New tools for studying osteoarthritis genetics in zebrafish.

OBJECTIVE: Increasing evidence points to a strong genetic component to osteoarthritis (OA) and that certain changes that occur in osteoarthritic cartilage recapitulate the developmental process of endochondral ossification. As zebrafish are a well validated model for genetic studies and developmental biology, our objective was to establish the spatiotemporal expression pattern of a number of OA susceptibility genes in the larval zebrafish providing a platform for functional studies into the role of these genes in OA. DESIGN: We identified the zebrafish homologues for Mcf2l, Gdf5, PthrP/Pthlh, Col9a2, and Col10a1 from the Ensembl genome browser. Labelled probes were generated for these genes and in situ hybridisations were performed on wild type zebrafish larvae. In addition, we generated transgenic reporter lines by modification of bacterial artificial chromosomes (BACs) containing full length promoters for col2a1 and col10a1. RESULTS: For the first time, we show the spatiotemporal expression pattern of Mcf2l. Furthermore, we show that all six putative OA genes are dynamically expressed during zebrafish larval development, and that all are expressed in the developing skeletal system. Furthermore, we demonstrate that the transgenic reporters we have generated for col2a1 and col10a1 can be used to visualise chondrocyte hypertrophy in vivo. CONCLUSION: In this study we describe the expression pattern of six OA susceptibility genes in zebrafish larvae and the generation of two new transgenic lines marking chondrocytes at different stages of maturation. Moreover, the tools used demonstrate the utility of the zebrafish model for functional studies on genes identified as playing a role in OA.

Size and asymmetry: are there costs to winning the royalty race?

Body size and morphology are key fitness-determining traits that can vary genotypically. They are likely to be important in social insect queens, which mate in swarms and found colonies independently, but genetic influences on queen morphology have been little investigated. Here, we show that the body size and morphology of queens are influenced by their genotype in the leaf-cutting ant Acromyrmex echinatior, a species in which certain lineages (patrilines) bias their development towards reproductive queens rather than sterile workers. We found no relationship between the queen-worker skew of patrilines and the size or morphology of queens, but there was a significant relationship with fluctuating asymmetry, which was greater in more queen-biased patrilines. Our results suggest that queen-biased patrilines do not incur a fitness cost in terms of body size, but may face more subtle costs in developmental stability. Such costs may constrain the evolution of royal cheating in social insects.

Study of exclusive charmless semileptonic B decays and |Vub|.

We study semileptonic B decay to the exclusive charmless states pi, rho/omega, eta, and eta;{'} using the 16 fb(-1) CLEO Upsilon(4S) data sample. We find B(B0-->pi-l+nu)=(1.37+/-0.15stat+/-0.11sys)x10(-4) and B(B0-->rho-l+nu)=(2.93+/-0.37stat+/-0.37sys)x10(-4) and find evidence for B+-->eta'l+nu, with B(B+-->eta'l+nu)=(2.66+/-0.80stat+/-0.56sys)x10(-4). From our B-->pilnu rate for q2>16 GeV2 and lattice QCD, we find |Vub|=(3.6+/0.4stat+/0.2syst-0.4thy+0.6)x10(-3) [corrected]

Comparative evaluation of treated bovine pericardium as a xenograft for hernia repair.

Two forms of bovine pericardium (BPC) were assessed as hernia repair materials: non-cross-linked (lyophilized) and cross-linked through treatment with glutaraldehyde (GA). These were compared with polypropylene mesh (Marlex) in a rabbit model. Over 52 wk implantation, the GA BPC grafts developed a strong, stable, fibrous tissue replacement with good incorporation into the abdominal muscle wall. The lyophilized BPC grafts were substantially resorbed within 12 wk of implantation, however the thin, fibrous replacement tissue was inadequate for abdominal wall support. Marlex grafts provided sufficient abdominal support, however these grafts were associated with extensive adhesion formation and, in this model, fat deposition around the perimeter of the graft. Control (ungrafted) rabbit abdominal muscle in the transverse orientation had an ultimate tensile load (UTL) of 11.4 +/- 5.1 N (x +/- s.d.) and a strain at UTL of 35 +/- 12% (n = 169). At 52 weeks the UTL of the repair sites was 7.3 +/- 4.5 N (n = 6), 5.1 +/- 3.5 N (n = 6) and 5.6 +/- 2.7 N (n = 6) for GA BPC, lypophilized BPC and Marlex grafts, respectively.

The relationship of exercise and diet on high-density lipoprotein cholesterol levels in women.

The relationship of exercise and diet on high-density lipoprotein (HDL) cholesterol was investigated in 45 long-distance runners (LD), 49 joggers (J), and 47 inactive (I) women. Fasting plasma triglycerides (TG), HDL cholesterol, total cholesterol (TC), and percent body fat (%BF) were measured in women ages 24-58 yr. TG levels were significantly lower in LD compared to I (p less than 0.02). Although TC was not significantly different among groups, HDL-cholesterol was higher in LD (78 mg/dl) compared to J (70 mg/dl) or I (62 mg/dl) (p less than 0.001). Multiple regression analyses indicated that alterations of plasma lipids and lipoprotein levels could not be attributed to intake differences of nutrients. Distance run and %BF were the strongest predictors of HDL-cholesterol in women. LD (23 %BF) were leaner than J (26 %BF) or I (30 %BF); however, when results were adjusted for %BF, significant differences between exercise groups remained for HDL cholesterol.

The toxicity of cyanobacterial toxins in the mouse: I microcystin-LR.

Blooms of cyanobacteria or blue-green algae are known to have caused poisoning in fish, waterfowl, animals and man. One of the toxins responsible for this is the hepatotoxin microcystin-LR which has been found to occur in blooms present intermittently in sources used for domestic water supplies. Three sets of experiments were undertaken to investigate the acute toxicity of microcystin-LR in mice and rats by the oral and intraperitoneal routes, the potential for effects on foetal development in the mouse, and the effects of repeated oral dosing over 13 weeks in the mouse. The results of this work were as follows: (1) Microcystin-LR is 30-100 times less toxic via oral ingestion than via intraperitoneal injection; (2) Microcystin-LR is not a selective developmental toxicant in the mouse. There was a No Observed Adverse Effect Level (NOAEL) of 600 microg kg(-1) bodyweight per day given on days 6-15 of pregnancy for any form of developmental toxicity; (3) There was a clear NOAEL for tissue damage in the liver of 40 microg kg(-1) bodyweight per day of microcystin-LR. Using this data, a value of 1 microg l(-1) microcystin-LR would be an appropriate guideline value for drinking water.

The toxicity of cyanobacterial toxins in the mouse: II anatoxin-a.

Blooms of cyanobacteria are known to have caused poisoning in fish, waterfowl, animals and man. One of the low molecular weight toxins responsible for this is the neurotoxin anatoxin-a which has been detected in reservoirs used for domestic water supplies. While the acute behaviour of this alkaloid is clear, there is uncertainty regarding the effects on man of ingestion of anatoxin-a at low levels over longer periods. In order to assess this risk, a series of in vitro and in vivo experiments were undertaken to investigate the pharmacology, subacute toxicity, and the teratogenicity of anatoxin-a in the mouse. The results of this work were as follows: (1) Pharmacological screening studies confirmed that anatoxin-a is a potent nicotinic agonist which can produce neuromuscular blockade and death by respiratory arrest. Recovery from a single sub-lethal dose is rapid and complete; (2) Repeated sub-lethal oral administration over 28 days in the mouse did not produce any reliable evidence of treatment-related toxicity; (3) From a preliminary screening study anatoxin-a does not appear to be a developmental toxicant in the mouse. These results indicate that a guideline value for anatoxin-a in drinking water of 1 microg l(-1) would provide an adequate margin of safety.

Cardiovascular epidemiology, exercise, and health: 40-year followup of the U.S. Navy's "1000 aviators".

The interrelationship of aging, performance, and stress modification has been the subject of investigations in the U.S. Navy. Beginning in 1940, a study of 1,056 student and instructor pilots lowered previously high attrition rates in training by emphasizing both physical and psychological screening. After World War II, when 208 pilots in the group died, followup studies of the survivors were conducted in 1951, 1957, 1963, 1969-71, 1977, and 1980-81. In February 1981, 715 questionnaires were mailed to known survivors, with 500 replies subsequently analyzed. Additionally, 114 of the respondents who had previously been examined during 1969, were again examined in 1980-81; those individuals were markedly different in their lifestyle, particularly in exercising regularly, abstaining from cigarette smoking, and drinking alcoholic beverages moderately, as contrasted to 28 aviators also examined in 1969 who died in the interim. Healthy lifestyle may alter cardiovascular risk, preventing premature death.

Longevity in military pilots: 37-year followup of the Navy's "1000 aviators".

The 37-year nonmilitary mortality rate for initially healthy aviators was determined in a followup program on the U.S. Navy's "1000 Aviator" cohort. Of the 800 survivors of World War II and the Korean conflict, 95 were found to have died from nonmilitary causes over this followup period. This is markedly less than the 208 that would be expected from a random sample of white American men over a similar period (p less than 0.005). It is also significantly less than the 143 that would have been expected from a group of men who had passed an initial insurance physical (p less than 0.005). Lower-than-expected death rates occurred in all three major categories of cause of death in this age group: cardiovascular, neoplastic, and accidental. The generally good socioeconomic background, the positive genetic influence of long-lived parents, the above average intelligence, and the health and fitness orientation of the military aviator are all thought to be factors contributing to this increased longevity.

Eight-year follow-up of exercise electrocardiograms in healthy, middle-aged aviators.

To study to prognostic capabilities of the exercise electrocardiogram (ECG) in a fit, healthy, middle-aged population, 548 members of the U.S. Navy's "1000 Aviator" cohort were exercised to 85% predicted maximum heart rate in 1969 and then followed-up in 1977 for the development of clinically evident coronary artery disease (CAD). Of these subjects, 23 (4.2%) had significant ST depression during their exercise test in 1969. At the end of the 8-year follow-up period, 38 of the 548 subjects (6.9%) had developed clinically evident CAD. The sensitivity (percent of disease predicted by an abnormal test) and predictive value (percent of abnormal tests predictive of disease) of an abnormal exercise test were 15.7% and 26%, respectively. We conclude that even in a carefully screened aviator population with a low risk for CAD, a single normal exercise ECG does not exclude the presence of latent CAD. Furthermore, in this population, a single abnormal exercise ECG should not be a disqualifying defect without further work-up.

Long-term follow-up of lung volume measurements in initially healthy young aviators.

Lung volume measurements on a large number of initially healthy young military aviators (the U.S. Navy's "1000 Aviator" cohort) were recorded periodically in follow-up from 1940-69. Vital capacities were measured spirometrically and total lung capacities were measured planimetrically from chest roentgenograms. Residual volumes were calculated by subtracting the vital capacity from the total lung capacity in each subject. Additional variables available for analysis were cigarette smoking histories, family histories, aviation career patterns, pulmonary symptoms, cardiac disease diagnoses, and anthropometric measurements. Multiple linear regression techniques were used on these variables to construct prediction equations for each lung volume in 1969. From these longitudinal analyses, cigarette smoking and pulmonary symptoms were found to be associated with an "obstructive lung volume pattern in 1969, while coronary artery disease and weight gain were found to be associated with a "restrictive" lung volume pattern in 1969. A career in military aviation had no significant association with lung volumes.

Orthopedic injuries experienced by U.S. prisoners of war during Operation Desert Storm: a descriptive analysis.

U.S. prisoners of war from Operation Desert Storm suffered significant orthopedic injuries. The repatriated prisoners of war (RPOWs) have been medically evaluated over a 3-year period with orthopedic follow-up. A significant proportion of the musculoskeletal injuries were located around the neck and spine, shoulder, and upper extremity. Aircraft ejection was the cause of the majority of these injuries. Lower extremity injuries after ejection, aside from the knee, were not reported. Flail injuries of the lower extremities were absent as well. These results were examined with reference to Vietnam RPOW data.