Cross-sectional study of the factors associated with the number of teeth in middle-aged and older persons with intellectual disabilities.

BACKGROUND: We investigated the number of teeth in middle-aged and older individuals with intellectual disability and evaluated the related factors. METHODS: A questionnaire survey on oral health was administered to 604 persons over 40 years old with intellectual disabilities in Aichi Prefecture, Japan. The questionnaire designed for this study included items on diagnosis of disability, severity of intellectual disability and type of residence. As oral-health-related factors, the proxy-reported number of teeth, family dentist, regular dental check-ups, problems during dental treatment and tooth-brushing frequency were evaluated. A multinomial logistic regression analysis was performed using the proxy-reported number of teeth (≥20, 10-19 or 0-9) as the dependent variable, and the odds ratios (ORs) and 95% confidence intervals (CIs) of independent variables were calculated. In addition, linear regression analysis was performed using the proxy-reported number of teeth as the dependent variable. RESULTS: In the multivariate multinomial logistic regression analysis, age, diagnosis of disability and type of residence were significantly associated with having 10-19 versus ≥20 teeth. Participants with autism spectrum disorder had a significantly lower OR for 10-19 teeth compared with those with intellectual disability [OR (95% CI): 0.49 (0.22-1.08)]. Age, diagnosis of disability, regular dental check-ups and tooth-brushing frequency were significantly associated with having 0-9 versus ≥20teeth. Participants with Down syndrome had a significantly higher OR for 0-9 teeth compared with those with intellectual disability [OR (95% CI): 3.17 (1.09-9.23)]. The ORs for 0-9 teeth of participants who did not attend regular dental check-ups and who brushed their teeth 1 time/day compared with ≥3 times/day were significantly high, and the OR (95% CI) was 2.37 (1.06-5.30) and 4.76 (1.09-20.77), respectively. [Corrections made on 22 August 2022, after first online publication: in the previous sentence, "who attend" has been changed to "who did not attend".] In the multivariate linear regression analysis, age, diagnosis of disability and regular dental check-ups were significantly associated with the proxy-reported number of teeth. The proxy-reported number of teeth was -0.42 less with each 1-year increase in age. With autism spectrum disorder, the proxy-reported number of teeth was 0.74 more compared with intellectual disability. In Down syndrome, the proxy-reported number of teeth was -0.93 less compared with intellectual disability. The proxy-reported number of teeth was -2.12 less for those who did not have regular dental check-ups. CONCLUSIONS: The number of teeth in middle-aged and older individuals with intellectual disability was related to age and the type of disability. Regular dental visits may be effective at preventing tooth loss in middle-aged and older persons with intellectual disability.

Erythrocyte glucose-6-phosphate dehydrogenase from Brazilian opossum Didelphis marsupialis.

In a comparative study of erythrocyte metabolism of vertebrates, the specific activity of glucose-6-phosphate dehydrogenase (G6PD) of the Brazilian opossum Didelphis marsupialis in a hemolysate was shown to be high, 207 +/- 38 IU g-1 Hb-1 min-1 at 37 degrees C, compared to the human erythrocyte activity of 12 +/- 2 IU g-1 Hb-1 min-1 at 37 degrees C. The apparent high specific activity of the mixture led us to investigate the physicochemical properties of the opossum enzyme. We report that reduced glutathione (GSH) in the erythrocytes was only 50% higher than in human erythrocytes, a value lower than expected from the high G6PD activity since GSH is maintained in a reduced state by G6PD activity. The molecular mass, determined by G-200 Sephadex column chromatography at pH 8.0, was 265 kDa, which is essentially the same as that of human G6PD (260 kDa). The Michaelis-Menten constants (Km: 55 microM) for glucose-6-phosphate and nicotinamide adenine dinucleotide phosphate (Km: 3.3 microM) were similar to those of the human enzyme (Km: 50-70 and Km: 2.9-4.4, respectively). A 450-fold purification of the opossum enzyme was achieved and the specific activity of the purified enzyme, 90 IU/mg protein, was actually lower than the 150 IU/mg protein observed for human G6PD. We conclude that G6PD after purification from the hemolysate of D. marsupialis does not have a high specific activity. Thus, it is quite probable that the red cell hyperactivity reported may be explained by increased synthesis of G6PD molecules per unit of hemoglobin or to reduced inactivation in the RBC hemolysate.

Erythrocyte glucose-6-phosphate dehydrogenase from Brazilian opossum Didelphis marsupialis

In a comparative study of erythrocyte metabolism of vertebrates, the specific activity of glucose-6-phosphate dehydrogenase (G6PD) of the Brazilian opossum Didelphis marsupialis in a hemolysate was shown to be high, 207 ± 38 IU g-1 Hb-1 min-1 at 37°C, compared to the human erythrocyte activity of 12 ± 2 IU g-1 Hb-1 min-1 at 37°C. The apparent high specific activity of the mixture led us to investigate the physicochemical properties of the opossum enzyme. We report that reduced glutathione (GSH) in the erythrocytes was only 50 percent higher than in human erythrocytes, a value lower than expected from the high G6PD activity since GSH is maintained in a reduced state by G6PD activity. The molecular mass, determined by G-200 Sephadex column chromatography at pH 8.0, was 265 kDa, which is essentially the same as that of human G6PD (260 kDa). The Michaelis-Menten constants (Km: 55 æM) for glucose-6-phosphate and nicotinamide adenine dinucleotide phosphate (Km: 3.3 æM) were similar to those of the human enzyme (Km: 50-70 and Km: 2.9-4.4, respectively). A 450-fold purification of the opossum enzyme was achieved and the specific activity of the purified enzyme, 90 IU/mg protein, was actually lower than the 150 IU/mg protein observed for human G6PD. We conclude that G6PD after purification from the hemolysate of D. marsupialis does not have a high specific activity. Thus, it is quite probable that the red cell hyperactivity reported may be explained by increased synthesis of G6PD molecules per unit of hemoglobin or to reduced inactivation in the RBC hemolysate.

Otimização da técnica para coloração de ferro (PERLS)

A coloração de Perls é de grande utilidade para identificar ferro em forma de grânulos no interior dos EritrobLastos (denominados sideroblastos), nos histiócitos e macrófagos, além do ferro intersticial. É muito útil no diagnóstico diferencial de várias patologias hematológicas. Assim, propusemos-nos a modificar algumas fases desta técnica a fim de otimizar a coloração do ferro nas células e interstício. Na primeira fase, o esfregaço sanguíneo que era fixado em vapor de formol por 30 minutos e lavado várias vezes em água, passou a ser fixado por 40 minutos sem lavagem subsequente em água, diminuindo o risco de danos (lise) ao esfregaço. A fase de coloração com ferrocianeto ácido contracoloração com Fast Red Nuclear, aumentamos a concentração...

The calcium-dependent protease of Loxosceles gaucho venom acts preferentially upon red cell band 3 transmembrane protein.

Eighty micrograms red blood cell (RBC) ghosts from patients who had previously exhibited the cutaneous form of loxoscelism (presenting localized dermonecrosis) and the viscerocutaneous form of loxoscelism (presenting dermonecrosis, hemoglobinuria, hematuria, and jaundice) and from controls were incubated with 2.5 microg crude Loxosceles gaucho venom in 5 mM phosphate buffer, pH 7.4, at 37 C. Among all membrane proteins, quantitative proteolysis of the important integral transmembrane protein 3 increased with venom dose and with incubation time from 30 to 120 min, as demonstrated by gel densitometry. Similar quantitative data were obtained for RBC ghosts from patients and from control subjects, a fact that argues against the possibility of genetic factors favoring the hemolytic viscerocutaneous form. These data suggest that the clinical forms may be different types of the same disease, with the viscerocutaneous form being the result of large amounts of intravascularly injected venom and the superficial form being the result of in situ venom action. Since protein 3 is a housekeeping integral membrane protein, whose genetic deficiency leads to hemolytic anemia, it is reasonable to relate it to the hemolysis which occurs in the viscerocutaneous form of loxoscelism. The venom protease responsible for the process was not inhibited after 120-min incubation by 0.2 mM paramethylsulfonyl fluoride or by 0.2 mM N-ethylmaleimide but was inhibited by 25 mM ethylenediaminetetraacetic acid (a calcium-chelating agent) in 5 mM phosphate buffer at pH 7.4, which suggests that the enzyme is a calcium-dependent metalloprotease.

The calcium-dependent protease of Loxosceles gaucho venom acts preferentially upon red cell band 3 transmembrane protein

Eighty micrograms red blood cell (RBC) ghosts from patients who had previously exhibited the cutaneous form of loxoscelism (presenting localized dermonecrosis) and the viscerocutaneous form of loxoscelism (presenting dermonecrosis, hemoglobinuria, hematuria, and jaundice) and from controls were incubated with 2.5 æg crude Loxosceles gaucho venom in 5 mM phosphate buffer, pH 7.4, at 37ºC. Among all membrane proteins, quantitative proteolysis of the important integral transmembrane protein 3 increased with venom dose and with incubation time from 30 to 120 min, as demonstrated by gel densitometry. Similar quantitative data were obtained for RBC ghosts from patients and from control subjects, a fact that argues against the possibility of genetic factors favoring the hemolytic viscerocutaneous form. These data suggest that the clinical forms may be different types of the same disease, with the viscerocutaneous form being the result of large amounts of intravascularly injected venom and the superficial form being the result of in situ venom action. Since protein 3 is a housekeeping integral membrane protein, whose genetic deficiency leads to hemolytic anemia, it is reasonable to relate it to the hemolysis which occurs in the viscerocutaneous form of loxoscelism. The venom protease responsible for the process was not inhibited after 120-min incubation by 0.2 mM paramethylsulfonyl fluoride or by 0.2 mM N-ethylmaleimide but was inhibited by 25 mM ethylenediaminetetraacetic acid (a calcium-chelating agent) in 5 mM phosphate buffer at pH 7.4, which suggests that the enzyme is a calcium-dependent metalloprotease

Increased fetal hemoglobin levels in patients infected with human immunodeficiency virus (HIV1/2)

Fetal hemoglobin was measured in HIV1/2 patients under treatment with combined therapy (zidovudine and a protease inhibitor). A total of 143 patients and 103 normal individuals were investigated by the quantitative method of Betke and the semi-quantitative acid elution method of Kleihauer. In the normal person, hemoglobin F makes up less than 1 percent and an increase higher than 1.5 percent was observed in 21.4 percent of HIV patients by the method of Betke and in 24.8 percent of HIV-infected patients by the method of Kleihauer. The quantitative biochemical method of Betke showed that the populations were significantly different (two-tailed Mann-Whitney test). The reason for this hemoglobin F increase might be ascribed to the effect of zidovudine or to direct viral action on gamma chain expression. The finding of a higher F cell frequency indicated by the method of Kleihauer rather suggests that there is an increased F cell clone proliferation rather than an increase in hemoglobin F level in every cell

Increased fetal hemoglobin levels in patients infected with human immunodeficiency virus (HIV1/2).

Fetal hemoglobin was measured in HIV1/2 patients under treatment with combined therapy (zidovudine and a protease inhibitor). A total of 143 patients and 103 normal individuals were investigated by the quantitative method of Betke and the semi-quantitative acid elution method of Kleihauer. In the normal person, hemoglobin F makes up less than 1% and an increase higher than 1.5% was observed in 21.4% of HIV patients by the method of Betke and in 24.8% of HIV-infected patients by the method of Kleihauer. The quantitative biochemical method of Betke showed that the populations were significantly different (two-tailed Mann-Whitney test). The reason for this hemoglobin F increase might be ascribed to the effect of zidovudine or to direct viral action on gamma chain expression. The finding of a higher F cell frequency indicated by the method of Kleihauer rather suggests that there is an increased F cell clone proliferation rather than an increase in hemoglobin F level in every cell.

Enzymes and membrane proteins of ADSOL-preserved red blood cells.

CONTEXT: The preservative solution ADSOL (adenine, dextrose, sorbitol, sodium chloride and mannitol) maintains red cell viability for blood trans-fusion for 6 weeks. It would be useful to know about its preservation qualities over longer periods. OBJECTIVE: To determine some red cell biochemical parameters for peri-ods of up to 14 weeks in order to determine whether the red cell metabo-lism integrity would justify further studies aiming at increasing red cell preservation and viability. DESIGN: Biochemical evaluation designed to study red cell preservation. SETTING: São Paulo University erythrocyte metabolism referral center. SAMPLE: Six normal blood donors from the University Hospital of the Universidade Federal do Paraná, Curitiba, Brazil. MAIN MEASUREMENTS: Weekly assay of erythrocyte adenosine-5;-triphosphate (ATP), 2, 3-diphosphoglycerate (2,3DPG), hexokinase (HX), phosphofructokinase (PFK), pyruvate kinase (PK), glucose-6-phosphate dehydrogenase (G-6-PD), 6-phosphogluconic dehydrogenase (6-PGD), glyceraldehyde-3-phosphate dehydrogenase (GAPD), glutathione reduc-tase (GR), glutathione peroxidase (GSHPx), plasma sodium and potas-sium, blood pH, and membrane proteins of red cells preserved in ADSOL were studied during storage for 14 weeks storage. RESULTS: During ADSOL preservation, erythrocyte ATP concentration decreased 60% after 5 weeks, and 90% after 10 weeks; the pH fell from 6.8 to 6. 4 by the 14th week. 2,3-DPG concentration was stable during the first week, but fell 90% after 3 weeks and was exhausted after 5 weeks. By the end of the 5th week, an activity decrease of 16-30% for Hx, GAPD, GR, G-6-PD and 6-PGD, 35% for PFK and GSHPx, and 45% for PK were observed. Thereafter, a uniform 10% decay was observed for all enzymes up to the 14th week. The red blood cell membrane pro-teins did not show significant alterations in polyacrylamide gel electro-phoresis (SDS-PAGE) during the 14 weeks. CONCLUSION: Although the blood viability was shown to be poor from the 6th week up to the 14th week of storage due to ATP and 2,3-DPG depletion, the other biochemical parameters remained in fairly good condition for longer storage. As there is a gradual and uniform decay in activity throughout these 14 weeks, it seems that ADSOL-preserved red cells may be used as red cell enzyme standards and membrane proteins as well.

Fecal microflora in a patient with short-bowel syndrome and identification of dominant lactobacilli.

Fecal microflora and lactate concentrations in blood and feces obtained from a patient (a 5 year-old boy) with short-bowel syndrome (SBS) were compared during acidosis to results for the normal condition (no SBS symptoms). The taxonomical position of the lactobacilli found predominantly in the feces sample obtained 2 days before the fifth attack was also studied. The D-lactate level in serum obtained 1 day after the fourth attack was 10-fold higher than that for the normal condition, although there was not a great difference in L-lactate levels. D-Lactate (3.91 mM) and L-lactate (2.86 mM) were also detected in the feces samples collected 2 days before the fifth attack, while no lactate was detected in the feces sample for the normal condition. The counts of total fecal bacteria, especially anaerobic bacteria such as members of the family Bacteroidaceae, were found to be low. The counts of lactobacilli and the total population of lactobacilli relative to total fecal bacteria in the feces 2 days before the fifth attack (40.4%) were extremely high. In this case, a majority of the lactobacilli were D-lactate producers as determined by homolactic fermentation. These lactobacilli were identified as Lactobacillus delbrueckii subsp. lactis. The percentages of bifidobacteria relative to total fecal bacteria in feces samples obtained both 2 days before the fifth attack (50.9%) and for normal condition (61.9%) were also high, although these bacteria were L-lactate producers. In the feces samples for the normal condition, the D-lactate producers decreased to less than 10(9) per g, while the counts of L- or DL-lactate producers were 100-fold higher than the numbers in feces samples obtained 2 days before the fifth attack. These results suggested that an increase in the level of D-lactate producers, such as L. delbrueckii subsp. lactis, in the colon may be associated with the clinical expression of metabolic acidosis.

Correlation of discocyte frequency and ATP concentration in preserved blood. A morphological indicator of red blood cell viability.

Red blood cells (RBC) are viable if kept in an adequate preservative solution, although gradual changes in morphology and metabolism may occur. There is a gradual decrease in adenosine-5'-triphosphate (ATP) concentration, pH, glucose consumption, and enzyme activity during preservation. The normal discocyte shapes are initially replaced by echinocytes and stomatocytes and, at final stages, by spherocytes, the last step before splenic sequestration. Post-transfusional survival has been correlated with the ATP concentration. RBC preserved in ADSOL, a solution containing adenine, dextrose, sodium chloride, and mannitol, are viable for transfusion for up to 6 weeks. Erythrocytes from 10 blood units taken from healthy adult donors were preserved for 12 weeks in ADSOL at 4 degrees C. We now report a significant correlation (r2 = 0.98) between the percentage of discocytes (89 to 7%) and ATP (100 to 10%) concentration in ADSOL-preserved RBC. The results suggest that the percent of discocyte shapes used as an indicator of ATP concentration may be a useful indicator for quality control of RBC viability in centers which have limited assay facilities.

Correlation of discocyte frequency and ATP concentration in preserved blood: a morphological indicator of red cell viability

Red blood cells (RBC) are viable if kept in an adequate preservative solution, although gradual changes in morphology and metabolism may occur. There is a gradual decrease in adenosine-5'-triphosphate (ATP) cocentration, pH, glucose consumption, and enzyme activity during preservation. The normal discocyte shapes are initially replaced by echimocytes and stomatocytes and, at final stages, by spherocytes, the last before eplenic sequestration. Post-transfusional survival has been correlated with the ATP concentration. RBC preserved in ADSOL, a solution containing adenine, dextrose, sodium chloride, and mannitol, are viable for transfusion for up to 6 weeks. Erythrocytes from 10 blood units taken from healthy adult donors were preserved for 12 weeks in ADSOL at 4 graus Celsius. We now report a significant correlation (r2 = 0.98) between the percentage of discocytes (89 to 7 percent) and ATP (100 to 10 percent) concentration in ADSOL-preserved RBC. The results suggest that the percent of discocyte shapes used as an indicator OF ATP concentration may be a useful indicator for quality control of RBC viability in centers which have limited assay facilites.

Vitamin dependent erythrocyte enzymes in newborns in relation to gestational age and birth weight.

In order to search for any difference in riboflavin and pyridoxine nutrition between term appropriate for gestational age (TAGA), term small for gestational age (TSGA) and preterm appropriate for gestational age (PTAGA) newborns, cord blood from 23, 19 and 20 infants, respectively, were studied, and red cell glutathione reductase (riboflavin-dependent) and glutamic oxaloacetic transaminase (pyridoxine-dependent) activities were measured, as well as their respective activity coefficients (GRase AC and GOT AC). Red cell enzymes were assayed according to BEUTLER [4] in a Gilford recording spectrophotometer model 2451 at 37 degrees C. Values of 1.42, 1.39 and 1.32 of GRase AC and values of 1.17, 1.02 and 1.08 GOT AC, respectively were obtained. These data indicate that there is no significant difference between the three categories of newborns with regard to riboflavin and pyridoxine nutrition, and suggest that neither maturity nor adequacy of intrauterine growth were related to differences in the availability of these vitamins. All groups demonstrate a uniform mild deficiency of riboflavin and an adequate level of pyridoxine, probably the effect of maternal diet.

Physiological red cell, 2,3-diphosphoglycerate increase by the sixth hour after birth.

As the available hemoglobin A1 at birth ranges from 20 to 30% a possible mechanism to favor oxygen release to the tissues could be a decrease of hemoglobin A1 affinity to oxygen. This may be accomplished by an increase in blood pH soon after birth and by an elevation in red cell 2,3-diphosphoglycerate (2,3-DPG). This hypothesis is supported by Valleri and Hirsch, who described a rapid 2,3-DPG recovery of transfused depleted 2,3-DPG red cells. That being so, we carried out this current study by assaying the 2,3-DPG of cord blood from 22 newborns and at 6, 24 and 72 hours after birth, as well as those enzymes assumed to be envolved in the 2,3-DPG levels regulation. 2,3-DPG (nmoles g-1 Hb) demonstrated the following values: cord blood: 9,770 +/- 1,026; 6h: 12,773 +/- 1,726; 72 h: 11,990 +/- 728, unveiling a distinct behavior of a sharp increase of 30% by the sixth hour. This confirmed our hypothesis. Regarding the metabolic mechanisms which can account for the 2,3-DPG increase, besides the rise of blood pH, we detected a significant decrease of the 2,3-DPG phosphatase activity, which might diminish the 2,3-DPG breakdown.

Search of intravascular hemolysis in patients with the cutaneous form of loxoscelism.

Haptoglobin assay, a highly sensitive method to detect intravascular hemolysis was carried out in the sera of 19 patients referred to Hospital Vital Brazil with the cutaneous form of loxoscelism in order to investigate the occurrence of mild intravascular hemolysis. Data from this series did not show decreased levels haptoglobin, ruling out intravascular hemolysis in these patients with cutaneous form of loxoscelism.

Alteraçoes bioquimicas durante a preservaçao de eritrocitos in vitro em meio CE com vitamina E / Biochemical changes during red blood cells preservation in CE medium plus vitamin E in vitro

Ja foi demonstrado que o meio CE com vitamina E mantem estaveis durante 100 dias os valçores hematimetricos de eritrocitos parcialmente fixados em glutaraldeido (Leonart,M.S.S. et al. Rev. Bras. Anal. Clin., 21:111, 1989). Neste trabalho, partiu-se de 20 unidades de sangue nenoso de individuos saudaveis e estudou-se o comportamento bioquimico dos eritrocitos durante 12 semanas de preservaçao a 4 C em meio CE com vitamina E (CE), em meio CE com vitamina E apos fixaçao parcial em glutaraldeido (GACE), e em ADSOL (Heaton, A et al. Transfusion, 21:600, 1981). Os valores de adenosina trifosfato (ATP), que se mantiveram acima de 80% dos iniciais durante 2 semanas de estoque, decairam rapidamente, e se exauriram em cerca de 4 semanas, sem diferenças significativas entre a s tres formas de preservaçao. As concentraçoes de 2,3-difosfoglicerato (2,3-DPG) permaneceram acima de 90% das iniciais na 1 semana. Nas amostras em ADSOL, os valores de 2,3-DPG foram significativamente mais baixos entre 14 e 28 dias (p< 0,05) e se exauriram ao final da 3 semana; nas CE e GACE, ao final da 5 semana. Observou-se decaimento na concentraçao extracelular de ions sodio e aumento na de ions potassio em 12 semanas de preservaçao. A entrada de sodio e a saida de potassio ocorreram mais lentamente nos eritrocitos preservados em ADSOL que em meio CE (p<0.05). A percentagem de hemolise observada durante o periodo de preservaçao foi significativamente diferente entre os tres grupos (p<0,05). Ao final de 12 semanas, ocorreu menos de 1% de hemolise em GACE, cerca de 2% em CE, e cerca de 7% em ADSOL. Concluiu-se que eritrocitos humanos conservados em meio CE mais vitaminas E, com ou sem fixaçao parcial em glutaraldeido, mantem o sistema de transporte de sodio e potassio e o metabolismo ativo de consumo de ATP e 2,3-DPG, e baixo grau de hemolise durante 12 semanas; portanto, adequados ao uso em laboratorio

X-linked syndrome: mental retardation, hip luxation, and G6PD variant [Gd(+) Butantan].

An apparently new X-linked syndrome is presented. It occurred in four male first cousins. The main manifestations of this syndrome are severe mental retardation, bilateral congenital hip luxation, and short stature. Three of the affected males showed a new glucose-6-phosphate dehydrogenase variant.