ER stress transforms random olfactory receptor choice into axon targeting precision.

Olfactory sensory neurons (OSNs) convert the stochastic choice of one of >1,000 olfactory receptor (OR) genes into precise and stereotyped axon targeting of OR-specific glomeruli in the olfactory bulb. Here, we show that the PERK arm of the unfolded protein response (UPR) regulates both the glomerular coalescence of like axons and the specificity of their projections. Subtle differences in OR protein sequences lead to distinct patterns of endoplasmic reticulum (ER) stress during OSN development, converting OR identity into distinct gene expression signatures. We identify the transcription factor Ddit3 as a key effector of PERK signaling that maps OR-dependent ER stress patterns to the transcriptional regulation of axon guidance and cell-adhesion genes, instructing targeting precision. Our results extend the known functions of the UPR from a quality-control pathway that protects cells from misfolded proteins to a sensor of cellular identity that interprets physiological states to direct axon wiring.

Intraoperative Hypotension-Physiologic Basis and Future Directions.

Intraoperative hypotension (IOH) is a major concern to the anesthesiologist. Its appropriate identification and management require an understanding of the physiology of blood pressure regulation, prudent blood pressure monitoring, and treatment. Even short durations of low mean arterial pressure have been associated with adverse postoperative clinical outcomes. The challenge is for the clinician to respond proactively, address the specific etiology of IOH, and keep in mind any changes to the patient's physiology. Predictive technology, such as the Hypotension Prediction Index, offers the clinician new insight into IOH. It has been shown to predict hypotension up to 15 minutes before occurrence. It also calculates stroke volume variation, dynamic arterial elastance, and left ventricular contractility, which can inform the anesthesiologist of the etiology of IOH to direct management. This new technology has the potential to reduce duration or even prevent IOH. In the authors' opinion, it is an example of how human-machine interaction will contribute to future advances in medicine. Additional studies should evaluate the effects of its use on postoperative outcomes.

An Examination of a Novel Weight Loss Supplement on Anthropometry and Indices of Cardiovascular Disease Risk.

PURPOSE: This study examined whether adding Dichrostachys glomerata (DG; 300 mg/d) to thermogenic supplements with (DG + C) and without (DG) caffeine and other nutrients affects weight loss, changes in body composition, and/or markers of health. METHODS: Sixty-eight participants (female, 54%) were grouped in a double-blind, parallel, stratified random, placebo-controlled manner to supplement their diet with a placebo, DG, or DG + C for 12 weeks while maintaining their normal diet and physical activity. Diet, physical activity, body weight, body composition, anthropometric measures, resting energy expenditure, fasting blood samples, and questionnaires were obtained at 0, 4, 8, and 12 weeks and analyzed using general linear models with repeated measures. Data are reported as mean (±SD) and change from baseline (mean, 95% confidence interval) for weeks 4, 8, and 12, respectively, with p values showing changes from baseline. RESULTS: DG treatment promoted significant but minor reductions in fat mass (-0.56 [-1.02, -0.14], p = 0.01; -0.63 [-1.23, -0.02], p = 0.04; -0.71 [-1.47, 0.09] kg, p = 0.08) and percent body fat (-0.46 [-0.96, -0.04], p = 0.07; -0.63 [-1.16, -0.10], p = 0.02; -0.78 [-1.45, 0.07] %, p = 0.03). There was some evidence that DG + C increased resting energy expenditure, decreased hunger, increased satiety, and improved sleep quality (diminished in DG + C). No other significant effects were observed. CONCLUSIONS: Ingestion of thermogenic supplements containing DG (300 mg/d) with and without caffeine and other nutrients in overweight but otherwise healthy participants who did not alter diet or physical activity promoted clinically insignificant changes in body weight and composition.

Hematological and Hemodynamic Responses to Acute and Short-Term Creatine Nitrate Supplementation.

In a double-blind, crossover, randomized and placebo-controlled trial; 28 men and women ingested a placebo (PLA), 3 g of creatine nitrate (CNL), and 6 g of creatine nitrate (CNH) for 6 days. Participants repeated the experiment with the alternate supplements after a 7-day washout. Hemodynamic responses to a postural challenge, fasting blood samples, and bench press, leg press, and cycling time trial performance and recovery were assessed. Data were analyzed by univariate, multivariate, and repeated measures general linear models (GLM). No significant differences were found among treatments for hemodynamic responses, clinical blood markers or self-reported side effects. After 5 days of supplementation, one repetition maximum (1RM) bench press improved significantly for CNH (mean change, 95% CI; 6.1 [3.5, 8.7] kg) but not PLA (0.7 [-1.6, 3.0] kg or CNL (2.0 [-0.9, 4.9] kg, CNH, p = 0.01). CNH participants also tended to experience an attenuated loss in 1RM strength during the recovery performance tests following supplementation on day 5 (PLA: -9.3 [-13.5, -5.0], CNL: -9.3 [-13.5, -5.1], CNH: -3.9 [-6.6, -1.2] kg, p = 0.07). After 5 days, pre-supplementation 1RM leg press values increased significantly, only with CNH (24.7 [8.8, 40.6] kg, but not PLA (13.9 [-15.7, 43.5] or CNL (14.6 [-0.5, 29.7]). Further, post-supplementation 1RM leg press recovery did not decrease significantly for CNH (-13.3 [-31.9, 5.3], but did for PLA (-30.5 [-53.4, -7.7] and CNL (-29.0 [-49.5, -8.4]). CNL treatment promoted an increase in bench press repetitions at 70% of 1RM during recovery on day 5 (PLA: 0.4 [-0.8, 1.6], CNL: 0.9 [0.35, 1.5], CNH: 0.5 [-0.2, 0.3], p = 0.56), greater leg press endurance prior to supplementation on day 5 (PLA: -0.2 [-1.6, 1.2], CNL: 0.9 [0.2, 1.6], CNH: 0.2 [-0.5, 0.9], p = 0.25) and greater leg press endurance during recovery on day 5 (PLA: -0.03 [-1.2, 1.1], CNL: 1.1 [0.3, 1.9], CNH: 0.4 [-0.4, 1.2], p = 0.23). Cycling time trial performance (4 km) was not affected. Results indicate that creatine nitrate supplementation, up to a 6 g dose, for 6 days, appears to be safe and provide some ergogenic benefit.

Short-Term Effects of a Ready-to-Drink Pre-Workout Beverage on Exercise Performance and Recovery.

In a double-blind, randomized and crossover manner, 25 resistance-trained participants ingested a placebo (PLA) beverage containing 12 g of dextrose and a beverage (RTD) containing caffeine (200 mg), ß-alanine (2.1 g), arginine nitrate (1.3 g), niacin (65 mg), folic acid (325 mcg), and Vitamin B12 (45 mcg) for 7-days, separated by a 7-10-day. On day 1 and 6, participants donated a fasting blood sample and completed a side-effects questionnaire (SEQ), hemodynamic challenge test, 1-RM and muscular endurance tests (3 × 10 repetitions at 70% of 1-RM with the last set to failure on the bench press (BP) and leg press (LP)) followed by ingesting the assigned beverage. After 15 min, participants repeated the hemodynamic test, 1-RM tests, and performed a repetition to fatigue (RtF) test at 70% of 1-RM, followed by completing the SEQ. On day 2 and 7, participants donated a fasting blood sample, completed the SEQ, ingested the assigned beverage, rested 30 min, and performed a 4 km cycling time-trial (TT). Data were analyzed by univariate, multivariate, and repeated measures general linear models (GLM), adjusted for gender and relative caffeine intake. Data are presented as mean change (95% CI). An overall multivariate time × treatment interaction was observed on strength performance variables (p = 0.01). Acute RTD ingestion better maintained LP 1-RM (PLA: -0.285 (-0.49, -0.08); RTD: 0.23 (-0.50, 0.18) kg/kgFFM, p = 0.30); increased LP RtF (PLA: -2.60 (-6.8, 1.6); RTD: 4.00 (-0.2, 8.2) repetitions, p = 0.031); increased BP lifting volume (PLA: 0.001 (-0.13, 0.16); RTD: 0.03 (0.02, 0.04) kg/kgFFM, p = 0.007); and, increased total lifting volume (PLA: -13.12 (-36.9, 10.5); RTD: 21.06 (-2.7, 44.8) kg/kgFFM, p = 0.046). Short-term RTD ingestion maintained baseline LP 1-RM (PLA: -0.412 (-0.08, -0.07); RTD: 0.16 (-0.50, 0.18) kg/kgFFM, p = 0.30); LP RtF (PLA: 0.12 (-3.0, 3.2); RTD: 3.6 (0.5, 6.7) repetitions, p = 0.116); and, LP lifting volume (PLA: 3.64 (-8.8, 16.1); RTD: 16.25 (3.8, 28.7) kg/kgFFM, p = 0.157) to a greater degree than PLA. No significant differences were observed between treatments in cycling TT performance, hemodynamic assessment, fasting blood panels, or self-reported side effects.

Effects of acute ingestion of a pre-workout dietary supplement with and without p-synephrine on resting energy expenditure, cognitive function and exercise performance.

BACKGROUND: The purpose of this study was to examine the effects of acute ingestion of a pre-workout dietary supplement (PWS) with and without p-synephrine (S) on perceptions of readiness to perform, cognitive function, exercise performance, and markers of safety. METHODS: In a randomized, double-blind, and counterbalanced manner; 25 healthy and recreationally active male and female participants ingested a flavored maltodextrin placebo (PLA), a PWS containing beta-alanine (3 g), creatine nitrate as a salt (2 g), arginine alpha-ketoglutarate (2 g), N-Acetyl-L-Tyrosine (300 mg), caffeine (284 mg), Mucuna pruiriens extract standardized for 15% L-Dopa (15 mg), Vitamin C as Ascorbic Acid (500 mg), niacin (60 mg), folate as folic acid (50 mg), and Vitamin B12 as Methylcobalamin (70 mg) with 2 g of maltodextrin and flavoring; or, the PWS with Citrus aurantium (PWS + S) extract standardized for 30% p-synephrine (20 mg). Participants had heart rate (HR), blood pressure, resting energy expenditure (REE), 12-lead electrocardiograms (ECG), perceptions about readiness to perform, cognitive function (Stroop Color-Word test), bench and leg press performance (2 sets of 10 repetitions at 70% of 1RM and 1 set to failure), and Wingate anaerobic capacity (WAC) sprint performance determined as well as donated blood samples prior to and/or following exercise/supplementation. Data were analyzed by MANOVA with repeated measures as well as mean changes from baseline with 95% confidence intervals (CI). RESULTS: No clinically significant differences were observed among treatments in HR, blood pressure, ECG, or general clinical blood panels. There was evidence that PWS and PWS + S ingestion promoted greater changes in REE responses. Participants reported higher perception of optimism about performance and vigor and energy with PWS and PWS + S ingestion and there was evidence that PWS and PWS + S improved changes in cognitive function scores from baseline to a greater degree than PLA after 1 or 2 h. However, the scores in the PWS + S treatment did not exceed PLA or PWS responses at any data point. No statistically significant differences were observed among treatments in total bench press lifting volume, leg press lifting volume or WAC sprint performance. CONCLUSIONS: Within the confines of this study, ingestion of PWS and/or PWS + S prior to exercise appears to be well-tolerated when consumed by young, healthy individuals. The primary effects appear to be to increase REE responses and improve perceptions about readiness to perform and cognitive function with limited to no effects on muscular endurance and WAC. The addition of 20 mg of p-synephrine to the PWS provided limited to no additive benefits. TRIAL REGISTRATION: This trial (NCT02952014) was retrospectively registered on September 13th 2016.

Efficacy of a randomized trial examining commercial weight loss programs and exercise on metabolic syndrome in overweight and obese women.

While commercial dietary weight-loss programs typically advise exercise, few provide actual programing. The goal of this study was to compare the Curves Complete 90-day Challenge (CC, n = 29), which incorporates exercising and diet, to programs advocating exercise (Weight Watchers Points Plus (WW, n = 29), Jenny Craig At Home (JC, n = 27), and Nutrisystem Advance Select (NS, n = 28)) or control (n = 20) on metabolic syndrome (MetS) and weight loss. We randomized 133 sedentary, overweight women (age, 47 ± 11 years; body mass, 86 ± 14 kg; body mass index, 35 ± 6 kg/m2) into respective treatment groups for 12 weeks. Data were analyzed using chi square and general linear models adjusted for age and respective baseline measures. Data are means ± SD or mean change ± 95% confidence intervals (CIs). We observed a significant trend for a reduction in energy intake for all treatment groups and significant weight loss for all groups except control: CC (-4.32 kg; 95% CI, -5.75, -2.88), WW (-4.31 kg; 95% CI, -5.82, -2.96), JC (-5.34 kg; 95% CI, -6.86, -3.90), NS (-5.03 kg; 95% CI, -6.49, -3.56), and control (0.16 kg, 95% CI, -1.56, 1.89). Reduced MetS prevalence was observed at follow-up for CC (35% vs. 14%, adjusted standardized residuals (adjres.) = 3.1), but not WW (31% vs. 28% adjres. = 0.5), JC (37% vs. 42%, adjres. = -0.7), NS (39% vs. 50% adjres. = -1.5), or control (45% vs. 55% adjres. = -1.7). While all groups improved relative fitness (mL·kg-1·min-1) because of weight loss, only the CC group improved absolute fitness (L/min). In conclusion, commercial programs offering concurrent diet and exercise programming appear to offer greater improvements in MetS prevalence and cardiovascular function after 12 weeks of intervention.

Effects of powdered Montmorency tart cherry supplementation on acute endurance exercise performance in aerobically trained individuals.

BACKGROUND: The purpose of this study was to determine whether short-term supplementation of a powdered tart cherry supplement prior to and following stressful endurance exercise would affect markers of muscle damage, inflammation, oxidative stress, and/or muscle soreness. METHODS: 27 endurance-trained runners or triathlete (21.8 ± 3.9 years, 15.0 ± 6.0 % body fat, 67.4 ± 11.8 kg) men (n = 18) and women (n = 9) were matched based on average reported race pace, age, body mass, and fat free mass. Subjects were randomly assigned to ingest, in a double-blind manner, capsules containing 480 mg of a rice flour placebo (P, n = 16) or powdered tart cherries [CherryPURE®] (TC, n = 11). Subjects supplemented one time daily (480 mg/day) for 10-d, including race day, up to 48-hr post-run. Subjects completed a half-marathon run (21.1 km) under 2-hr (111.98 ± 11.9 min). Fasting blood samples and quadriceps muscle soreness ratings using an algometer with a graphic pain rating scale were taken pre-run, 60-min, 24 and 48-h post-run and analyzed by MANOVA with repeated measures. RESULTS: Subjects in the TC group averaged 13 % faster half-marathon race finish times (p = 0.001) and tended to have smaller deviations from predicted race pace (p = 0.091) compared to P. Attenuations in TC muscle catabolic markers were reported over time for creatinine (p = 0.047), urea/blood urea nitrogen (p = 0.048), total protein (p = 0.081), and cortisol (p = 0.016) compared to P. Despite lower antioxidant activity pre-run in TC compared to P, changes from pre-run levels revealed a linear increase in antioxidant activity at 24 and 48-h of recovery in TC that was statistically different (16-39 %) from P and pre-run levels. Inflammatory markers were 47 % lower in TC compared to P over time (p = 0.053) coupled with a significant difference between groups (p = 0.017). Soreness perception between the groups was different over time in the medial quadriceps (p = 0.035) with 34 % lower pre-run soreness in TC compared to P. Over the 48-h recovery period, P changes in medial quadriceps soreness from pre-run measures were smaller compared to TC. CONCLUSION: Results revealed that short-term supplementation of Montmorency powdered tart cherries surrounding an endurance challenge attenuated markers of muscle catabolism, reduced immune and inflammatory stress, better maintained redox balance, and increased performance in aerobically trained individuals.

Acute and chronic safety and efficacy of dose dependent creatine nitrate supplementation and exercise performance.

BACKGROUND: Creatine monohydrate (CrM) and nitrate are popular supplements for improving exercise performance; yet have not been investigated in combination. We performed two studies to determine the safety and exercise performance-characteristics of creatine nitrate (CrN) supplementation. METHODS: Study 1 participants (N = 13) ingested 1.5 g CrN (CrN-Low), 3 g CrN (CrN-High), 5 g CrM or a placebo in a randomized, crossover study (7d washout) to determine supplement safety (hepatorenal and muscle enzymes, heart rate, blood pressure and side effects) measured at time-0 (unsupplemented), 30-min, and then hourly for 5-h post-ingestion. Study 2 participants (N = 48) received the same CrN treatments vs. 3 g CrM in a randomized, double-blind, 28d trial inclusive of a 7-d interim testing period and loading sequence (4 servings/d). Day-7 and d-28 measured Tendo™ bench press performance, Wingate testing and a 6x6-s bicycle ergometer sprint. Data were analyzed using a GLM and results are reported as mean ± SD or mean change ± 95 % CI. RESULTS: In both studies we observed several significant, yet stochastic changes in blood markers that were not indicative of potential harm or consistent for any treatment group. Equally, all treatment groups reported a similar number of minimal side effects. In Study 2, there was a significant increase in plasma nitrates for both CrN groups by d-7, subsequently abating by d-28. Muscle creatine increased significantly by d-7 in the CrM and CrN-High groups, but then decreased by d-28 for CrN-High. By d-28, there were significant increases in bench press lifting volume (kg) for all groups (PLA, 126.6, 95 % CI 26.3, 226.8; CrM, 194.1, 95 % CI 89.0, 299.2; CrN-Low, 118.3, 95 % CI 26.1, 210.5; CrN-High, 267.2, 95 % CI 175.0, 359.4, kg). Only the CrN-High group was significantly greater than PLA (p < 0.05). Similar findings were observed for bench press peak power (PLA, 59.0, 95 % CI 4.5, 113.4; CrM, 68.6, 95 % CI 11.4, 125.8; CrN-Low, 40.9, 95 % CI -9.2, 91.0; CrN-High, 60.9, 95 % CI 10.8, 111.1, W) and average power. CONCLUSIONS: Creatine nitrate delivered at 3 g was well-tolerated, demonstrated similar performance benefits to 3 g CrM, in addition, within the confines of this study, there were no safety concerns.

Effects of powdered Montmorency tart cherry supplementation on an acute bout of intense lower body strength exercise in resistance trained males.

BACKGROUND: The purpose of this study was to examine whether short-term ingestion of a powdered tart cherry supplement prior to and following intense resistance-exercise attenuates muscle soreness and recovery strength loss, while reducing markers of muscle damage, inflammation, and oxidative stress. METHODS: Twenty-three healthy, resistance-trained men (20.9 ± 2.6 yr, 14.2 ± 5.4% body fat, 63.9 ± 8.6 kg FFM) were matched based on relative maximal back squat strength, age, body weight, and fat free mass. Subjects were randomly assigned to ingest, in a double blind manner, capsules containing a placebo (P, n = 12) or powdered tart cherries [CherryPURE(®)] (TC, n = 11). Participants supplemented one time daily (480 mg/d) for 10-d including day of exercise up to 48-h post-exercise. Subjects performed ten sets of ten repetitions at 70% of a 1-RM back squat exercise. Fasting blood samples, isokinetic MVCs, and quadriceps muscle soreness ratings were taken pre-lift, 60-min, 24-h, and 48-h post-lift and analyzed by MANOVA with repeated measures. RESULTS: Muscle soreness perception in the vastus medialis (») (p = 0.10) and the vastus lateralis (») (p = 0.024) was lower in TC over time compared to P. Compared to pre-lift, TC vastus medialis (») soreness was significantly attenuated up to 48-h post-lift with vastus lateralis (») soreness significantly lower at 24-h post-lift compared to P. TC changes in serum creatinine (p = 0.03, delta p = 0.024) and total protein (p = 0.018, delta p = 0.006) were lower over time and smaller from pre-lift levels over time compared to P Significant TC group reductions from pre-lift levels were found for AST and creatinine 48-h post-lift, bilirubin and ALT 60-min and 48-h post-lift. No significant supplementation effects were observed for serum inflammatory or anti-inflammatory markers. None of the free radical production, lipid peroxidation, or antioxidant capacity markers (NT, TBARS, TAS, SOD) demonstrated significant changes with supplementation. Changes in TC whole blood lymphocyte counts (p = 0.013) from pre-lift were greater compared to P, but TC lymphocyte counts returned to pre-lift values quicker than P. CONCLUSION: Short-term supplementation of Montmorency powdered tart cherries surrounding a single bout of resistance exercise, appears to be an effective dietary supplement to attenuate muscle soreness, strength decrement during recovery, and markers of muscle catabolism in resistance trained individuals.

Chemosensory receptor specificity and regulation.

The senses provide a means by which data on the physical and chemical properties of the environment may be collected and meaningfully interpreted. Sensation begins at the periphery, where a multitude of different sensory cell types are activated by environmental stimuli as different as photons and odorant molecules. Stimulus sensitivity is due to expression of different cell surface sensory receptors, and therefore the receptive field of each sense is defined by the aggregate of expressed receptors in each sensory tissue. Here, we review current understanding on patterns of expression and modes of regulation of sensory receptors.

Co-opting the unfolded protein response to elicit olfactory receptor feedback.

Olfactory receptor (OR) expression requires the transcriptional activation of 1 out of 1,000s of OR alleles and a feedback signal that preserves this transcriptional choice. The mechanism by which olfactory sensory neurons (OSNs) detect ORs to signal to the nucleus remains elusive. Here, we show that OR proteins generate this feedback by activating the unfolded protein response (UPR). OR expression induces Perk-mediated phosphorylation of the translation initiation factor eif2α causing selective translation of activating transcription factor 5 (ATF5). ATF5 induces the transcription of adenylyl cyclase 3 (Adcy3), which relieves the UPR. Our data provide a role for the UPR in defining neuronal identity and cell fate commitment and support a two-step model for the feedback signal: (1) OR protein, as a stress stimulus, alters the translational landscape of the OSN and induces Adcy3 expression; (2), Adcy3 relieves that stress, restores global translation, and makes OR choice permanent.

Affinity purification of microRNA-133a with the cardiac transcription factor, Hand2.

Predictions of microRNA-mRNA interactions typically rely on bioinformatic algorithms, but these algorithms only suggest the possibility of microRNA binding and may miss important interactions as well as falsely predict others. We developed an affinity purification approach to empirically identify microRNAs associated with the 3'UTR of the mRNA encoding Hand2, a transcription factor essential for cardiac development. In addition to miR-1, a known regulator of Hand2 expression, we determined that the Hand2 3'UTR also associated with miR-133a, a microRNA cotranscribed with miR-1 in cardiac and muscle cells. Using a sequential binding assay, we showed that miR-1 and miR-133a could occupy the Hand2 3'UTR concurrently. miR-133a inhibited Hand2 expression in tissue culture models, and miR-133a double knockout mice had elevated levels of Hand2 mRNA and protein. We conclude that Hand2 is regulated by miR-133a in addition to miR-1. The affinity purification assay should be generally applicable for identifying other microRNA-mRNA interactions.

Genetic contribution to variation in larval development time, adult size, and longevity of starved adults of Anopheles gambiae.

The variation in mosquito life-history traits such as adult size has been studied with respect to environmental factors, but the genetic contribution to such variation has received almost no consideration. Using a full-sib design of F1s produced by wild caught Anopheles gambiae (M molecular form) females, we estimated broad-sense heritability of larval developmental time, adult size (based on dry weight and wing length), and longevity of starved adults. These traits were correlated (at the phenotypic level) with each other in females and males (|r(p)|>0.5, P<0.001). Longevity of starved adults increased with adult size, and both traits (adult longevity and size) decreased with longer larval development. Genetic correlations were lower (|r(g)|>0.45, P<0.05) but provided consistent evidence against a trade off between adult size and larval development time predicting that a mosquito can develop faster into a smaller adult or be a larger adult by a longer development. Estimates of heritability of the three traits were moderate to high (range: 0.05-0.48) and statistically significant (P<0.05), indicating substantial genetic contribution to the phenotypic variation in these traits. These results suggest that adaptive differences are likely to be found in these traits between A. gambiae populations.

Larval development of the molecular forms of Anopheles gambiae (Diptera: Culicidae) in different habitats: a transplantation experiment.

We compared the development of the molecular forms of Anopheles gambiae s.s. in different larval habitats. First stage larvae (L1s) of wild-caught females were placed into cages in natural habitats of the M form (rice fields) or the S form (puddles/ quarries). Each cage was covered with cloth, allowing exchange of water, solutes, and small particles, including microorganisms, and was seeded with 100 L1s of a single form (M or S) or by a mixture of 50:50 of M and S forms. Emergence success of both forms in puddles and quarries was three-fold higher than in the rice fields. The emergence rate of the S form was higher than that of the M form in both habitats, but the form x habitat interaction was not significant. In temporary larval sites such as puddles, emergence success of the M form was lower in mixed cages than in single form cages, whereas the reverse was true for the S form, suggesting competition between the forms. The median developmental time was not significantly different between forms. Although these findings demonstrate differences between forms, they do not suggest that their spatial segregation is determined by differences in their exploitation of the physical and chemical conditions in these environments. These results should be regarded with caution because small numbers of first stage larvae could pass through the cloth of the cages.