Project DyAdd: Fatty acids and cognition in adults with dyslexia, ADHD, or both

Both attention deficit hyperactivity disorder (ADHD) and dyslexia are suggested to co-occur with altered fatty acid (FA) metabolism, but it is unknown how FAs are associated with the cognitive domains that characterize these disorders. In the project DyAdd, we investigated the associations between FAs in serum phospholipids and phonological processing, reading, spelling, arithmetic, executive functions, and attention. Healthy controls (n=36), adults with ADHD (n=26), dyslexia (n=36), or both (n=9) were included in the study. FAs included saturated, monounsaturated, total polyunsaturated, n-3, and n-6 FAs, together with n-6/n-3, AA/EPA, and LA/ALA ratios. When all the study subjects were included in the analyses, especially polyunsaturated FAs (PUFAs) were positively associated with cognition, but reading was least associated with FAs. These associations were modulated by gender, intelligence, n-3 PUFA intake, and group. Accordingly, within the ADHD group, only few associations emerged with PUFAs, n-6 PUFAs, and cognitive domains, whereas in the dyslexia group the more prevalent associations appeared with PUFAs and n-3 PUFAs.     

An Exploratory Study on the Effects of Tele-neurofeedback and Tele-biofeedback on Objective and Subjective Sleep in Patients with Primary Insomnia

Insomnia is a sleeping disorder, usually studied from a behavioural perspective, with a focus on somatic and cognitive arousal. Recent studies have suggested that an impairment of information processes due to the presence of cortical hyperarousal might interfere with normal sleep onset and/or consolidation. As such, a treatment modality focussing on CNS arousal, and thus influencing information processing, might be of interest. Seventien insomnia patients were randomly assigned to either a tele-neurofeedback (n = 9) or an electromyography tele-biofeedback (n = 8) protocol. Twelve healthy controls were used to compare baseline sleep measures. A polysomnography was performed pre and post treatment. Total Sleep Time (TST), was considered as our primary outcome variable. Sleep latency decreased pre to post treatment in both groups, but a significant improvement in TST was found only after the neurofeedback (NFB) protocol. Furthermore, sleep logs at home showed an overall improvement only in the neurofeedback group, whereas the sleep logs in the lab remained the same pre to post training. Only NFB training resulted in an increase in TST. The mixed results concerning perception of sleep might be related to methodological issues, such as the different locations of the training and sleep measurements.     

A genome-wide association study identifies a new variant associated with word reading fluency in Chinese children

Reading disability exhibited defects in different cognitive domains, including word reading fluency, word reading accuracy, phonological awareness, rapid automatized naming and morphological awareness. To identify the genetic basis of Chinese reading disability, we conducted a genome-wide association study (GWAS) of the cognitive traits related to Chinese reading disability in 2284 unrelated Chinese children. Among the traits analyzed in the present GWAS, we detected one genome-wide significant association (p < 5 × 10⁻⁸) on word reading fluency for one SNP on 4p16.2, within EVC genes (rs6446395, p = 7.33 × 10⁻¹⁰). Rs6446395 also showed significant association with Chinese character reading accuracy (p = 2.95 × 10⁻⁴), phonological awareness (p = 7.11 × 10⁻³) and rapid automatized naming (p = 4.71 × 10⁻³), implying multiple effects of this variant. The eQTL data showed that rs6446395 affected EVC expression in the cerebellum. Gene-based analyses identified a gene (PRDM10) to be associated with word reading fluency at the genome-wide level. Our study discovered a new candidate susceptibility variant for reading ability and provided new insights into the genetics of developmental dyslexia in Chinese children.     

Modulation of attention in healthy children using a course of EEG-feedback sessions

We studied changes in the power spectra of EEG in the course of sessions of feedback by EEG characteristics (neurofeedback sessions) and estimated the effects of neurofeedback on psychological and EEG correlates of voluntary attention. Indices of the latter were estimated using Bourdon’s test (a correcture test) and Schulte’s tables. Twenty-nine reasonably healthy 10-to 13-year-old children took part in the study; they were divided into two groups, an experimental group (n = 12) and a control group (n = 15). The results obtained support the statement on noticeable changes in the functional state of the brain both immediately in the course of a neurofeedback session and after a course of such trainings. Changes in the ratios of the spectral powers of the beta₁ vs theta rhythms and the low-frequency beta vs theta rhythms were found in EEG recorded from the sensorimotor zone of the right hemisphere (C4). The observed changes in the spectral characteristics of EEG induced by the course of neurofeedback sessions were accompanied by the improvement of a few indices of voluntary attention. Neirofiziologiya/Neurophysiology, Vol. 38, Nos. 5/6, pp. 458–465, September–December, 2006.     

Cortical Signatures of Dyslexia and Remediation: An Intrinsic Functional Connectivity Approach

This observational, cross-sectional study investigates cortical signatures of developmental dyslexia, particularly from the perspective of behavioral remediation. We employed resting-state fMRI, and compared intrinsic functional connectivity (iFC) patterns of known reading regions (seeds) among three dyslexia groups characterized by (a) no remediation (current reading and spelling deficits), (b) partial remediation (only reading deficit remediated), and (c) full remediation (both reading and spelling deficits remediated), and a group of age- and IQ-matched typically developing children (TDC) (total N = 44, age range = 7–15 years). We observed significant group differences in iFC of two seeds located in the left posterior reading network – left intraparietal sulcus (L.IPS) and left fusiform gyrus (L.FFG). Specifically, iFC between L.IPS and left middle frontal gyrus was significantly weaker in all dyslexia groups, irrespective of remediation status/literacy competence, suggesting that persistent dysfunction in the fronto-parietal attention network characterizes dyslexia. Additionally, relative to both TDC and the no remediation group, the remediation groups exhibited stronger iFC between L.FFG and right middle occipital gyrus (R.MOG). The full remediation group also exhibited stronger negative iFC between the same L.FFG seed and right medial prefrontal cortex (R.MPFC), a core region of the default network These results suggest that behavioral remediation may be associated with compensatory changes anchored in L.FFG, which reflect atypically stronger coupling between posterior visual regions (L.FFG-R.MOG) and greater functional segregation between task-positive and task-negative regions (L.FFG-R.MPFC). These findings were bolstered by significant relationships between the strength of the identified functional connections and literacy scores. We conclude that examining iFC can reveal cortical signatures of dyslexia with particular promise for monitoring neural changes associated with behavioral remediation.     

Project DyAdd: Fatty acids in adult dyslexia, ADHD, and their comorbid combination

In project DyAdd, we compared the fatty acid (FA) profiles of serum phospholipids in adults with attention deficit hyperactivity disorder (ADHD) (n=26), dyslexia (n=36), their comorbid combination (n=9), and healthy controls (n=36). FA proportions were analyzed in a 2×2 design with Bonferroni corrected post hoc comparisons. A questionnaire was used to assess dietary fat quality and use of supplements. Results showed that ADHD and dyslexia were not associated with total saturated FAs, monounsaturated FAs, or n-3 polyunsaturated FAs (PUFAs). However, those with ADHD had elevated proportions of total n-6 PUFAs (including γ-linolenic and adrenic acids) as compared to those without ADHD. Dyslexia was related to a higher proportion of monounsaturated nervonic acid and a higher ratio of n-6/n-3 PUFAs. Among females none of the associations were significant. However in males, all the original associations observed in all subjects remained and ADHD was associated with elevated nervonic acid and n-6/n-3 PUFA ratio like dyslexia. Controlling for poorly diagnosed reading difficulties, education, dietary fat quality, or use of FA supplements did not generally remove the originally observed associations.     

Prevalence of <Emphasis Type="Italic">emm</Emphasis> types of Group A streptococci recovered from school children and hospital patients in Bangalore City,India

A total of sixty non repetitive isolates of group A streptococci (GAS) recovered from throat cultures of asymptomatic school children (n = 36), school children with clinical pharyngitis (n = 9) and hospital patients with clinical pharyngitis (n = 4) as well as diverse clinical samples from invasive GAS disease (n = 11) were subjected to emm typing. A total of 35 emm types including 3 new subtypes were identified among them. Types emm 12 (16.6%) and emm 71 (6.6%) were the most prevalent. The distribution of emm types identified among school children were similar to those encountered among invasive isolates but was significantly different from those reported elsewhere in India. Our study confirms the high diversity among Indian GAS isolates as well as significant regional differences among the distribution of emm types.     

Reading Performance Is Enhanced by Visual Texture Discrimination Training in Chinese-Speaking Children with Developmental Dyslexia

Background

High order cognitive processing and learning, such as reading, interact with lower-level sensory processing and learning. Previous studies have reported that visual perceptual training enlarges visual span and, consequently, improves reading speed in young and old people with amblyopia. Recently, a visual perceptual training study in Chinese-speaking children with dyslexia found that the visual texture discrimination thresholds of these children in visual perceptual training significantly correlated with their performance in Chinese character recognition, suggesting that deficits in visual perceptual processing/learning might partly underpin the difficulty in reading Chinese.

Methodology/Principal Findings

To further clarify whether visual perceptual training improves the measures of reading performance, eighteen children with dyslexia and eighteen typically developed readers that were age- and IQ-matched completed a series of reading measures before and after visual texture discrimination task (TDT) training. Prior to the TDT training, each group of children was split into two equivalent training and non-training groups in terms of all reading measures, IQ, and TDT. The results revealed that the discrimination threshold SOAs of TDT were significantly higher for the children with dyslexia than for the control children before training. Interestingly, training significantly decreased the discrimination threshold SOAs of TDT for both the typically developed readers and the children with dyslexia. More importantly, the training group with dyslexia exhibited significant enhancement in reading fluency, while the non-training group with dyslexia did not show this improvement. Additional follow-up tests showed that the improvement in reading fluency is a long-lasting effect and could be maintained for up to two months in the training group with dyslexia.

Conclusion/Significance

These results suggest that basic visual perceptual processing/learning and reading ability in Chinese might at least partially rely on overlapping mechanisms.     

Analysis of dyslexia candidate genes in the Raine cohort representing the general Australian population

Several genes have been suggested as dyslexia candidates. Some of these candidate genes have been recently shown to be associated with literacy measures in sample cohorts derived from the general population. Here, we have conducted an association study in a novel sample derived from the Australian population (the Raine cohort) to further investigate the role of dyslexia candidate genes. We analysed markers, previously reported to be associated with dyslexia, located within the MRPL19/C2ORF3, KIAA0319, DCDC2 and DYX1C1 genes in a sample of 520 individuals and tested them for association with reading and spelling measures. Association signals were detected for several single nucleotide polymorphisms (SNPs) within DYX1C1 with both the reading and spelling tests. The high linkage disequilibrium (LD) we observed across the DYX1C1 gene suggests that the association signal might not be refined by further genetic mapping.     

Age-related changes of muscle and plasma amino acids in healthy children

The aim of the study was to explore if changes in muscle and plasma amino acid concentrations developed during growth and differed from levels seen in adults. The gradient and concentrations of free amino acids in muscle and plasma were investigated in relation to age in metabolic healthy children. Plasma and specimens from the abdominal muscle were obtained during elective surgery. The children were grouped into three groups (group 1: < 1 year, n = 8; group 2: 1–4 years, n = 13 and group 3: 5–15 years, n = 15). A reference group of healthy adults (21–38 years, n = 22) was included in their comparisons and reflected specific differences between children and adults. In muscle the concentrations of 8 out of 19 amino acids analysed increased with age, namely taurine, aspartate, threonine, alanine, valine, isoleucine, leucine, histidine, as well as the total sums of branched chain amino acids (BCAA), basic amino acids (BAA) and total sum of amino acids (P < 0.05). In plasma the concentrations of threonine, glutamine, valine, cysteine, methionine, leucine, lysine, tryptophane, arginine, BCAA, BAA and the essential amino acids correlated with age (P < 0.05). These results indicate that there is an age dependency of the amino acid pattern in skeletal muscle and plasma during growth.     

Different fertility and meiotic regularity in allohexaploids derived from trigenomic hybrids between three cultivated <Emphasis Type="Italic">Brassica</Emphasis> allotetraploids and <Emphasis Type="Italic">B</Emphasis>. <Emphasis Type="Italic">maurorum</Emphasis>

The wild species Brassica maurorum Durieu (MM, 2n = 16) is useful for the improvement of Brassica crops. Herein, interspecific reciprocal crosses between B. maurorum and three cultivated Brassica allotetraploids were carried out with the aid of embryo rescue. Trigenomic hybrids with Brassica napus (AACC, 2n = 38) and Brassica carinata (BBCC, 2n = 34) were produced from reciprocal crosses, but the hybrids with Brassica juncea (AABB, 2n = 36) were obtained only when B. maurorum was used as female. All the hybrids were morphologically intermediate between their parents, and were male and female sterile. By in vitro chromosome doubling of the trigenomic hybrids, the allohexaploids (AACC.MM/MM.AACC, 2n = 54; BBCC.MM, 2n = 50; MM.AABB, 2n = 52) were established and characterized for their phenotype and cytology. The fertilities of three allohexaploids were different, for AACC.MM and MM.AACC failed to produce seeds by selfing, but BBCC.MM showed low seed-set and MM.AABB had good seed-set. They also expressed variable extents of male meiotic regularity as to chromosome pairing and segregation, with MM.AABB > BBCC.MM > AACC.MM/MM.AACC, the same order as their fertility. So their meiotic behavior contributed to the fertility. Finally, the potential of these allohexaploids as a bridge for genetic improvement of Brassica crops was discussed.     

Spatial properties of a forest buffalo herd and individual positioning as a response to environmental cues and social behaviour

Many animals aggregate into organized temporary or stable groups under the influence of biotic and abiotic factors, and some studies have shown the influence of habitat features on animal aggregation. This study, conducted from 2002 to 2004 in the Dzanga-Ndoki National Park, Central African Republic, studied a herd of forest buffaloes (Syncerus caffer nanus) to determine whether spatial aggregation patterns varied by season and habitat. Our results show that both habitat structure and season influenced spatial aggregation patterns. In particular, in open habitats such as clearings, the group covered a larger area when resting and was more rounded in shape compared to group properties noted in forest during the wet season. Moreover, forest buffaloes had a more aggregated spatial distribution when resting in clearings than when in the forest, and individual positions within the herd in the clearing habitat varied with age and sex. In the clearings, the adult male (n = 24) was generally, on most occasions, located in the centre of the herd (n = 20), and he was observed at the border only four times. In contrast, females (n = 80) occupied intermediate (n = 57), peripheral (n = 14) and central positions (n = 9) within the group. Juveniles (n = 77) also occurred in intermediate (n = 64) and peripheral positions (n = 13). Based on these results, we concluded that habitat characteristics and social behaviour can have relevant effects on the spatial distribution of animals within a group.     

Additions to the cytologically investigated species of <Emphasis Type="Italic">Potentilla</Emphasis> L. (Rosaceae) from India

At present 14 species of Potentilla L. have been cytologically worked out from different geographical areas of Kashmir and Himachal Pradesh in the Western Himalayas. New chromosome numbers in nine species—Potentilla argyrophylla (n = 14), P. atrosanguinea (n = 7, 14), P. desertorum (n = 7), P. gerardiana (n = 14), P. indica (n = 14), P. micropetala (n = 14), P. nepalensis (n = 14), P. sibbaldia (n = 14) and P. thomsonii (n = 7)—have been reported on a worldwide basis for the first time. Additional chromosomal races of polyploid cytotypes for P. argyrophylla (n = 28) and P. desertorum (n = 14) along with a diploid cytotype for P. micropetala (n = 7) plus diploid cytotypes for the five species as P. fulgens (n = 7), P. gelida (n = 7), P. kleiniana (n = 7), P. sibbaldia (n = 7) and P. sundaica (n = 7) as well as a tetraploid cytotype for P. fruticosa (n = 14) all have been cytologically worked out from India for the first time. The course of meiosis varies from normal to abnormal in different populations of the majority of the species, such as P. argyrophylla, P. atrosanguinea, P. desertorum, P. fruticosa, P. fulgens, P. gelida, P. indica, P. nepalensis, P. sibbaldia and P. sundaica, except for normal meiosis observed in P. gerardiana, P. kleiniana, P. micropetala and P. thomsonii. The anomalous taxa are marked with meiotic abnormalities in the form of cytomixis, chromosomal stickiness, unoriented bivalents, formation of laggards and bridges resulting in abnormal microsporogenesis, and production of heterogenous-sized fertile pollen grains along with reduced pollen fertility. All the taxa with normal meiotic courses show nearly one hundred percent pollen fertility.     

Emergence,propagation or disappearance of novel behavioral patterns in the habituated chimpanzees of Mahale: a review

Each local population of chimpanzees shows cultural variation, but little is known about how behavioral variations first emerge, and how often variants spread to other individuals and then become fixed as a local culture in chimpanzee society. Although field studies of chimpanzees are still too short to answer these questions definitively, it may stimulate further study in various sites to summarize the developments observed over the past 40 years at Mahale, Tanzania. Innovative patterns were operationally defined as new behavioral patterns performed by M group chimpanzees from 1981 onwards. Innovations included patterns of feeding (n = 8), human-directed behavior (n = 3), hygiene behavior (n = 4), maternal carrying of infants (n = 2), courtship (n = 2), play (n = 6), intimidation displays (n = 3), and quasi-grooming (n = 4). Although most patterns were repeated later by other individuals, six patterns were never seen performed by another individual, and eight patterns were performed by one or a few individuals but social transmission was unlikely. Thus, innovation was not rare, but emergence of fashion or establishment of traditions seems to occur rarely in chimpanzee society.     

Effect of Autogenic Training on General Improvement in Patients with Irritable Bowel Syndrome: A Randomized Controlled Trial

Autogenic training (AT) is a useful and comprehensive relaxation technique. However, no studies have investigated the effects of AT on irritable bowel syndrome (IBS). In this study we tested the hypothesis that AT improves symptoms of IBS. Twenty-one patients with IBS were randomly assigned to AT (n = 11, 5 male, 6 female) or control therapy (n = 10, 5 male, 5 female). AT patients were trained intensively, while the control therapy consisted of discussions about patients’ meal habits and life styles. All patients answered a question related to adequate relief (AR) of IBS symptoms and four questionnaires: Self-induced IBS Questionnaire (SIBSQ), Self-reported Depression Scale (SDS), State-Trait Anxiety Inventory (STAI), and Medical Outcome Short Form 36 Health Survey (SF-36). The proportion of AR in the last AT session in the AT group (9/11, 81.8%) was significantly higher than that in the controls (3/10, 30.0%, Chi-square test, p = 0.048). Two subscales of the SF-36, i.e., social functioning and bodily pain, were significantly improved in the AT group (p < 0.05) as compared to the control group. Role emotional (p = 0.051) and general health (p = 0.068) showed a tendency for improvement in the AT group. AT may be useful in the treatment of IBS by enhancing self-control.     

Genetic diversity in Iranian indigenous sheep vis-à-vis selected exogenous sheep breeds and wild mouflon

The heterogeneity of climate and different agro-ecological conditions in Iran have resulted in development of 27 indigenous sheep breeds. Wild Asiatic mouflon (Ovis orientalis) is believed to be the ancestor of Iranian sheep. Evaluation of genetic diversity and population structure within and among domestic breeds has important implications for animal breeding programs and genetic resources management. Based on 50K SNP genotype data, we studied the genetic diversity of five indigenous Iranian sheep breeds: Afshari (n = 37), Moghani (n = 34), Qezel (n = 35), Zel (n = 46) and Lori-Bakhtiari (n = 46), and Asiatic mouflon (n = 8) sampled from Iran. Furthermore, genetic diversity and the breed admixture of Iranian sheep were assessed on a larger geographic scale using a reference panel comprising: three indigenous Afghan breeds – Arabi (n = 15), Balouchi (n = 15) and Gadik (n = 15); three indigenous breeds from Turkey and Cyprus – Cyprus Fat Tail (n = 30), Karakas (n = 18) and Norduz (n = 20); and three commercial European breeds – Suffolk (n = 19), Comisana (n = 24) and Engadine Red Sheep (n = 24). The results revealed that the investigated breeds are divided into five genetically distinct clusters according to their geographic origin. Afshari was closest to the local mouflon population and showed signs of mouflon admixture. Qezel was identified as a hybrid sheep breed. Much evidence supported the Afghan breeds being identical. Inbreeding values, which were estimated based on ROHs, were highest for Suffolk (F_ROH = 0.0544) and lowest for Balouchi (F_ROH = 0.0078). In conclusion, analysis of selected breeds from neighboring countries along with Asiatic mouflon gave a deeper insight into the evolutionary history and origin of Iranian sheep with important implications for future breed management.     

Effects of Age and Region on Fecal Microflora in Elderly Subjects Living in Bama,Guangxi, China

Intestinal microflora analysis was performed on 52 healthy elderly subjects of different ages and in different regions in Bama County, Guangxi, China. The participants were assigned to three groups depending on their age and location: longevous (group M; mean age = 98 years; n = 21); rural younger elderly (group S; mean age = 70 years; n = 18); and urban elderly (group C; mean age = 82 years; n = 13). Ten groups of bacteria were quantified using real-time PCR. Age-related differences were observed in the number of Clostridium coccoides–Eubacterium rectale—there were more in longevous participants. Region affected the numbers of Bacteroides—Prevotella and Clostridium perfringens subgroup, and longevous participants had significantly more of the two bacterial groups than urban elderly participants. Region-related effects were also observed for the relative abundance of E. coli, and rural elderly participants had a lower proportion. Both age and regional effects were observed in the amount of total bacteria, and longevous participants had higher numbers than urban elderly participants. A significantly higher proportion of lactobacilli was observed in rural younger elderly participants than urban elderly participants, but independent age or regional effects did not contribute to this difference. This study suggests that age and region can affect the intestinal microflora of elderly people.     

Exploring the Effectiveness of a Computer-Based Heart Rate Variability Biofeedback Program in Reducing Anxiety in College Students

Given the pervasiveness of stress and anxiety in our culture it is important to develop and implement interventions that can be easily utilized by large numbers of people that are readily available, inexpensive and have minimal side effects. Two studies explored the effectiveness of a computer-based heart rate variability biofeedback program on reducing anxiety and negative mood in college students. A pilot project (n = 9) of highly anxious students revealed sizable decreases in anxiety and negative mood following utilizing the program for 4 weeks. A second study (n = 35) employing an immediate versus delayed treatment design replicated the results, although the magnitude of the impact was not quite as strong. Despite observing decreases in anxiety, the expected changes in psychophysiological coherence were not observed.     

Elevated serum levels of IL-1ra in children with Plasmodium falciparum malaria are associated with increased severity of disease

Animal models suggest that cytokines and chemokines play a role in cerebral malaria (CM) pathogenesis, but levels of a number of cytokines and chemokines thought to be important in the pathogenesis of other infectious diseases are not well characterized in children with CM. Serum levels of granulocyte-colony stimulating factor (G-CSF), interleukin-1 receptor antagonist (IL-1ra), interleukin-8 (IL-8) and monocyte chemoattractant protein-1 (MCP-1) were measured in 77 children with CM, 70 children with uncomplicated malaria (UM) and 63 healthy community children (CC) in Uganda. Children with CM had elevated serum levels of IL-1ra and IL-8 as compared to children with UM (median levels in pg/ml, 11,891 vs. 6510, P = 0.05, and 63 vs. 41, P = 0.01, respectively). Children with CM who died (n = 4) had higher serum levels than survivors of IL-1ra (median levels in pg/ml, 65,757 vs. 10,355, P = 0.02), G-CSF (709 vs. 117, P = 0.02), and MCP-1 (1275 vs. 216, P = 0.03) but not IL-8 (76 vs. 62, P = NS). Elevated IL-1ra levels are associated with increased disease severity in children with malaria, and very elevated levels of IL-1ra, G-CSF and MCP-1 are seen in children who die of CM.     

Evaluation of long‐term effects of artificial sweeteners on rat brain: a biochemical,behavioral, and histological study

The aim of the present study was to compare the effects of artificial sweeteners (aspartame, saccharin, and sucralose) on rat brain. Twenty‐four adult male Sprague–Dawley rats were included in the study. The control group (n = 6) received regular tap water, whereas other groups received aspartame (3 mg/kg/day, n = 6,) or saccharin (3 mg/kg/day, n = 6) or sucralose (1.5 mg/kg/day, n = 6) in the drinking water. Following 6 weeks, the passive avoidance learning (PAL) test was performed to evaluate the neurobehavioral effects of sweeteners. The brains were assessed for lipid peroxides, neuron count, and Glial fibrillary acidic protein (GFAP) immunohistochemistry. Our results demonstrated that chronic intake of sweeteners significantly impaired PAL performance in all groups. Hippocampal CA1–CA3 areas revealed significantly lower neuronal count in aspartame and increased GFAP expression in all groups. Brain lipid peroxides were significantly higher in all groups. Our findings suggest that long‐term consumption of artificial sweeteners may have harmful effects on cognition and hippocampal integrity in rats.