash, and sweet corn. Overhead sprinkler irrigation was used in 1973, and trickle irrigation under the film mulch was used on sweet corn in 1974. Soil was assayed for nematodes, and roots of plants were evaluated for damage by root-knot nematodes. Nematode populations were reduced by soil treatment with an organic phosphate or carbamate nematicide-herbicide-fungicide combination (NHF), DD-MENCS, methyl bromide-chloropicrin (MBR-CP), ethoprop, carbofuran, and sodium azide + ethoprop or carbofuran. Sodium azide, sodium azide + ethoprop or carbofuran, ethoprop, and carbofuran were less effective than DD-MENCS, MBR-CP, attd the NHF combination. The NHF combination controlled grasses and broadteaf weeds as effectively as the herbicide alone. Growth and yield were greatest when nematodes and weeds were controlled. Yields of marketable vegetables were highest from plants in plots treated with DD-MENCS with a film mulch.

The frequent correspondence of exons to structural or functional domains in proteins has suggested that many proteins have evolved by modular assembly. This idea is supported by examples of apparent exon duplication and by shared domains among both alternatively spliced and completely separate genes. During this process it is probable that some combinations of exons would not prove advantageous and would therefore be lost. Here we report that within the active single-copy human gene for alpha A-crystallin there is a 'pseudo-exon' in the early stages of being extinguished, perhaps the result of a failed experiment in the evolution of this specialized, lens-specific protein.

Okra was grown in field plots of Tifton loamy sand naturally infested with the nematodes Meloidogyne incognita and Criconemoides ornalus and the pathogenic fungi Fusarium oxysporum, F. solani, F. roseum, and Pythium spp. Plots were treated with various soil pesticides and left exposed or covered with biodegradable paper film mulch under trickle irrigation. Soil was assayed for nematodes and fungi, and plant roots were examined for root-rot and insect damage. Fewer nematodes and fungi generally were recovered from soil treated with DD-MENCS (with and without film mulch) or methyl bromide-chloropicrin (2:1) (MBC) and film mulch than from nontreated soil. Funfigation with DD-MENCS or MBC suppressed populations of M. incognita, C. ornatus, F. oxysporum, F. solani, F. roseum, and Pythium spp. Ethoprop (alone or combined with other pesticides), sodium azide, and chloroneb were less effective than DD-MENCS and MBC. Plant growth anti yield were greatest when nematodes and pathogenic fungi were controlled. Yield was increased 3-fold by DD-MENCS + film mulch or MBC + film mulch in comparison with the average yield of okra produced in Georgia. The root-knot nematode-Fusarium wilt complex was most severe in nonfuntigated soil.

The intracellular concentration of cyclic AMP regulates many cellular properties of normal rat, mouse, and human fibroblasts in culture. Thus it is important to elucidate how the activity of adenylate cyclase of fibroblasts might be altered and regulated by both intracellular and extracellular agents and events. In studies with several virally transformed fibroblast lines, as well as those transformed spontaneously or by 3-methylcholanthrene, a common feature of each type of transformation is a defective adenylate cyclase system. However, the means by which adenylate cyclase activity is altered differs with the cell system and the type of transformation. Here we concentrate on efforts to understand the regulatory properties of the adenylate cyclase from normal rat kidney fibroblasts. The modulation (increase or decrease) of the hormonal responsiveness of this enzyme may play an important role in its regulation. Of substantial interest is the isolation from serum of a high-molecular weight factor that selectively decreases the GTP and hormone-stimulated activities of adenylate cyclase. These findings are of value in our attempts at elucidation of the reasons for altered cyclase activity following virus transformation and during various stages of growth.

In a 6-year cover crop sequence study, nematode population densities varied with different cover crops. Millet favored rapid increase of Belonolaimus longicaudatus and supported relatively large numbers of Pratylenchus brachyurus. Beggarweed and 'Coastal' bermudagrass favored rapid increase of B. Iongicaudatus and to a lesser extent P. brachyurus and Trichodorus christiei. Hairy indigo and Crotalaria supported relatively large numbers of P. brachyurus but suppressed B. longicaudatus. Hairy indigo also supported increases of T. christiei and Xiphinema americanum. Marigold did not favor development of any parasitic nematode species present. Tomato transplant yield was inversely related to nematode population, particularly to B. Iongicaudatus. Largest yields were obtained from plots with smallest numbers of B. longicaudatus and smallest yields were from plots with largest numbers of B. longicaudatus.

Five nematode species were studied for ability to develop on seven summer cover crops in rotation with tomato transplants grown every third year. Increase of Tylenchorhynchus claytoni, Trichodorus christiei, Pratylenchus brachyurus, Helicotylenchus dihystera, and Xiphinema americanum in newly cleared soil varied with different cover crops. No substantial nematode population increases occurred until the third summer of crop growth. All species except X. americanum and H. dihystera developed best on sudangrass and millet. Crotalaria caused substantial increase of H. dihystera and P. brachyurus but suppressed the other species. Marigold suppressed all species except X. americanum which increased substantially on marigold during the 5th year. Cotton favored rapid increase of T. christiei, and moderate increases of all species except T. claytoni which was suppressed. Beggarweed favored moderate increases of T. christiei and H. dihystera but suppressed the other species. Hairy indigo favored rapid increase of H. dihystera, moderate increases of T. christiei and X. americanum, and suppressed the other species. Number of marketable transplants was reduced after 2 years of sudangrass and cotton; these crops favored increases of T. christiei and T. claytoni. The better cover crops prevented increases of most plant parasitic nematodes in land cropped to tomato, a suitable host.