A light and scanning electron microscopic study of enamel decalcification in children living in a water-fluoridated area.

Eleven children, each having one or two pairs of premolars to be extracted for orthodontic purposes, participated in the study. The model involved placement of a special orthodontic band that allowed the accumulation of plaque in a defined area between the band and the buccal enamel. Examination of enamel changes was carried out in experimental teeth that had been exposed to local plaque accumulation for one, two, four, eight or 14 days. The specimens were examined under the light (LM) and the scanning electron microscope (SEM). All teeth had signs of very mild dental fluorosis. No indications of demineralization were noted after one day. SEM examination showed signs of crystal dissolution in some of the two-day specimens. Six of eight four-day specimens exhibited surface dissolution. All eight- and 14-day specimens showed signs of surface demineralization in the LM as well as in the SEM. These observations documented that undisturbed bacterial deposits are capable of initiating enamel demineralization within short time periods, even in children living in a water-fluoridated area.

The microflora associated with the development of initial enamel decalcification below orthodontic bands in vivo in children living in a fluoridated-water area.

Thirty-four caries-free teeth destined for orthodontic extraction were banded to provide a protected area for the accumulation of plaque. The teeth were extracted at one, two, four, eight, and 14 days after being banded, and samples of the flora below the band were analyzed for the presence of Streptococcus sanguis, Streptococcus 'mitior', 'mutans streptococci', Actinomyces viscosus, Actinomyces naeslundii, Actinomyces odontolyticus, Lactobacillus species, and Veillonella. After plaque sampling, the teeth were sent to the Royal Dental College (Copenhagen) for histological analysis. The results showed that S. mutans could colonize the area below the band after one day, but that colonization was only 100% at 14 days. Lactobacillus was only isolated from 2/8 samples at four days and from 4/8 samples at 14 days. S. sanguis and 'S. mitior' were regularly isolated at all banding times, and Veillonella was isolated from all samples. A. viscosus was the most commonly isolated Actinomyces. The numbers of Streptococcus and Veillonella were significantly higher at day 1 than at day 14 (p less than 0.05). S. mutans and A. viscosus were isolated more frequently at day 14 than at day 1 (p less than 0.01). Histological examination revealed that dissolution of the enamel below a band could occur after two days, but that even after 14 days dissolution could be questionable. Dissolution was detected in areas where S. mutans was not isolated (8/34), but S. mutans was also present in samples showing dissolution (12/34). There was no relationship between dissolution and the numbers of S. mutans; however, the isolation frequency of S. mutans was associated with dissolution (p less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Influence of pH and fluoride on properties of an oral strain of Lactobacillus casei grown in continuous culture.

A freshly isolated oral strain, Lactobacillus casei RB1014, was grown in continuous culture to compare the effects of pH and fluoride on growth and metabolism. The cells were grown at pH 7.0 to 3.2 in the absence of fluoride and from pH 7.0 to 5.4 with 20 mM NaF. Cell numbers varied from 3 X 10(9) to 30 X 10(9)/ml on blood agar during alterations in the growth pH from 7.0 to 4.27. Only when the culture was stressed by lowering the pH to 3.2 were cell numbers drastically reduced. Cells growing at pH 7.0 without fluoride were unable to grow when plated on fluoride agar (10.5 mM) at pH 5.5; however, when the growth pH was allowed to decrease to 4.94, cells grew on the fluoride plates in numbers equal to those growing on blood agar. This fluoride tolerance trait appeared rapidly once pH control was removed and was lost when the culture was returned to pH 7.0. The addition of 20 mM NaF to the culture medium did not adversely affect growth, provided that the pH was maintained at 6.0 or above; cells tolerant to 10.5 and 16 mM NaF appeared on pH 5.5 plates during this phase. In cells removed from the chemostat throughout the experiment and incubated at the pH of growth in a pH stat, glycolytic activity was optimum at pH 5.5 in the absence of NaF. Fluoride stimulated glycolytic activity by cells incubated at pH 7.0 and by cells growing with 20 mM NaF, provided that the pH of growth remained at or above 6.0. A more detailed examination of the adaptation to fluoride tolerance during shifts to acidic pH values revealed that cells capable of growth on acidic fluoride agar plates appeared within 2 h of the start of the fall in pH of the chemostat culture. Estimation of the intracellular pH during the period of the initial pH fall revealed that the intracellular pH was identical to the extracellular pH (i.e., no pH gradient [delta pH]), indicating that fluoride would not be transported into the cells to inhibit metabolism. However, once the pH of the medium was stabilized, delta pHs were generated, with the delta pH increasing as the pH declined. The inhibition of glycolysis by fluoride increased in proportion to the delta pH. Cells grown at pH 5.5 generated larger delta pHs than did cells grown at pH 7.0, although the values were normally small (approximately 0.9 U). The data suggest that the inherent fluoride tolerance of L. casei RB1014 was associated with relatively small delta pHs.

The 24-hour secretory pattern of LH and the response to LHRH in transsexual men.

Ten separate aspects of hypothalamic and pituitary function were studied in 13 male-to-female transsexuals and compared to the results of 7 heterosexual adult men. In 4 of 5 transsexuals, the 24-hour mean serum concentration of LH, the LH pulse frequency or amplitude, or the apparent half-life of disappearance of serum LH were greater than the 95% confidence limit of normal men. The maximum concentration of LH or FSH following the administration of 100 micrograms LHRH, the area under the response curve of LH or FSH following LHRH, or both were significantly greater than normal in 5 of 13 male-to-female transsexuals. The response of LH following the administration of LHRH was repeated in 3 subjects during estrogen therapy, and in one there was a paradoxical increase in the response of LH. Transsexualism may be associated with a neuroendocrine defect in the hypothalamus or pituitary that is characterized by high-frequency, high-amplitude pulsatile secretion of pituitary LH.

Anterior-pituitary adrenal function of gold-treated patients with juvenile rheumatoid arthritis.

The hypothesis that gold therapy enhances endogenous cortisol secretion was tested in juvenile rheumatoid arthritis patients. The secretion of cortisol, dehydroepiandrosterone-sulfate (DHEA-S) and several peptide hormones was determined from plasma samples collected every 20 min for 24 hours. In random patients cortisol secretion was significantly greater in gold treated patients than in similar patients not receiving gold. However, when untreated patients were restudied after initiation of gold therapy, there was no significant change in cortisol secretion. Gold therapy also did not significantly alter secretion of the peptide hormones or DHEA-S. Thus, gold does not appear to influence endogenous hormone secretion.

Daily rhythms in cortisol and melatonin in primate cerebrospinal fluid. Effects of constant light and dark.

Cerebrospinal fluid was continuously collected from the cisternal-cervical subarachnoid space of chair-restrained rhesus monkeys. The concentrations of melatonin and cortisol were measured in the cerebrospinal fluid. Under diurnal lighting (light:dark, 12:12 h) melatonin concentrations were elevated during darkness and low during illumination. The melatonin rhythm persisted in constant darkness but was suppressed in constant illumination. Under diurnal lighting, cortisol concentrations were elevated in the early portion of the light period. This daily rhythmicicty of cortisol secretion was not altered by constant illumination or constant darkness. The differential response of the two hormones to constant light suggest that the daily fluctuation of melatonin secretion was not responsible for the daily rhythmicity of cortisol secretion in the rhesus monkey.

Gonadal function in trisomy 21.

Endocrinologic evaluation of 39 patients with trisomy 2 and associated hypogonadism demonstrated elevations of follicle-stimulating hormone and luteinizing hormone; consequently, it can be concluded that both germinal and Leydig cell function are affected. A negative correlation between testicular size and plasma follicle-stimulating hormone was documented. Plasma testosterone levels were found to be normal in male patients as were estradiol levels in female patients with trisomy 21. On the basis of these findings, the simplest and most practical diagnostic approach to evaluate germinal cell function appears to be a single plasma follicle-stimulating hormone determination supplemented by an accurate measurement of testicular volume in males.

Effects of clonidine on 24-hour hormonal secretory patterns, cardiovascular hemodynamics, and central nervous function in hypertensive adolescents.

To assess the potential of antihypertensive drugs for interference with somatic growth and sexual development in hypertensive children, the effect of clonidine therapy on various endocrine, cardiovascular, and neuromuscular functions has been examined in five male adolescents with idiopathic hypertension. In studies done before and at the end of 4 weeks of twice-daily clonidine therapy, in an average daily dose of 0.31 mg, no significant effects were noted in the secretory patterns of growth hormone, luteinizing hormone, follicle-stimulating hormone, prolactin, cortisol, aldosterone, or testosterone, measured in blood obtained every 20 minutes for 24 hours. In blood obtained while the patients were supine and then erect, plasma renin activity and norepinephrine levels were significantly lowered after clonidine therapy. Cardiovascular responses to dynamic exercise were little altered beyond a 17% decrease in maximal oxygen consumption. The performance of fine motor skills was minimally altered. These data provide preliminary evidence that clonidine, an antihypertensive drug that affects the adrenergic nervous system, may not interfere with normal growth and maturation in adolescent males.

Circadian cortisol secretory rhythms in Cushing's disease.

Plasma cortisol was measured at 20-min intervals for 24 h in eight patients with Cushing's disease and ACTH-secreting pituitary tumors. The 24-h mean (+/- SD) cortisol level was 25.6 +/- 11.3 microgram/dl (range, 15.5--40.6), which was significantly higher than the level of normal control subjects (P less than 0.01). The 24-h mean ACTH level varied between 22--107 pg/ml, with a mean +/- SD of 63.4 +/- 27.2. The mean ACTH level was higher than that of the control subjects but the difference was not statistically significant. The 24-h cortisol secretory pattern was characterized by an absence of the normal circadian variation and a failure of the plasma cortisol level to fall to less than 2 microgram/dl between 2300--0300 h. The coefficient of variation, an expression of the amplitude of cortisol secretory episodes, was significantly decreased in patients with Cushing's disease compared to normal control subjects; there was no significant difference in the number of cortisol secretory episodes in the patients vs. control subjects. Three of the patients were restudied after successful resection of their ACTH-secreting pituitary tumors. Two showed normalization of their 24-h circadian cortisol patterns and normal metyrapone responses. In the third, the 24-h mean cortisol level was normal, but the circadian cortisol rhythm remained abnormal. This patient had diminished ACTH reserve, demonstrated by a subnormal response to metyrapone. Additional studies will be required to determine if normalization of the circadian cortisol rhythm occurs in all patients with Cushing's disease who are cured after transsphenoidal microsurgery and who also show normal ACTH reserve.

[Melatonin and LH secretion patterns in pubertal boys (author's transl)]. / Dosage radioimmunologique de la mélatonine au cours du nycthémère chez le garçon pubère.

Plasma melatonin and LH were measured at 20 minute intervals for 24 hours in four normal pubertal boys. All four subjects showed a significant augmentation of LH and melatonin during noctural sleep. There was also a significant correlation between the LH and melatonin levels (p less than 0.001). There were periods of episodic secretion of melanin during the diurnal waking period which seemed related to "stress". These data indicate that the peripheral concentrations of melatonin which occur during sleep are insufficient to prevent spontaneous LH secretion during puberty.

LH and melatonin secretion patterns in pubertal boys.

Four normal pubertal boys had plasma LH and melatonin measured at 20-minute intervals for 24-hours. All four subjects showed a significant augmentation of LH and melatonin during nocturnal sleep. There was also a significant correlation between the LH and melatonin levels (P less than 0.001). These data indicate that the peripheral concentrations of melatonin which occur during sleep are insufficient to prevent spontaneous LH secretion during puberty.

Luteinizing hormone and follicle-stimulating hormone secretory dynamics in Turner's syndrome.

In eight teenage patients with Turner's syndrome, LH and FSH were measured at 20-min intervals for 24 h. The 24-h mean LH and FSH levels ranged from 20.2-70.5 mIU/ml and 60.4-229 mIU/ml, respectively. There was a significant positive correlation between the individual LH and FSH levels in the eight patients; the common correlation coefficient was 0.449 (P less than 0.001). The 24-h mean estradiol level was measurable in only two of the patients and the 24-h mean testosterone level for the eight patients was 0.10 ng/ml. The mean LH concentration during sleep was significantly higher (P less than .01) than during waking. The mean FSH concentration during sleep was also significantly higher (P less than 0.05) than during waking. The LH and FSH peak levels after LRH were significantly correlated with the 24-h mean LH (r = 0.918; P less than 0.01) and FSH concentrations (r = 0.754; P less than 0.05), respectively.

Studies of gonadotropin-gonadal dynamics in patients with androgen insensitivity.

Four patients with androgen insensitivity had plasma LH and FSH measured at 20-min intervals for 24 h and at 15- to 30-min intervals for 3 h after the injection of LRH. Twenty-four-hour mean testosterone (T), estradiol, and androstenedione (delta 4) levels were also measured. Patients with androgen insensitivity had significantly elevated LH levels (P less than 0.05) and an increase in the number of LH secretory episodes (P less than 0.001) compared to normal subjects. The amplitude of the LH secretory episodes, expressed as the absolute increment, was significantly higher than normal controls (P less than 0.005). The LH response to LRH (absolute increment) was twice that of normal, but was not significantly different from normal subjects. The 24-h mean FSH levels were normal in three of the patients and elevated in one. This patient had the mildest degree of androgen insensitivity on clinical exam and the greatest degree of testicular atrophy. The 24-h mean T, estradiol, and delta 4 levels were higher than normal, but only the delta 4 was significantly increased (P less than 0.05). To determine if the elevated LH levels were in response to a decrease in the free T level, we measured T-binding capacity (TBG), TBG was higher than normal controls but was not significantly different, suggesting that elevated LH levels were probably in response to a decrease in T action at the hypothalamic-pituitary level. This was further supported by the inability of prolonged dihydrotestosterone administration to affect LH secretion in one of the patients with the Reifenstein syndrome.

Twenty-four-hour plasma prolactin patterns in prepubertal and adolescent boys.

The concentration of PRL was measured every 20 min for 24 h in six prepubertal and three adolescent boys. In both groups, PRL secretory episodes occurred throughout the 24-h period. In all subjects, the mean concentration of PRL was significantly higher during sleep than during wakefulness; the mean concentration during the entire 24-h period, during sleep or during wakefulness, was not different between the prepubertal subjects and the adolescents. These data suggest the absence of an ontogenetic change for PRL secretion in boys. During acute sleep-wake reversal, two of three pubertal boys showed significantly higher PRL during daytime sleep than during nocturnal wakefulness. This suggest that PRL release in adolescent boys is linked with sleep, rather than with clock time.

The effect of oral contraceptive treatment on the serum concentration of dehydroisoandrosterone sulfate.

Dehydroisoandrosterone sulfate (DS), the major C19-steroid in the human circulation, was measured in serum obtained from blood samples collected daily (8 to 10 A.M.) throughout the menstrual cycles of eight normal, presumably ovulatory women and daily throughout the treatment cycles in four women taking an oral contraceptive (norethindrone, 1 mg., plus mestranol, 80 mcg.). The serum concentrations of DS in the ovulatory women ranged from 1,025 to 4,200 ng. per milliliter; mean, 2,062 +/- 137 ng. per milliliter (mean and standard error; n = 213). Serum DS concentrations during the follicular and luteal phases of the menstrual cycles of these women were similar. In women taking the oral contraceptive, the plasma DS concentrations ranged from 475 to 1,400 ng. per milliliter (mean, 895 +/- 83; n = 119). The 24 hour secretory pattern of DS was evaluated in one subject during a nontreatment cycle and again after 20 days of oral contraceptive treatment. In this subject, the mean serum DS level was 34 per cent lower during oral contraceptive treatment than the level before treatment. The decrease in the serum concentration of DS during oral contraceptive treatment likely results from a reduction in adrenal DS secretion since DS secretion by the normal human ovary is negligible and ovarian dehydroisoandrosterone secretion is small. Therefore, it is likely that the reduced serum DS levels in women taking oral contraceptives are the consequence of reduced adrenal secretion of DS resulting from reduced release of adrenocorticotropic hormone.

PIP: Serum levels of dehydroisoandrosterone sulfate (DS), the major adrenal C19-steroid in humans, were measured daily in 4 women taking an oral contraceptive (OC; 1 mg norethindrone plus 80 mcg mestranol) and in 8 controls. Serum DS concentration averaged 2.062 ng/ml among the controls and 895 among the OC users (p .001). Serum levels remained fairly constant throughout the menstrual cycles of the controls. The 24-hour secretory pattern of DS was examined in 1 woman during a nontreatment cycle and again after 20 days of OC use; the mean serum DS level was 34% lower after OC use than it was before. Because the amount of DS and dehydroisoandrosterone secretion by the ovary is negligible and because reduced serum DS cannot be accounted for by an increase in its metabolic clearance rate, the reduction probably results from reduced adrenal secretion of DS, which in turn results from reduced release of adrenocorticotropic hormone.

Effect of dopamine agonist (Lergotrile mesylate) therapy on twenty-four hour secretion of prolactin in treated Parkinson's disease.

Plasma PRL was measured at 20-min intervals in six patients with Parkinson's disease under various treatment protocols. In addition, 24-h mean GH levels were measured. The results of these studies showed that two untreated patients with Parkinson's disease had normal 24-h mean PRL levels with the normal increase during sleep. During chronic treatment with L-dopa-carbidopa (Sinemet), the 24-h PRL level was 12.8 +/- 4.9 ng/ml (mean +/- SD) and there was persistence of augmented PRL secretion during sleep. The 24-h mean GH level ranged from 1.5-4.4 ng/ml, with a mean of 2.5 ng/ml. The addition of a dopamine agonist (Lergotrile mesylate) resulted in a significant (P less than 0.01) suppression of the 24-h mean PRL levels and abolition of the normal sleep augmentation after 2 weeks of therapy. This suppression was maintained in one patient who was restudied 4 months after the addition of dopamine agonist therapy to L-dopa-carbidopa. The 24-h mean GH levels did not change significantly after the addition of the dopamine agonist when compared to L-dopa-carbidopa alone. These results suggest a dichotomy between the PRL and GH responses to combined L-dopa-carbidopa and dopamine agonist therapy. In addition, the preservation of normal PRL regulation in the two untreated patients with Parkinson's disease suggests that dopaminergic neurons are not universally affected in this disorder.

Analysis of secretory patterns of prolactin and gonadotropins during twenty-four hours in a lactating woman before and after resumption of menses.

Twenty-four hour secretory patterns of prolactin, luteinizing hormone (LH), and follicle-stimulating hormone (FSH) were obtained on two separate occasions from a woman with late physiologic lactation. The studies were performed 26 and 34 months after her child's birth. During the initial study, she had amenorrhea, and her child suckled 13 per cent of the 24 hour period (Study 1). At the time of the second study, she had resumed regular menses and her child suckled for 7 per cent of the 24 hour period (Study 2). The average concentrations of prolactin during Studies 1 and 2 were 40 +/- 1.0 (mean and standard error; No. = 72) and 31 +/- 1.4 ng. per milliliter, respectively. The mean plasma prolactin concentration in Study 1 was significantly greater than that in Study 2 (p less than 0.001). The plasma concentrations of LH and FSH were significantly less in Study 1 than in Study 2 (p less than 0.001 and less than 0.01, respectively). It is concluded that hyperprolactinemia and hypogonadotropinemia were endocrinologic correlates of the amenorrhea of late physiologic lactation in this woman.