In vivo axial dynamization of canine tibial fractures using the Securos external skeletal fixation system.

Bilateral transverse mid-shaft tibial osteotomies, with a 4-mm gap, were performed in purpose-bred research dogs and stabilized using a Securos Type 2 external skeletal fixotor (ESF). Full (100%) axial dynamization of one randomly selected ESF in each dog was performed at 31 days postoperatively. Caudo-cranial radiographs were obtained at weekly intervals, which were qualitatively and quantitatively evaluated (densitometry and ImageJ analysis). The dogs were euthanatized 13 weeks postoperatively, at which time dual energy x-ray absorptiometry (DEXA), peripheral quantitative computed tomography (pQCT), mechanical testing in torsion, and qualitative histological analysis were performed. A two-tailed paired Student's t-test was performed for statistical analysis of all parameters of interest, with significance set at p < 0.05. Three of five dynamized bones bridged quicker, and four of five dynamized bones appeared to have greater callus formation, however, statistically significant differences could not be definitively determined. Statistically significant differences were not found with densitometry (any time frame), DEXA, pQCT, torsional stiffness or maximum torque. Despite the lack of statistically relevant data, trends were observed with larger callus size and density in the dynamized tibiae. The dynamized tibiae appeared to fracture more consistently outside of the area of the healing callus as compared to the control tibiae. Histological evaluation showed greater remodelling in four of five control limbs when compared to the dynamized limb. Dynamization at 31 days post-operatively may delay bone remodelling, despite a trend towards a larger callus size. The results of this study failed to show a definitive role for early full axial dynamization.

Leptin, body composition and bone mineral density in premenopausal women.

Body weight is known to be associated with bone mass, however, it is unclear whether body composition, as reflected by the percent of total weight that is fat tissue (%fat), is associated with bone mass independently of weight. Fat tissue is metabolically active, and hormonal factors may mediate an association of %fat with bone mass. Leptin, a hormone produced in fat tissue, has recently been shown to be inversely related to bone mass in mice, but whether it is related to human bone mass is uncertain. We sought to investigate the associations of %fat and of serum leptin concentration with bone mineral density (BMD) in a cohort of 153 premenopausal women. BMD measurements of the total hip, lumbar spine and total body as well as body composition were measured by dual energy X-ray absorptiometry (DXA). Serum leptin levels were established using a commercial competitive binding assay. Individually, body weight, %fat and leptin were each positively associated with BMD at all three sites. However, when we examined BMD either as a function of both body weight and %fat together, or as a function of both body weight and leptin together, we found that for a given body weight, BMD appeared to be inversely associated with %fat and similarly appeared to be inversely associated with leptin. When BMD was examined as a function of %fat and leptin together, we found that for a given %fat, leptin appeared to be inversely associated with BMD. In summary, the results of this study suggest that for a given body weight, a higher proportion of fat and a higher serum leptin concentration have negative associations with bone mass in premenopausal women.

Household tobacco smoke exposure is negatively associated with premenopausal bone mass.

Subjects exposed to environmental tobacco smoke have been found to be at increased risk for several health problems. Whether exposure to passive tobacco smoke is associated with reduced bone mineral density (BMD) is unknown. In order to examine this, we measured BMD in 154 healthy premenopausal women (age range 40-45 years). BMD of the total hip, femoral neck, lumbar spine and total body was measured by dual-energy X-ray absorptiometry (DXA). Data were collected on exposure to household tobacco smoke from age 10 years to the present as well as on other lifestyle factors related to bone mass. We found that 67.5% of the subjects had a history of household tobacco smoke exposure. Subjects exposed to household tobacco smoke had a mean adjusted BMD that was significantly lower at the total hip ( p = 0.021) and femoral neck ( p = 0.018) compared with subjects who were not exposed. In addition, duration of household tobacco smoke exposure was negatively associated with BMD at the total hip ( p = 0.010), femoral neck ( p = 0.004), lumbar spine ( p = 0.037) and total body ( p = 0.031). Subjects exposed to household tobacco smoke for 15 years or more had mean adjusted BMD that was 4% lower at the total body, and more than 8% lower at the total hip, femoral neck and lumbar spine, compared with subjects who were not exposed. In conclusion, household tobacco smoke exposure during adolescence and young adulthood was found to be negatively associated with BMD at the total hip and femoral neck, and duration of exposure was negatively associated with BMD at the total hip, femoral neck, lumbar spine and total body in premenopausal women.

Recall of early menstrual history and menarcheal body size: after 30 years, how well do women remember?

The validity of recall of early menstrual characteristics is of interest because of their putative role in the etiology of breast cancer and other diseases. A retrospective follow-up of the Newton Girls Study (1965-1975) provided an opportunity to assess the accuracy and precision of recall of several early menstrual characteristics. In 1998-1999, 57 percent of the original 793 Newton Girls Study participants completed a mailed questionnaire to assess the accuracy of recall for age and body size at menarche, usual cycle length during the first 2 years, and age at regularity. Recalled and original age at menarche were highly correlated (r = 0.79, p < 0.001). The body mass index percentile at menarche was well correlated with recalled body size at menarche (r = 0.61, p < 0.001), but with some evidence of systematic bias. Overall, a woman's recall of menarcheal age and body size was better than recall of cycle length and occurrence of regularity. The failure to identify certain menstrual characteristics as exposures for subsequent disease may reflect limitations in the accuracy and precision of the recalled measures.

Peripheral vascular endothelial function testing as a noninvasive indicator of coronary artery disease.

OBJECTIVES: We studied whether assessment of endothelium-dependent vasomotion (EDV) with brachial artery ultrasound (BAUS) imaging predicts the presence or absence of coronary artery disease (CAD) as defined by exercise myocardial perfusion imaging (ExMPI). BACKGROUND: Abnormalities in EDV can be detected in arteries before the development of overt atherosclerosis, and its presence may predict poor long-term prognosis. Brachial artery ultrasound during reactive hyperemia is a noninvasive method of assessing peripheral EDV. METHODS: Clinically-indicated ExMPI along with BAUS were performed in 94 subjects (43 women, 51 men). Coronary artery disease was defined by myocardial ischemia or infarction on single photon emission computed tomography images. Flow-mediated dilation (FMD) after upper arm occlusion was defined as the percent change in arterial diameter during reactive hyperemia relative to the baseline. RESULTS: Subjects with CAD by ExMPI (n = 23) had a lower FMD (6.3 +/- 0.7%) than those without CAD by ExMPI (n = 71) (10.5 +/- 0.6%; p = 0.0004). Flow-mediated dilation was highly predictive for CAD with an odds ratio of 1.32 for each percent decrease in FMD (p = 0.001). Based on a receiver-operator analysis, an FMD of 10% was used as a cut-point for further analysis. Twenty-one of 23 subjects who were positive for ExMPI had an FMD < 10% (sensitivity 91%), whereas only two of 40 subjects with an FMD > or =10% were ExMPI-positive (negative predictive value: 95%). There was a correlation between the number of cardiac risk factors and FMD. Individuals with an FMD < 10% exercised for a shorter duration than those with an FMD > or =10% (456 +/- 24 vs. 544 +/- 31 s, respectively; p = 0.02). CONCLUSIONS: Assessment of EDV with BAUS has a high sensitivity and an excellent negative predictive value for CAD and, thus, has the potential for use as a screening tool to exclude CAD in low-risk subjects. Further standardization of BAUS is required, however, before specific cut-points for excluding CAD can be established.

Reduced kidney function and anemia as risk factors for mortality in patients with left ventricular dysfunction.

OBJECTIVES: We sought to evaluate the relationship between the level of kidney function, level of hematocrit and their interaction on all-cause mortality in patients with left ventricular (LV) dysfunction. BACKGROUND: Anemia and reduced kidney function occur frequently in patients with heart failure. The level of hematocrit and its relationship with renal function have not been evaluated as risk factors for mortality in patients with LV dysfunction. METHODS: We retrospectively examined the Studies Of LV Dysfunction (SOLVD) database. Glomerular filtration rate (GFR) was predicted using a recently validated formula. Kaplan-Meier survival analyses were used to compare survival times between groups stratified by level of kidney function (predicted GFR) and hematocrit. Cox proportional-hazards regression was used to explore the relationship of survival time to level of kidney function, hematocrit and their interaction. RESULTS: Lower GFR and hematocrit were associated with a higher prevalence of traditional cardiovascular risk factors. In univariate analysis, reduced kidney function and lower hematocrit, in men and in women, were risk factors for all-cause mortality (p < 0.001 for both). After adjustment for other factors significant in univariate analysis, a 10 ml/min/1.73 m(2) lower GFR and a 1% lower hematocrit were associated with a 1.064 (95% CI: 1.033, 1.096) and 1.027 (95% CI: 1.015, 1.038) higher risk for mortality, respectively. At lower GFR and lower hematocrit, the risk was higher (p = 0.022 for the interaction) than that predicted by both factors independently. CONCLUSIONS: Decreased kidney function and anemia are risk factors for all-cause mortality in patients with LV dysfunction, especially when both are present. These relationships need to be confirmed in additional studies.

Weight and body mass index at menarche are associated with premenopausal bone mass.

Adolescence is a critical time for skeletal growth and mineralization. Exposure to protective or detrimental factors during this period may influence peak bone mass attainment and subsequent development of osteoporosis. In order to evaluate the association of body size during adolescence with subsequent adult bone mass, we conducted a follow-up study of a community-based cohort of girls who participated in a growth and sexual maturation study 30 years ago. Data from the original study included age at menarche, height at menarche and weight at menarche. Follow-up evaluation of 119 subjects, now premenopausal women ages 40-45 years, included bone mineral density (BMD) measurements of the total body, lumbar spine, femoral neck, total hip, and ultradistal radius by dual-energy X-ray absorptiometry. After adjustment for current adult weight and other factors related to bone mass, weight at menarche was found to be positively associated with subsequent adult BMD. Similarly, body mass index (BMI) at menarche was positively associated with adult BMD. In contrast, age at menarche was not found to predict adult BMD. When the subjects were divided into quartiles based on their BMI at menarche, subjects in the lowest quartile of BMI at menarche had adult mean BMD that was 8-15% lower at the measured sites compared with subjects in the highest quartile of BMI at menarche. In conclusion, low body weight and low BMI at menarche appear to be significant predictors of reduced bone mass in healthy premenopausal women ages 40-45 years.

Evaluation of a discontinuous treatment protocol (VELCAP-S) for canine lymphoma.

Eighty-two dogs with lymphoma received a single 15-week course of chemotherapy, after which treatment was ceased until relapse. Fifty-six dogs (68%) achieved complete remission for a median 1st remission duration of 20 weeks. Forty-eight dogs relapsed, of which 30 repeated the induction cycle. In 22 of these dogs, 1st remission had been short, and they received maintenance chemotherapy; the other 8 dogs received 2 or 3 cycles of induction chemotherapy. Second remission rate for these 30 dogs was 87% (26 dogs). Overall disease control for the 38 dogs that remained on protocol was 44 weeks, which was not markedly shorter than for dogs treated with a previously reported protocol in which maintenance chemotherapy was instituted in all dogs after an identical 1st induction (VELCAP-L). Dogs that were febrile and dogs that were dyspneic were less likely to achieve a complete remission to induction chemotherapy. Of dogs that achieved a complete remission, those that were thrombocytopenic at entry had a shorter 1st remission, and dogs that were anorexic at entry had shorter overall disease control. There was a correlation between 1st remission duration and length of any subsequent remission obtained. The incidence of toxicity was high, particularly after the combination of doxorubicin and vincristine. Dose reductions because of toxicity did not markedly reduce remission duration. We conclude that discontinuous chemotherapy may reduce patient visits in a small number of patients because of long-term disease control. Delaying maintenance chemotherapy until after 2nd remission is achieved does not markedly affect overall disease control.

Neuropeptide Y has a central inhibitory action on the hypothalamic-pituitary-thyroid axis.

Recent evidence suggests that neuropeptide Y (NPY), originating in neurons in the hypothalamic arcuate nucleus, is an important mediator of the effects of leptin on the central nervous system. As these NPY neurons innervate hypophysiotropic neurons in the hypothalamic paraventricular nucleus (PVN) that produce the tripeptide, TRH, we raised the possibility that NPY may be responsible for resetting of the hypothalamic-pituitary-thyroid (HPT) axis during fasting. To test this hypothesis, the effects of intracerebroventricularly administered NPY on circulating thyroid hormone levels and proTRH messenger RNA in the PVN were studied by RIA and in situ hybridization histochemistry, respectively. NPY administration suppressed circulating levels of thyroid hormone (T(3) and T(4)) and resulted in an inappropriately normal or low TSH. These alterations were associated with a significant suppression of proTRH messenger RNA in the PVN, indicating that NPY infusion had resulted in a state of central hypothyroidism. Similar observations were made in NPY-infused animals pair fed to the vehicle-treated controls. These data are reminiscent of the effect of fasting on the thyroid axis and indicate that NPY may play a major role in the inhibition of HPT axis during fasting.

Single agent chemotherapy with doxorubicin for feline lymphoma: a retrospective study of 19 cases (1994-1997).

Medical records of 21 cats with confirmed lymphoma treated with single-agent doxorubicin were reviewed. Nineteen cats met the inclusion criteria for this retrospective study. Doxorubicin was given at a dosage of 25 mg/m2 (n = 8) or 1 mg/kg (n = 11) IV, every 3 weeks for a total of 5 treatments. Four of 16 tested cats were positive for feline leukemia virus (FeLV) and all 16 cats tested negative for feline immunodeficiency virus. Eight of the 19 cats (42%) responded to doxorubicin for a median duration of 64 days (range, 35-575 days). Five cats (26%) achieved a complete response (CR) to doxorubicin for a median duration of 92 days (range, 54-575 days). Partial response was observed in 3 cats. Institution was the only significant prognostic indicator for response, with cats treated at Colorado State University being more likely to achieve CR than cats treated at Tufts University. Cats that achieved CR to doxorubicin and FeLV-negative cats had significantly longer survival times. Loss of appetite was the most common toxicity, observed in 9 cats (47%), and was severe in 5 cats (26%). Other toxicoses were less frequent and included vomiting, diarrhea, and myelosuppression. Doxorubicin was not very effective at inducing and maintaining remission in the cats in this study. Therefore, if doxorubicin is used for the treatment of feline lymphoma, it should be combined with other effective chemotherapeutic drugs in a combination protocol.

Prognostic significance of serum alkaline phosphatase activity in canine appendicular osteosarcoma.

Sixty-one dogs with appendicular osteosarcoma were treated with amputation and chemotherapy of cisplatin and doxorubicin. Serum samples were obtained before and after treatment for determination of total alkaline phosphatase (TALP) activity as well as the activities of the constituent bone (BALP), liver (LALP), and corticosteroid-induced (CALP) isoenzymes. The relationship between alkaline phosphatase activities and survival was examined by Cox proportional hazards regression analysis and Kaplan-Meier log rank analysis. Mean activity of TALP, BALP, and LALP decreased significantly after treatment (P < .001). TALP and LALP activities before treatment were significantly correlated with survival (P = .006 and .001, respectively). The correlation between BALP activity before treatment and survival approached significance (P = .054). CALP activity and TALP, BALP, and LALP activities after treatment were not significantly correlated with survival. Dogs with normal pretreatment TALP and BALP activities survived significantly longer than dogs with increased pretreatment activities (P = .001 and .003, respectively). Median survival times for dogs with normal or increased TALP activities before treatment were 12.5 and 5.5 months, respectively; and median survival times for dogs with normal or increased BALP activities before treatment were 16.6 and 9.5 months, respectively. In the design of future clinical trials involving dogs with osteosarcoma, consideration should be given to stratifying the randomization according to alkaline phosphatase activity. In addition, alkaline phosphatase activity should be a factor considered by clinicians attempting to tailor the aggressiveness of adjuvant chemotherapy to the needs of individual patients or owners.

Effect of eye rubbing on signs and symptoms of allergic conjunctivitis in cat-sensitive individuals.

OBJECTIVE: To evaluate the effect of eye rubbing on signs and symptoms of allergic conjunctivitis in cat-sensitive individuals. DESIGN: Two prospective, nonrandomized comparative studies. PARTICIPANTS: Thirteen patients in the first study and 20 patients in the second study with a documented history of acute allergic conjunctivitis induced by exposure to cats were enrolled. INTERVENTION: In the first trial, all patients had one eye rubbed 15 times by the investigator without exposure to airborne allergens. Both eyes were evaluated after 5, 15, 30, and 60 minutes using subject questionnaires and slit-lamp examination. At least 1 week later, each patient was exposed to cat dander for 75 minutes; 15 minutes after entering the cat room, each patient had one eye rubbed 15 times by the examiner. Subjects' eyes were then evaluated using questionnaires and slit-lamp examination. In the second trial, the visits were identical to the first trial, except that the rubbed eye in each visit was rubbed 20 times and with more force, and that patients wore masks during exposure to cat dander. MAIN OUTCOME MEASURES: In both studies, the difference between patients' rubbed and nonrubbed eyes with respect to ocular itching, chemosis, and hyperemia was noted 5, 15, 30, and 60 minutes after controlled eye rubbing. RESULTS: Without exposure to the cat room, rubbed eyes exhibited increased itching at 5 minutes in both studies and at 15 minutes in the second study (P < 0.05), increased chemosis at 5 and 15 minutes in the second study (P < 0.05), and increased hyperemia at 5 minutes in the second study (P < 0.05) compared with nonrubbed eyes. During exposure to cat dander, rubbed eyes consistently exhibited increased itching at 5, 15, and 30 minutes in the first and second study as well as at 60 minutes in the second study compared with nonrubbed eyes. Furthermore, during exposure to cat dander, rubbed eyes consistently exhibited increased chemosis at 5 and 15 minutes (P < 0.05) and increased hyperemia at 5, 15, and 30 minutes (P < 0.05) in the second study compared with nonrubbed eyes. CONCLUSIONS: Firm eye rubbing causes a mild and transient increase in ocular itching, chemosis, and hyperemia. However, after exposure to cat allergens in cat-sensitive individuals, the effects of eye rubbing are longer and more dramatic. Eye rubbing may play a role in ocular signs and symptoms of allergic conjunctivitis in cat-sensitive individuals, especially after exposure to cat dander.

Effect of dietary protein content and tryptophan supplementation on dominance aggression, territorial aggression, and hyperactivity in dogs.

OBJECTIVE: To evaluate the effect of high- and low-protein diets with or without tryptophan supplementation on behavior of dogs with dominance aggression, territorial aggression, and hyperactivity. DESIGN: Prospective crossover study. ANIMALS: 11 dogs with dominance aggression, 11 dogs with territorial aggression, and 11 dogs with hyperactivity. PROCEDURE: In each group, 4 diets were fed for 1 weeks each in random order with a transition period of not < 3 days between each diet. Two diets had low protein content (approximately 18%), and 2 diets had high protein content (approximately 30%). Two of the diets (1 low-protein and 1 high-protein) were supplemented with tryptophan. Owners scored their dog's behavior daily by use of customized behavioral score sheets. Mean weekly values of 5 behavioral measures and serum concentrations of serotonin and tryptophan were determined at the end of each dietary period. RESULTS: For dominance aggression, behavioral scores were highest in dogs fed unsupplemented high-protein rations. For territorial aggression, [corrected] tryptophan-supplemented low-protein diets were associated with significantly lower behavioral scores than low-protein diets without tryptophan supplements. CONCLUSIONS AND CLINICAL RELEVANCE: For dogs with dominance aggression, the addition of tryptophan to high-protein diets or change to a low-protein diet may reduce aggression. For dogs with territorial aggression, tryptophan supplementation of a low-protein diet may be helpful in reducing aggression.

alpha-Melanocyte stimulating hormone prevents fasting-induced suppression of corticotropin-releasing hormone gene expression in the rat hypothalamic paraventricular nucleus.

During fasting, corticotropin-releasing hormone (CRH) mRNA decreases in the hypothalamic paraventricular nucleus (PVN), but the mechanism by which this takes place is not well understood. To test the hypothesis that the melanocortin system may be involved in the regulation of CRH mRNA in the PVN during fasting, the effect of intracerebroventricularly administered alpha-melanocyte stimulating hormone (MSH) on CRH mRNA in the PVN was studied in fasted animals by in situ hybridization histochemistry. Whereas fasting suppressed CRH mRNA levels in the PVN, alpha-MSH at doses of 150 and 300 ng every 6 h for 64 h prevented the fasting-induced suppression of CRH gene expression in the PVN. These data indicate that the suppression of alpha-MSH synthesis may be responsible for the decreased CRH gene expression in the PVN during fasting.

The role of the community teaching hospital in surgical undergraduate education.

BACKGROUND: Significant changes in surgical practice have resulted in a reexamination of surgical undergraduate education. The increasing emphasis toward ambulatory procedures positions the community hospital as an excellent alternative site for surgical education. This study compares the quality of one medical school's surgical education at a principal teaching hospital to that of affiliated teaching hospitals. METHODS: Surgical undergraduate education offered through four programs was evaluated for 1993 to 1997. Students' performance was objectively rated by the National Board Examination in surgery, an oral examination, and a clinical appraisal. A subjective appraisal was determined via students' clerkship evaluation. RESULTS: There was a significant difference (P <0.01) in National Board Examination scores and clerkship evaluations that favored some affiliated teaching hospitals over the principal teaching hospital. CONCLUSION: The quality of surgical undergraduate education, documented by objective testing and subjective perception, indicated that the education obtained at the affiliated hospitals was at least equivalent to the principal teaching hospital.

Survivors of catastrophic illness: outcome after direct transfer from intensive care to extended care facilities.

OBJECTIVE: To describe outcomes of adult survivors of prolonged critical illness after direct transfer to extended care facilities. DESIGN: A retrospective cohort study. SETTING: All adult intensive care units (ICUs) in a tertiary care university hospital. PATIENTS: A consecutive series of 97 adult survivors with an ICU stay of > or =7 days transferred directly from intensive care to extended care facilities between 1990 and 1996. INTERVENTIONS: None. METHODS AND MAIN RESULTS: Hospital and extended care facility charts were reviewed for patient characteristics, resource utilization, and survival. Survivors were for a minimum of 1 yr and a maximum of 6 yrs, and were interviewed to assess quality of life and functionality. The mean age of the patients was 66+/-16 (range, 19-93) yrs. The median length of ICU stay for these patients was 39 (range, 7-276) days. Only 18 of the 71 ventilator-assisted patients were weaned from mechanical ventilation after transfer to the extended care facility. Survival for the study period, at 1 yr after discharge from the ICU, was 49.5%. One year after discharge from the ICU, 11.5% of all patients had returned home, were breathing spontaneously, had a fair or better quality of life, and had good physical functionality. Each successive year, an increasing proportion of patients underwent direct transfer to an extended care facility. This strategy decreased the patients' length of stay (p<.002) in the ICU from year to year, but was significantly associated with an increase in readmissions to acute care hospitals (p<.002). CONCLUSIONS: Survivors of catastrophic illness who are so debilitated that they require transfer to an extended care facility have a low likelihood of achieving both survival and functional independence 1 yr after discharge from the ICU. Aggressive cost-conscious strategies to accelerate the transfer of these patients successfully reduced the length of ICU stay and hospital costs, but were associated with a high rate of readmission to tertiary care facilities.

alpha-Melanocyte-stimulating hormone is contained in nerve terminals innervating thyrotropin-releasing hormone-synthesizing neurons in the hypothalamic paraventricular nucleus and prevents fasting-induced suppression of prothyrotropin-releasing hormone gene expression.

The hypothalamic arcuate nucleus has an essential role in mediating the homeostatic responses of the thyroid axis to fasting by altering the sensitivity of prothyrotropin-releasing hormone (pro-TRH) gene expression in the paraventricular nucleus (PVN) to feedback regulation by thyroid hormone. Because agouti-related protein (AGRP), a leptin-regulated, arcuate nucleus-derived peptide with alpha-MSH antagonist activity, is contained in axon terminals that terminate on TRH neurons in the PVN, we raised the possibility that alpha-MSH may also participate in the mechanism by which leptin influences pro-TRH gene expression. By double-labeling immunocytochemistry, alpha-MSH-IR axon varicosities were juxtaposed to approximately 70% of pro-TRH neurons in the anterior and periventricular parvocellular subdivisions of the PVN and to 34% of pro-TRH neurons in the medial parvocellular subdivision, establishing synaptic contacts both on the cell soma and dendrites. All pro-TRH neurons receiving contacts by alpha-MSH-containing fibers also were innervated by axons containing AGRP. The intracerebroventricular infusion of 300 ng of alpha-MSH every 6 hr for 3 d prevented fasting-induced suppression of pro-TRH in the PVN but had no effect on AGRP mRNA in the arcuate nucleus. alpha-MSH also increased circulating levels of free thyroxine (T4) 2.5-fold over the levels in fasted controls, but free T4 did not reach the levels in fed controls. These data suggest that alpha-MSH has an important role in the activation of pro-TRH gene expression in hypophysiotropic neurons via either a mono- and/or multisynaptic pathway to the PVN, but factors in addition to alpha-MSH also contribute to the mechanism by which leptin administration restores thyroid hormone levels to normal in fasted animals.

Association of cocaine- and amphetamine-regulated transcript-immunoreactive elements with thyrotropin-releasing hormone-synthesizing neurons in the hypothalamic paraventricular nucleus and its role in the regulation of the hypothalamic-pituitary-thyroid axis during fasting.

Because cocaine- and amphetamine-regulated transcript (CART) coexists with alpha-melanocyte stimulating hormone (alpha-MSH) in the arcuate nucleus neurons and we have recently demonstrated that alpha-MSH innervates TRH-synthesizing neurons in the hypothalamic paraventricular nucleus (PVN), we raised the possibility that CART may also be contained in fibers that innervate hypophysiotropic thyrotropin-releasing hormone (TRH) neurons and modulate TRH gene expression. Triple-labeling fluorescent in situ hybridization and immunofluorescence were performed to reveal the morphological relationships between pro-TRH mRNA-containing neurons and CART- and alpha-MSH-immunoreactive (IR) axons. CART-IR axons densely innervated the majority of pro-TRH mRNA-containing neurons in all parvocellular subdivisions of the PVN and established asymmetric synaptic specializations with pro-TRH neurons. However, whereas all alpha-MSH-IR axons in the PVN contained CART-IR, only a portion of CART-IR axons in contact with pro-TRH neurons were immunoreactive for alpha-MSH. In the medial and periventricular parvocellular subdivisions of the PVN, CART was co-contained in approximately 80% of pro-TRH neuronal perikarya, whereas colocalization with pro-TRH was found in <10% of the anterior parvocellular subdivision neurons. In addition, >80% of TRH/CART neurons in the periventricular and medial parvocellular subdivisions accumulated Fluoro-Gold after systemic administration, suggesting that CART may serve as a marker for hypophysiotropic TRH neurons. CART prevented fasting-induced suppression of pro-TRH in the PVN when administered intracerebroventricularly and increased the content of TRH in hypothalamic cell cultures. These studies establish an anatomical association between CART and pro-TRH-producing neurons in the PVN and demonstrate that CART has a stimulatory effect on hypophysiotropic TRH neurons by increasing pro-TRH gene expression and the biosynthesis of TRH.

Statistical analysis of nitrogen balance data with reference to the lysine requirement in adults.

Statistical analysis of nitrogen balance data is an important approach to the estimation of human nutrient requirement. The usual procedure is to regress nitrogen (N) balance on intake and to define the requirement as intake that would produce zero balance. In the actual application of this methodology, there are a number of options, and in the present study we explore the sensitivity of the regression approach to those options. To put this problem into a realistic context, we examine the current controversy over the lysine requirements of healthy adults. From early N balance studies, investigators concluded that the mean requirement was generally less than 10 mg. kg(-1). d(-1), whereas based on recent (13)C-tracer and metabolic studies, we propose a tentative mean requirement of approximately 30 mg. kg(-1). d(-1). Jones et al. (1956) conducted careful N balance studies from which they derived an estimate of lysine requirement of less than about 8 mg. kg(-1). d(-1). We reanalyzed these data with different choices of modes of analysis, mathematical models, and different assumptions concerning the magnitude of miscellaneous N losses. We find that for these data the choice of a specific mathematical model has only a small effect on resultant estimates of requirement, while estimated requirements are very sensitive to amount of unmeasured losses that are assumed and how the model is applied (whether the aggregate data are fitted in one pass to a single model, or the data for each individual subject are fitted to that individual's unique model). Moreover, our reanalysis suggests that the population requirement for lysine is in the range of 17 to 36 mg. kg(-1). d(-1) and strongly supports a lysine requirement value of about 30 mg. kg(-1). d(-1). In general, our results indicate that whenever possible, N balance data should be analyzed using a square root model fitted to individual data and that the median of the individual requirements so derived be used as the estimate of population requirement. Moreover, clearly any statistical analyses of N balance data should include a sensitivity analysis to determine the influence of underlying assumptions. Finally, the finding that these estimates are highly dependent on the assumed amount of N miscellaneous losses recommends that further studies on these losses and of the factors that influence them are essential.