Blinking and superficial punctate keratopathy in patients with diabetes mellitus.

AIM: To evaluate blinking patterns in patients with diabetes mellitus and whether blinking contributes to the formation of superficial punctate keratopathy in diabetic patients. METHODS: We examined 163 patients with type II diabetes mellitus and 76 without diabetes. Blinks were recorded, analysed using six parameters, and compared between patients with and without diabetes. Multivariate regression analysis was performed to assess the influence of other ocular factors, such as status of tear lipid layer, tear breakup time, corneal sensitivity, the result of cotton thread test, or blinking rate related to superficial punctate keratopathy. RESULTS: In patients with diabetes, the average mean and maximum interblinking times were longer, the average coefficient of variation of interblinking time was higher, and the average blinking rates were lower than those in patients without diabetes. Multivariate regression analysis revealed that the status of tear lipid layer and tear breakup time were significantly relevant to superficial punctate keratopathy (P < 0.01). CONCLUSION: Interblinking time was longer in diabetic patients, resulting in a decreased blinking rate. The prevalence of superficial punctate keratopathy cannot be predicted from blinking patterns in patients with diabetes.

Abnormal blinking of newscasters.

When compared with a control group, blink rates of Japanese TV newscasters were increased, suggesting a possible reaction to reading the news under stressful studio conditions. Frequent blinking may be distractive to the viewing audience.

Efficacy and safety of infrared warming of the eyelids.

PURPOSE: To evaluate our newly developed infrared heater (IRH) and compare it to a broad-spectrum heater (BSH) for warming the eyelids. MATERIALS AND METHODS: Ten normal subjects were enrolled in this study. All measurements were recorded in a room with temperature 23 degrees C, 40% humidity, and no wind. The IRH is composed of two hard eye patches that have light-emitting diodes (LEDs) emitting near-infrared radiation. We first compared the temperature rises in the cornea, lacrimal gland, and eyelids after warming through closed eyelids with the IRH for 5 and 10 min. Next, we compared warming with the IRH or BSH for 30 min. We then used the IRH for 5 min with the eyes open to confirm its safety. Finally, we determined subjective feeling after warming the eyes. RESULTS: Direct comparison of 5 versus 10 min of warming with the IRH showed no significant differences in temperature rises in the upper eyelid (p = 0.09). The IRH caused significantly more heating (p < 0.05) than did the BSH everywhere except the cornea. The temperatures never rose above 37.7 degrees C for either heater during 30 min or with the IRH with the eyes open for 5 min. The subjects' comfort level rose significantly (p < 0.05) after treatment with the IRH. CONCLUSIONS: Our study showed the efficacy and safety of warming the eyelids with a newly developed IRH. Only 5 min is necessary to increase ocular temperature and enhance comfort.

Quantitative videographic analysis of blinking in normal subjects and patients with dry eye.

OBJECTIVE: To study patterns of eye blinking in normal subjects and patients with dry eye. METHODS: We developed an automated, noninvasive blink monitor that permits quantitative analysis of 6 parameters of blinking. We used this method under normal conditions and then examined the effects on the patterns of blinking in patients with dry eye; several steps in this method were designed to exacerbate or ameliorate ocular surface desiccation. RESULTS: The mean (+/-SD), maximum, and coefficient of variation of the interblinking time in normal subjects and patients with dry eye were 4.0 +/- 2.0 and 1.5 +/- 0.9 seconds, 8.9 +/- 4.0 and 4.2 +/- 2.4 seconds, and 55% +/- 21% and 65% +/- 24%, respectively. Those values for the blinking time were 0.20 +/- 0.04 and 0.27 +/- 0.16 seconds, 0.35 +/- 0.12 and 0.99 +/- 1.30 seconds, and 23% +/- 9% and 46% +/- 34%, respectively. The use of artificial tears or spectacles with moist panels and moist inserts tended to normalize the patterns of blinking in the patients with dry eye, whereas exposure to wind made them more abnormal. CONCLUSIONS: Our technique permitted a rigorous analysis of blinking that was previously unavailable. We have shown that local ocular surface conditions alone can significantly affect patterns of blinking. This method should be applicable to studying psychologic and any other factors that may influence blinking.

[Treatment of a refractory chronic osteomyelitis of the jaw].

Two cases of refractory chronic osteomyelitis of the jaw were presented, the first in a patient with pycnodysostosis, the second that had been receiving radiotherapy. Both cases were primary chronic osteomyelitis with a markedly protracted course of recovery, in spite of the antibiotic and local irrigation therapy. However the lesion was cured by the removal of sequesters and granulation tissues without radical treatment such as a decorticotomy. Radical surgery, decortication, is an effective procedure in the treatment of chronic osteomyelitis of the jaw, but treatment with the surgical procedures which remove only the necrotic bones and granulation tissues followed by antibiotic therapy is also possible.

Novel formation of alpha-amino acids from alpha-oxo acids and ammonia in an aqueous medium.

In the course of a study of possible mechanisms for chemical evolution in the primeval sea, we found the novel formation of alpha-amino acids and N-acylamino acids from alpha-oxo acids and ammonia in an aqueous medium. Glyoxylic acid reacted with ammonia to form N-oxalylglycine, which gave glycine in a 5-39% yield after hydrolysis with 6N HC1. Pyruvic acid and ammonia reacted to give N-acetylalanine, which formed alanine in a 3-7% overall yield upon hydrolysis. The pH optima in these reactions were between pH 3 and 4. These reactions were further extended to the formation of other amino acids. Glutamic acid, phenylalanine and alanine were formed from alpha-ketoglutaric acid, phenylpyruvic acid and oxaloacetic acid, respectively, under similar conditions. N-Succinylglutamic acid was obtained as an intermediate in glutamic acid synthesis. Phenylacetylphenylalanineamide was also isolated as an intermediate in phenylalanine synthesis. Alanine, rather than aspartic acid, was produced from oxaloacetic acid. These reactions provide a novel route for the prebiotic synthesis of amino acids. A mechanism for the reactions will be proposed.

A novel way for the formation of alpha-amino acids and their derivatives in an aqueous medium.

In the course of a study of possible mechanism for chemical evolution in the primeval sea, we observed the formation of alpha-amino acids and N-acylamino acids from alpha-oxo acids and ammonia in an aqueous medium. Glyoxylic acid reacted with ammonia to form N-oxalylglycine, which gave glycine in a 5-39% yield after hydrolysis with 6N HCl. Similarly when glyoxylic acid was treated with methylamine it yielded N-oxalylsarcosine, which could be hydrolyzed to sarcosine with 17-25% overall yield upon hydrolysis. Pyruvic acid and ammonia reacted to give N-acetylalanine, which formed alanine in a 3-7% overall yield upon hydrolysis. The pH optima in these reactions were pH 3-4. These reactions were further extended to the formation of other amino acids. Glutamic acid, phenylalanine and serine were formed from alpha-ketoglutaric acid, phenylpyruvic acid and hydroxypyruvic acid, respectively, under similar conditions. N-Succinylglutamic acid was obtained as an intermediate for glutamic acid synthesis. Phenylacetylphenylalanineamide was also isolated as an intermediate for phenylalanine synthesis. Alanine, rather than aspartic acid, was produced from oxaloacetic acid. These reactions provide a novel route for the prebiotic synthesis of amino acids. A mechanism for the reactions is proposed.

Novel formation of alpha-amino acids and their derivatives from oxo acids and ammonia in an aqueous medium.

In the course of a study of chemical evolution of the primeval sea, a novel reaction of carbonyl compounds with ammonia was found. Glyoxylic acid reacted with ammonia to afford N-oxalylglycine, which gave glycine in a 3-20% yield after 6 N HCl hydrolysis. Similarly, glyoxylic acid was treated with methylamine to give N-oxalylsarcosine, which afforded sarcosine in a 9-12% yield upon hydrolysis. Pyruvic acid reacted with ammonia to give N-acetylalanine, which gave alanine in a 1-4% yield upon hydrolysis. These reactions provide a novel and facile route to alpha-amino acids and their derivatives. A mechanism for the reactions is proposed.

DNase A, a poly(dA) and poly(dT)-specific deoxyribonuclease from Achatina fulica. Purification and characterization.

DNases A1 and A2 have been purified to homogeneity from the hepatopancreas of Achatina fulica by a series of steps: acetate buffer extraction, ammonium sulfate precipitation and column chromatography on hydroxylapatite, phosphocellulose, Blue-Sepharose, and poly(A)-Sepharose. The purified enzymes are free of acidic phosphomonoesterase, phosphodiesterase, and RNase activities. They are slightly acidic glycoproteins with identical isoelectric point (6.90). On 0.1% SDS gel electrophoresis, DNase A2 had a molecular weight of 30,000 when dissolved in 1% SDS, but it had molecular weights of 17,500, 8,000, and 4,800 when dissolved in 1% SDS and 1% 2-mercaptoethanol. This was evidence that the enzyme consists of three different subunits joined by interchain disulfide bonds. DNases A1 and A2 are endonucleases working at acidic pH (3.5--6.0) and do not require divalent cations for their activities. The enzymes degrade poly(dA) 5 times faster and poly(dT) 3 times faster than heat-denatured DNA under optimal conditions but do not appreciably digest poly(dG) and poly(dC). We developed an analytical procedure for oligodeoxynucleotides by high-performance liquid chromatography. The phosphomonoester end group and the mode of degradation were examined by the method. The termini produced by the enzymes have 3'-phosphoryl and 5'-hydroxy end groups. The products of exhaustive hydrolysis contain di-, tri-, tetra-, and pentanucleotides and mononucleotide was barely detected. The hydrolyzing activities of DNases A1 and A2 are stimulated by polyamines such as spermine, spermidine, and putrescine, but are inhibited by synthetic polynucleotides and various drugs. Adenosine deaminase highly active on oligoadenylic acids was found in a crude DNase A fraction. The enzyme preparation has higher activity on 3'-adenylic acid than on 5'-adenylic acid. The first adenosine residue of oligoadenylic acids was deaminated considerably more rapidly than the second or succeeding ones.

DNase A, a poly(dA) and poly(dT)-specific deoxyribonuclease from Achatina fulica. Further investigation on the specificity.

The degradation of heat-denatured and native calf thymus DNAs, poly(dA-dT), poly(dA-dC) . poly(dG-dT), and poly(dG-dC) by DNase A has been investigated with the main aim of providing background information for studying the specificity of the enzyme. The specificity of the DNase A was studied by determining the base compositions of 5'- and 3'-terminal nucleotides of oligonucleotides released by the enzyme. The 5'- and 3'-terminal nucleotide compositions were found to vary in the average chain length (Pn) range of 45 to 7 for degradations of heat-denatured calf thymus DNA at pH 4.0, 4.5, 5.0, and 5.5. When the heat-denatured DNA was digested at the optimal pH (pH 5.0), at the Pn = 45 dAdo was predominant (39%) and dThd was minor (9%) in 5'-terminals, whereas dThd was predominant (43%) and dAdo was minor (9%) in 3'-terminals. At Pn = 7, dAdo was even more predominant (49%) and there was very little dThd (7%) in 5'-terminals. No preference was seen in 3'-terminals. This finding indicates that change in the specificity takes place during digestion. The compositions of 5'- and 3'-terminal nucleotides of oligonucleotides released from poly(dA-dT) by exhaustive digestion of the enzyme showed pronounced preferences for dAdo in 5'-terminals (81%) and dThd in 3'-terminals (78%). The hexamer or above deoxyadenylate and the nonamer or above of thymidylate were good substrates of the enzyme. It can be concluded that DNase A is a novel cluster-specific DNase which recognizes and cleaves sequences of oligodeoxyadenylate and/or sequences of oligothymidylate.

3',5'-Cyclic nucleotide phosphodiesterase activity of the sulphatase A of ox liver.

The sulphatase A (aryl-sulphate sulphohydrolase, EC 3.1.6.1) of ox liver hydrolyses adenosine 3',5'-monophosphate (cyclic AMP) to adenosine 5'-phosphate at an optimum pH of approx. 4.3, close that for the hydrolysis of cerebroside sulphate, a physiological substrate for sulphatase A. The Km is 11.6 mM for cyclic AMP. On polyacrylamide gel electrophoresis sulphatase A migrates as a single protein band which coincides with both the arylsulphatase and phosphodiesterase activities, suggesting that these are due to a single protein. Cyclic AMP competitively inhibits the arylsulphatase activity of sulphatase A, showing that both activities are associated with a single active site on the enzyme. sulphatase A also hydrolyses guanosine 3',5'-monophosphate, but not uridine 3',5'-monophosphate nor adenosine 2',3'-monophosphate.

Genesis of amino acids in the primeval sea. Formation of amino acids from sugars and ammonia in a modified sea medium.

Sugars such as glycolaldehyde, glyceraldehyde, erythrose, ribose, and glucose were heated with ammonia in a modified sea medium. Amino acids such as glycine, alanine, serine, threonine, aspartic acid, and glutamic acid were obtained from the reaction mixture. This provides the basis for a model of the genesis of amino acids in the primeval sea.

Polymers produced by heating an amino acid mixture in sea water enriched with transition elements.

A mixture of eighteen protein amino acids was heated in sea water medium enriched with transition metal ions. Small granules were obtained as precipitates. Both dialyzable polymers and undialyzable polymers were obtained from the supernatant. Dialyzable polymers yielded mainly Glu, Asp, Ser, and Thr on hydrolysis; undialyzable polymers (C, 29.45; H, 3.87; N, 4.87; and ash, 31.5 wt%) yielded Thr, Asp, Glu, Gly, Leu, Ser, Lys, Pro, His, Phe, and a few unidentified ninhydrin positive peaks after acid hydrolysis. Five wt% of the undialyzable polymers con-ist of acid-hydrolyzable protein amino acids.

Formation of molecules of biological interest from formaldehyde and hydroxylamine in a modified sea medium.

As reported previously, glycine, serine, aspartic acid, and beta-alanine are the predominant amino acids produced from equimolecular formaldehyde and hydroxylamine in a modified sea medium enriched with essential transition metal ions. The time course of formation of the amino acids, and related substances was studied. Among the amino acids, glycine was produced earlier. Urea was produced initially and disappeared rapidly. Formation of cyanide, glycinenitrile, and glycinamide preceded the formation of amino acids. Probable pathways for the formation of amino acids and urea are suggested.