Genetic Rodent Models of Huntington Disease.

The monogenic nature of Huntington disease (HD) has led to the development of a spectrum of useful genetically modified models. In particular, rodents have pioneered as the first HD model being generated and have since been the most widely used animal model for HD in both basic research and preclinical therapeutic studies. Based on the generation strategies, these rodent models can be classified into 3 major groups, the transgenic fragment models, the transgenic full-length models and the knock-in models. These models display a range of HD-like characteristics which resemble the clinical symptoms of HD patients. Their applications in research are thus regarded as an invaluable approach to speeding up the unraveling of the underlying pathological mechanisms of HD and for finding a disease-modifying treatment for this devastating disease. In this chapter, the similarities and differences of the most commonly used rodent HD models and their relevance to human HD will be compared and discussed. This also serves to guide the selection of an appropriate rodent HD model according to the nature of investigation.

First Report of Stubby Root Caused by Trichodorus obtusus on Zoysiagrass and Bermudagrass in South Carolina.

In September 2011, diagnostic samples were taken from 'Tifway' Bermudagrass (Cynodon dactylon × C. transvaalensis) tees and from 'Emerald' Zoysia (Zoysia japonica) roughs of a golf course in Charleston, SC. Additional samples were taken from a sod farm located near Charleston, SC from a field of 'Empire' Zoysia. The soil was sandy loam and the samples were taken at a depth of 10 to 15 cm from symptomatic turf. Symptoms on bermudagrass and zoysiagrass included stubby roots and lightly to severely chlorotic or dead patches of irregular sizes and shapes. Nematodes were extracted by sugar centrifugal-flotation and counted. The predominant nematode species recovered was Trichodorus obtusus Cobb, 1913: syn. T. proximus Allen, 1957, n.syn. (3). Nematode densities (per 100 cm3 of soil) were 30 to 170 (average 94, n = 5) at the sod farm, and 30 to 230 (average 107, n = 7) at the golf course. This nematode has been reported as a pathogen of bermudagrass in Florida, where it is more damaging than Paratrichodorus minor, the other stubby root nematode commonly associated with turfgrass (1). In Florida, 120 T. obtusus individuals per 100 cm3 is considered high risk (2). We have encountered several additional samples from across South Carolina with comparable densities since our first diagnosis. Infested soil (94 individuals per 100 cm3) collected from the sod farm was put into columns and planted with 'Empire' sod and maintained in the greenhouse. After 140 days, the population density increased to an average of 230 individuals per 100 cm3. Plants were prone to wilting and new root growth showed symptoms similar to those observed in the field. Morphologic and morphometric identification of T. obtusus was made by examining male and female specimens in temporary water mounts. Males had ventrally curved spicules with three ventral precloacal papillae, with the posterior papilla just anterior to the head of the retracted spicules, one ventromedian cervical papilla anterior to the excretory pore, and tail with non-thickened terminal cuticle. Females had a deep, barrel-shaped, pore-like vulva, and one or two postadvulvar lateral body pores on each side. Males and females had distinctly offset esophagus. Females (n = 10) were 1,100 to 1,440 (1,250) µm long, body width 40 to 53 (45) µm, onchiostyle 63 to 75 (67) µm, and V 583 to 770 (673) µm. Males (n = 10) were 1,076 to 1,353 (1,222) µm long, body width 33 to 45 (39) µm, onchiostyle 62 to 69 (65) µm, and spicule 55 to 63 (59) µm. From individuals representing the two locations, an 898-bp section of the 18S rDNA region was sequenced using primers 37F (5'-GCCGCGAAAAGCTCATTACAAC-3') and 932R (5'-TATCTGATCGCTGTCGAACC-3') (4). A BLASTn search revealed no similar sequences to those of our two populations (Accessions JX289834 and JX279930). As such, it appears that these are the first sequences of this portion of the 18S rDNA for T. obtusus, although a different, non-overlapping portion of 18S was found in GenBank (AY146460) under the synonym T. proximus. To our knowledge, this is the first report of T. obtusus on zoysiagrass and the first report of the species on bermudagrass in South Carolina. References: (1) W. T. Crow and J. K. Welch. Nematropica 34:31, 2004. (2) W. T. Crow et al. Florida Nematode Management Guide. SP-54. University of Florida, Gainesville, 2003. (3) W. Decraemer. The Family Trichodoridae: Stubby Root and Virus Vector Nematodes. Kluwer Academic Publishers, Dordrecht, The Netherlands. Pp. 27-30, 1995. (4) I. Duarte et al. Nematology 12:171, 2010.

The impact of market integration on arranged marriages in Matlab, Bangladesh.

Success in marriage markets has lasting impacts on women's wellbeing. By arranging marriages, parents exert financial and social powers to influence spouse characteristics and ensure optimal marriages. While arranging marriages is a major focus of parental investment, marriage decisions are also a source of conflict between parents and daughters in which parents often have more power. The process of market integration may alter parental investment strategies, however, increasing children's bargaining power and reducing parents' influence over children's marriage decisions. We use data from a market integrating region of Bangladesh to (a) describe temporal changes in marriage types, (b) identify which women enter arranged marriages and (c) determine how market integration affects patterns of arranged marriage. Most women's marriages were arranged, with love marriages more recent. We found few predictors of who entered arranged vs. love marriages, and family-level market integration did not predict marriage type at the individual level. However, based on descriptive findings, and findings relating women's and fathers' education to groom characteristics, we argue that at the society-level market integration has opened a novel path in which daughters use their own status, gained via parental investments, to facilitate good marriages under conditions of reduced parental assistance or control.

Religiosity is associated with greater size, kin density, and geographic dispersal of women's social networks in Bangladesh.

Human social relationships, often grounded in kinship, are being fundamentally altered by globalization as integration into geographically distant markets disrupts traditional kin based social networks. Religion plays a significant role in regulating social networks and may both stabilize extant networks as well as create new ones in ways that are under-recognized during the process of market integration. Here we use a detailed survey assessing the social networks of women in rural Bangladesh to examine whether religiosity preserves bonds among kin or broadens social networks to include fellow practitioners, thereby replacing genetic kin with unrelated co-religionists. Results show that the social networks of more religious women are larger and contain more kin but not more non-kin. More religious women's networks are also more geographically diffuse and differ from those of less religious women by providing more emotional support, but not helping more with childcare or offering more financial assistance. Overall, these results suggest that in some areas experiencing rapid social, economic, and demographic change, religion, in certain contexts, may not serve to broaden social networks to include non-kin, but may rather help to strengthen ties between relatives and promote family cohesion.

Massard Prairie Restoration and Soil Microbiome Succession.

We have initially sequenced soil microbial DNA from 4 restored and 3 virgin tallgrass prairie soil samples from Ben Geren Park and Massard Prairie (Fort Smith, AR), respectively. As expected, the soil microbiomes are distinct, with several lineages of nitrogen-fixing bacteria more common in virgin tallgrass prairie. However, we predict that as restoration of tallgrass prairie in Ben Geren Park progresses, the soil microbiome of restored prairie will more closely mirror those of the virgin prairie.

Ouabain binding to renal tubules of the rabbit.

It is well known that ouabain, a specific inhibitor of Na-K ATPase-dependent transport, interferes with renal tubular salt reabsorption. In this study, we employed radiochemical methods to measure the kinetics of [3H]ouabain binding to slices of rabbit renal medulla and high resolution quantitative autoradiography to determine the location and number of cellular binding sites. The kinetics obeyed a simple bimolecular reaction with an association constant of 2.86 +/- 0.63 SD x 10(3) M-1 min-1 and a dissociation constant of 1.46 x 10(-3) min-1, yielding an equilibrium binding constant of 0.51 x 10(-6) M. Binding was highly dependent upon temperature. At a concentration of 10(-6) M, the rate of accumulation between 25 degrees C and 35 degrees C exhibited a Q10 of 1.8. At 0 degree C the rate of ouabain dissociation was negligible. The specificity of binding was demonstrated with increasing potassium concentrations. At a concentration of 1 microM, 6 mM, and 50 mM K+ produced a 2.5- and 7-fold decrease, respectively, in the rate of ouabain accumulation observed at zero K+. Binding was completely inhibited by 1 mM strophanthin K. The major site of ouabain binding was the thick ascending limb; little or no binding was observed in thin limbs and collecting ducts. Moreover, binding was confined to the basolateral membranes. From autoradiographic grain density measurements, it was estimated that each cell contains over 4 x 10(6) ouabain binding sites or Na-K ATPase molecules. These results taken together with physiological and biochemical observations suggest that Na-K ATPase plays a key role in salt reabsorption by this segment.

Achieving synaptically relevant pulses of neurotransmitter using PDMS microfluidics.

Fast synaptic transmission is mediated by post-synaptic ligand-gated ion channels (LGICs) transiently activated by neurotransmitter released from pre-synaptic vesicles. Although disruption of synaptic transmission has been implicated in numerous neurological and psychiatric disorders, effective and practical methods for studying LGICs in vitro under synaptically relevant conditions are unavailable. Here, we describe a novel microfluidic approach to solution switching that allows for precise temporal control over the neurotransmitter transient while substantially increasing experimental throughput, flexibility, reproducibility, and cost-effectiveness. When this system was used to apply ultra-brief ( approximately 400micros) GABA pulses to recombinant GABA(A) receptors, members of the cys-loop family of LGICs, the resulting currents resembled hippocampal inhibitory post-synaptic currents (IPSCs) and differed from currents evoked by longer, conventional pulses, illustrating the importance of evaluating LGICs on a synaptic timescale. This methodology should therefore allow the effects of disease-causing mutations and allosteric modulators to be evaluated in vitro under physiologically relevant conditions.

The effect of steady-state increases in systemic arterial pressure on the duration of left ventricular ejection time.

The effect of steady-state increases in systemic arterial pressure on the duration of left ventricular ejection time was studied in 11 normal male subjects. Methoxamine, a pressor amine of predominantly vasoconstrictor activity but lacking significant inotropic effect, was administered intravenously resulting in an average increase in mean arterial pressure of 27 mm Hg. Heart rate was held constant by high right atrial pacing, and there was no significant change in cardiac output. During methoxamine infusion, when stroke volume, heart rate, and inotropic state were held constant, left ventricular ejection time increased as mean arterial pressure increased. There was a highly significant correlation between the increase in mean systolic blood pressure and the prolongation of left ventricular ejection time (r = 0.870). In one subject, an increase in mean systolic pressure of 75 mm Hg prolonged left ventricular ejection time 55 msec, producing paradoxical splitting of the second heart sound. The prolongation of left ventricular ejection time during infusion was not blocked by the prior intravenous administration of atropine sulfate or propranolol hydrochloride, thus ruling out both vagal inhibition of the left ventricle and reflex withdrawal of sympathetic tone as its cause. In three subjects, left ventricular end diastolic pressure was measured and found to be significantly increased. This finding suggests that the normal left ventricle maintains a constant stroke volume in the presence of an increased pressure load by the Frank Starling mechanism. This study concludes that arterial pressure must be included as a prime determinant of left ventricular ejection time along with stroke volume, heart rate, and inotropic state in intact man.

Left atrial booster function in valvular heart disease.

This study was designed to assess atrial booster pump action in valvular heart disease and to dissect booster pump from reservoir-conduit functions. In five patients with aortic stenosis and six with mitral stenosis, sequential atrioventricular (A-V) pacing was instituted during the course of diagnostic cardiac catheterization. Continuous recording of valvular gradient allowed estimation of flow for each cardiac cycle by transposition of the Gorlin formula. Left ventricular ejection time and left ventricular stroke work in aortic stenosis or left ventricular mean systolic pressure in mitral stenosis were also determined. Control observations were recorded during sequential A-V pacing with well-timed atrial systole. Cardiac cycles were then produced with no atrial contraction but undisturbed atrial reservoir function by intermittently interrupting the atrial pacing stimulus during sequential A-V pacing. This intervention significantly reduced valvular gradient, flow, left ventricular ejection time, and left ventricular mean systolic pressure or stroke work. Cardiac cycles were then produced with atrial booster action eliminated by instituting synchronous A-V pacing. The resultant simultaneous contraction of the atrium and ventricle not only eliminated effective atrial systole but also placed atrial systole during the normal period of atrial reservoir function. This also significantly reduced all the hemodynamic measurements. However, comparison of the magnitude of change from these two different pacing interventions showed no greater impairment of hemodynamic state when both booster pump action and reservoir function were impaired than when booster pump action alone was impaired. The study confirms the potential benefit of well placed atrial booster pump action in valvular heart disease in man.

Autonomic mechanisms in hemodynamic responses to isometric exercise.

Selective autonomic blockade with intravenous propranolol, practolol, atropine, and combined atropine-propranolol was utilized to elucidate the role of the autonomic nervous system in the hemodynamic responses in young adult male volunteers to handgrip sustained at 30% of maximal voluntary contraction for 3 min. The initial 30 s of the tachycardia response was found to be mediated by withdrawal of vagal dominance, as evidenced by blockade of this response by prior atropinization. The mid and late portion of the heart rate response curve was demonstrated to be sympathetic in origin, since it was unaffected by atropine, but was suppressed by combined atropine-propranolol blockade. Sympathetic stimulation appears to be a secondary mechanism for increasing the heart rate, however, as it becomes operative only after the first mechanism of vagal withdrawal has been utilized. This was confirmed by the finding that beta adrenergic receptor blockade alone had little effect on the heart rate response curve. The pressor response to handgrip was accompanied by increased cardiac output and no change in calculated systemic vascular resistance. After propranolol, handgrip resulted in increased peripheral resistance and an equivalent rise in arterial pressure, but no increase in cardiac output. It was concluded that the increase in resistance was the result of sympathetically induced vasoconstriction. This response was shown to be independent of peripheral beta adrenergic receptor blockade by the use of practolol, a cardio-selective beta adrenergic receptor-blocking drug which caused identical hemodynamic responses to those observed after propranolol. Left ventricular ejection time (corrected for heart rate) was prolonged by handgrip. The increased afterload imposed on the left ventricle by sustained handgrip may explain the prolongation of ejection time index. Preejection period was prolonged by SHG after propranolol and shortened after atropine. In addition to confirming the previously defined role of the parasympathetic nervous system, this study delineates the role of the sympathetic nervous system in the heart rate and pressor responses to sustained handgrip.

Impact of ankle-foot orthoses on static foot alignment in children with cerebral palsy.

BACKGROUND: Children with cerebral palsy who are able to walk are often managed with an ankle-foot orthosis to assist with walking. Previous studies have shown kinematic, kinetic, and energetic benefits during gait with the addition of an ankle-foot orthosis, although the mechanism of this gait improvement is unknown. The ability of orthoses to correct foot malalignment in children with cerebral palsy is not known. The current study was performed to determine the impact of orthoses on static foot alignment in children with cerebral palsy. METHODS: A retrospective radiographic review was performed for 160 feet (102 patients). All patients had a diagnosis of cerebral palsy. Standing anteroposterior and lateral radiographs of the foot and ankle were made with the patient barefoot and while wearing the prescribed orthosis and were compared with use of the technique of quantitative segmental analysis of foot and ankle alignment. RESULTS: Analysis of the foot and ankle radiographs made with the patient barefoot and while wearing the brace revealed significant changes in all measurements of segmental alignment (p < 0.05). The magnitudes of these differences were small (<6 degrees or <10%) and would be considered clinically unimportant. The coupled malalignment of equinoplanovalgus (clinical flatfoot) showed radiographic correction of at least one segment (hindfoot, midfoot, or forefoot) to within the normal range in 24% to 44% of the feet. The coupled malalignment of equinocavovarus (clinical high arched foot) showed correction of at least one segment to within the normal range in 5% to 20% of feet. CONCLUSIONS: The present study demonstrates that the use of the ankle-foot orthoses failed to improve the static foot alignment in the majority of feet in children with cerebral palsy who were able to walk. LEVEL OF EVIDENCE: Therapeutic Level IV. See Instructions to Authors for a complete description of levels of evidence.

Reversed closure sequence of the mitral and tricuspid valves in congestive heart failure.

OBJECTIVES: The purpose of this study was to evaluate functional and hemodynamic factors that determine the mitral-tricuspid and aortic-pulmonary valve closure sequence in patients with dilated cardiomyopathy. BACKGROUND: The physiologic factors determining closure sequence of cardiac valves in various forms of heart disease have been found to be complex. Few data exist for dilated cardiomyopathy, particularly for differentiating the effects of a conduction delay versus changes in ventricular performance. METHODS: A group of 64 patients were compared with 36 control subjects. Timing of valve closure and electromechanical intervals were determined by combined M-mode echocardiography, phonocardiography and apexcardiography. Hemodynamic data from right heart catheterization were available in 46 patients. RESULTS: In all control subjects, the aortic valve closed before the pulmonary valve and the mitral valve closed before the tricuspid valve. In the study group, 30 patients (49%) had reversed aortic-pulmonary valve closure and 27 (90%) of these had a left-sided conduction delay. There were 38 patients (60%) who had reversed mitral-tricuspid valve closure, but this was unrelated to the presence of a left-sided conduction delay. The presence of high ventricular filling pressures and poor systolic function was associated with delayed closure of both the mitral and the tricuspid valve. This caused the closure sequence to be related to the size of the difference between mean pulmonary artery wedge pressure and mean central venous pressure and also the magnitude of right ventricular dysfunction. Patients with a low wedge pressure (< 16 mm Hg) and a low central venous pressure (< 10 mm Hg) had a low prevalence of mitral-tricuspid valve closure reversal (30%). Those with a high wedge pressure (> 16 mm Hg) but a low central venous pressure had a high prevalence (86%) of reversal of mitral-tricuspid valve closure. Patients with high wedge and central venous pressures had a moderate prevalence (47%) of mitral-tricuspid valve closure reversal. Similar findings were observed for right ventricular dysfunction. If the right ventricle was normal or severely dysfunctional, a reversed closure sequence was less common (52% and 41%, respectively) than if moderate dysfunction was present (78%). CONCLUSIONS: Aortic-pulmonary valve closure sequence is strongly related to the presence of a left-sided conduction delay. The mitral-tricuspid valve closure sequence is unrelated to a conduction delay but can be reversed by relative differences in the severity of systolic dysfunction and filling pressures between the two ventricles.

Isovolumic relaxation period in hypertrophic cardiomyopathy.

Previous reports have demonstrated that patients with hypertrophic cardiomyopathy have a prolonged isovolumic relaxation period as a result of a delay in mitral valve opening, reflecting a reduced rate of fall of left ventricular pressure. This period as measured from the aortic closure sound (A2 on phonocardiogram) to the opening of the mitral valve (on echocardiogram) was determined in 84 patients with hypertrophic cardiomyopathy and compared with findings in 31 normal volunteers. The duration of the isovolumic relaxation period in the 84 patients had a wide range from 0 to 160 ms (mean 71 +/- 32) that was not significantly different from that in normal subjects (63 +/- 11 ms). However, it was possible to identify a group of 15 patients with an extremely short isovolumic relaxation period, 2 standard deviations below the normal range. This shortening was due to a marked delay in aortic closure sound (A2) due to late left ventricular-aortic pressure crossover, as well as early opening of the mitral valve secondary to elevated left atrial pressure, which was confirmed by hemodynamic correlations and digitized echocardiographic data. In this subset of patients, A2 is a poor marker of the onset of rapid left ventricular pressure decline and, thus, the interval from A2 to mitral valve opening is not a valid reflection of left ventricular relaxation. It is concluded that in hypertrophic cardiomyopathy, both the timing and sequence of relaxation are abnormal, as is the rate of relaxation. Furthermore, the isovolumic relaxation period is multifactorially determined and depends not only on the rate of left ventricular pressure decline, but also on the magnitude of the pressure drop from A2 to mitral valve opening. All of these determinants must be kept in mind when the isovolumic relaxation period is used as a measure of left ventricular relaxation.

Atrial septal defect: attenuation of respiratory variation in systolic and diastolic time intervals.

The effects of pathologic states on right and left ventricular function have been studied extensively. However, there have been few studies on the interrelations between right and left ventricular function in normal human subjects and in patients with disease. Respiratory effects on ventricular interrelations reflected by diastolic time, right or left ventricular systolic time and ventricular performance (pre-ejection period/ejection time ratio) were studied in 12 normal subjects and 15 patients with a normal pressure-large shunt atrial septal defect. Simultaneous pulmonary artery (intracardiac manometer recordings) and left ventricular external recordings were performed in both groups. Left ventricular diastolic time increased with inspiration in the normal subjects and decreased in the patients with atrial septal defect (12.6 +/- 2.39 [1 SE] versus -13.4 +/- 3.48 ms, p less than 0.001). Left ventricular systolic time and ejection time decreased with inspiration in the normal group and remained unchanged in the patient group (-7.6 +/- 0.95 versus -0.9 +/- 0.77 ms, p less than 0.001 and -10.4 +/- 1.09 versus -1.7 +/- 0.80 ms, p less than 0.001, respectively). Left ventricular pre-ejection period/ejection time ratio increased with inspiration in the normal subjects and remained unchanged in the patients with atrial septal defect (0.03 +/- 0.008 versus 0 +/- 0.01, p less than 0.01). Right ventricular diastolic time decreased with inspiration in normal and patient groups (-8.8 +/- 1.6 versus -17 +/- 3.87 ms).(ABSTRACT TRUNCATED AT 250 WORDS)

Venous systolic thrill and murmur in the neck: a consequence of severe tricuspid insufficiency.

A palpable venous systolic thrill and murmur at the base of the neck are described as new physical findings in five patients with severe tricuspid regurgitation. In two of these patients, the tricuspid valve had been resected as treatment for infective endocarditis related to intravenous drug abuse. The third patient had severe chronic pulmonary disease with right heart failure. The fourth patient had a complex congenital defect in which the mitral valve served as the venous atrioventricular valve and was severely incompetent. The fifth patient suffered from long-standing rheumatic mitral and tricuspid disease with pulmonary hypertension 10 years after placement of a mitral prosthesis. From these observations, it is apparent that pulsatile retrograde flow in the cervical veins resulting from severe right-sided atrioventricular valve incompetence can produce a palpable systolic thrill and murmur at the base of the neck.

Desensitization of adenylate cyclase in Chinese hamster ovary cells transfected with human thyroid-stimulating hormone receptor.

Desensitization or decreased response to the same (homologous) or other stimuli (heterologous) is a well known process. Homologous desensitization to TSH has been demonstrated in normal thyroid tissue. Chinese hamster ovary cells (CHO) transfected with normal human TSH receptor (hTSHR) DNA, in contrast, have been reported not to desensitize. The purpose of our investigation was to determine whether CHO cells transfected with hTSHR desensitize in response to TSH and postreceptor stimulation. CHO cells were stably transfected with plasmid DNA containing hTSHR; nontransfected CHO cells served as the control. TSH (10 mU/ml), 5'-beta,gamma-imido-triphosphate [Gpp(NH)p; 0.1 mM], sodium fluoride (NaF; 10 mM), forskolin (10 microM), and (Bu)2cAMP (100 microM) were used to determine whether homologous or postreceptor heterologous desensitization of adenylate cyclase activity occurred in CHO-transfected cells. Intracellular cAMP accumulation was determined by RIA. Cells were incubated with TSH (to stimulate TSH receptor), Gpp(NH)p, NaF (to stimulate G-protein), forskolin (to stimulate adenylate cyclase activity), and (Bu)2cAMP (nonmetabolized cAMP analog). A second incubation was carried out with TSH (10 mU/ml). Maximal desensitization to either TSH or postreceptor stimulation was observed at 2 h. When transfected CHO cells were preexposed to TSH (10 mU/ml) for 4 h, even the smallest dose of TSH (0.001 mU/ml) caused desensitization. All substances that increased the intracellular cAMP concentration, such as TSH, Gpp(NH)p, NaF, forskolin, and (Bu)2cAMP, caused desensitization. The decrease in the cAMP response to TSH added in the second incubation was 63% less than the initial response to TSH or to postreceptor stimulation (P = 0.0001). In conclusion, desensitization of hTSHR-transfected CHO cells occurs in response to both receptor and postreceptor stimulation that increase cAMP levels. Because hTSHR transfected CHO cells desensitize, no specific thyroid factor(s) other than increased levels of cAMP is required.

Decreased nocturnal levels of prolactin and growth hormone in women with fibromyalgia.

Fibromyalgia (FM) is a complex syndrome, primarily of women, characterized by chronic pain, fatigue, and sleep disturbance. Altered function of the somatotropic axis has been documented in patients with FM, but little is known about nocturnal levels of PRL. As part of a laboratory study of sleep patterns in FM, we measured the serum concentrations of GH and PRL hourly from 2000--0700 h in a sample of 25 women with FM (mean, 46.9 +/- 7.6 yr) and in 21 control women (mean, 42.6 +/- 8.1 yr). The mean (+/-SEM ) serum concentrations (micrograms per L) of GH and of PRL during the early sleep period were higher in control women than in patients with FM [GH, 1.6 +/- 0.4 vs. 0.6 +/- 0.2 (P < 0.05); PRL, 23.2 +/- 2.2 vs. 16.9 +/- 2.0 (P < 0.025)]. The mean serum concentrations of GH and PRL increased more after sleep onset in control women than in patients with FM [GH, 1.3 +/- 0.4 vs. 0.3 +/- 0.2 (P < 0.05); PRL, 16.2 +/- 2.4 vs. 9.7 +/- 1.5 (P < 0.025)]. Sleep efficiency and amounts of sleep or wake stages on the blood draw night were not different between groups. There was a modest inverse relationship between sleep latency and PRL and a direct relationship between sleep efficiency and PRL in FM. There was an inverse relationship between age and GH most evident in control women. Insulin-like growth factor I levels were not different between the groups. These data demonstrate altered functioning of both the somatotropic and lactotropic axes during sleep in FM and support the hypothesis that dysregulated neuroendocrine systems during sleep may play a role in the pathophysiology of FM.

Immunocytochemical detection of p53 in human thyroid carcinomas is associated with mutation and immortalization of cell lines.

Mutations in the tumor suppressor gene p53 are the most-common mutations found in human cancers. In thyroid cancers, p53 mutations generally are found only in poorly differentiated and undifferentiated tumors and in cell lines. To determine the prevalence of p53 mutations in thyroid neoplasms and thyroid cell lines, we screened 58 thyroid tissues and 3 thyroid cell lines, p53 primers bracketing exons 4, 5/6, 7, and 8 were used to amplify genomic DNA using the PCR. Mutations were screened by denaturing gradient gel electrophoresis and confirmed by sequencing. The two papillary thyroid cancer cell lines and the follicular thyroid carcinoma cell line (positive control) had transitions (CGT->CAT) in exon 8, codon 273, resulting in the replacement of arginine with histidine. No normal thyroid tissues or primary tumors from which the cell lines were derived demonstrated exon 8 mutations, using this technique. p53 immunocytochemistry demonstrated a progression of p53 immunopositivity between synchronous and metachronous neoplasms, paralleling the neoplastic progression from a benign adenoma to primary carcinoma, regional, and distant metastasis and ultimately, the cell lines, where intense immunopositivity is noted. In addition, fluorescence in situ hybridization, using probes specific for the p53 locus, revealed the presence of 3 homologues of p53 in the follicular cell line and 2 homologues in the papillary and Hürthle cell lines. These results suggest that a point mutation present in a small number of original tumor cells and amplification of the mutant allele may be responsible for immortalizing well-differentiated thyroid cancer cells into cell lines.

Undesirable cardiac arrhythmias associated with rate hysteresis pacemakers.

The introduction of the ventricular inhibited pulse generator with the feature of rate hysteresis has been associated with a variety of rhythm disturbances, some causing serious concern. This pulse generator has two different pacing rates: (1) the automatic rate, which is the interval between two successive paced beats (usually 860 msec or 70/min), and (2) the hysteresis interval, which results in a 1,000 msec delay between a sensed cardiac contraction and the next pacemaker discharge. The hysteresis interval after a sensed signal may result in long pauses that may predispose to the development of serious cardiac arrhythmias. Two examples of this type of complication were recently observed. One patient had bigeminal rhythm with mechanically ineffective cardiac contractions and an effective cardiac rate of 35/min; he experienced dyspnea and weakness during these prolonged episodes. Another patient had repeated episodes of ventricular fibrillation. The cardiac arrhythmias were not controlled by antiarrhythmic agents, and both patients required replacement of the pulse generator. The proposed advantages of pulse generator hysteresis are (1) prolongation of battery life, and (2) maintenance of effective atrial transport; these advantages may be outweighed by undesirable cardiac arrhythmias that may be associated with this mode of pacemaker function. Rate hysteresis cardiac pacemakers should be reserved for patients having predominantly sinus rhythm without ventricular irritability. In patients with frequent ectopic ventricular activity, atrial fibrillation or high degree atrioventricular block, the rate hysteresis pacemaker offers no advantage over the conventional demand pacemaker. For patients with frequent ectopic ventricular activity not easily controlled by antiarrhythmic agents, consideration should be given to the use of a permanent demand pacemaker with external rate control, which may provide greater flexibility in arrhythmia management.

Effect of heart rate alterations produced by atrial pacing on the pattern of diastolic filling in normal subjects.

To determine the effect of heart rate alterations on diastolic timing intervals and filling parameters, 10 normal patients were paced from the right atrium at 30 and 50 beats/min above their baseline rates. M-mode echocardiograms of the aortic valve, mitral valve and left ventricle were obtained and digitized at baseline and with each pacing rate. With increased atrial pacing, left ventricular systolic time became an increasingly greater proportion of cycle length while the diastolic filling period occupied a lesser proportion of the cycle length. The time to peak filling rate and the rapid filling period occupied a greater proportion of the diastolic filling period. The peak filling rate increased progressively with increased atrial pacing (baseline 128 +/- 19 mm/s, first paced rate 146 +/- 27 mm/s, p less than 0.05 vs baseline; second paced rate 167 +/- 23 mm/s, p less than 0.01 vs baseline and first paced rate). The early diastolic filling fraction and rapid filling fraction also increased with pacing. Increasing the heart rate resulted in an alteration of the time course of diastolic filling and extent of diastolic filling during the rapid filling period. Interventions that improve diastolic filling and increase heart rate may in part be due to heart rate changes.