Effect of cerebrocrast on body and organ weights,food and water intake,and urine output of normal rats

Type 2 diabetes is associated with obesity, insulin resistance, hyperglycemia, hyperphagia, polyuria, body weight gain, excessive secretion of glucocorticoids (GCs), thymus involution, adrenal gland hypertrophy, diabetic nephropathy, etc. We examined the effect of cerebrocrast, a new antidiabetic agent (synthesized in the Latvian Institute of Organic Synthesis), on body weight, food and water intake, urine output, and on changes of organ weight: that is, kidney, thymus, adrenal gland of normal rats. Cerebrocrast was administered at doses of 0.05 and 0.5 mg kg⁻¹ per os (p.o.) once a day for three consecutive days, and its effects were observed from 3 to 27 days after the last administration. Cerebrocrast, during the experimental period, decreased body weight by an average of approximately 32.3%, food intake by about 10–15% at the beginning of the experiments and by 22.6% at the end of the experiments, especially at a dose of 0.5 mg kg⁻¹. Water intake and urine output in comparison with controls were decreased. The daily food intake decreased about 1.0 and 2.1 g by administering single cerebrocrast doses of 0.05 and 0.5 mg kg⁻¹ body weight (b.w.), respectively, but by administering for three consecutive days, food intake decreased by about 2.2 and 3.4 g, respectively. The weekly body weight gain decreased by administering a single dose of cerebrocrast by 2.61 and 2.51 g, respectively, and by triple administration it decreased by 4.36 and 3.07 g, respectively. Cerebrocrast has long‐lasting effects on these parameters and on thymus and adrenal gland weight. As cerebrocrast decreased glucose levels in normal and streptozotocin (STZ)‐induced diabetic rats, it also promoted glucose uptake by the brain, intensified insulin action and formation de novo of insulin receptors. We can conclude that cerebrocrast may regulate food intake and body weight through glucose sensing by proopiomelanocortin (POMC) neurons, that are involved in control of glucose homeostasis, stimulation of α‐melanocyte‐stimulating hormone (α‐MSH) secretion, activation of MC4‐Rs and inhibition of neuropeptide Y (NPY) in the ARC of the hypothalamus, affecting the kidney, and causing decreased urine output and water intake. Moreover, it could stimulate secretion of vasopressin. By administration of cerebrocrast thymus mass was increased, thereby preventing the action of GCs. As cerebrocrast inhibited L‐ and T‐type calcium channels, it can prevent vasoconstriction of kidney arterioles and aldosterone secretion that have significant roles in the development of hypertension and diabetic nephropathy. These properties of cerebrocrast are important for treatment of Type 2 diabetes and its consequent development of hypertension and diabetic nephropathy. Copyright © 2008 John Wiley & Sons, Ltd.     

The relationship between organ weights and body weights, facial dimensions, and dental dimensions in a population of olive baboons (Papio cynocephalus anubis)

Questions concerned with the relationship between organ weights and body weight on an intraspecific level are best answered by using a sample of animals collected in the wild from a single locale during a single season. The organ weights and body weights for this study were obtained from the necropsy reports prepared in the field (Athi Plain, Kenya) by H. C. McGill, Jr. on 36 adult animals (18 males and 18 females). Dental and facial measurements were taken by M.I. Siegel. In order to avoid erroneous results produced by statistical treatment of combined sex samples of sexually dimorphic species, data on the sexes were analyzed separately. Means and standard deviations are reported for selected organ weights and body dimensions (heart, lung, liver, spleen, kidney, adrenal, brain, and pancreas weights, crown–rump length, crown–heel length, head circumference, chest circumference, and body weight). All of the above measures were significantly (p<0.05) different between the sexes. Logarithms of these measures were significantly correlated with the logarithm of body weight in males for heart, lung, liver, spleen, kidney, and adrenal weights, crown–heel length, and head and chest circumference, and in females for crown–heel length, heart, liver, and kidney weights. Partial correlational analysis, removing the effects of body size (weight), showed mostly negative correlational relationships between dental and visceral dimensions. Most of the correlations between facial and visceral dimensions were negative. Allometric equations were calculated for the dental, facial, and visceral dimensions versus body weight, and are compared with prior published results.     

High throughput screening of rooting depth in rice using buried herbicide

Root research requires high throughput phenotyping methods that provide meaningful information on root depth if the full potential of the genomic revolution is to be translated into strategies that maximise the capture of water deep in soils by crops. A very simple, low cost method of assessing root depth of seedlings using a layer of herbicide (TRIK or diuron) buried 25 or 30 cm deep in soil‐filled boxes of varying size is described that is suitable for screening hundreds or thousands of rice accessions in controlled environment conditions. Variation in cultivar sensitivity to the herbicide when injected into pots was detected but considered small in relation to the variation detected when the herbicide was buried. Using 32 rice cultivars previously characterised for root traits in rhizotron and hydroponic systems, 80% of variation in herbicide score at 35 days was explained by cultivar and herbicide score correlated strongly with rooting depth traits. Using 139 genotypes of the Bala × Azucena mapping population, heritability for herbicide symptoms reached 55% and quantitative trait loci were detected which match those previously reported in this population. In repeated experiments using different soils, the method did not always perform to its maximum potential (in terms of speed of symptom development or discrimination between cultivars). This was not due to degradation or reduced bio‐availability of the herbicide in the soil but is believed to be due to the soil water content and water release characteristics as it relates to plant water use. Therefore, when using this technique, thorough preliminary experiments to determine the best water application regime for the particular combination of soil and environmental conditions are required. The method should be applicable to seedling stage screening of rice and other crops.     

Internal organ weights of the blue penguin Eudyptula minor

Abstract

This work presents a database of weights of normal internal organs for the blue penguin, Eudyptula minor. During a necropsy study of 213 adult and fledged immature blue penguins, external body measurements and internal organ weights were recorded. The relationship between organ weights and body weights shows that the liver and abdominal fat pad are most closely correlated with the body mass index, a quantitative indication of adequacy of condition.     

The relationship between body and gonadal weights of the dusky leaf monkey (Presbytis obscura)

Data from 25 male and 44 female free-ranging dusky leaf monkeys, Presbytis obscura,are analyzed. In a male the mean total testicular weight, 5.82 g, represents 0.078% of the mean adult body weight, 7.39 kg. The mean total ovarian weight, 0.52 g, represents 0.008% of the mean female adult body weight, 6.47 kg. A moderate degree of sexual dimorphism thus exists in this species, female body weight representing 88% that of the male. No seasonal variation in the adult body weight or total gonadal weight of either sex occurs. Only in males is there a strong positive correlation between left and right gonadal weight, and no correlation exists between the total gonadal weight and the estimated age of either male or female adults. Among adults, only females evince a significant positive correlation between total gonadal weight and body weight. No marked change in total ovarian weight occurs during the menstrual cycle, pregnancy, or lactation.     

Annual changes in blood parameters,organ weights,and total body lipid in Peromyscus maniculatus and Microtus ochrogaster from a prairie environment

Abstract

Annual cycles in hematological parameters, organ weights, and total body fat were measured in two species of mice from a prairie environment. Hematocrit was higher in winter in both Microtus ochrogaster and Peromyscus maniculatus. Plasma protein concentration decreased in winter in both species. Liver, kidneys, and adrenal glands were larger in female P. maniculatus than in males. Greatest liver and kidney size occurred during lactation. Total body lipid was about 5 to 7% of live body weight throughout the year, providing enough energy to survive only about one day of fasting. Adaptive shifts in bodily functions can be expected to coincide with periods of unusual energy expenditure such as lactation and winter. In these mice conserving energy seems to be a more important survival strategy than storing it.     

Beyond the response—High throughput behavioral analyses to link genome to phenome in Caenorhabditis elegans

The development and application of methods for automated behavioral analysis have revolutionized behavioral genetics across model organisms. In this review we summarize the history of automated behavioral analysis in the nematode Caenorhabditis elegans. We highlight recent studies of learning and memory to exemplify just how complex the genetic and neural circuit mechanisms underlying a seemingly simple single behavioral response can be. We finish by looking forward at the exciting prospects of combing genomic technologies with connectomic and phenomic level measurements.     

Mapping QTL for growth and shank traits in chickens divergently selected for high or low body weight

An F₂ population (695 individuals) was established from broiler chickens divergently selected for either high (HG) or low (LG) growth, and used to localize QTL for developmental changes in body weight (BW), shank length (SL9) and shank diameter (SD9) at 9 weeks. QTL mapping revealed three genome‐wide QTL on chromosomes (GGA) 2, 4 and 26 and three suggestive QTL on GGA 1, 3 and 5. Most of the BW QTL individually explained 2–5% of the phenotypic variance. The BW QTL on GGA2 explained about 7% of BW from 3 to 7 weeks of age, while that on GGA4 explained 15% of BW from 5 to 9 weeks. The BW QTL on GGA2 and GGA4 could be associated with early and late growth respectively. The GGA4 QTL also had the largest effect on SL9 and SD9 and explained 7% and 10% of their phenotypic variances respectively. However, when SL9 and SD9 were corrected with BW9, a shank length percent QTL was identified on GGA2. We identified novel QTL and also confirmed previously identified loci in other chicken populations. As the foundation population was established from commercial broiler strains, it is possible that QTL identified in this study could still be segregating in commercial strains.     

Quantitative trait loci for organ weights and adipose fat composition in Jersey and Limousin back‐cross cattle finished on pasture or feedlot

A QTL study of live animal and carcass traits in beef cattle was carried out in New Zealand and Australia. Back‐cross calves (385 heifers and 398 steers) were generated, with Jersey and Limousin backgrounds. This paper reports on weights of eight organs (heart, liver, lungs, kidneys, spleen, gastro‐intestinal tract, fat, and rumen contents) and 12 fat composition traits (fatty acid (FA) percentages, saturated and monounsaturated FA subtotals, and fat melting point). The New Zealand cattle were reared and finished on pasture, whilst Australian cattle were reared on grass and finished on grain for at least 180 days. For organ weights and fat composition traits, 10 and 12 significant QTL locations (P < 0.05), respectively, were detected on a genome‐wide basis, in combined‐sire or within‐sire analyses. Seven QTL significant for organ weights were found at the proximal end of chromosome 2. This chromosome carries a variant myostatin allele (F94L), segregating from the Limousin ancestry, and this is a positional candidate for the QTL. Ten significant QTL for fat composition were found on chromosomes 19 and 26. Fatty acid synthase and stearoyl‐CoA desaturase (SCD1), respectively, are positional candidate genes for these QTL. Two FA QTL found to be common to sire groups in both populations were for percentages of C14:0 and C14:1 (relative to all FAs) on chromosome 26, near the SCD1 candidate gene.     

Detection of quantitative trait loci for body weights and conformation traits in Beijing ducks

Quantitative trait loci (QTL) for body weights and conformation traits were detected in Beijing ducks. Traits included body weights (BW) at hatching and at 1-7 weeks of age; lengths of the body (BL), keel bone (KBL), shank (SL) and neck (NL) at 7 weeks of age; width of breast (BTW) at 7 weeks; and girths of shank (SG) and breast (BG) at 7 weeks. Using a half-sib analysis with a multiple-QTL model, linkage between the phenotypic traits and 95 microsatellite markers was studied. Six genome-wide suggestive QTL for three body weights and two conformation traits were identified in CAU1, CAU2, CAU6 and CAU12. Chromosome-wide significant QTL influencing one body weight trait and one conformation trait were located in CAU4 and CAU10 respectively. Twelve chromosome-wide suggestive QTL for six body weight traits and four conformation traits were found in seven linkage groups (CAU1, CAU2, CAU3, CAU4, CAU6, CAU10 and CAU12). In addition, the QTL in CAU6 at 21 and 73 cM jointly affected SG and explained 10.6% of the phenotypic variation. This study provides the first evidence for QTL involved in body weights and conformation traits in ducks, and will stimulate further investigations into the genetic architecture of these traits in this species.     

High throughput automated microbial bioreactor system used for clone selection and rapid scale‐down process optimization

High throughput automated fermentation systems have become a useful tool in early bioprocess development. In this study, we investigated a 24 x 15 mL single use microbioreactor system, ambr 15f, designed for microbial culture. We compared the fed‐batch growth and production capabilities of this system for two Escherichia coli strains, BL21 (DE3) and MC4100, and two industrially relevant molecules, hGH and scFv. In addition, different carbon sources were tested using bolus, linear or exponential feeding strategies, showing the capacity of the ambr 15f system to handle automated feeding. We used power per unit volume (P/V) as a scale criterion to compare the ambr 15f with 1 L stirred bioreactors which were previously scaled‐up to 20 L with a different biological system, thus showing a potential 1,300 fold scale comparability in terms of both growth and product yield. By exposing the cells grown in the ambr 15f system to a level of shear expected in an industrial centrifuge, we determined that the cells are as robust as those from a bench scale bioreactor. These results provide evidence that the ambr 15f system is an efficient high throughput microbial system that can be used for strain and molecule selection as well as rapid scale‐up. © 2017 The Authors Biotechnology Progress published by Wiley Periodicals, Inc. on behalf of American Institute of Chemical Engineers Biotechnol. Prog., 34:58–68, 2018     

Influence of gel dimensions on resolution and sample throughput on two-dimensional gels

To achieve high throughput and economical format of 2-D PAGE, comparison between gel size and resolution was conducted on human breast carcinoma cell line (MCF-7/AZ) proteins. SDS gel length showed a weaker influence of separation length on resolution in the second dimension, and there was little benefit of separation distances greater than 15 to 19 cm. IPG strip separation distances were very important with dramatic increase in resolution of longer gels compared with smaller gels, and maximal resolution was obtained using 18- and 24-cm IPG strips. Loading optimal amount of proteins on 2-D gels can also increase the number of detected spots. Therefore, taken together, compromise 2-D gels are crucial for higher capacity and higher throughput.     

Analysis of genetic association and epistasis interactions between circadian clock genes and symptom dimensions of bipolar affective disorder

Bipolar affective disorder (BD) is a severe psychiatric disorder characterized by periodic changes in mood from depression to mania. Disruptions of biological rhythms increase risk of mood disorders. Because clinical representation of disease is heterogeneous, homogenous sets of patients are suggested to use in the association analyses. In our study, we aimed to apply previously computed structure of bipolar disorder symptom dimension for analyses of genetic association. We based quantitative trait on: main depression, sleep disturbances, appetite disturbances, excitement and psychotic dimensions consisted of OPCRIT checklist items. We genotyped 42 polymorphisms from circadian clock genes: PER3, ARNTL, CLOCK and TIMELSSS from 511 patients BD (n?=?292 women and n?=?219 men). As quantitative trait we used clinical dimensions, described above. Genetic associations between alleles and quantitative trait were performed using applied regression models applied in PLINK. In addition, we used the Kruskal–Wallis test to look for associations between genotypes and quantitative trait. During second stage of our analyses, we used multidimensional scaling (multifactor dimensionality reduction) for quantitative trait to compute pairwise epistatic interactions between circadian gene variants. We found association between ARNTL variant rs11022778 main depression (p?=?0.00047) and appetite disturbances (p?=?0.004). In epistatic interaction analyses, we observed two locus interactions between sleep disturbances (p?=?0.007; rs11824092 of ARNTL and rs11932595 of CLOCK) as well as interactions of subdimension in main depression and ARNTL variants (p?=?0.0011; rs3789327, rs10766075) and appetite disturbances in depression and ARNTL polymorphism (p?=?7?×?10^?4; rs11022778, rs156243).     

High throughput solution-based measurement of antibody-antigen affinity and epitope binning

Advances in human antibody discovery have allowed for the selection of hundreds of high affinity antibodies against many therapeutically relevant targets. This has necessitated the development of reproducible, high throughput analytical techniques to characterize the output from these selections. Among these characterizations, epitopic coverage and affinity are among the most critical properties for lead identification. Biolayer interferometry (BLI) is an attractive technique for epitope binning due to its speed and low antigen consumption. While surface-based methods such as BLI and surface plasmon resonance (SPR) are commonly used for affinity determinations, sensor chemistry and surface related artifacts can limit the accuracy of high affinity measurements. When comparing BLI and solution equilibrium based kinetic exclusion assays, significant differences in measured affinity (10-fold and above) were observed. KinExA direct association (k_a) rate constant measurements suggest that this is mainly caused by inaccurate k_a measurements associated with BLI related surface phenomena. Based on the kinetic exclusion assay principle used for KinExA, we developed a high throughput 96-well plate format assay, using a Meso Scale Discovery (MSD) instrument, to measure solution equilibrium affinity. This improved method combines the accuracy of solution-based methods with the throughput formerly only achievable with surface-based methods.     

High‐throughput mAb expression and purification platform based on transient CHO

A high‐cell‐density transient transfection system was recently developed in our laboratory based on a CHO‐GS‐KO cell line. This method yields monoclonal antibody titers up to 350 mg/L from a simple 7‐day process, in volumes ranging from 2 mL to 2 L. By performing transfections in 24‐deep‐well plates, a large number of mAbs can be expressed simultaneously. We coupled this new high‐throughput transfection process to a semiautomated protein A purification process. Using a Biomek FX^p liquid handling robot, up to 72 unique mAbs can be simultaneously purified. Our primary goal was to obtain >0.25 mg of purified mAb at a concentration of >0.5 mg/mL, without any concentration or buffer‐exchange steps. We optimized both the batch‐binding and the batch elution steps. The length of the batch‐binding step was important to minimize mAb losses in the flowthrough fraction. The elution step proved to be challenging to simultaneously maximize protein recovery and protein concentration. We designed a variable volume elution strategy based on the average supernatant titer. Finally, we present two case studies. In the first study, we produced 56 affinity maturation mAb variants at an average yield of 0.33 ± 0.05 mg (average concentration of 0.65 ± 0.10 mg/mL). In a second study, we produced 42 unique mAbs, from an early‐stage discovery effort, at an average yield of 0.79 ± 0.31 mg (average concentration of 1.59 ± 0.63 mg/mL). The combination of parallel high‐yielding transient transfection and semiautomated high‐throughput protein A purification represents a valuable mAb drug discovery tool. © 2014 American Institute of Chemical Engineers Biotechnol. Prog., 31:239–247, 2015