Cohort-level disease prediction by extrapolation of individual-level predictions in transition dairy cattle.

Dairy cattle experience metabolic stress during the transition from late gestation to early lactation resulting in higher risk for several economically important diseases (e.g. mastitis, metritis, and ketosis). Metabolic stress is described as a physiological state composed of 3 processes: nutrient metabolism, oxidative stress, and inflammation. Current strategies for monitoring transition cow nutrient metabolism include assessment of plasma non-esterified fatty acids and beta-hydroxybutyrate concentrations around the time of calving. Although this method is effective at identifying cows with higher disease risk, there is often not enough time to implement intervention strategies to prevent health disorders from occurring around the time of calving. Previously, we published predictive models for early lactation diseases at the individual cow level at dry-off. However, it is unknown if predictive probabilities from individual-level models can be aggregated to the cohort level to predict cohort-level incidence. Therefore, our objective was to test different data aggregation methods using previously published models that represented the 3 components of metabolic stress (nutrient metabolism, oxidative stress, and inflammation). We included 277 cows from five Michigan dairy herds for this prospective cohort study. On each farm, two to four calving cohorts were formed, totaling 18 cohorts. We measured biomarker data at dry-off and followed the cows until 30 days post-parturition for cohort disease incidence, which was defined as the number of cows: 1) having one or more clinical transition disease outcome, and/or 2) having an adverse health event (abortion or death of calf or cow) within each cohort. We tested three different aggregation methods that we refer to as the p-central, p-dispersion, and p-count methods. For the p-central method, we calculated the averaged predicted probability within each cohort. For the p-dispersion method, we calculated the standard deviation of the predicted probabilities within a cohort. For the p-count method, we counted the number of cows above a specified threshold of predicted probability within each cohort. We built four sets of models: one for each aggregation method and one that included all three aggregation methods (p-combined method). We found that the p-dispersion method was the only method that produced viable predictive models. However, these models tended to overestimate incidence in cohorts with low observed counts and underestimate risk in cohorts with high observed counts.

Cohort-level disease prediction using aggregate biomarker data measured at dry-off in transition dairy cattle: A proof-of-concept study.

During the transition from late gestation to early lactation, dairy cattle are at increased risk for disease. Herd-level monitoring for disease risk involves evaluating multiple factors, including food intake, cow density, and biomarkers of nutrient metabolism. Biomarkers that are measured include non-esterified fatty acids (NEFA) and beta-hydroxybutyrate (BHB), which are usually measured in a subset of the herd (i.e. cohort). If a certain proportion of cows in the cohort are above a specific threshold for a biomarker, the cohort is considered at high risk of disease. Few previous studies have investigated other methods to aggregate individual cow-level data to the cohort level. We designed a proof-of-concept study to determine if biomarker aggregation methods may be useful to predict cohort incidence of adverse health events including 1) clinical diseases: mastitis, metritis, retained placenta, ketosis, lameness, pneumonia, milk fever, displaced abomasum, 2) and abortion or death of the calf or the cow. The study design was a prospective cohort study that used cows (N = 277) from five Michigan commercial dairy herds. Multiple cohorts of cows (two to four cohorts per farm, 18 total) were enrolled that shared the same dry-off date. We tested three different methods (central, dispersion, and count) to aggregate individual cow data (i.e. biomarkers and covariates) measured at dry-off. The central method consisted of calculating the average value of each variable within a cohort, and the dispersion method involved taking the standard deviation or mean absolute deviation about the median of each variable within a cohort. The count method consisting of counting the number of cows above a specific threshold for each variable within a cohort. We used best subsets selection to select a bouquet of candidate models for each aggregation method and averaged the predictions over the model set. We built 4 sets of Poisson regression models: one for each aggregation method and a combined model that included all three methods. We evaluated the models based on goodness-of-fit, model calibration using scoring rules, and comparison of observed versus predicted counts. The central and the combined method produced models that had good fit and model calibration. These results indicate that it may be possible to use aggregate measures to predict cohort disease incidence as early as dry-off. The next step is to test biomarker aggregation methods in studies with larger sample sizes.

Predictive models for early lactation diseases in transition dairy cattle at dry-off.

During the transition period, dairy cattle undergo tremendous metabolic and physiological changes to prepare for milk synthesis and secretion. Failure to sufficiently regulate these changes may lead to metabolic stress, which increases risk of transition diseases. Metabolic stress is defined as a physiological state consisting of 3 components: aberrant nutrient metabolism, oxidative stress, and inflammation. Current monitoring methods to detect cows experiencing metabolic stress involve measuring biomarkers for nutrient metabolism. However, these biomarkers, including non-esterified fatty acids, beta-hydroxybutyrate, and calcium are typically measured a few weeks before to a few days after calving. This is a retroactive approach, because there is little time to integrate interventions that remediate metabolic stress in the current cohort. Our objective was to determine if biomarkers of metabolic stress measured at dry-off are predictive of transition disease risk. We designed a prospective cohort study carried out on 5 Michigan dairy farms (N = 277 cows). We followed cows from dry-off to 30 days post-calving. Diseases and adverse outcomes were grouped in an aggregate outcome that included mastitis, metritis, retained placenta, ketosis, lameness, pneumonia, milk fever, displaced abomasum, abortion, and death of the calf or the cow. We used best subsets selection to select candidate models for four different sets of models: one set for each component of metabolic stress (nutrient metabolism, oxidative stress, and inflammation), and a combined model that included all 3 components. We used model averaging to obtain averaged predicted probabilities across each model set. We hypothesized that the averaged predictions from the combined model set with all 3 components of metabolic stress would be more effective at predicting disease than each individual component model set. The area under the curve estimated using receiver operator characteristic curves for the combined model set (0.93; 95% confidence interval [CI] = 0.90-0.96) was significantly higher compared with averaged predictions from the inflammation (0.87; 95% CI = 0.83-0.91), oxidative stress (0.78; 95% CI = 0.72-0.84), and nutrient metabolism (0.73; 95% CI = 0.67-0.79) model sets (p < 0.05). Our results indicate that it may be possible to detect cattle at risk for some transition diseases as early as dry-off. This has important implications for disease prevention, as earlier identification of cows at risk of health disorders will allow for earlier implementation of intervention strategies. A limitation of the current study is that we did not perform external validation. Future validation studies are needed to confirm our findings.

Population structure and residency of whale sharks Rhincodon typus at Utila, Bay Islands, Honduras.

There were 479 reported whale shark Rhincodon typus encounters between 1999 and 2011 at the island of Utila, which forms part of the Meso-American Barrier Reef System (MBRS) in the western Caribbean Sea. The majority of R. typus were found to feed on small bait fish associated with various tuna species. Ninety-five individual R. typus, ranging from 2 to 11 m total length (LT ), were identified through their unique spot patterns. A significant male bias (65%) was present. There was no significant difference between the mean ± s.d. LT of female (6·66 ± 1·65 m) and male (6·25 ± 1·60 m) R. typus. Most R. typus were transient to Utila, with 78% sighted only within a single calendar year, although some individuals were sighted in up to 5 years. Mean residency time was modelled to be 11·76 days using maximum likelihood methods.

Unusually high levels of n-6 polyunsaturated fatty acids in whale sharks and reef manta rays.

Fatty acid (FA) signature analysis has been increasingly used to assess dietary preferences and trophodynamics in marine animals. We investigated FA signatures of connective tissue of the whale shark Rhincodon typus and muscle tissue of the reef manta ray Manta alfredi. We found high levels of n-6 polyunsaturated fatty acids (PUFA), dominated by arachidonic acid (20:4n-6; 12-17 % of total FA), and comparatively lower levels of the essential n-3 PUFA-eicosapentaenoic acid (20:5n-3; ~1 %) and docosahexaenoic acid (22:6n-3; 3-10 %). Whale sharks and reef manta rays are regularly observed feeding on surface aggregations of coastal crustacean zooplankton during the day, which generally have FA profiles dominated by n-3 PUFA. The high levels of n-6 PUFA in both giant elasmobranchs raise new questions about the origin of their main food source.

Biology, ecology and conservation of the Mobulidae.

The Mobulidae are zooplanktivorous elasmobranchs comprising two recognized species of manta rays (Manta spp.) and nine recognized species of devil rays (Mobula spp.). They are found circumglobally in tropical, subtropical and temperate coastal waters. Although mobulids have been recorded for over 400 years, critical knowledge gaps still compromise the ability to assess the status of these species. On the basis of a review of 263 publications, a comparative synthesis of the biology and ecology of mobulids was conducted to examine their evolution, taxonomy, distribution, population trends, movements and aggregation, reproduction, growth and longevity, feeding, natural mortality and direct and indirect anthropogenic threats. There has been a marked increase in the number of published studies on mobulids since c. 1990, particularly for the genus Manta, although the genus Mobula remains poorly understood. Mobulid species have many common biological characteristics although their ecologies appear to be species-specific, and sometimes region-specific. Movement studies suggest that mobulids are highly mobile and have the potential to rapidly travel large distances. Fishing pressure is the major threat to many mobulid populations, with current levels of exploitation in target fisheries unlikely to be sustainable. Advances in the fields of population genetics, acoustic and satellite tracking, and stable-isotope and fatty-acid analyses will provide new insights into the biology and ecology of these species. Future research should focus on the uncertain taxonomy of mobulid species, the degree of overlap between their large-scale movement and human activities such as fisheries and pollution, and the need for management of inter-jurisdictional fisheries in developing nations to ensure their long-term sustainability. Closer collaboration among researchers worldwide is necessary to ensure standardized sampling and modelling methodologies to underpin global population estimates and status.

The use and abuse of photographic identification in sharks and rays.

The use of photography to discriminate between individuals in a population using natural markings or aberrations is increasingly being utilized to support field research on elasmobranchs. This non-intrusive method has facilitated investigation of a wide variety of subjects including population composition, abundance estimates, residency and movement, demography and social behaviours. Here the first detailed review of photo-identification as a research technique for sharks and rays is provided, and its assumptions, current applications and potential highlighted. The limitations and practical considerations of photographic studies are also investigated with recommendations on initial survey design and ongoing data collection using current technology. Future directions are also explored with an emphasis on a move towards standardized approaches and automated recognition programmes to facilitate global collaborative work.

How large is the world's largest fish? Measuring whale sharks Rhincodon typus with laser photogrammetry.

Laser photogrammetry was found to be a promising new cost-effective technique for measuring free-swimming whale sharks Rhincodon typus. Photogrammetric measurements were more precise than visual size estimates by experienced researchers, with results from the two methods differing by 9· 8 ± 1· 1% (mean ±s.e.). A new metric of total length and the length between the fifth gill and first dorsal fin (r² = 0· 93) is proposed to facilitate easy, accurate length measurements of whale sharks in the field.

Dispersal of Agrilus planipennis (Coleoptera: Buprestidae) from discrete epicenters in two outlier sites.

Emerald ash borer, Agrilus planipennis (Fairmaire) (Coleoptera: Buprestidae), a phloem-feeding beetle native to Asia, has become one of the most destructive forest pests in North America. Since it was first identified in 2002 in southeast Michigan and Windsor, Ontario, dozens of isolated A. planipennis populations have been discovered throughout Michigan and Ontario, and in 12 other states and the province of Quebec. We assessed realized A. planipennis dispersal at two discrete outlier sites that originated 1 yr and 3 yr earlier from infested nursery trees. We systematically sampled ash trees within an 800 m radius of the origin of each infestation to locate galleries constructed by the progeny of dispersing A. planipennis adults. Our sampling identified eight trees at the 1 yr site infested with a mean +/- SE of 11.6 +/- 8.4 A. planipennis larvae and 12 trees at the 3 yr site with 25.8 +/- 11.1 larvae per meter squared. Dendroentomological analysis indicated that A. planipennis populations were predominantly undergoing a 2 yr (semivoltine) life cycle at both sites. Colonized trees were found out to 638 and 540 m from the epicenters at the 1 yr and 3 yr sites, respectively. Logistic regression was used to determine whether the likelihood of A. planipennis colonization was affected by wind direction, ash phloem abundance, distance from the epicenter, or land-use type (i.e., wooded, residential, agricultural, or urban). Results show that the probability of A. planipennis colonization was significantly affected by ash phloem abundance and decreased with distance from the epicenter.

Deep-diving behaviour of a whale shark Rhincodon typus during long-distance movement in the western Indian Ocean.

A whale shark Rhincodon typus satellite tagged off the coast of Mozambique showed a highly directional movement across the Mozambique Channel and around the southern tip of Madagascar, a minimum distance of 1200 km in 87 days. Dives to depths well into the mesopelagic and bathypelagic zones (1286 m maximum depth) were recorded in a bathymetrically non-constraining habitat. The water temperature range recorded during the fish's movement was 3.4-29.9 degrees C.

Reproduction of the blue-spotted maskray Neotrygon kuhlii (Myliobatoidei: Dasyatidae) in south-east Queensland, Australia.

This study examined the reproduction and population structure of the blue-spotted maskray Neotrygon kuhlii within Moreton Bay, a subtropical embayment in south-east Queensland, Australia. Mean sizes at maturity were 314 mm disc width (W(D)) in females (n = 140, 115-465 mm W(D)) and 294 mm W(D) in males (n = 123, 129-381 mm W(D)). Female N. kuhlii had a synchronous annual reproductive cycle, with one litter of one to three pups (mean +/-s.d. 1.67 +/- 0.71) produced per year. Mating behaviours were observed in October and November, and ovulation occurred early in the Austral summer, overlapping with the start of embryonic development. Gestation took c. 4 months with parturition occurring in late February and March. Size at birth was 115-170 mm W(D). The population showed a significant female bias, particularly in larger size classes. Tagging studies produced a total recapture rate of 16.1% and indicated that N. kuhlii were site resident for up to 1081 days.

Validated annual band-pair periodicity and growth parameters of blue-spotted maskray Neotrygon kuhlii from south-east Queensland, Australia.

Age and growth parameters were derived for blue-spotted maskray Neotrygon kuhlii from Moreton Bay in subtropical eastern Australia. Maximum age estimates of 13 and 10 years were obtained from female (n = 76) and male (n = 44) N. kuhlii, respectively. Estimated ages at maturity for 50% of females and males were 6.32 and 3.95 years, respectively. A three-parameter power function provided the best statistical fit to size at age data in both sexes, providing parameter estimates of y(0) = 163.13, a = 58.52 and b = 0.58 for females and y(0) = 165.13, a = 59.02 and b = 0.54 in males. The two-parameter von Bertalanffy growth function was used to estimate biological parameters based on disc width (W(D)) for both female (W(Dinfinity) = 465.81 mm, K = 0.13 year(-1), b = 0.63) and male N. kuhlii (W(Dinfinity) = 385.19 mm, K = 0.20 year(-1), b = 0.54). Annual band-pair deposition was observed in three calcein-injected N. kuhlii after periods of liberty ranging from 631 to 1081 days. Centrum edge analysis indicated that annual band-pair formation was generally consistent within this population, with translucent bands formed over spring and summer and opaque bands formed in autumn and winter. Individual growth rates obtained from tagged specimens were similar to power function growth predictions. These results support previous characterizations of this common trawl by-catch species as comparatively resilient to non-targeted catches, although higher catch rates outside Australia infer a need for cautious management.

Molecular typing, serotyping and cytotoxicity testing of Campylobacter jejuni strains isolated from commercial broilers in Puerto Rico.

AIMS: Thirty Campylobacter jejuni strains isolated from fecal samples (n = 94; 32%) from 13 positive farms (n = 17; 76%) from commercial broiler chickens in Puerto Rico were analysed by molecular methods. METHODS AND RESULTS: Isolates were identified with multiplex polymerase chain reaction assays, tested for their antimicrobial susceptibility and characterized with pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), serotyping and bacterial cytotoxicity in mammalian cells. Isolates exhibited high resistance to vancomycin (minimum inhibitory concentration, MIC of >256 microg ml(-1)) and trimethoprim (MIC of >32 microg ml(-1)); few were resistant to clindamycin (MIC(90) 4 microg ml(-1)), erythromycin (MIC(90) 8 microg ml(-1)) and tetracycline (MIC(90) 8 microg ml(-1)); but none was resistant to azithromycin (MIC(90) 4 microg ml(-1)), ciprofloxacin (MIC(90) 1 microg ml(-1)) or gentamycin (MIC(90) 4 microg ml(-1)). Most strains restricted with SmaI, but a combination of SmaI-KpnI digestion was more discriminatory. MLST analysis yielded four sequence types (ST), and ST-2624 was the predominant one. Phylogenetic analysis revealed a high degree of recombination for glnA and pgm genes. The predominant serotypes were O:3 and O:5. Most strains had lowest cytotoxicity potential with Caco-2 cells, medium cytotoxicity with INT-407 and Hep-2 cells and high cytotoxicity with CHO cells. CONCLUSION: A low degree of antimicrobial resistance, 13 PFGE profiles, 4 ST and a large variability in cytotoxicity assays were found for these strains. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first characterization of C. jejuni strains isolated from broilers in Puerto Rico. The genetic diversity of these strains suggests that several techniques are needed for strain characterization.

The effects of metabolic inhibition on intracellular calcium and contractility of human myometrium.

OBJECTIVE: Hypoxia occurs in the uterus during labour and may contribute to dysfunctional labours. We wanted to establish its effects on pregnant human myometrium and elucidate the mechanisms involved. DESIGN: Scientific study. SETTING: University Hospital and laboratories. POPULATION OR SAMPLE: Term pregnant women. METHODS: We measured contractions and intracellular [Ca(2+)] ([Ca(2+)](i)), in biopsies from term pregnant women undergoing elective caesarean section, and used cyanide to block oxidative phosphorylation. MAIN OUTCOME MEASURES: Changes in contractility and calcium. RESULTS: Although basal levels of [Ca(2+)](i) and tone rose, spontaneous and agonist-induced Ca(2+) transients and phasic contractions were rapidly reduced and abolished by cyanide. Neither stimulation of the uterus with oxytocin nor the Ca channel agonist, Bay K8644, prevented the changes produced by cyanide. The tonic force produced by depolarising the myometrium was also decreased by cyanide, but slowly recovered towards control levels, whereas [Ca(2+)](i) was maintained throughout. Similar data were obtained when nitrogen, rather than cyanide, was applied to the depolarised uterus. CONCLUSIONS: Impairment of oxidative phosphorylation is a potent depressor of phasic activity in human myometrium, irrespective of how it is produced, and our data suggest its effects lie at and beyond the surface membrane. Stimulation of the hypoxic uterus was not effective, which may explain the unpredictability of oxytocin application in some dysfunctional labours.

Calcium signaling and uterine contractility.

Changes in Ca(2+) signals within the myometrium have important functional consequences, as they determine contractility. We show that the basic phasic nature of uterine contractions, which is essential for successful labor, is critically dependent on Ca(2+) influx through voltage-gated L-type Ca(2+) channels, and hence in turn, on membrane potential. Thus changes in ion channel expression around term will play an important role in governing uterine excitability and contractility. There remains uncertainty about which channels are present in human myometrium and the nature of the pacemaker mechanism that initiates the action potential. The sarcoplasmic reticulum may augment, to a small extent, the necessary increase in [Ca(2+)] for contraction when agonists stimulate the uterus, but its main role appears to be to control excitability, acting as a negative feedback mechanism to limit contractions. Myosin light chain kinase activity and phosphorylation of myosin are essential components in the pathway of uterine contraction, once Ca(2+) has been elevated. Modulation of myosin light chain phosphatase activity can also influence contractions, but the effects are small compared with those modulating myosin light chain kinase. Ca(2+)-sensitizing pathways may not be utilized much in modulating normal phasic uterine activity, and caution is needed in extrapolating from in vitro experiments to in vivo conditions, especially because there may be redundant pathways. There is a need to study appropriate physiologic preparations, but these are not always available (eg, preterm laboring myometrium) and to combine functional studies with modern molecular approaches, to advance our understanding to a new level, from which better therapeutics will be developed.

Improving depressive symptoms among caregivers of patients with cancer: results of a randomized clinical trial.

PURPOSE/OBJECTIVES: Determine the impact of a 16-week supportive nursing intervention on caregivers of patients with newly diagnosed cancer. DESIGN: Randomized clinical trial. SETTING: Two midwestern cancer treatment sites. SAMPLE: Caregivers of newly diagnosed patients. Patients' mean age was 55.73 years; 55% had breast cancer, and 76% were female. Caregivers' mean age was 52.44 years, and 50% were female. 125 dyads consented to participate; 89 dyads completed the study. METHODS: A nursing intervention was delivered to the experimental group that emphasized symptom monitoring/management, education, emotional support, coordination of services, and caregiver preparation to care. Nurses made a total of nine contacts, five in person and four by telephone, over 16 weeks. Centers for Epidemiological Studies-Depression (CES-D) and a symptom inventory were used. Medical record audits were conducted retrospectively. MAIN RESEARCH VARIABLES: Patient and caregiver depression scores and patient symptom experience. FINDINGS: Baseline caregiver depression and the number of patient symptoms at baseline, 9, and 24 weeks were significant predictors of caregiver depression at 9 and 24 weeks. However, no main effect of the experimental condition existed on caregiver depression. At the final observation, a nonsignificant inverse relationship was found between the number of interventions and depression scores for caregivers. CONCLUSIONS: The intervention appeared to be more effective in slowing the rate of deterioration of depressive symptoms than in decreasing levels of depression in this sample of caregivers. Determining the effectiveness of this intervention in decreasing caregiver depression was difficult because caregivers with higher levels of depression were more likely to withdraw from the study. IMPLICATIONS FOR NURSING PRACTICE: Nurses must be vigilant in monitoring caregivers for signs of depression and must intervene to provide emotional support and make appropriate referrals for follow-up care to promote positive outcomes for patients and caregivers.

Long-term use of gonadotropin-releasing hormone analogs and hormone replacement therapy in the management of endometriosis: a randomized trial with a 6-year follow-up.

OBJECTIVE: To identify the effects of long-term GnRH agonist use (6-24 months), with and without add-back therapy, and spontaneous reversibility of bone mass density (BMD) up to 6 years after treatment. DESIGN: A prospective, randomized, long-term follow-up study. SETTING: Obstetrics and gynecology department in a university hospital in the United Kingdom. PATIENT(S): Forty-nine symptomatic women with a laparoscopic diagnosis of endometriosis who had been identified for treatment with long-acting GnRH agonist and volunteered to participate in the study. INTERVENTION(S): Women were randomly allocated to receive hormone replacement therapy (HRT) as a daily oral dose of estradiol, 2 mg, and norethisterone acetate, 1 mg, or no treatment in addition to monthly subcutaneous implants of goserelin acetate for up to 2 years, until cessation of symptoms. Bone mineral density (BMD) at the lumbar spine (C2-C4) and hip (Ward triangle) was measured every 6 months. MAIN OUTCOME MEASURE(S): BMD changes in both groups. RESULT(S): 45 women were followed up for 6 years, at the end of which the groups did not differ significantly in the reduction in mean BMD at the lumbar spine or hip. CONCLUSION(S): BMD reduction occurs during long-term GnRH agonist use and is not fully recovered by up to 6 years after treatment. Use of HRT does not affect this process.

Prolonged incremental tests do not necessarily compromise VO2max in well-trained athletes.

Existing literature suggests that tests for maximal oxygen uptake (VO2max) should last 8-12 minutes and that prolonged tests do not produce valid measurements. The research underlying this suggestion has been performed with non-athletic populations and trained athletes may be more tolerant of longer protocols. Eleven rowers (8 males, 3 females) each underwent four different incremental tests on a standard rowing ergometer in randomised counterbalanced order. One of the tests was continuous with workload increments each minute (IT1MIN). This test lasted an average of 12 min+/-4 s (SEM). The other three tests were discontinuous and consisted of 7 stages separated by 1-minute recovery intervals. Stage durations of 3, 4 and 5 min were used in the different tests (IT3MIN, IT4MIN and IT5MIN). Mean values for VO2max were almost identical for IT1MIN (4.56+/-0.22 L x min(-1)), IT3MIN (4.60+/-0.23 L x min(-1)) and IT4MIN (4.60+/-0.21 L x min(-1)), while IT5MIN produced a significantly lower value (4.47+/-0.21 L x min(-1), p<0.05). There was no significant difference between protocols in peak post-exercise blood lactate concentration (approx 13 mmol x L(-1) in each case), but IT1MIN produced lower peak heart rates and higher respiratory exchange ratios. We conclude that with well trained rowing athletes discontinuous test protocols involving 7 stages of 3-4 minutes duration can provide valid measurements of VO2max.

Cross-sectional studies of grammatical morphemes in autistic and mentally retarded children.

The frequency of occurrence of functors in obligatory contexts was studied in verbal autistic and mentally retarded children matched for nonverbal mental age, and the percentages of correct use of functors were rank-ordered. The grammatical complexity of their language was also described using a transformational grammar. The data were compared to those obtained in a normal group matched for mental age and to the data presented by Brown (1973) and deVilliers and deVilliers (1973) in younger children. The autistic subjects omitted functors frequently and independently of the grammatical complexity of their language. The rank ordering of morphemes was consistent within both the autistic and mentally retarded groups but showed no correlation between the two groups or to the rank ordering described by deVilliers and deVilliers. It is suggested that functors in autistic subjects may develop in an atypical but consistent order and that this may be due to specific semantic deficits, particularly in the areas of person and time deixis.