Inhibition of apical domain formation does not block blastocyst development in bovine embryos.

The first event of cellular differentiation consists of the segregation of the trophectoderm and the inner cell mass. Studies in mice suggest that cell contractility and the formation of an apical domain play important roles in this event; however, this remains unknown in the bovine. We tested the hypothesis that blocking apical domain formation would halt subsequent trophectoderm differentiation in bovine embryos. We first assessed the formation of an apical domain by the presence of Par-6 Family Cell Polarity Regulator Beta (PARD6B) and Ezrin (EZR), which appeared after the 8-cell stage. We inhibited apical domain formation by blocking cell contractility with 25µM (-)-blebbistatin. Treatment from 90 to 186h after insemination did not reduce blastocyst development compared with the untreated control group or the group treated with inactive (+)-blebbistatin. Immunofluorescence staining after blebbistatin treatment revealed the absence of EZR and the trophectoderm marker Caudal Type Homeobox 2 (CDX2). Following blebbistatin treatment, Yes1 Associated Transcriptional Regulator (YAP), which is involved in the Hippo signalling pathway, exhibited cytoplasmic staining instead of nuclear localisation. Despite changes in protein expression and localisation, no difference in trophectoderm or total cell numbers was observed. In conclusion, inhibition of cell contractility inhibited apical domain formation without impairing blastocyst formation, suggesting that a different biological mechanism is involved in trophectoderm and inner cell mass differentiation in bovine embryos.

Blood pressure regulation in the three-toed sloth, Bradypus variegatus.

The aim of this study was to elucidate the role of the baroreflex in blood pressure control in sloths, Bradypus variegatus, since these animals show labile levels in this parameter. Unanesthetized cannulated sloths were positioned in an experimental chair and the arterial catheter was coupled to a strain gauge pressure transducer. Blood pressure was monitored before, during and after the administration of phenylephrine (0.0625 to 4 microg/kg) and sodium nitroprusside (0.0625 to 2 microg/kg), bringing about changes in mean blood pressure from +/-30 mmHg in relation to control values. The relation between heart rate changes due to blood pressure variation was estimated by linear regression analysis. The slope was considered the reflex baroreceptor gain. The results (means+/-SD) showed that the reflex baroreceptor gain was -0.3+/-0.1 bpm/mmHg (r=0.88) to phenylephrine and -0.5+/-0.1 bpm/mmHg (r=0.92) to sodium nitroprusside, denoting a reduced reflex baroreceptor gain when compared with other mammals, suggesting that in sloths the baroreceptors are minimally involved in the buffering reflex response to these drugs. These findings suggest that the labile blood pressure could be influenced or be a result of this lowering in the reflex baroreceptor gain.

Fecal estradiol and progesterone metabolite levels in the three-toed sloth (Bradypus variegatus)

The present study was carried out to assess the possibility of measuring fecal steroid hormone metabolites as a noninvasive technique for monitoring reproductive function in the three-toed sloth, Bradypus variegatus. Levels of the estradiol (E2) and progesterone (P4) metabolites were measured by radioimmunoassay in fecal samples collected over 12 weeks from 4 captive female B. variegatus sloths. The validation of the radioimmunoassay for evaluation of fecal steroid metabolites was carried out by collecting 10 blood samples on the same day as defecation. There was a significant direct correlation between the plasma and fecal E2 and P4 levels (P < 0.05, Pearson's test), thereby validating this noninvasive technique for the study of the estrous cycle in these animals. Ovulation was detected in two sloths (SL03 and SL04) whose E2 levels reached 2237.43 and 6713.26 pg/g wet feces weight, respectively, for over four weeks, followed by an increase in P4 metabolites reaching 33.54 and 3242.68 ng/g wet feces weight, respectively. Interestingly, SL04, which presented higher levels of E2 and P4 metabolites, later gave birth to a healthy baby sloth. The results obtained indicate that this is a reliable technique for recording gonadal steroid secretion and thereby reproduction in sloths.

Electrocardiographic studies of the three-toed sloth, Bradypus variegatus

Electrocardiograms (ECG) obtained with standard limb leads and augmented unipolar limb leads were recorded from 17 unanesthetized adult sloths. The animals were held in their habitual position in an experimental chair. We determined heart rate and rhythm from the R-R intervals, the amplitude and duration of each wave, and the duration of the segments and intervals of the ECG. The mean electrical axes of P and T waves and QRS complex were calculated on the basis of the amplitude of these waves in leads I, II, III, aV R, aV L, and aV F. The P wave appeared positive in most tracings with low amplitude in lead II, the QRS complex was generally negative in leads aV R, III and aV F, and no arrhythmias were observed. With a mean ± SD heart rate for all recordings of 81 ± 18 bpm, the duration of P and T waves, QRS complex, and PR, QT and RR intervals averaged 0.05 ± 0.02, 0.15 ± 0.05, 0.07 ± 0.02, 0.13 ± 0.02, 0.38 ± 0.04, and 0.74 ± 0.17 s, respectively. The ECG shape had a definite configuration on each lead. The angles of the mean ± SD electrical axes for atrial and ventricular depolarization and ventricular repolarization in the horizontal plane were +34 ± 68°, -35 ± 63°, and -23 ± 68°, respectively. All electrical axes showed great variations and their mean values suggest that, when the sloth is in a seated position, the heart could be displaced by the diaphragm to a semi-horizontal position.

Electrocardiographic studies of the three-toed sloth, Bradypus variegatus.

Electrocardiograms (ECG) obtained with standard limb leads and augmented unipolar limb leads were recorded from 17 unanesthetized adult sloths. The animals were held in their habitual position in an experimental chair. We determined heart rate and rhythm from the R-R intervals, the amplitude and duration of each wave, and the duration of the segments and intervals of the ECG. The mean electrical axes of P and T waves and QRS complex were calculated on the basis of the amplitude of these waves in leads I, II, III, aV R, aV L, and aV F. The P wave appeared positive in most tracings with low amplitude in lead II, the QRS complex was generally negative in leads aV R, III and aV F, and no arrhythmias were observed. With a mean +/- SD heart rate for all recordings of 81 +/- 18 bpm, the duration of P and T waves, QRS complex, and PR, QT and RR intervals averaged 0.05 +/- 0.02, 0.15 +/- 0.05, 0.07 +/- 0.02, 0.13 +/- 0.02, 0.38 +/- 0.04, and 0.74 +/- 0.17 s, respectively. The ECG shape had a definite configuration on each lead. The angles of the mean +/- SD electrical axes for atrial and ventricular depolarization and ventricular repolarization in the horizontal plane were +34 +/- 68 masculine, -35 +/- 63 masculine, and -23 +/- 68 masculine, respectively. All electrical axes showed great variations and their mean values suggest that, when the sloth is in a seated position, the heart could be displaced by the diaphragm to a semi-horizontal position.

Cardiovascular responses to locomotor activity and feeding in unrestrained three-toed sloths, Bradypus variegatus

Heart rate (HR) and systolic (SBP), diastolic (DBP) and mean (MBP) blood pressure were recorded by biotelemetry in nine conscious unrestrained sloths for 1 min every 15 min over a 24-h period. The animals were allowed to freely move in an acoustically isolated and temperature-controlled (24 ± 1ºC) experimental room with light-dark cycle (12/12 h). Behavior was closely monitored through a unidirectional visor and classified as resting (sitting or suspended), feeding (chewing and swallowing embauba leaves, Cecropia adenops), or locomotor activity around the tree trunk or on the room floor. Locomotor activity caused statistically significant increases in SBP (+8 percent, from 121 ± 22 to 131 ± 18 mmHg), DBP (+7 percent, from 86 ± 17 to 92 ± 10 mmHg), MBP (+8 percent, from 97 ± 19 to 105 ± 12 mmHg), and HR (+14 percent, from 84 ± 15 to 96 ± 15 bpm) compared to resting values, indicating a possible major influence of the autonomic nervous system on the modulation of cardiac function during this behavior. During feeding, the increase in blood pressure was even higher (SBP +27 percent, from 119 ± 21 to 151 ± 21 mmHg; DBP +21 percent, from 85 ± 16 to 103 ± 15 mmHg; MBP +24 percent, from 96 ± 17 to 119 ± 17 mmHg), while HR remained at 14 percent (from 84 ± 15 to 96 ± 10 bpm) above resting values. The proportionally greater increase in blood pressure than in HR during feeding suggests an increase in peripheral vascular resistance as part of the overall response to this behavior.

Cardiovascular responses to locomotor activity and feeding in unrestrained three-toed sloths, Bradypus variegatus.

Heart rate (HR) and systolic (SBP), diastolic (DBP) and mean (MBP) blood pressure were recorded by biotelemetry in nine conscious unrestrained sloths for 1 min every 15 min over a 24-h period. The animals were allowed to freely move in an acoustically isolated and temperature-controlled (24 +/- 1 degree C) experimental room with light-dark cycle (12/12 h). Behavior was closely monitored through a unidirectional visor and classified as resting (sitting or suspended), feeding (chewing and swallowing embauba leaves, Cecropia adenops), or locomotor activity around the tree trunk or on the room floor. Locomotor activity caused statistically significant increases in SBP (+8%, from 121 +/- 22 to 131 +/- 18 mmHg), DBP (+7%, from 86 +/- 17 to 92 +/- 10 mmHg), MBP (+8%, from 97 +/- 19 to 105 +/- 12 mmHg), and HR (+14%, from 84 +/- 15 to 96 +/- 15 bpm) compared to resting values, indicating a possible major influence of the autonomic nervous system on the modulation of cardiac function during this behavior. During feeding, the increase in blood pressure was even higher (SBP +27%, from 119 +/- 21 to 151 +/- 21 mmHg; DBP +21%, from 85 +/- 16 to 103 +/- 15 mmHg; MBP +24%, from 96 +/- 17 to 119 +/- 17 mmHg), while HR remained at 14% (from 84 +/- 15 to 96 +/- 10 bpm) above resting values. The proportionally greater increase in blood pressure than in HR during feeding suggests an increase in peripheral vascular resistance as part of the overall response to this behavior.

Circadian rhythms in blood pressure in free-ranging three-toed sloths (Bradypus variegatus).

Blood pressure (BP) profiles were monitored in nine free-ranging sloths (Bradypus variegatus) by coupling one common carotid artery to a BP telemetry transmitter. Animals moved freely in an isolated and temperature-controlled room (24 degrees C) with 12/12-h artificial light-dark cycles and behaviors were observed during resting, eating and moving. Systolic (SBP) and diastolic (DBP) blood pressures were sampled for 1 min every 15 min for 24 h. BP rhythm over 24 h was analyzed by the cosinor method and the mesor, amplitude, acrophase and percent rhythm were calculated. A total of 764 measurements were made in the light cycle and 721 in the dark cycle. Twenty-four-hour values (mean +/- SD) were obtained for SBP (121 +/- 22 mmHg), DBP (86 +/- 17 mmHg), mean BP (MBP, 98 +/- 18 mmHg) and heart rate (73 +/- 16 bpm). The SBP, DBP and MBP were significantly higher (unpaired Student t-test) during the light period (125 +/- 21, 88 +/- 15 and 100 +/- 17 mmHg, respectively) than during the dark period (120 +/- 21, 85 +/- 17 and 97 +/- 17 mmHg, respectively) and the acrophase occurred between 16:00 and 17:45 h. This circadian variation is similar to that observed in cats, dogs and marmosets. The BP decreased during "behavioral sleep" (MBP down from 110 +/- 19 to 90 +/- 19 mmHg at 21:00 to 8:00 h). Both feeding and moving induced an increase in MBP (96 +/- 17 to 119 +/- 17 mmHg at 17:00 h and 97 +/- 19 to 105 +/- 12 mmHg at 15:00 h, respectively). The results show that conscious sloths present biphasic circadian fluctuations in BP levels, which are higher during the light period and are mainly synchronized with feeding.

Circadian rhythms in blood pressure in free-ranging three-toed sloths (Bradypus variegatus)

Blood pressure (BP) profiles were monitored in nine free-ranging sloths (Bradypus variegatus) by coupling one common carotid artery to a BP telemetry transmitter. Animals moved freely in an isolated and temperature-controlled room (24ºC) with 12/12-h artificial light-dark cycles and behaviors were observed during resting, eating and moving. Systolic (SBP) and diastolic (DBP) blood pressures were sampled for 1 min every 15 min for 24 h. BP rhythm over 24 h was analyzed by the cosinor method and the mesor, amplitude, acrophase and percent rhythm were calculated. A total of 764 measurements were made in the light cycle and 721 in the dark cycle. Twenty-four-hour values (mean ± SD) were obtained for SBP (121 ± 22 mmHg), DBP (86 ± 17 mmHg), mean BP (MBP, 98 ± 18 mmHg) and heart rate (73 ± 16 bpm). The SBP, DBP and MBP were significantly higher (unpaired Student t-test) during the light period (125 ± 21, 88 ± 15 and 100 ± 17 mmHg, respectively) than during the dark period (120 ± 21, 85 ± 17 and 97 ± 17 mmHg, respectively) and the acrophase occurred between 16:00 and 17:45 h. This circadian variation is similar to that observed in cats, dogs and marmosets. The BP decreased during "behavioral sleep" (MBP down from 110 ± 19 to 90 ± 19 mmHg at 21:00 to 8:00 h). Both feeding and moving induced an increase in MBP (96 ± 17 to 119 ± 17 mmHg at 17:00 h and 97 ± 19 to 105 ± 12 mmHg at 15:00 h, respectively). The results show that conscious sloths present biphasic circadian fluctuations in BP levels, which are higher during the light period and are mainly synchronized with feeding

The effect of feeding on the respiratory activity of the sloth.

The aim of the present study was to confirm whether feeding influences the resting breathing rate and to observe possible alterations in blood gas and pH levels produced by feeding in unanesthetized sloths (Bradypus variegatus). Five adult male sloths (4.1 +/- 0.6 kg) were placed daily in an experimental chair for a period of at least 4 h for sitting adaptation. Five measurements were made for each sloth. However, the sloths one, two and five were studied once and the sloths three and four were studied twice. Breathing rate was determined with an impedance meter and the output signal was digitized. Arterial blood samples were collected for blood gas analysis with a BGE electrolytes analyzer and adjusted for the animal's body temperature and hemoglobin content. The data are reported as mean +/- SD and were collected during the resting period (8:00-10:00 h) and during the feeding period (16:00-18:00 h). The mean breathing rate increased during mastication of ymbahuba leaves (rest: 5.0 +/- 1, feeding: 10 +/- 1 bpm). No significant alterations were observed in arterial pH (rest: 7.42 +/- 0.05, feeding: 7.45 +/- 0.03), PCO2 (rest: 35.2 +/- 5.3, feeding: 33.3 +/- 4.4 mmHg) or PO2 (rest: 77.5 +/- 8.2, feeding: 78.4 +/- 5.2 mmHg) levels. These results indicate that in unanesthetized sloths 1) feeding evokes an increase in breathing rate without a significant change in arterial pH, PCO2 or PO2 levels, and 2) the increase in breathing rate produced by feeding probably is due to the act of mastication.

The effect of feeding on the respiratory activity of the sloth

The aim of the present study was to confirm whether feeding influences the resting breathing rate and to observe possible alterations in blood gas and pH levels produced by feeding in unanesthetized sloths (Bradypus variegatus). Five adult male sloths (4.1 ± 0.6 kg) were placed daily in an experimental chair for a period of at least 4 h for sitting adaptation. Five measurements were made for each sloth. However, the sloths one, two and five were studied once and the sloths three and four were studied twice. Breathing rate was determined with an impedance meter and the output signal was digitized. Arterial blood samples were collected for blood gas analysis with a BGE electrolytes analyzer and adjusted for the animal's body temperature and hemoglobin content. The data are reported as mean ± SD and were collected during the resting period (8:00-10:00 h) and during the feeding period (16:00-18:00 h). The mean breathing rate increased during mastication of ymbahuba leaves (rest: 5.0 ± 1, feeding: 10 ± 1 bpm). No significant alterations were observed in arterial pH (rest: 7.42 ± 0.05, feeding: 7.45 ± 0.03), PCO2 (rest: 35.2 ± 5.3, feeding: 33.3 ± 4.4 mmHg) or PO2 (rest: 77.5 ± 8.2, feeding: 78.4 ± 5.2 mmHg) levels. These results indicate that in unanesthetized sloths 1) feeding evokes an increase in breathing rate without a significant change in arterial pH, PCO2 or PO2 levels, and 2) the increase in breathing rate produced by feeding probably is due to the act of mastication

Treatment of Chlamydia pneumoniae infection with roxithromycin and effect on neointima proliferation after coronary stent placement (ISAR-3): a randomised, double-blind, placebo-controlled trial.

BACKGROUND: Vascular infection with Chlamydia pneumoniae might boost inflammatory responses that play a pivotal part in neointima formation, which is the main cause of restenosis after stenting. Our aim was to investigate whether or not treatment of C pneumoniae infection with antibiotics prevents restenosis after coronary stent placement. METHODS: We enrolled 1010 consecutive patients with successful coronary stenting into a randomised, double-blind trial. Patients received the macrolide antibiotic roxithromycin 300 mg once daily for 28 days (506), or placebo (504). Primary endpoint was frequency of restenosis (diameter stenosis >50%) at follow-up angiography, and secondary endpoint was rate of target vessel revascularisation during the year after stenting. A prespecified secondary analysis addressed treatment effect with respect to titre of C pneumoniae in serum. Analysis was by intention to treat. FINDINGS: Rate of angiographic restenosis was 31% (157 lesions) in the roxithromycin group and 29% (148) in the placebo group (relative risk 1.08 [95% CI 0.92-1.26]; p50.43), corresponding to a rate of target vessel revascularisation of 19% (120) and 17% (105), respectively (1.13 [0.95-1.36]; p50.30). The combined 1-year rates of death and myocardial infarction were 7% (36) in the roxithromycin group and 6% (30) in the placebo group (p50.45). We showed a significant interaction between treatment and C pneumoniae antibody titre (p50.038 for restenosis, p50.006 for revascularisation), favouring roxithromycin at high titres (adjusted odds ratios at a titre of 1/512 were 0.44 [0.19-1.06] and 0.32 [0.13-0.81], respectively). INTERPRETATION: Non-selective use of roxithromycin is inadequate for prevention of restenosis after coronary stenting. There is, however, a differential effect dependent on C pneumoniae titres. In patients with high titres, roxithromycin reduced the rate of restenosis.

Sloth biology: an update on their physiological ecology, behavior and role as vectors of arthropods and arboviruses.

This is a review of the research undertaken since 1971 on the behavior and physiological ecology of sloths. The animals exhibit numerous fascinating features. Sloth hair is extremely specialized for a wet tropical environment and contains symbiotic algae. Activity shows circadian and seasonal variation. Nutrients derived from the food, particularly in Bradypus, only barely match the requirements for energy expenditure. Sloths are hosts to a fascinating array of commensal and parasitic arthropods and are carriers of various arthropod-borne viruses. Sloths are known reservoirs of the flagellate protozoan which causes leishmaniasis in humans, and may also carry trypanosomes and the protozoan Pneumocystis carinii.

Metal ion-carbonyl oxygen recognition in complexes of acetyl phosphate.

Studies on acetyl phosphate (AcP2-), one of the so-called 'energy-rich' mixed-acid anhydrides, are summarized. Based on stability constants determined by potentiometric pH titrations in aqueous solution, it is shown that the M(AcP) complexes of Ca2+, Mg2+, Mn2+, Cu2+, and Zn2+ are more stable than is expected from the basicity of the phosphate group of AcP2-. This observed stability increase is attributed to an additional interaction of the already phosphate-coordinated metal ion (M2+) with the carbonyl oxygen of the anhydride unit. These conclusions are corroborated by the properties of the complexes of the hydrolysis-stable acetonylphosphonate (AnP2-). The formation degrees of the various six-membered chelates occurring in the M(AcP) and M(AnP) systems are presented and evidence is given that these chelates persist in mixed ligand complexes and that their formation degree is promoted by a low solvent polarity. The biological relevance of these results regarding carbonyl oxygen-metal ion interactions is briefly indicated.

An update on the physiology of two- and three-toed sloths.

Physiological and pharmacological research undertaken on sloths during the past 30 years is comprehensively reviewed. This includes the numerous studies carried out upon the respiratory and cardiovascular systems, anesthesia, blood chemistry, neuromuscular responses, the brain and spinal cord, vision, sleeping and waking, water balance and kidney function and reproduction. Similarities and differences between the physiology of sloths and that of other mammals are discussed in detail.

An update on the physiology of two- and three-toed sloths

Physiological and pharmacological research undertaken on sloths during the past 30 years is comprehensively reviewed. This includes the numerous studies carried out upon the respiratory and cardiovascular systems, anesthesia, blood chemistry, neuromuscular responses, the brain and spinal cord, vision, sleeping and waking, water balance and kidney function and reproduction. Similarities and differences between the physiology of sloths and that of other mammals are discussed in detail

Lead(II)-binding properties of the 5'-monophosphates of adenosine (AMP2-), inosine (IMP2-), and guanosine (GMP2-) in aqueous solution. Evidence for nucleobase-lead(II) interactions.

The stability constants of the 1:1 complexes formed between Pb2+ and the nucleosides (Ns), adenosine and guanosine, as well as between the nucleotides (NMP2-), AMP2-, IMP2-, and GMP2-, were determined by potentiometric pH titrations in aqueous solution (25 degrees C; I = 0.1 M, NaNO3). Based on previously established log KPb(R-PO3)Pb versus pKH(R-PO3)H straight-line plots (R-PO3(2-) = simple phosphate monoester or phosphonate ligands where R is a noninteracting site), it is shown that the Pb(IMP) and Pb(GMP) complexes are more stable than is expected on the basis of the basicity of the phosphate group of IMP2- and GMP2-. This means that macrochelates are formed, where the phosphate-coordinated Pb2+ also interacts with N7 of the nucleobase residue. In contrast, the stability of the Pb(AMP) complex is governed by the basicity of the AMP2- phosphate group. These results agree with the observations made for the Pb(Ns)2+ complexes: Pb(adenosine)2+ is very unstable in contrast to Pb(guanosine)2+, the stability of which is very similar to the one of Pb(cytidine)2+ studied previously. The stability constants of the Pb(Ns)2+ complexes also allowed an evaluation of the structure in solution of the monoprotonated Pb(H;NMP)+ complexes, the stabilities of which were also determined. We were able to show that the proton is located at the phosphate group and Pb2+ at the N7/(C6)O site of H(GMP)-; in the case of H(AMP)- Pb2+ is probably about equally distributed between the adenine residue and the monoprotonated phosphate group. On the basis of the stability constants of these complexes and their structures in solution, it is possible to provide a series which reflects the decreasing affinity for Pb2+ of nucleobase residues in single-stranded nucleic acids: guanine approximately equal to cytosine > (hypoxanthine) > adenine > uracil approximately equal to thymine. The Pb2+ affinity of the phosphodiester linkage, -PO3(-)-, is similar to the one of the adenine residue, but is expected to be more significant due to its larger abundance. The relevance of these results for lead-activated ribozymes is briefly discussed.

Stabilities of complexes formed between lead(II) and simple phosphonate or phosphate monoester ligands including some pyrimidine-nucleoside 5'-monophosphates (CMP2-, UMP2-, dTMP2-).

The stability constants of the 1:1 complexes formed between Pb2+ and several simple phosphate monoesters (4-nitrophenyl phosphate, phenyl phosphate, D-ribose 5-monophosphate, n-butyl phosphate) or phosphonate ligands (methylphosphonate, ethylphosphonate) (R-PO3(2-)) were determined by potentiometric pH titrations in aqueous solution (25 degrees C; I = 0.1 M, NaNO3). The construction of a logK(Pb)Pb(R-PO3) versus pK(H)H(R-PO3) plot for the mentioned ligand systems results in a straight line on which the data pairs (the corresponding equilibrium constants were also measured) for uridine 5'-monophosphate (UMP2-) and thymidine 5'-monophosphate (dTMP2-) also fall; this result shows that in the Pb2+ complexes of UMP2-and dTMP2- the nucleobase residues do not interfere, in neither a positive nor a negative way, with the binding of Pb2+ and that the stability of all these complexes is determined by the basicity of the phosph(on)ate group. The mentioned straight-line correlation (as defined by the least-squares procedure) allowed us to demonstrate (via constants determined now) that the stability of the Pb2+ complex of cytidine 5'-monophosphate (CMP2-) is also solely determined by the basicity of its phosphate group. A similar evaluation, based on literature data, for the Pb(HPO4) complex reveals that its stability corresponds closely to the expectations based on the Pb(R-PO3) data, though there is a slight hint that Pb(HPO4) may be somewhat more stable [which would be in agreement with previous observations of other M(HPO4) complexes]; clearly, more such comparisons are possible with the reference line given now. Based on the stability constants of the monoprotonated Pb(H;CMP)+ complex and the Pb(cytidine)2+ species (which was also measured now), it is concluded that in Pb(H;CMP)+ the proton is located at the phosphate group and Pb2+ mainly at the N3/(C2)O site of the cytosine residue. Regarding nucleic acids in solution, it is further concluded that the affinity of Pb2+ towards the negatively mono-charged phosphate unit, -O--P(O)2- -O-, of a nucleic acid backbone is comparable to that of the cytosine moiety, the affinity towards other nucleobase residues being smaller. This information may prove helpful regarding the properties of lead ribozymes.

Immunological analysis of serum for buckwheat fed celiac patients.

Buckwheat (Fagopyrum esculentum, Moench), a herbaceous plant of the Polygonaceae family, does not have any affinity for the Gramineae family. It is commercially rated as a cereal, which has led to some misunderstanding concerning its use in gluten induced enteropathy or celiac disease. The effect of buckwheat flour ingestion by celiac patients was evaluated through the indirect immunofluorescence technique. Samples of serum were collected 30 days after the flour ingestion. Cryostat performed cuts of wheat and buckwheat grains were accomplished in order to compare the reactivity of their proteins and serum of celiac and normal patients. The assay revealed that the buckwheat flour presents no toxicity for the celiac patient and no anti-protein antibodies formation in the grain was observed for serum dilutions of 1:2 up to 1:32 with 0.15 M NaCl solution in 0.01 M phosphate buffer, pH 7.2 throughout the 30 day assay period. The results of the immunological assays showed that the buckwheat flour does not present toxic prolamines to celiac disease patients.

Responses of the three-toed sloth, Bradypus tridactylus, to some commonly used pharmacologic agents. II. Chloralose and reserpine.

1. Chloralose, 50 mg/kg i.v., is a safe effective anesthetic for sloths and reduces incidence of cardiac arrhythmias. 2. However, chloralose blocks baroreflexes and may reduce the sensitivity of beta 1 cardiac receptors. 3. Reserpine, 0.70 mg/kg given i.v. in divided doses, blocks the hypertensive effect of 100 micrograms/kg of tyramine in sloths. 4. Reserpine in this dosage materially reduces arterial pressure and heart rate; these effects last at least 7 days. 5. Reserpine potentiates the hypertensive effects of epinephrine and norepinephrine materially. 6. In sloths reserpine increases cardiac irritability but does not block baroreflexes. 7. As is true with most other drugs sloths are more sensitive to chloralose and reserpine than most common laboratory animals.