Total chemical synthesis of a D-enzyme: the enantiomers of HIV-1 protease show reciprocal chiral substrate specificity [corrected].

The D and L forms of the enzyme HIV-1 protease have been prepared by total chemical synthesis. The two proteins had identical covalent structures. However, the folded protein-enzyme enantiomers showed reciprocal chiral specificity on peptide substrates. That is, each enzyme enantiomer cut only the corresponding substrate enantiomer. Reciprocal chiral specificity was also evident in the effect of enantiomeric inhibitors. These data imply that the folded forms of the chemically synthesized D- and L-enzyme molecules are mirror images of one another in all elements of the three-dimensional structure. Enantiomeric proteins are expected to display reciprocal chiral specificity in all aspects of their biochemical interactions.

Stimulation of gonadotropin release by a non-GnRH peptide sequence of the GnRH precursor.

The human gonadotropin-releasing hormone (GnRH) precursor comprises the GnRH sequence followed by an extension of 59 amino acids. Basic amino acid residues in the carboxyl terminal extension may represent sites of processing to biologically active peptides. A synthetic peptide comprising the first 13 amino acids (H X Asp-Ala-Glu-Asn-Leu-Ile-Asp-Ser-Phe-Gln-Glu-Ile-Val X OH) of the 59-amino acid peptide was found to stimulate the release of gonadotropic hormones from human and baboon anterior pituitary cells in culture. The peptide did not affect thyrotropin or prolactin secretion. A GnRH antagonist did not inhibit gonadotropin stimulation by the peptide, and the peptide did not compete with GnRH for GnRH pituitary receptors, indicating that the action of the peptide is independent of the GnRH receptor. The GnRH precursor contains two distinct peptide sequences capable of stimulating gonadotropin release from human and baboon pituitary cells.

Receptor binding and gonadotropin-releasing activity of a novel chicken gonadotropin-releasing hormone ([His5, Trp7, Tyr8]GnRH) and a D-Arg6 analog.

Receptor binding and gonadotropin-releasing activity was compared for mammalian GnRH, [Gln8]GnRH (chicken I GnRH), [His5, Trp7, Tyr8]GnRH (chicken II GnRH), [Trp7, Leu8]GnRH (salmon GnRH), and [D-Arg6] chicken II GnRH. The mean ED50 values for mammalian GnRH, chicken I GnRH, chicken II GnRH, and salmon GnRH in stimulating LH release from dispersed chicken pituitary cells were 0.27 nM, 0.28 nM, 0.055 nM, and 0.11 nM, respectively. The relative potencies of the peptides compared in the same assay were 0.93, 1.0, 5.6, and 2.5. The ED50 values for chicken I GnRH, chicken II GnRH, and salmon GnRH in stimulating FSH release were 0.37 nM, 0.034 nM, and 0.18 nM, and the relative potencies were 1.0, 13.5, and 1.8. Chicken II GnRH was, therefore, more potent than chicken I GnRH and mammalian GnRH in releasing LH and appeared to have an even greater relative FSH-releasing activity than chicken I GnRH or mammalian GnRH. Introduction of D-Arg6 into chicken II GnRH enhanced the activity of this analog 4- and 2-fold relative to chicken II GnRH in LH- and FSH-releasing activity, respectively. The ED50 values of mammalian GnRH, chicken I GnRH, chicken II GnRH, and salmon GnRH in releasing LH from cultured sheep pituitary cells were 2.9 nM, 96 nM, 22 nM, and 104 nM, respectively. The relative potencies were 1.0, 0.016, 0.084, and 0.047. Introduction of D-Arg6 into chicken II GnRH enhanced activity 9-fold. In a rat pituitary receptor binding assay the ED50 values of mammalian GnRH, chicken I GnRH, chicken II GnRH, and salmon GnRH were 2.9 nM, 1480 nM, 19 nM, and 258 nM, respectively. [D-Arg6]Chicken II GnRH was 46 times more active than the natural chicken II GnRH peptide. The results show: 1) chicken II GnRH is more potent than chicken I GnRH, which is equipotent with mammalian GnRH in releasing LH from chicken pituitary cells. Chicken II GnRH is even more potent at releasing FSH. 2) Salmon GnRH is also more potent than chicken I GnRH and mammalian GnRH in stimulating gonadotropin release from chicken pituitary cells. It appears, therefore, that Trp in the 7 position contributes to the enhanced activity of salmon and chicken II GnRH. 3) The low activity of chicken I GnRH, chicken II GnRH, and salmon GnRH in the sheep pituitary cell bioassay and rat pituitary receptor binding assay confirms that Arg8 in mammalian GnRH is important for activity.(ABSTRACT TRUNCATED AT 400 WORDS)

Mild vitamin A deficiency and childhood morbidity--an Indian experience.

Over 1500 preschool urban Indian children were followed weekly for morbidity from 12 to 18 mo. Examination for mild xerophthalmia (Bitot's spots and night blindness) was done initially and at 6 and 12 mo. Children with mild xerophthalmia at the start of a 6-mo interval developed respiratory disease in the interval twice as often as children with normal eyes at the start of the interval. No association was found between mild xerophthalmia and incidence of diarrhea.

Diarrhea, respiratory infections, and growth are not affected by a weekly low-dose vitamin A supplement: a masked, controlled field trial in children in southern India.

Incidence, duration, and severity of diarrhea and respiratory symptoms were monitored weekly for 1 y in 15,419 children 6-60 mo of age in a randomized, placebo-controlled, masked clinical trial conducted in southern India. Half the children received weekly doses of 8.7 mumol (2500 microgram) vitamin A and 46 mumol (20 mg) vitamin E (treated) and the other half, 46 mumol vitamin E (control). Medical and ocular examinations and anthropometric measurements were obtained before and after 52 wk of intervention. Ocular examinations also were obtained after 26 wk. Supplements were delivered weekly from calibrated dispenser bottles by community health volunteers who also recorded each mother's recall of daily morbidity of her child during the previous week. Baseline characteristics of treated and control subjects were similar and documented a prevalence of 11% xerophthalmia and 72% undernutrition. Weekly treatment with the low-dose vitamin A supplement did not influence the incidence, severity, or duration of diarrhea or respiratory infections and did not influence linear or ponderal growth.

Relationship between measles, malnutrition, and blindness: a prospective study in Indian children.

A prospective study was conducted in slum children to determine the incidence of post-measles corneal disease and to clarify its relationship with nutritional status. A total of 318 cases of measles were identified over a period of 15 mo; maximum incidence was observed for children between 1-2 yr. Most of the children showed weight loss and serum proteins decrease during the acute stage of measles. Corneal lesions were observed in 3% of the children, and the lesions responded well to treatment. Serum vitamin A and RBP levels were significantly depressed during the acute stage of measles but were restored to normal 8 wk after recovery. There were no significant differences in the serum levels for those with and without eye lesions, which suggests that these lesions may not be mediated simply through the effect of infection on serum concentration of vitamin A.

Location and severity of cortical opacities in different regions of the lens in age-related cataract.

PURPOSE: To examine the distribution of cortical opacities across the lens in the Italian-American Natural History Study of Age-Related Cataract and to study the association between an index of sunlight exposure and the location of cortical cataract within the lens. METHODS: Lens photographs of one eye of 731 persons with cortical opacities (503 with pure and 228 with mixed types of opacity) were included in the analysis. A radial grid superimposed on the photographs was used to assess presence, location, and severity of wedge-shaped cortical opacities. RESULTS: Both the prevalence and the extent of cortical opacities were highest in the inferior-nasal quadrant and lowest in the superior-nasal quadrant of the lens. In polychotomous logistic regression, persons with the greatest excess areal involvement in the inferior half of the lens were more likely to have high exposure to sunlight, as measured by a sunlight index, than persons with excess involvement in the superior half of the lens (odds ratio, 1.73; 95% confidence interval 1.03, 2.93). Excess areal involvement of the inferior lens also was associated with the pure type of cortical cataract and with the total extent of the opacity. CONCLUSIONS: Age-related cortical opacities occur more frequently inferiorly than superiorly and, to a lesser extent, nasally than temporally. Possibly higher exposure of these lens segments to sunlight may explain this preferential location of cortical opacities.

A dose-response effect between a sunlight index and age-related cataracts. Italian-American Cataract Study Group.

To explore the existence of a dose-response relationship between sunlight exposure and risk of age-related cataracts, we analyzed data collected from 1008 patients with cataracts and 469 control subjects enrolled in the Italian-American Case-Control Study of Age-Related Cataracts. Fourteen variables related to sunlight exposure history were included in the questionnaire administered to the study participants. A sunlight index was constructed and its relationship to the presence of cataracts was modeled by logistic regression. After adjustments for potential confounding variables and for age and sex, a significant dose-response effect (P = 0.01) was detected between the sunlight exposure index and the presence of pure cortical cataracts. With the exception of corticonuclear cataracts, all the other mixed types of opacity also showed a dose-response association with the sunlight index. These data support the hypothesis that sunlight exposure is a risk factor in the development of cortical cataracts, and demonstrate the existence of a dose-response relationship in this association.

Statistical computing in the United States.

Recent history and developments related to the increase in statistical computing activities in the United States and by U.S. participants in international efforts are reviewed, with emphasis on important events, organizations, references, and products which contribute to informed selection and use of statistical programs. Three features matrices for major statistical packages are included as potential aids to Japanese statisticians in assessing the utility of these packages in biostatistical applications.

Impaired vision and hip fracture. The Framingham Study.

Falls affect a large proportion of the elderly and can result in a variety of injuries, including hip fractures. Several studies have suggested that visual impairment contributes to falls, but studies have not used standardized definitions of visual impairment and have not examined injurious falls or fractures. We looked at the risk of hip fracture associated with visual impairment in those members of the Framingham Study Cohort who took part in the Framingham Eye Study in 1973-75. Of 2,633 subjects followed for 10 years after the eye exam, 110 sustained hip fractures. The fracture rates in those with moderately impaired (20/30 to 20/80) vision (8.5%) and poor (20/100 or worse) vision (11.3%) were higher than in those with good (20/25 or better) vision (3.0%). After adjustment for age, sex, weight, alcohol consumption, and (in women) estrogen use, the relative risk of fracture in those with moderate impairment was 1.54 (95% CI = 0.95-2.49), while for those with poor vision, the relative risk was 2.17 (95% CI = 1.24-3.80). Of note, those with moderately impaired vision in one eye and good vision in the other had a higher risk of fracture (relative risk = 1.94) than those with a similar degree of binocular impairment (relative risk = 1.11). Poor vision in one or both eyes was linked to an elevated fracture risk. This suggests that good stereoscopic vision may be necessary to prevent falls. The risk of fracture with poor and moderately impaired vision combined was increased in women (relative risk = 1.96, 95% CI = 1.23-3.11) but not in men (relative risk = 0.79, 95% CI = 0.23-2.72).(ABSTRACT TRUNCATED AT 250 WORDS)

Alternative definitions of open-angle glaucoma. Effect on prevalence and associations in the Framingham eye study.

Framingham Eye Study data were used to examine the effect of alternative definitions of open-angle glaucoma (OAG) on prevalence and on associations with selected variables from the Framingham Heart Study. Definitions were based on various combinations of history of glaucoma, types of visual field defects, and functions of intraocular pressure and cup-disc ratio. Visual field defect irrespective of blind spot enlargement is used as a standard for comparison. Prevalence of OAG by this definition is higher for men than for women and increases with age, more markedly than for most other definitions. Alcohol consumption is directly related to glaucoma defined either by the reference standard or by visual field defects including blind spot enlargement. Systemic blood pressure, ventricular rate, and diagnosis of diabetes are directly related to IOP.

Age-related cataract in the Tibet Eye Study.

The Tibet Eye Study was designed to estimate the prevalence of age-related cataract in Duilong-Deqing County, west of Lhasa, China (altitude, 4000 m). Previous reports have suggested an unusually high prevalence of age-related cataract in Tibet. A two-stage probability sample of persons aged 20 years or older from the 35 townships of the county targeted 2884 persons for inclusion in the study; 2665 (92.4%) were examined. Age-related cataract was diagnosed when (1) visual acuity was worse than 6/12 (20/40) because of nuclear or cortical (including posterior subcapsular) opacities, or (2) aphakia associated with a history of age-related cataract was present in either eye. The prevalence of age-related cataract among persons aged 20 to 39 years was 0.2%; among persons 40 years old or older, the prevalence was 11.8%. Cortical cataracts were by far the most common type of cataract diagnosed. Age- and sex-adjusted prevalence in Tibet was 60% higher than the prevalence in a similar, previously conducted study of 6951 person in Shunyi County, northeast of Beijing (altitude, 50 m). A second, independent slit-lamp classification of lens status was conducted in the Tibet Eye Study using standard photographs previously described. Age-specific cataract prevalence was similar with the two examination techniques. Results from the Tibet Eye Study support previous suggestions of a high prevalence of age-related cataract in Tibet.

Biochemical factors in the lens opacities. Case-control study. The Lens Opacities Case-Control Study Group.

OBJECTIVE: To evaluate associations with biochemical indicators of nutritional and other risk factors in the Lens Opacities Case-Control Study. DESIGN: Case-control study. SETTING AND PARTICIPANTS: The Lens Opacities Case-Control Study determined risk factors for cortical, nuclear, and posterior subcapsular opacities among 1380 participants aged 40 to 79 years. DATA COLLECTION: Vitamin E, selenium, and biochemistry profile determinations were performed on all patients; red blood cell enzymes and amino acids were measured in systematic samples of about 25% of the Lens Opacities Case-Control Study population. OUTCOME: Laboratory test values in cases and controls were compared and expressed as odds ratios and 95% confidence intervals. RESULTS: In polychotomous logistic regression analyses controlling for age and sex, the risk of opacities was reduced to less than one half in persons with higher levels of vitamin E (odds ratio, 0.44 for nuclear opacities), albumin-globulin ratio (odds ratio, 0.41 for mixed opacities), or iron (odds ratio, 0.43 for cortical opacities); higher uric acid levels increased risk (odds ratio, 1.74 for mixed opacities). Persons with opacities were twice as likely to have high glutathione reductase activity (with flavin adenine dinucleotide), which suggests low riboflavin status (odds ratio, 2.13). Most odds ratios for amino acids were under unity and were significantly decreased for glycine (0.36) and aspartic acid (0.31). CONCLUSIONS: Lens opacities were associated with lower levels of riboflavin, vitamin E, iron, and protein nutritional status. Higher levels of uric acid increased risk of mixed opacities. The findings for riboflavin, vitamin E, iron, and uric acid are compatible with the dietary intake and medical history results of the Lens Opacities Case-Control Study.

The Linxian cataract studies. Two nutrition intervention trials.

OBJECTIVE: To determine whether the vitamin/mineral supplements used in two cancer intervention trials affected the risk of developing age-related cataracts. DESIGN: Two randomized, double-masked trials with a duration of 5 to 6 years and end-of-trial eye examinations. SETTING: Rural communes in Linxian, China. PARTICIPANTS: In trial 1, 2141 participants aged 45 to 74 years, and, in trial 2, 3249 participants aged 45 to 74 years. INTERVENTIONS: Multivitamin/mineral supplement or matching placebo in trial 1; factorial design to test the effect of four different vitamin/mineral combinations in trial 2 (retinol/zinc, riboflavin/niacin, ascorbic acid/molybdenum, and selenium/alpha-tocopherol/beta carotene). MAIN OUTCOME MEASURES: Prevalence of nuclear, cortical, and posterior subcapsular cataracts in treatment groups at end of trials. RESULTS: In the first trial, there was a statistically significant 36% reduction in the prevalence of nuclear cataract for persons aged 65 to 74 years who received the supplements. In the second trial, the prevalence of nuclear cataract was significantly lower in persons receiving riboflavin/niacin compared with persons not receiving these vitamins. Again, persons in the oldest group, 65 to 74 years, benefited the most (44% reduction in prevalence). No treatment effect was noted for cortical cataract in either trial. Although the number of posterior subcapsular cataracts was very small, there was a statistically significant deleterious effect of treatment with riboflavin/niacin. CONCLUSIONS: Findings from the two trials suggest that vitamin/mineral supplements may decrease the risk of nuclear cataract. Additional research is needed in less nutritionally deprived populations before these findings can be translated into general nutritional recommendations.

Cigarette smoking and the risk of development of lens opacities. The Framingham studies.

OBJECTIVE: To examine the association between cigarette smoking and the incidence of nuclear and non-nuclear lens opacities in members of the Framingham Eye Study Cohort. PARTICIPANTS AND METHODS: Eye examinations were conducted on surviving members of the Framingham Heart Study Cohort from 1973 to 1975 (Framingham Eye Study I) and again from 1986 to 1989 (Framingham Eye Study II). Smoking data, collected biennially since 1948 in the Heart Study, were used to examine the relationship between cigarette smoking and the incidence of lens opacities. Two thousand six hundred seventy-five persons were examined in the Framingham Eye Study I. Our analysis included 660 persons, aged 52 to 80 years, who were free of lens opacities at the first eye examination. RESULTS: During the approximately 12.5 years between eye examinations, lens opacities developed in a total of 381 persons, with nuclear opacities constituting the most frequent type. In logistic regression analyses that controlled for age, sex, education, and diabetes, a significant positive association with increasing duration of smoking and number of cigarettes smoked daily was found for nuclear lens opacities, alone or in combination (test for trend, P < or = .002), but not for nonnuclear opacities (test for trend, P = .62). Among the heavier smokers (persons who smoked > or = 20 cigarettes per day according to 6 or more biennial Framingham Heart Study examinations), 77% were still smoking at the time of the first eye examination. Persons who smoked 20 or more cigarettes per day at the time of the first eye examination were at substantially increased risk for the development of nuclear opacities than nonsmokers (odds ratio, 2.84; 95% confidence interval, 1.46-5.51). There was no apparent excess risk for persons with nonnuclear lens opacities (odds ratio, 1.42; 95% confidence interval, 0.65-3.07). CONCLUSION: This study provides further evidence that cigarette smokers have an increased risk of developing nuclear lens opacities. The risk was greatest for heavier smokers, who tended to be current smokers and who smoked more cigarettes and for a longer duration.

India-US case-control study of age-related cataracts. India-US Case-Control Study Group.

In a hospital-based case-control study of 1441 patients with age-related cataracts and 549 controls, we studied associations between types of cataract--nuclear, cortical, posterior subcapsular, and mixed--and a number of physiologic, behavioral, environmental, and biochemical variables. Using polychotomous logistic regression analysis, we found an increased risk of cataract with lower educational achievement (all types of cataract), decreased cloud cover at place of residence (all types), use of aspirin less than once a month (posterior subcapsular and mixed), diets low in selected nutrients (posterior subcapsular, nuclear, and mixed), higher blood pressure (nuclear and mixed), lower body mass index (nuclear and mixed), use of cheaper cooking fuels (cortical, nuclear, and mixed), and lower levels of an antioxidant index based on red blood cell levels of glutathione peroxidase and glucose-6-phosphate dehydrogenase and plasma levels of ascorbic acid and vitamin E (posterior subcapsular and mixed). All risks cited were significantly different from those for the other cataract types, a finding that emphasizes the need to investigate the epidemiology of specific types of cataract.

The Framingham eye study: introduction to the monograph.

This article briefly summarizes the monography, The Framingham Eye Study Monograph: an Ophthalmological and Epidemiological Study of Cataract, Glaucoma, Diabetic Retinopathy, Macular Degeneration, and Visual Acuity in a General Population of 2631 Adults, 1973-1975. This 275-page monograph, authored by H. Liebowitz, D. Krueger, C. Maunder et al, has recently been published by Survey of Ophthalmology (Surv Ophthalmol 24 (suppl):335-610, 1980). The purpose of the study, undertaken by the National Eye Institute, was to provide a description of the prevalence and severity of four diseases that are believed to be the major causes of severe visual handicap among adults in the United States. The monography presents the detailed protocols and record forms, definitions of the specific abnormalities and characteristics, criteria for suspicion and diagnosis of disease, detailed tables of the basic data, evaluation of quality of the data, and discussion of selected findings.

The Framingham Eye Study monograph: An ophthalmological and epidemiological study of cataract, glaucoma, diabetic retinopathy, macular degeneration, and visual acuity in a general population of 2631 adults, 1973-1975.

Ophthalmologic examinations for cataract, glaucoma, diabetic retinopathy, macular degeneration and visual acuity were performed on 2631 of the 3977 members of the Framingham (Massachusetts) Heart Study population still living in 1973-1975. The subjects ranged in age from 52 to 85 years. This monograph presents the detailed protocols and record forms for screening and diagnostic examinations, definitions of the specific abnormalities and characteristics used to screen for each disease, criteria for suspicion and diagnosis of diseases, detailed tables of the basic data from the study, evaluation of quality of the data, and discussion of selected findings. The tables provide data on the number and proportion of persons and of eyes with each type of abnormality and each disease, by age and sex. Where appropriate, the data are further classified by location of abnormality, severity, bilaterality and associated visual acuity limitation. The study was sponsored by the National Eye Institute.

Activity of vertebrate gonadotropin-releasing hormones and analogs with variant amino acid residues in positions 5, 7 and 8 in the goldfish pituitary.

All non-mammalian vertebrates as well as marsupial mammals have two or more forms of gonadotropin-releasing hormone (GnRH) in the brain. Goldfish brain and pituitary contains two molecular forms of GnRH, salmon GnRH ([Trp7, Leu8]m-GnRH; s-GnRH) and chicken GnRH-II ([His5, Trp7, Tyr8]m-GnRH; cII-GnRH). Both sGnRH and cII-GnRH stimulate gonadotropin (GtH) as well as growth hormone (GH) release from the goldfish pituitary. The purpose of the present study was to study the activity of the five known forms of GnRHs as well as analogs of mammalian GnRH (m-GnRH) with variant amino acid residues in positions 5, 7 and 8 in terms of binding to GnRH receptors, and release of GTH and GH from the perifused fragments of goldfish pituitary in vitro. All five vertebrate GnRH peptides stimulated both GtH and GH release in a dose-dependent manner, although their potencies were very different. cII-GnRH was somewhat more active than s-GnRH in releasing GtH, whereas s-GnRH tended to have a greater potency than cII-GnRH in terms of GH release. Both chicken GnRH-I (cI-GnRH) and lamprey GnRH (l-GnRH) were significantly less potent than mGnRH, s-GnRH and cII-GnRH in releasing GtH and GH. cII-GnRH binds with higher affinity for the high affinity binding sites compared to all other native peptides. The activity of [Trp7]-GnRH was similar to both s-GnRH and cII-GnRH in releasing GtH and GH. Substitution of His5 resulted in a significant decrease in GtH releasing potencies compared to mGnRH, sGnRH and cII-GnRH. [His5]-GnRH also had lower GH releasing potency than mGnRH and sGnRH. Tyr8, His8 and Leu8 substitutions caused significant decreases in GtH releasing potencies compared to mGnRH, s-GnRH and cII-GnRH, but did not cause a significant change in GH releasing potency. The combination of [His5, Trp7]-GnRH had GtH and GH releasing activities similar to m-GnRH, s-GnRH and cII-GnRH.(ABSTRACT TRUNCATED AT 400 WORDS)

Relative letter and position difficulty on visual acuity charts from the Early Treatment Diabetic Retinopathy Study.

Ten Sloan letters were used in the visual acuity charts developed for use in the Early Treatment Diabetic Retinopathy Study. We used the data from the 3,710 Early Treatment Diabetic Retinopathy Study subjects to investigate the relative difficulty of the ten Sloan letters and to evaluate whether the position of a letter on a line affected its relative difficulty. In general, our findings were consistent with those of the previous study. The four letters with curved contours (C, O, S, and D) were more difficult to discern at threshold than the six letters (Z, N, H, V, R, and K) composed of straight lines. Our data demonstrate that under these test conditions, letters at the end of a line are more likely to be read incorrectly than letters at the beginning of the line. This finding indicates that these data are probably not useful for evaluating possible crowding phenomena.