Chronobiological Analysis of Sudden Death Observed in an Emergency Department

The objective of this study was to evaluate the epidemiological characteristics of sudden death (S.D.), with particular reference to the presence of chronobiological rhythms. One hundred eighty-five fatal events were observed during one solar year at the Emergency Department of a northern Italian town. Twenty-three cases were due to accidents or violence, while the remaining 162 corresponded to the operative definition of S.D. we adopted. Investigation by autopsy permitted to recognize that 36% of total deaths were due to cardiovascular diseases (in 2/3 of the cases secondary to ischemic heart diseases). A significant circadian periodicity was observed in S.D. (p=0.01, with two peaks, at 9.26 a.m. and 9.26 p.m.) and also a circannual rhythm, peaking in late autumnDearly winter (p=0.036). The latter was particularly evident in elderly people. A relationship with cardiovascular event rate (of which S.D. recalls the circadian periodicity) is a plausible explanation, while the frequent occurrence of respiratory distress during winter may be at least partly responsible for the high incidence of unexpected deaths seen in this period of the year.     

Chronobiology of Symptomatic Supraventricular Paroxysmal Tachycardia

The circadian, circaseptenal and circaannual occurrence of supraventricular paroxysmal tachycardias (SVPT) were examined in patients addressing an Emergency Department. Sixty-one patients, 22 men and 39 women aged 48.3+13.0 y, were eligible and investigated during a solar year. Cosinor method indicated a significant circadian SVPT variation, with an acrophase at about 6 p.m. A similar periodicity was also detected in supraventricular premature beat prevalence (acrophase at 3 h 31 min), concordant with heart rate peaking. Monday was the most frequently affected day, while the circa-annual analysis disclosed a peak in summer and autumn. Our findings provide further evidence that SVPT shows the highest occurrence during daytime, in possible connection with adrenergic influence. Moreover there are clues that cyclic stressors, e.g. the ones connected with working-activity resumption or climate influence, may partly condition SVPT appearance during the week or the year.     

Chronobiology of Symptomatic Supraventricular Paroxysmal Tachycardia

The circadian, circaseptenal and circaannual occurrence of supraventricular paroxysmal tachycardias (SVPT) were examined in patients addressing an Emergency Department. Sixty-one patients, 22 men and 39 women aged 48.3+13.0 y, were eligible and investigated during a solar year. Cosinor method indicated a significant circadian SVPT variation, with an acrophase at about 6 p.m. A similar periodicity was also detected in supraventricular premature beat prevalence (acrophase at 3 h 31 min), concordant with heart rate peaking. Monday was the most frequently affected day, while the circa-annual analysis disclosed a peak in summer and autumn. Our findings provide further evidence that SVPT shows the highest occurrence during daytime, in possible connection with adrenergic influence. Moreover there are clues that cyclic stressors, e.g. the ones connected with working-activity resumption or climate influence, may partly condition SVPT appearance during the week or the year.     

Availability of vitamin D photoconversion weighted UV radiation in southern South America

Ultraviolet radiation (UVR) plays a key role in several biological functions, including human health. Skin exposure to UVR is the main factor in vitamin D photoconversion. There is also evidence relating low levels of vitamin D with certain internal cancers, mainly colon, breast and prostate, as well as other diseases. Several epidemiological studies have shown an inverse relationship between the above-mentioned diseases and latitude, in accordance with the ultraviolet radiation latitudinal gradient. The aim of this study is to determine whether UV irradiance levels in the southern South America are sufficient to produce suitable levels of vitamin D year around. For this purpose, vitamin D photoconversion weighted-irradiance was analyzed between S.S. de Jujuy (24.17°S, 65.02°W) and Ushuaia (54° 50'S, 68° 18'W). In addition to irradiance, skin type and area of body exposed to sunlight are critical factors in vitamin D epidemiology. Due to a broad ethnic variability, it was assumed that the skin type in this region varies between II and V (from the most to the less sensitive). All sites except South Patagonia indicate that skin II under any condition of body area exposure and skin V when exposing head, hands, arms and legs, would produce suitable levels of vitamin D year round (except for some days in winter at North Patagonian sites). At South Patagonian sites, minimum healthy levels of vitamin D year round can be reached only by the more sensitive skin II type, if exposing head, hands, arms and legs, which is not a realistic scenario during winter. At these southern latitudes, healthy vitamin D levels would not be obtained between mid May and beginning of August if exposing only the head. Skin V with head exposure is the most critical situation; with the exception of the tropics, sun exposure would not produce suitable levels of vitamin D around winter, during a time period that varies with latitude. Analyzing the best exposure time during the day in order to obtain a suitable level of vitamin D without risk of sunburn, it was concluded that noon is best during winter, as determined previously. For skin type II when exposing head, exposure period in winter varies between 30 and 130 min, according to latitude, except for South Patagonian sites. During summer, noon seems to be a good time of day for short periods of exposure, while during leisure times, longer periods of exposure without risk of sunburn are possible at mid-morning and mid-afternoon. At 3 h from noon, solar zenith angles are almost the same for sites between the tropics and North Patagonia, and at 4 h from noon, for all sites. Then, in these cases, the necessary exposure periods varied slightly between sites, only due to meteorological differences.     

Seasonal and circadian fluctuations of the biochemical indices of the blood in mice

Seasonal (in January, April, July, October) changes of aspartate aminotransferase (AST), alanine aminotransferase (ALT), protein, bilirubin, glucose, cholesterol, creatinine, blood urea nitrogen, Cl-, K+, Na+ content were studied in the blood plasma of mice at different time of day (6 p. m., midnight, 6 a. m., midday). The analysis of the average daily indices has shown that the most expressed variations were the following: AST (spring maximum is 3.7 times higher than autumn minimum), ALT (winter maximum is 2.9 times higher than autumn minimum), creatinine (summer maximum is 2.5 times higher than winter minimum), blood urea nitrogen (summer maximum is 2.5 times higher than autumn minimum). Bilirubin and protein content in spring is insignificant, but it is significantly higher than in other seasons. Cholesterol content is lower in winter. No differences in glucose, Cl-, K+, Na+ content in different months have been revealed. The largest circadian synchronization was observed in winter in AST, glucose, cholesterol, protein, Cl-, K+, Na+ (the level observed at 6 p. m. and at midday is higher than that observed at midnight and 6 a. m.) and in autumn in AST, ALT, glucose, cholesterol, blood urea nitrogen, with the circadian curves inverse as compared to the winter period. In spring practically no circadian synchronization was observed.     

The nutritional status and eating habits of shift workers: a chronobiological approach

The eating habits of workers may vary according to the season of the year and corresponding work schedule. A study aiming at verifying the changes in their diet in summer and winter, as well as the nutritional status of those who work fixed shifts, was conducted. The distribution during the 24h in the quantity of calories and macronutrients ingested and the circadian rhythm of calories consumed were also analyzed. The study was conducted on 28 workers subject to three fixed work (morning, afternoon, and night) shifts at a transport company in the city of S?o Paulo, Brazil. The mean age of the workers was 32.8 (SD+/-5.3) yrs. Their food intake was ascertained by the use of a 3-day dietary record, and their nutritional status was evaluated by their body mass index (BMI), both in winter and summer. Two-way ANOVA (shift and season) showed food consumption--measured in calories/24 h--was significantly higher in winter than summer (F(1.25)=11.7; p<0.001). No statistically significant differences were found among shifts (F(2.25)=0.85; p<0.44), and the interaction effect between shift and season was also not significant (F(2.25) = 0.15; p < 0.86). No seasonal difference in BMI was detected (Kruskal-Wallis test). Cosinor analyses showed circadian rhythmicity in calories consumed by morning (p < 0.01) as well as afternoon shift workers (p < 0.001), both in the winter and summer. Circadian rhythmicity in calories consumed by night workers was found only in summer (p < 0.01). The changes observed in the workers' eating habits from one season to another and during the 24h period show the need for further studies to help develop educational programs to improve the nutrition of shift employees taking into consideration shift schedule and season of the year when work is performed.     

The seasonal variation of thermophilic campylobacters in beef cattle, dairy cattle and calves

The epidemiology of clinical cases of campylobacter in temperate climates shows a striking seasonality. In the search for a seasonal environmental reservoir changes in the carriage rate and population size of campylobacters in bovine hosts with time have been measured. Most probable number (MPN) methodology was used to enumerate thermophilic campylobacters in samples taken from the small intestines of beef cattle at slaughter and the fresh faeces of four dairy herds and new-born calves. Statistical analyses revealed significant evidence for seasonal periodicity in the data from dairy herds ( P = 0·044). Not only was there a departure from constancy within a 12-month interval but these data revealed a true seasonality, that is, the same periodicity in numbers from one year to the next. Each herd had two peaks per year, in approximately spring and autumn. Peaks coincided in herds on neighbouring farms but those on farms in the north preceded those on farms in the south by 2 and 1 months, respectively ( P = 0·0057). Intestinal carriage by beef cattle at slaughter was 89·4% ( n = 360) with an average MPN campylobacters per gram fresh weight (MPN gfw⁻¹) of 6·1 × 10². Average MPN gfw⁻¹ in faeces from the dairy herds and calves were 69·9 ( S.D. 3) and 3·3 × 10⁴ ( S.D. 1·7 × 10²). There was no evidence of seasonal periodicity in the size of the campylobacter population in beef cattle at slaughter. Calves were campylobacter free at birth but became colonized within a few days.     

Circadian variation in cortisol reactivity to an acute stressor

To investigate the role of the circadian pacemaker in cortisol reactivity to a cold pressor challenge, 26 diurnally subjects participated in a constant-routine protocol and were divided into two groups. Group 1 started immediately after a monitored sleep period at 09:00 h, while group 2 started 12 h later. After 2 h of adaptation, a cold pressor test was presented every 3 h. The cortisol response was assessed by means of saliva samples that were taken before and after the test. The pretest samples were considered to be base-rate measures and base-rate values as subtracted from post-test values were considered as reactivity measures. Both measures showed distinct Time-of-Day variations (respectively: F(7,168) = 16.92, p < 0.001, epsilon = 0.383; and F(7,175) = 8.01, p < 0.001, epsilon = 0.523). These findings are interpreted as evidence for the existence of an endogenous circadian periodicity underlying the sensitivity of the hypothalamus-pituitary-adrenal (HPA)-axis to acute stress.     

Effects of bilateral striatal 6-OHDA lesions on circadian rhythms in the rat: a radiotelemetric study

The present work aims to document, in a previously described animal model of Parkinson (double bilateral striatal injection of 6-OHDA), the possible modifications of circadian markers rhythms i.e. temperature (T), heart rate (H) and locomotor activity (A) registered continuously by telemetry, in order to evaluate a possible perturbation of the circadian rythmicity. H, T and A were measured by radiotelemetry after surgical implantation of the transmitters (Data Sciences). After a recovery period, the study was divided into a control period (C) for baseline measurements of T, H and A daily rhythms. Then, the stereotaxic 6-OHDA striatal lesion was done to a group of 4 rats while the control rats were injected into striata with saline; a second period of four registration weeks was observed [D 1-7 (W1), D 8-14 (W2), D 15-21 (W3), D 22-28 (W4)]. Finally, at the end of this period the seven rats were decapited in order to determine their striatal dopamine (DA) content. Our data document that the circadian rhythms of H, T and A were differently affected according to time. Thus, a temporary loss of circadian periodicity was observed particularly for heart rate. 6-OHDA-induced modifications of H, T and A circadian rhythm characteristics were also observed: a significant decrease of the mesor was observed for the three rhythms as well as a phase advance. Concerning the amplitude of these rhythms, only H was significantly decreased. These perturbations were observed during the four weeks following the intervention, never reaching the initial control levels. Such observed perturbations would supply a basis for the future study of the chronopharmacology of antiparkinsonian drugs.     

The same molecular mechanism at the maternal meiosis I produces mono- and dicentric 8p duplications.

We studied 16 cases of 8p duplications, with a karyotype 46,XX or XY,dup(8p), associated with mental retardation, facial dysmorphisms, and brain defects. We demonstrate that these 8p rearrangements can be either dicentric (6 cases) with the second centromere at the tip of the short arm or monocentric (10 cases). The distal 8p23 region, from D8S349 to the telomere, including the defensin 1 locus, is deleted in all the cases. The region spanning from D8S252 to D8S265, at the proximal 8p23 region, is present in single copy, and the remaining part of the abnormal 8 short arm is duplicated in the dicentric cases and partially duplicated in the monocentric ones. The distal edge of the duplication always spans up to D8S552 (8p23.1), while its proximal edge includes the centromere in the dicentric cases and varies from case to case in the monocentric ones. The analysis of DNA polymorphisms indicates that the rearrangement is consistently of maternal origin. In the deleted region, only paternal alleles were present in the patient. In the duplicated region, besides one paternal allele, some loci showed two different maternal alleles, while others, which were duplicated by FISH analysis, showed only one maternal allele. We hypothesize that, at maternal meiosis I, there was abnormal pairing of chromosomes 8 followed by anomalous crossover at the regions delimited by D8S552 and D8S35 and by D8S252 and D8S349, which presumably contain inverted repeated sequences. The resulting dicentric chromosome, 8qter-8p23.1(D8S552)::8p23.1-(D8S35)-8q ter, due to the presence of two centromeres, breaks at anaphase I, generating an inverted duplicated 8p, dicentric if the breakage occurs at the centromere or monocentric if it occurs between centromeres.     

Rhythms in the ovulatory cycle. 3rd: Cortisol and dehydroepiandrosterone sulphate (DHEA-S)

The circadian profiles of cortisol and dehydroepiandrosterone sulphate (DHEA-S) were analyzed in a homogeneous group of 15 young normally cycling women, at 4 times of the menstrual cycle: early follicular (EF), late follicular (LF), early luteal (EL) and late luteal (LL) stages. The circatrigintan variations of the same hormones were also evaluated. Population-mean cosinor analysis allowed the demonstration of a highly significant circadian periodicity for both variables in any of the 4 stages of the menstrual cycle; on the other hand, the same computation failed to demonstrate a significant circatrigintan periodicity. In each of the stages considered, the circadian acrophase of DHEA-S was delayed in comparison to that of cortisol, being located in the early afternoon hours. The demonstration of a clear-cut circadian oscillation in serum DHEA-S prompts studies on possible chrono-abnormalities of the steroid production in women with hyperandrogenic diseases.     

Annual change in insulin sensitivity

The incidence of both type 2 diabetes and cardiac events is reported to be higher during winter, indicating a putative annual periodic change in insulin sensitivity (IS). Annual differences in IS - quantified as HOMA-%S and Matsuda-Sensitivity Index - were analyzed using a cosine wave-fitting algorithm in a cross-sectional study group including 2?385 participants. Additionally, semi-annual differences in IS were compared. We found periodicity for HOMA-%S and Matsuda-Sensitivity Index (p=0.02 or 0.006), which was strengthened after restriction to participants without diabetes (p=0.009 or 0.004). The rhythm amplitude of 0.08 indicated moderate changes in IS throughout the year. IS was significantly higher when participants were enrolled during the second vs. the first half of the year (HOMA-%S 112.0±3.0% vs. 97.4±2.4%, p<0.001). The impact of the half-year on IS, which remained significant after adjustment for confounders, was again moderate and explained only 0.5% of the variation. IS showed a significant moderate annual periodicity, which may affect the interpretation of studies reporting small changes in IS.     

Rubidium Chloride Fuses Split Circadian Activity Rhythms in Hamsters Housed in Bright Constant Light

Chronotypic effects of rubidium (Rb) were examined in hamsters whose circadian activity rhythms had split into two components while they were housed in bright constant light. Seven of 12 hamsters receiving RbCl in drinking water for 10 weeks showed fusing of the components into an intact rhythm compared with none of 7 control hamsters (p = 0.016). Rb may modify coupling between circadian oscillators via reduced photic input to the suprachiasmatic nuclei. Alternative mechanisms include changes in potassium metabolism or endocrine function or behavioral changes that in turn alter circadian function. This normalization of a circadian anomaly by a putative antidepressant suggests that Rb may be valuable in strengthening coupling between oscillators in cases of human chronopathology, including those implicated in the etiology of some affective disorders.     

Rubidium Chloride Fuses Split Circadian Activity Rhythms in Hamsters Housed in Bright Constant Light

Chronotypic effects of rubidium (Rb) were examined in hamsters whose circadian activity rhythms had split into two components while they were housed in bright constant light. Seven of 12 hamsters receiving RbCl in drinking water for 10 weeks showed fusing of the components into an intact rhythm compared with none of 7 control hamsters (p = 0.016). Rb may modify coupling between circadian oscillators via reduced photic input to the suprachiasmatic nuclei. Alternative mechanisms include changes in potassium metabolism or endocrine function or behavioral changes that in turn alter circadian function. This normalization of a circadian anomaly by a putative antidepressant suggests that Rb may be valuable in strengthening coupling between oscillators in cases of human chronopathology, including those implicated in the etiology of some affective disorders.     

Circadian variation affects the biology and digestive profiles of a nocturnal insect Spodoptera litura (Insecta: Lepidoptera)

The present study was conducted to decipher the impact of circadian rhythm on digestive enzymes of Spodoptera litura under three photoperiods (12L:12D, 0L:24D, and 24L:0D). Longer life cycle, higher developmental traits and significant food utilizing capability were observed in dark conditions (DD), while there was no effect on survival. The activity of lactate dehydrogenase (LDH), α and β-glucosidase depended on complete absence of light (DD) while LL had a significant effect on protease activity. The presence of polypeptides (35, 60 kDa) and lower protease inhibition by PMSF in 0L:24D, and 24L:0D indicated that serine proteases (trypsin) were the main proteases in larval midgut. Overall, zymography profiles suggested that circadian variation, particularly dark period influenced the S. litura development due to fluctuations in the midgut enzymes via food utilization. Although the effect of photoperiod on digestive processes of insects is still unclear, dark regime may underlie the midgut digestive enzymes in S. litura larvae.     

Circadian periodicity of plasma prolactin in some neurological diseases.

The circadian rhythmicity of plasma PRL has been studied in some neurological diseases in which hypothalamic involvement or abnormalities of brain neurotransmitters has been postulated. 11 patients with Steinert's myotonic dystrophy, 7 with cluster headache and 10 with Huntington's chorea have been studied. By the mean cosinor procedure, a significant circadian rhythm of plasma PRL has been observed both in Steinert's disease and in cluster headache, whereas a circadian periodicity is not detectable in Huntington's chorea, a degenerative disorder affecting the basal ganglia, the hypothalamus and many other areas of the CNS.     

Circadian rhythms of sterol 12alpha-hydroxylase, cholesterol 7alpha-hydroxylase and DBP involved in rat cholesterol catabolism

Circadian rhythms of important enzymes involved in the conversion of cholesterol to bile acids [sterol 12alpha-hydroxylase (12alpha-hydroxylase) and cholesterol 7alpha-hydroxylase (7alpha-hydroxylase)] and an albumin site D-binding protein (DBP) were examined in rats. When the animals were fed freely, they usually ate in the dark and the circadian rhythms of activities of 12alpha-hydroxylase and 7alpha-hydroxylase showed the same peaks (at 10 p.m.) and lows (at 2 p.m.). Their mRNA levels were determined at four timepoints: 3 a.m., 10 a.m., 3 p.m. and 10 p.m. A maximum of the rhythm of 12alpha-hydroxylase was observed at 3 p.m. and the minimum at 3 a.m. These results are distinct from those of 7alpha-hydroxylase, whose maximum point was at 10 p.m. and minimum at 3 p.m. When the rats were fed only in the day-time (from 9 a.m. to 5 p.m.), a marked shift of the activity and mRNA rhythms was observed with both enzymes. The circadian rhythms of the activities of both enzymes showed the same peaks (at 3 p.m.), but the mRNA levels of 12alpha-hydroxylase were distinct from those of 7alpha-hydroxylase, whose maximum point was at 3 a.m. and minimum at 10 p.m. Differences between the maximum and the minimum points of each enzyme mRNA level were statistically significant (P < 0.01 for 12alpha-hydroxylase and 0.05 for 7alpha-hydroxylase). Moreover, circadian rhythms of DBP were also markedly shifted with the change of feeding period. The maximum mRNA level was observed at 10 p.m. instead of 10 a.m. and the minimum was at 10 a.m. instead of 10 p.m.     

Autosomal dominant aniridia linked to the chromosome 11p13 markers catalase and D11S151 in a large Dutch family

In a large pedigree with autosomal dominant aniridia, we found close linkage between the aniridia locus AN2 and the markers catalase (CAT) (zeta = 7.27 at theta = 0.00) and D11S151 (zeta = 3.86 at theta = 0.10) flanking the AN2 locus on 11p13. Positive lod scores were also obtained for the 11p13----11p14 markers D11S16 and FSHB with the linkage group CAT/AN2/D11S151. We conclude that the autosomal dominant aniridia in this family is due to a mutation at the AN2 locus on 11p13. We have excluded linkage (zeta less than -2 at theta less than 0.18) between the aniridia and the chromosome 2p25 marker D2S1 (linked to ACP1).     

Altered Orcadian Rhythms of Blood Pressure and Heart Rate in Non-Hemodialysis Chronic Renal Failure

The extended use of ambulatory monitoring has permitted the identification of many conditions in which the circadian rhythm of blood pressure is altered. The common denominator seems to be an impairment of the autonomic nervous system function. We examined whether the circadian blood pressure rhythm is altered in chronic renal failure (where autonomic dysfunction is usually present) by using a standardized chronobiological inferential statistical method in hospitalized subjects. For this purpose, a group of 30 non-hemodialysis hypertensive patients with chronic renal failure was compared with a second group of 30 patients affected by uncomplicated mild-to-moderate essential hypertension. The two groups were matched by age, sex and circadian mesors of blood pressure. Diet, meal times, sleep and activity logs were standardized. Blood pressure and heart rate recordings were obtained by using an automatic oscillometric recorder and subsequently analyzed according to the cosinor method. A mean circadian rhythm of blood pressure was documented in both groups, but while the mean acrophases occurred between 2 and 3 p.m. in essential hypertension, in renal failure they were between 11 p.m. and midnight for blood pressure and around 7 p.m. for heart rate. In addition, the mean circadian amplitudes were significantly lower in renal failure, while the mean circadian mesor of heart rate was significantly higher. Our data demonstrate that the circadian rhythms of blood pressure and heart rate are altered also in hypertension due to chronic renal failure.     

Altered Orcadian Rhythms of Blood Pressure and Heart Rate in Non-Hemodialysis Chronic Renal Failure

The extended use of ambulatory monitoring has permitted the identification of many conditions in which the circadian rhythm of blood pressure is altered. The common denominator seems to be an impairment of the autonomic nervous system function. We examined whether the circadian blood pressure rhythm is altered in chronic renal failure (where autonomic dysfunction is usually present) by using a standardized chronobiological inferential statistical method in hospitalized subjects. For this purpose, a group of 30 non-hemodialysis hypertensive patients with chronic renal failure was compared with a second group of 30 patients affected by uncomplicated mild-to-moderate essential hypertension. The two groups were matched by age, sex and circadian mesors of blood pressure. Diet, meal times, sleep and activity logs were standardized. Blood pressure and heart rate recordings were obtained by using an automatic oscillometric recorder and subsequently analyzed according to the cosinor method. A mean circadian rhythm of blood pressure was documented in both groups, but while the mean acrophases occurred between 2 and 3 p.m. in essential hypertension, in renal failure they were between 11 p.m. and midnight for blood pressure and around 7 p.m. for heart rate. In addition, the mean circadian amplitudes were significantly lower in renal failure, while the mean circadian mesor of heart rate was significantly higher. Our data demonstrate that the circadian rhythms of blood pressure and heart rate are altered also in hypertension due to chronic renal failure.