Circadian and seasonal changes of the inducer:suppressor ratio (OKT4+:OKT8+) in venous blood of healthy adults

Circadian and seasonal variations in the T helper: T suppressor-cytotoxic ratio were investigated in peripheral blood from five healthy young men. Mononuclear cells were isolated on Ficoll-Paque gradient, then incubated with OKT4 and OKT8 monoclonal antibodies. Plasma cortisol was determined in four of these seven time series. Large interindividual differences were documented and statistically validated for the 24-hr.-means of total lymphocytes, OKT4+:OKT8+ ratio, and of plasma cortisol (both total and free). For a pooled data, a circadian rhythm was demonstrated by cosinor (p less than 0.001) for total lymphocytes (acrophase at 1.00 hr.), total plasma cortisol (acrophase at 10.30 hrs.) and free plasma cortisol (acrophase at 9.50 hrs.), but not for OKT4+:OKT8+ ratio. This index however exhibited a statistically significant circadian rhythm in April and August, but not in November. Its double-amplitude exceeded 80% of the 24-hour-mean and its acrophase was localized at 6.40 hrs. in April and at 22.30 hrs. in August. Its 24-hr-mean was higher in August as compared to April and November. The circadian rhythm in the OKT4+:OKT8+ ratio did not seem to be related to that of plasma cortisol. Both circadian and seasonal variations need to be taken into account when investigating the regulations of immune variables such as T helper: T suppressor-cytotoxic ratio.     

Monitoring Heparin Therapy with the Activated Partial Thromboplastin Time

Difficulties associated with the whole blood clotting time (W.B.C.T.) as a method of monitoring heparin therapy have led to the investigation of the activated partial thromboplastin time (A.P.T.T.) as an alternative. The conclusion is reached that the latter procedure possesses several advantages. Using the method described and a citrate-preserved blood sample collected just prior to the administration of the next serial dose of heparin, the suggested therapeutic duration of the A.P.T.T. is 70 seconds or twice the mean control value. A practical range for this method is 60 to 70 seconds.     

Axillary temperature rhythms: the predominance of ultradian periodicity in major affective disorders

Axillary temperature was recorded at least twice during a 48 hrs. span at 6 min. intervals in 10 hospitalized subjects with major affective disorders (DSM III 296. xx). During the clinical occurrence of acute symptoms 7 out of 10 subjects exhibited a prominent ultradian periodicity (period tau less than 20 hrs.) in their temperature time series. Whatever the used therapeutic mean (electroconvulsive therapy and/or chemotherapy) the improvement was associated with a circadian rhythmicity (20 hrs. less than or equal to tau less than or equal to 28 hrs.). A prominent temperature ultradian rhythm (which occurs only in the new born) could be the index of an internal desynchronization associated with major affective disorders.     

Light entrainment and stimulation of the secretion of prolactin in rams

Light entrainment and stimulation of PRL release were studied in rams submitted to artificial light regimes: In a first experiment four groups of animals were submitted to light regimes with a period of 6, 4, 3 or 2 months respectively (T6 to T2 groups) and amplitude of 8-16 h of daylength. In all four groups PRL level was significantly correlated with daylength (r greater than 0.51; P less than 0.01). However in July, when temperatures were indeed high, an additional PRL increase was observed in the T6, T4 and T3 groups, although the role of temperature appears debatable. In a second experiment, animals were exposed daily to 8 hrs. of light given in two photofractions. The first fraction of 7 hrs. began at the same time as "dawn", and the second of 1 hr ended at the same time as "dusk" of the T6 group in the first experiment which served as control group. Correlation of plasma prolactin in rams receiving 8 h light in one or two photofractions was highly significant (r = 0.66; P less than 0.001). This suggests that rams measured daylength between two limits considered as "dawn" and "dusk" even if lights were turned off during part of this interval.     

Optimization of isolation conditions for three types of pigment protein-lipid complexes from chloroplasts during solubilization with Triton X-100]

The maximal total release of pigment protein-lipid complexes (PPLC) during their isolation from pea chloroplasts was achieved by 1-hr solubilization with Triton X-100, the Triton:chlorophyll (T/Chl) ratio being 50 mg/mg/ml. The total yield of the reaction center complexes (sigma PPLC RC) was 22,3%, whereas that of the auxiliary light-accumulating complex (ALA-PPLC) was approximately 32% with respect to Chl. An increase in the solubilization time and of the T/Chl ratio resulted in dissociation of ALA-PPLC. On the contrary, the reaction center complexes steadily maintained their composition and high photochemical activity within a wide range of T/Chl during 24--28 hrs of solubilization. The purest preparations of PPLC RC of phostosystem I (PS-I) were obtained by 24 hr-incubation (T/Chl = 80); their Chl/P700 ratio after a single fractionation on DEAE-cellulose was equal to 36. A considerable increase of T/Chl and of the solubilization time hampered the chromatographical separation of PPLC RC of PS-I and PPLC RC of PS-II. The optimal conditions for isolation of PPLC RC of PS-I and PPLC RC of PS-II were: solubilization at T/Chl 80--120 and prolongation of incubation time from 5 to 7 hrs. The photochemical activity of the complexes obtained was maximal and correlated with the minimal content of admixture P700 (1 molecule of P700 per 450--500 molecules of Chl.).     

Phlebotomines of the Isle of Cyprus. III. Species inventory

Three surveys on canine leishmaniasis were carried out in Cyprus (1993, 1998 and 1999) emphasise the presence of eleven species of phlebotomine sandflies: Phlebotomus (Phlebotomus) papatasi, P. (Paraphlebotomus) alexandri, P. (Pa.) jacusieli (first mention in Cyprus), P. (Pa.) sergenti, P. (Larroussius) galilaeus, P. (L.) tobbi, P. (Transphlebotomus) economidesi, P. (T.) mascittii, Sergentomyia (Sergentomyia) azizi (its specific statute is validated by the authors), S. (S.) fallax et S. (S.) minuta. P. (Adlerius) kyreniae was not caught during the surveys. The authors propose hypothesis of settlement of the island by phlebotomine sandflies according this species inventory. A first migration period took probably place during the Miocene time and a second one during the Pleistocene time.     

Restoration of a normal phenotype, microtubular pattern and DNA synthesis in embryonic fibroblasts concanavalin A pretreated

The present research investigated the time required for reconstructing normal microtubular pattern and the phenotype of cells after short-term (30 min.) and long-term (24 hrs.) pretreatment with the lectin concanavalin A (Con A). Short-term pretreatment led to the formation of incomplete tubules within 2 and 24 hrs. in cells cultured in 199 medium alone. The addition of serum to the medium reversed the globular phenotype and allowed the formation of normal microtubules, even after prolonged pretreatment with Con A. Whereas long-term Con A treatment provoked a reduction in DNA synthesis in 199 alone, in serum-added 199 the percentage of 3H-thymidine incorporation of treated cells tended to reach that of controls over time.     

Changes in UsnRNA biosynthesis during rat liver regeneration

Partial hepatectomy (P.H.) induces a partially synchronized growth response of liver under normal regulation of growth. In this phase changes in cellular morphology, radial distribution pattern of cells and other biological as well as major biochemical changes are well documented [24]. Here, we have shown that the cellular content of UsnRNAs altered during this proliferative phase as well. The level of spliceosomal UsnRNAs (U1, U2, U4–U6) gradually decreased by 30–50% upto 48 hrs of P.H. followed by gradual increase to reach the normal level within one month of P.H. The U3 snRNA level on the other hand, was nearly equal to that in normal liver at 48 hrs of P.H. but in 24 and 72 hrs of P.H. its level was high (4 fold) in contrast to that in other UsnRNAs. Thus, it is clear from our data that the level of all the six UsnRNAs decreased during 48 hrs of P.H. compared to that after first 24 hrs. This has been correlated in the kinetics of UsnRNAs' synthesis (in terms of labelling) in isolated hepatocytes, where the rate of labelling of all the six UsnRNAs increased 20–30% in 24 hrs regenerating hepatocytes (R.H.) followed by sharp decrease by 30–50% within next 24 hrs, compared to that in the normal hepatocytes. But from 72 hrs onwards in R.H. the rate of labelling of all the six UsnRNAs again increased by 30–50% (compared to that in normal hepatocytes) followed by decrease of their labelling-rate to reach the normal level in R.H. within one month of P.H. Thus, it may be concluded that the changes in UsnRNAs' level during the proliferative phase of liver regeneration may be either due to the alteration in the rate of synthesis (in terms of labelling) or along with it differential turn over rate; this phenomenon may have some consequences with the regenerative process of liver.This paper was published in Molecular and Cellular Biochemistry131:67–73, 1994. Kluwer Academic Publishers regret the publication of the only partly corrected version.     

Thermal adaptability of large white pigs in the tropical environment

Twenty-four Large White weaners-twelve males and twelve females, were randomly divided into three groups of eight (four males and four females per two separate pens) and were assigned to three groups of two pens each for the males and the females weaners.One group of two pens was without wallow facility (control), one other group was provided with wallows (wet) and the third group was in air-conditioned room (cold). Twice a day, the respiratory rate and the rectal temperatures were measured early in the mornings at 8.00–9.00 hrs (A.M.) and late in the early evenings at 16.00–18.00 hrs.The mean respiratory frequency (A.M) ranged from 7 to 9; 6–12 and 8–13 breaths per minute for the cold, wet and control respectively while the mean respiratory frequency (P.M.) ranged from 6 to 9, 10 to 17 and 13 to 19 breaths per minute for the cold, wet and control respectively.The mean rectal temperatures (A.M.) ranged very slightly from 38.54° to 39.12°C; 38.50° to 39.05°C and 38.61°C to 39°C for the cold, wet as control respectively while the mean rectal temperatures (P.M.) ranged from 39.00° to 39.22°C; 38.97° to 39.29°C and 39.28° to 39.55°C for the cold, wet and control respectively.The animals were maintained for another seven to ten weeks and were slaughtered. The slaughter characteristics did not indicate an appreciable thermal stress except for the reproductive organs which showed weight increase indicating reduced efficiency of the thermally stressed animals as is the case in the tropical environment.     

肝功能障碍患者对罗库溴铵药效动力学的影响

目的:探讨不同肝功能分级患者对肌松药罗库溴铵药效动力学的影响。方法:纳入肝功能障碍的手术患者152例,根据肝功能Child-Pugh分级将其分为Child-Pugh A级组62例、Child-Pugh B级组55例、Child-Pugh C级组35例,同时纳入肝功能正常患者60例为对照组。以维库溴铵以动态靶控输注程序控制,0.15~0.2μg/m L之间,维持肌松水平T1/TC在25%以内。对比分析四组间一般情况(包括性别、年龄、体重和BMI指数)、麻醉起效时间、最大阻滞持续时间、T110%恢复时间、T125%恢复时间、T175%恢复时间、恢复指数。结果:Child-Pugh A、B、C组患者的麻醉起效时间逐渐延长,组间差异有统计学意义(P0.05)。Child-Pugh A、B、C三组间患者的麻醉最大阻滞程度逐渐下降,组间差异有统计学意义(P0.05)。Child-Pugh A、B、C三组间患者的麻醉恢复指数逐渐增大,组间差异有统计学意义(P0.05)。Child-Pugh A、B、C和对照组间T110%恢复时间、T125%恢复时间、T175%恢复时间无明显差异(P0.05)。Child-Pugh A级组女性麻醉起效时间明显快于男性,差异有统计学意义(P0.05);男性最大阻滞持续时间小于女性,性别间差异有统计学意义(P0.05)。肝功能Child-Pugh B级组女性麻醉起效时间明显快于男性,差异有统计学意义(P0.05)。肝功能Child-Pugh C级组性别间麻醉起效时间、最大阻滞持续时间、恢复指数差异无统计学意义(P0.05)。结论:肝功能障碍明显影响肌松药罗库溴铵麻醉起效时间,重度肝功能患者使用罗库溴铵时可不考虑性别的影响因素。     

Further evidence that prolactin does not affect gonadotropin release at pituitary level

In a primary monolayer cell culture of the anterior pituitary from mature male rats the effects of exogenous rPrl (rPrl exog.) and endogenously secreted rPrl (rPrl endog.) on basal and LHRH stimulated LH secretion were investigated. In pilot studies basal Prl- and LH secretion as well as influence of various LHRH concentrations (10(-1)-10(+3) ng/ml) on Prl- and LH release were observed. The influence of exogenous rPrl was studied at various concentrations (50-500 ng/ml) and with preincubation periods of 2 hrs and 6 hrs before starting LHRH stimulation. The dopamine agonist bromocriptine and the dopamine antagonist sulpirid were preferentially used to prove physiologic function of the cell system presented. Basal LH secretion started after a delay of 3 hrs, whereas basal Prl secretion began immediately showing a linear rise for 9 hrs. LHRH stimulation resulted in a non-linear dose and time dependent LH secretion. LHRH showed no influence on endogenous Prl (rPrl endog.) secretion of the mammotroph cells. Exogenous Prl (rPrl exog.) did not affect spontaneous Prl release excluding ultra short loop inhibition in this cell system. Furthermore, exogenous Prl had no effect on either basal or LHRH stimulated LH secretion even after a preincubation period of up to 6 hrs and at concentrations generally observed for prolactin secreting tumors. Bromocriptine suppressed endogenous Prl release and did not affect LH secretion. Sulpirid had no influence on either Prl or LH secretion.(ABSTRACT TRUNCATED AT 250 WORDS)     

Alteration of peripheral beta-adrenergic responsiveness in fasted rats

Total food deprivation for 72 hrs (3 day fast) in female rats resulted in a reduction in serum thyroid hormones as well as a reduced peripheral beta-adrenergic responsiveness to isoproterenol. Food deprivation for 48 or 72 hrs significantly decreased both serum T3 and T4 values as compared to non-fasted controls. There were no significant differences in either T3 or T4 levels as a result of a 24 hr fast. Rats deprived of food for 72 hr had significantly smaller increases in oxygen consumption, colonic and tail skin temperatures following administration of isoproterenol (100 micrograms/kg b.w., s.c.) when compared to non-fasted control rats. Arterial blood pressure and heart rates were measured in unrestrained, unanesthetized, chronically cannulated rats. Food deprivation for 72 hrs significantly attenuated the decrease in blood pressure and the increase in heart rate associated with administration of isoproterenol (10 micrograms/kg b.w., s.c.). Possible mechanisms for the reduced beta-adrenergic responsiveness associated fasting are discussed.     

Postprandial exchange of urea and ammonia nitrogen between the digestive tract and portal blood in the conscious pig after ingestion of a diet with or without urea

The concentrations of urea and ammonia were measured in the portal and arterial blood simultaneously to the blood flow rate in the portal vein during the postprandial period (8 hrs.) after ingestion of a normal protein diet with 3% urea (10 meals) or without urea (12 meals) in conscious pigs (mean body weight: 55.5 +/- 2.3 kg). When no urea was present in the diet, there was a slight and permanent uptake of blood urea by the gut (570 mg/h, i.e. 9,5 mmoles/h) as well as a permanent appearance of ammonia in the portal vein (258 mg/h i.e. 15.2 mmoles/h), increasing with time (P less than 0.05). The absorbed ammonia nitrogen represented a maximum of 70% of urea nitrogen taken up. 2. Addition of urea to the diet brought about a large absorption of that substance (73% of the ingested amount) followed by a rather large excretion (960 mg/h, i.e. 16 mmoles/h), 5-6 hrs. after the meal and led to an increase (P less than 0.05) in the absorption of ammonia.     

The effect of adrenaline pretreatment on the in vitro generation of 3,5,3'-triiodothyronine and 3,3',5'-triiodothyronine (reverse T3) in rat liver preparation

The effects of adrenaline (A) on liver T3 and rT3 neogenesis from T4 were studied in Wistar rats. The animals were implanted subcutaneously either with A or placebo (P) especially coated tablets which linearly released the hormone. The serum A values 6 hrs after implantation of 7.5, 15.0 and 45.0 mg tablets were 6.5 +/- 1.31, 6.8 +/- 1.8 and 16.4 +/- 1.9 ng/ml, respectively vs 4.4 +/- 2.5 ng/ml seen in P pretreated group. The output rates of A were 0.11 (7.5 mg), 0.18 (15 mg) and 0.52 microgram/ml (45 mg). The pretreatment with A led to hyperglycemia and the "low T3 syndrome". Neogenesis of T3 from T4 in medium containing liver microsomes of P pretreated rats was 5.49 +/- 0.25 pmol of T3/mg protein/min and decreased in A pretreated rats to 3.82 +/- 0.17, 3.12 +/- 0.27 and 3.06 +/- 0.11 pmol of T3/mg of protein/min. Neogenesis of rT3 from T4 in microsomes from P group was 1.52 +/- 0.09 pmol rT3/mg protein/min and increased after A to 2.71 +/- 0.11, 2.60 +/- 0.21 and 2.21 +/- 0.34 pmol of rT3/mg protein/min thus showing no dose dependency. Enrichment of microsomes medium with cytosol either from P or A pretreated rats had no effect on T3 generation thus excluding effect of A on cytosolic cofactor. Although cytosol further increased rT3 neogenesis this was seen regardless of whether cytosol was obtained from A or P implanted rats. It is concluded that A decreases the activity of T4-5'-deiodinase in liver, and possibly increases the activity of T4-5-deiodinase.     

Lung phospholipids. I. In vivo studies of the incorporation of 32P, [methyl-14C] choline, 1-14C-palmitic acid and 1-14C-oleic acid into phosphatidylethanolamine, phosphatidyl-N, N-dimethylethanolamine and phosphatidylcholine

³²P and 1-¹⁴C-palmitate were rapidly incorporated into lung phosphatidyl-N,N-dimethylethanolamine (PDME). The half-life of the PDME based on the decay of 1-¹⁴C-Palmitate was about 5 hrs. A greater half-life (>20 hrs.) occurred with PDME labelled with 1-¹⁴C-oleate. No evidence was found for the conversion of phosphatidylcholine to PDME or the recycling of phosphatidylcholine N-methyl groups to PDME. In light also of the known negligible contribution of methionine methyl transfer in the biosynthesis of dipalmitoylphosphatidylcholine, which is synthesized mainly by the CDP-choline mechanism, lung PDME should not be viewed primarly as an intermediate in the successive N-methylation of phosphatidylethanolamine. Rather, it appears to be an important biosynthetic product.     

Analysis of kinetics of consecutive metabolic cerebral activations induced by spatial discrimination testing, using (14C)-glucose, in an eight-arm radial maze in mice

Regional mapping of relative (14C)-glucose (GLU) uptake was analyzed in Balb/c mice at 3 time intervals (5 min., 1 hr., 3 hrs.) after either the first (Day 1) or the last (Day 9) daily sessions of a spatial discrimination testing procedure in an eight-arm radial maze. On Day 1, increased labelling was found 5 min. post-training in subcortical, hippocampal and cortical regions. Decreased GLU uptake was observed 1 hr. later in the same regions, followed at 3 hrs. post-training by a retarded activation in the above areas and particularly in thalamic and cortical structures. On Day 9, there was only an early (5 min.) post-training increase in metabolic activity followed by a subsequent monotonic decrease over 3 hrs. post-training period.     

The histopathology of Candida albicans invasion in neonatal rat tissues and in the human blood-brain barrier in culture revealed by light, scanning, transmission and immunoelectron microscopy

The present studies examined the effects of Candida albicans yeast and hyphal morphologies on tissue pathologies and transmigration properties of the fungus in two experimental models: 1) an in vivo, neonatal rat model, and 2) a cell culture model of human brain microvascular endothelial cells (ECs) (BMVEC). We inoculated a hyphae-producing strain (CAI4-URA3) and a non-hyphae-producing strain (CAI4) of C. albicans into 4-10 day old rats and BMVEC cultures. Animals were inoculated by intraperitonal (i.p.), intranasal (i.n.), oral (p.o.) and intracerebral (i.c.) routes and several tissues were examined after 24-48 hrs. Rats inoculated i.p. with the hyphae-producing strain showed pathology in the kidneys, liver, spleen, and other tissues associated with inoculation tracks of the nose, and muscle and connective tissues of the abdominal wall. Few animals inoculated i.p., however, presented evidence of meningitis. The non-hyphae phase yeast produced neither tissue pathology nor meningitis. Animals inoculated i.c. with the hyphae strain after 1 and 3 hrs expressed minimal meningitis, with an increasing neutrophillic meningitis between 4 and 18 hrs after inoculation. At 18 hrs after i.c. inoculation, however, the inflammatory foci and brain pathology were extensive and demonstrated mycelia within the lateral ventricles associated with necrosis of adjacent brain tissue. Neutrophillic meningitis at this time period was pronounced. BMVEC co-cultured 1-2 hrs with both C. albicans strains showed EC phagocytosis of hyphae and blastospores into intercellular adhesion molecule-1 (ICAM-1)-labeled caveolae suggesting a transcellular role for ICAM-1 in the internalization process of C. albicans.     

Pineal involvement in the diurnal rhythm of nociception in the rat

Male Wistar rats under cyclic lighting conditions (LD 12:12) were tested for tail flick latencies. A day-night rhythm of pain sensitivity was clearly demonstrated; response latencies were longest 2 hrs. before 'lights on' (-2 hrs.) and shortest 4 hours into the light phase (+4 hrs.). Hot plate data conformed to the tail flick results and supported the notion that the light-dark cycle cues were responsible for the observed diurnal rhythm of analgesia. The possible involvement of the pineal was studied on rats under LD 12:12 schedules, using two paradigms: (1) Functional pinealectomy by light induced suppression and (2) Surgical pinealectomy. The difference between hot plate response latencies measured at '-2 hrs.' and '+4 hrs.', was reduced when the analgesia tests were preceded by either functional pinealectomy or surgical removal of the pineal gland. The data indicates that the pineal gland is involved in modulation of the baseline diurnal rhythm of analgesia in the rat.     

Non-surgical embryo transfer in cattle embryo-recipient interactions

Results from 2286 non-surgical embryo transfers were analyzed to find relationships between embryo factors and recipient factors that affect pregnancy. Pregnancy rates from morulae (44%) and advanced morulae (53%) were reduced (P < .05) when compared to early blastocysts (65%), blastocysts (65%), blastocysts (66%) and advanced blastocysts (64%). Pregnancy rates from good quality Grade 1 embryos (64%) were higher (P < .05) than from Grade 2 (45%) or Grade 3 (33%) embryos. Pregnancy rates related to synchronization of estrus between donor and recipients are: -36 hrs. (before donor) (59%), -24 hrs. (61%), -12 hrs. (68%), 0 hr. (59%), +12 hrs. (61%), +24 hrs. (58%) and +36 hrs. (41%). Differences in pregnancy rate were significant for -12 hrs. (P < .01) and +36 hrs. (P < .01) recipients. Overall pregnancy rates in recipients on day 5 (48%), 5 1/2 (50%) and 6 (53%) were reduced (P < .02) when compared to day 6 1/2 (62%), 7 (63%), and 7 1/2 (66%) and 8 (62%). Pregnancy rates within each embryo stage of development were not related to synchronization with the donor cow (-36 hrs. to +36 hrs.) or the day of the recipient cycle (day 5 to 8).     

Disjunction between the daily duration of light and the plasma level of prolactin under hyperlong days in rams

Adult rams were submitted to an artificial light regime with increasing and decreasing daylengths between 6 and 20 hrs. over a period of 8 months. Blood samples occurred at different durations of daylengths for measuring plasma levels of prolactin. Levels were high for a daily duration of light of 14.20, 17 and 18.30 hrs. along the increasing light-phase and of 18.30 hrs. during the decreasing light-phase. On the other hand prolactin levels were low for short daylengths, but also for a daylength of 20 hrs. It clearly appears that hyperlong days (greater than 18 hrs.) do not stimulate (or do actively inhibit) the secretion of prolactin in ovines.