Long-term fasting induced basal thermogenesis flexibility in female Japanese quails.

Male Japanese quails (Coturnix japonica) have been found to exhibit a three-phase metabolic change when subjected to prolonged fasting, during which basal thermogenesis is significantly reduced. A study had shown that there is a significant difference in the body temperature between male and female Japanese quails. However, whether female Japanese quails also show the same characteristic three-phase metabolic change during prolonged fasting and the underlying thermogenesis mechanisms associated with such changes are still unclear. In this study, female Japanese quails were subjected to prolonged starvation, and the body mass, basal metabolic rate (BMR), body temperature, mass of tissues and organs, body fat content, the state-4 respiration (S4R) and cytochrome c oxidase (CCO) activity in the muscle and liver of these birds were measured to determine the status of metabolic changes triggered by the starvation. In addition, the levels of glucose, triglyceride (TG) and uric acid, and thyroid hormones (T3 and T4) in the serum and the mRNA levels of myostatin (MSTN) and avian uncoupling protein (av-UCP) in the muscle were also measured. The results revealed the existence of a three-phase stage similar to that found in male Japanese quails undergoing prolonged starvation. Fasting resulted in significantly lower body mass, BMR, body temperature, tissues masses and most organs masses, as well as S4R and CCO activity in the muscle and liver. The mRNA level of av-UCP decreased during fasting, while that of MSTN increased but only during Phase I and II and decreased significantly during Phase III. Fasting also significantly lowered the T3 level and the ratio of T3/T4 in the serum. These results indicated that female Japanese quails showed an adaptive response in basal thermogenesis at multiple hierarchical levels, from organismal to biochemical, enzyme and cellular level, gene and endocrine levels and this integrated adjustment could be a part of the adaptation used by female quails to survive long-term fasting.

Effect of Fungicide Application and Corn Hybrid Class on the Presence of Fusarium graminearum and the Concentration of Deoxynivalenol in Ear and Stalk Parts of Corn (Zea mays) Used for Silage.

In Wisconsin, the use of brown midrib (BMR) corn (Zea mays) hybrids for ensiling and subsequent feeding to dairy cows is quite common. The overall milk production from cows fed silage from BMR hybrids is typically higher than those fed silage made from dual-purpose hybrids. Gibberella diseases (ear and stalk rot) caused by Gibberella zeae (anamorph; Fusarium graminearum) and the accompanying accumulation of the mycotoxin deoxynivalenol (DON) can be significant issues during the field production of BMR hybrids. The work presented here aimed to understand the role of hybrid class on the distribution of F. graminearum DNA and DON in the ear and stalk parts of corn for silage. An ear and stalk partitioned sample experiment was conducted on silage corn from field trials in Arlington, Wisconsin, in 2020 and 2021. The trials were arranged in a randomized complete block design in both years, including one BMR hybrid, one dual-purpose hybrid, and seven fungicide application regimes. Paired ear and stalk samples were physically separated, dried, and ground at harvest before determining the concentration of F. graminearum DNA and DON in each sample. Across both years, the main effects of hybrid, treatment, and plant part were not significant (P > 0.1) on DON concentration. However, the hybrid-by-plant part interaction effect was significant (P < 0.01). Ears of the BMR hybrid accumulated the most DON, whereas the dual-purpose hybrid ears had the lowest DON concentration. The concentrations of DON and F. graminearum DNA were significantly (P < 0.01) and highly correlated in the ear (r = 0.73) but not in the stalk (r = 0.09, P = 0.33). These findings suggest that DON accumulation in the corn ear is a major contributor in the difference observed in the total DON between the hybrid classes. Therefore, growers and researchers are encouraged to focus production and breeding on hybrids in both classes that accumulate less DON in ears, resulting in lower total DON in corn chopped for silage.

Trends in measuring BMR and RMR after spinal cord injury: a comprehensive review.

Studying factors that contribute to our understanding of maintaining normal energy balance are of paramount significance following spinal cord injury (SCI). Accurate determination of energy needs is crucial for providing nutritional guidance and managing the increasing prevalence of malnutrition or obesity after SCI. BMR represents 75-80 % of the total energy expenditure in persons with SCI. Accurately measuring BMR is an important component for calculating total energetic needs in this population. Indirect calorimetry is considered the gold-standard technique for measuring BMR. However, technical challenges may limit its applications in large cohort studies and alternatively rely on prediction equations. Previous work has shown that BMR changes in response to disuse and exercise in the range of 15-120 %. Factors including sex, level of injury and type of assistive devices may influence BMR after SCI. RMR is erroneously used interchangeably for BMR, which may result in overestimation of energetic intake when developing nutritional plans. To address this concern, we comprehensively reviewed studies that conducted BMR (n=15) and RMR (n=22) in persons with SCI. The results indicated that RMR is 9 % greater than BMR in persons with SCI. Furthermore, the SCI-specific prediction equations that incorporated measures of fat-free mass appeared to accurately predict BMR. Overall, the current findings highlighted the significance of measuring BMR as well as encouraging the research and clinical community to effectively establish countermeasures to combat obesity after SCI.

Generation of human scFv-IgG1Fc antibodies for detection of lymphatic filarial recombinant antigens, BmR1 and BmSXP.

Lymphatic filariasis is a neglected parasitic disease that affects millions in tropical and subtropical countries and is caused by Wuchereria and Brugia species. Specific and sensitive detection methods are essential in mapping infected areas where rapid tests are needed to cover underdeveloped and remote regions, which facilitates eliminating the disease as a public health problem. A few commercialized rapid tests based on antigen or antibody detection are available, but the former only detects infection by Wuchereria species and cross-reacts with nonlymphatic filaria, whereas antibody detection might provide positive results of previous infection. Here, we report the production of three different recombinant immunoglobulin gamma (IgG)1 antibodies based on scFvs previously generated via human antibody phage display technology, that is, anti-BmR1 clone 4, anti-BmXSP clone 5B, and anti-BmXSP clone 2H2. The scFv sequences were cloned into a pCMV-IgG1 vector, then transfected into a HEK293F cell line. The generated antibodies were found to be able to bind to their respective targets even at relatively low concentration. Conjugation of Fc to scFv induces binder stability and provides multiple labeling sites for probes and signaling molecules that can be used in rapid tests.

The seasonal variation of basal metabolic rate is related to the expression of torpor among small mammals.

A variety of responses to climate seasonality have evolved by small mammals, including adjustments of the basal rate of metabolism (BMR) and the use of daily or seasonal torpor (here referred to as short-bout and long-bout torpor). The seasonal variation of their BMR is known to depend mainly on the concurrent variation of body mass, but it should also be affected by structural and functional changes occurring within the body that could depend on the expression of torpor. Thus it was hypothesized that BMR seasonality is related to the expression of torpor at an interspecific level. Seasonal BMR and body mass data were gathered from the literature and phylogenetic comparative analyses were done to test this hypothesis among mammals of less than 1 kg. BMR seasonality (dBMR) was quantified as the log-transformed ratio of the mean whole-animal BMR reported for the period P2 (autumn-winter) over that for the period P1 (spring-summer). Predictors were the seasonal body mass adjustment (dm), mean body mass (m) and torpor expression (TO, a three-level factor: no torpor, short-bout torpor, long-bout torpor). The seasonal variation of BMR was significantly related to dm but also to TO. Accounting for dm, species expressing long-bout torpor, but not those entering short-bout torpor, collectively exhibited a lower dBMR than species not entering torpor. Fat storage and use by species entering long-bout torpor, alone, could not explain their lower dBMR, as the TO:dm interaction was not significant. The low dBMR of species entering long-bout torpor may result from their collective tendency to down-regulate more strongly costly visceral organs during P2. The dBMR of the different TO categories overlapped appreciably, which highlights our still limited knowledge of the BMR seasonality among small mammals.

Molecular Lego of Human Cytochrome P450: The Key Role of Heme Domain Flexibility for the Activity of the Chimeric Proteins.

The cytochrome P450 superfamily are heme-thiolate enzymes able to carry out monooxygenase reactions. Several studies have demonstrated the feasibility of using a soluble bacterial reductase from Bacillus megaterium, BMR, as an artificial electron transfer partner fused to the human P450 domain in a single polypeptide chain in an approach known as 'molecular Lego'. The 3A4-BMR chimera has been deeply characterized biochemically for its activity, coupling efficiency, and flexibility by many different biophysical techniques leading to the conclusion that an extension of five glycines in the loop that connects the two domains improves all the catalytic parameters due to improved flexibility of the system. In this work, we extend the characterization of 3A4-BMR chimeras using differential scanning calorimetry to evaluate stabilizing role of BMR. We apply the 'molecular Lego' approach also to CYP19A1 (aromatase) and the data show that the activity of the chimeras is very low (<0.003 min−1) for all the constructs tested with a different linker loop length: ARO-BMR, ARO-BMR-3GLY, and ARO-BMR-5GLY. Nevertheless, the fusion to BMR shows a remarkable effect on thermal stability studied by differential scanning calorimetry as indicated by the increase in Tonset by 10 °C and the presence of a cooperative unfolding process driven by the BMR protein domain. Previously characterized 3A4-BMR constructs show the same behavior of ARO-BMR constructs in terms of thermal stabilization but a higher activity as a function of the loop length. A comparison of the ARO-BMR system to 3A4-BMR indicates that the design of each P450-BMR chimera should be carefully evaluated not only in terms of electron transfer, but also for the biophysical constraints that cannot always be overcome by chimerization.

Pathogen and drought stress affect cell wall and phytohormone signaling to shape host responses in a sorghum COMT bmr12 mutant.

BACKGROUND: As effects of global climate change intensify, the interaction of biotic and abiotic stresses increasingly threatens current agricultural practices. The secondary cell wall is a vanguard of resistance to these stresses. Fusarium thapsinum (Fusarium stalk rot) and Macrophomina phaseolina (charcoal rot) cause internal damage to the stalks of the drought tolerant C4 grass, sorghum (Sorghum bicolor (L.) Moench), resulting in reduced transpiration, reduced photosynthesis, and increased lodging, severely reducing yields. Drought can magnify these losses. Two null alleles in monolignol biosynthesis of sorghum (brown midrib 6-ref, bmr6-ref; cinnamyl alcohol dehydrogenase, CAD; and bmr12-ref; caffeic acid O-methyltransferase, COMT) were used to investigate the interaction of water limitation with F. thapsinum or M. phaseolina infection. RESULTS: The bmr12 plants inoculated with either of these pathogens had increased levels of salicylic acid (SA) and jasmonic acid (JA) across both watering conditions and significantly reduced lesion sizes under water limitation compared to adequate watering, which suggested that drought may prime induction of pathogen resistance. RNA-Seq analysis revealed coexpressed genes associated with pathogen infection. The defense response included phytohormone signal transduction pathways, primary and secondary cell wall biosynthetic genes, and genes encoding components of the spliceosome and proteasome. CONCLUSION: Alterations in the composition of the secondary cell wall affect immunity by influencing phenolic composition and phytohormone signaling, leading to the action of defense pathways. Some of these pathways appear to be activated or enhanced by drought. Secondary metabolite biosynthesis and modification in SA and JA signal transduction may be involved in priming a stronger defense response in water-limited bmr12 plants.

Energetic constraints on mammalian distribution areas.

Energy is a universal resource essential for all life functions. The rate of transformation of energy into an organism, and the energetic investment into reproduction, determines population and ecological-level processes. Several hypotheses predicted that the ecological expansion and size of the geographic distribution of a species are shaped by, among other factors, metabolic performance. However, how organismal energetic characteristics contribute to species geographic range size is poorly understood. With phylogenetic comparative methods whether energetic maintenance costs (basal metabolic rate, BMR), aerobic capacity (maximum exercise metabolic rate, VO2 max), summit thermoregulation (summit metabolic rate, VO2 sum) and the ability to sustain energy provisioning (daily energy expenditure, DEE) determine the distribution of mammalian species range sizes was tested. Both basal and maximum exercise metabolic rates (accounting for body mass), but not summit thermogenic metabolic rate, were positively associated with species range sizes. Furthermore, daily energy expenditure (accounting for body mass) was positively associated with species ranges. Body mass (accounting for energetic maintenance) was negatively related to range sizes. High aerobic exercise capacity, aiding mobility such as running and dispersal, and high sustained energy provisioning, aiding reproductive effort such as pregnancy, lactation and natal dispersal, can facilitate the establishment of large mammalian geographic ranges. Consequently, the pace of organismal physiological processes can shape important ecological and biodiversity patterns by setting limits to species' range sizes.

GLP-1 vasodilatation in humans with coronary artery disease is not adenosine mediated.

BACKGROUND: Incretin therapies appear to provide cardioprotection and improve cardiovascular outcomes in patients with diabetes, but the mechanism of this effect remains elusive. We have previously shown that glucagon-like peptide (GLP)-1 is a coronary vasodilator and we sought to investigate if this is an adenosine-mediated effect. METHODS: We recruited 41 patients having percutaneous coronary intervention (PCI) for stable angina and allocated them into four groups administering a specific study-related infusion following successful PCI: GLP-1 infusion (Group G) (n = 10); Placebo, normal saline infusion (Group P) (n = 11); GLP-1 + Theophylline infusion (Group GT) (n = 10); and Theophylline infusion (Group T) (n = 10). A pressure wire assessment of coronary distal pressure and flow velocity (thermodilution transit time-Tmn) at rest and hyperaemia was performed after PCI and repeated following the study infusion to derive basal and index of microvascular resistance (BMR and IMR). RESULTS: There were no significant differences in the demographics of patients recruited to our study. Most of the patients were not diabetic. GLP-1 caused significant reduction of resting Tmn that was not attenuated by theophylline: mean delta Tmn (SD) group G - 0.23 s (0.27) versus group GT - 0.18 s (0.37), p = 0.65. Theophylline alone (group T) did not significantly alter resting flow velocity compared to group GT: delta Tmn in group T 0.04 s (0.15), p = 0.30. The resulting decrease in BMR observed in group G persisted in group GT: - 20.83 mmHg s (24.54 vs. - 21.20 mmHg s (30.41), p = 0.97. GLP-1 did not increase circulating adenosine levels in group GT more than group T: delta median adenosine - 2.0 ng/ml (- 117.1, 14.8) versus - 0.5 ng/ml (- 19.6, 9.4); p = 0.60. CONCLUSION: The vasodilatory effect of GLP-1 is not abolished by theophylline and GLP-1 does not increase adenosine levels, indicating an adenosine-independent mechanism of GLP-1 coronary vasodilatation. TRIAL REGISTRATION: The local research ethics committee approved the study (National Research Ethics Service-NRES Committee, East of England): REC reference 14/EE/0018. The study was performed according to institutional guidelines, was registered on http://www.clinicaltrials.gov (unique identifier: NCT03502083) and the study conformed to the principles outlined in the Declaration of Helsinki.

Metabolic rate is negatively linked to adult survival but does not explain latitudinal differences in songbirds.

Survival rates vary dramatically among species and predictably across latitudes, but causes of this variation are unclear. The rate-of-living hypothesis posits that physiological damage from metabolism causes species with faster metabolic rates to exhibit lower survival rates. However, whether increased survival commonly observed in tropical and south temperate latitudes is associated with slower metabolic rate remains unclear. We compared metabolic rates and annual survival rates that we measured across 46 species, and from literature data across 147 species of birds in northern, southern and tropical latitudes. High metabolic rates were associated with lower survival but survival varied substantially among latitudinal regions independent of metabolism. The inability of metabolic rate to explain latitudinal variation in survival suggests (1) species may evolve physiological mechanisms that mitigate physiological damage from cellular metabolism and (2) extrinsic rather than intrinsic sources of mortality are the primary causes of latitudinal differences in survival.

Overexpression of ferulate 5-hydroxylase increases syringyl units in Sorghum bicolor.

Ferulate 5-hydroxylase (F5H) of the monolignol pathway catalyzes the hydroxylation of coniferyl alcohol, coniferaldehyde and ferulic acid to produce 5-hydroxyconiferyl moieties, which lead to the formation of sinapic acid and syringyl (S) lignin monomers. In contrast, guaiacyl (G) lignin, the other major type of lignin monomer, is derived from polymerization of coniferyl alcohol. In this study, the effects of manipulating S-lignin biosynthesis in sorghum (Sorghum bicolor) were evaluated. Overexpression of sorghum F5H (SbF5H), under the control of the CaMV 35S promoter, increased both S-lignin levels and the ratio of S/G lignin, while plant growth and development remained relatively unaffected. Maüle staining of stalk and leaf midrib sections from SbF5H overexpression lines indicated that the lignin composition was altered. Ectopic expression of SbF5H did not affect the gene expression of other monolignol pathway genes. In addition, brown midrib 12-ref (bmr12-ref), a nonsense mutation in the sorghum caffeic acid O-methyltransferase (COMT) was combined with 35S::SbF5H through cross-pollination to examine effects on lignin synthesis. The stover composition from bmr12 35S::SbF5H plants more closely resembled bmr12 stover than 35S::SbF5H or wild-type (WT) stover; S-lignin and total lignin concentrations were decreased relative to WT or 35S::SbF5H. Likewise, expression of upstream monolignol biosynthetic genes was increased in both bmr12 and bmr12 35S::SbF5H relative to WT or 35S::SbF5H. Overall, these results indicated that overexpression of SbF5H did not compensate for the loss of COMT activity. KEY MESSAGE: Overexpression of F5H in sorghum increases S-lignin without increasing total lignin content or affecting plant growth, but it cannot compensate for the loss of COMT activity in monolignol synthesis.

Not that hot after all: no limits to heat dissipation in lactating mice selected for high or low BMR.

Heat dissipation has been suggested as a limit to sustained metabolic effort, e.g. during lactation, when overheating is a possible risk. We tested this hypothesis using mice artificially selected for either high or low BMR that also differed with respect to parental effort. We used fixed size cross-fostered families and recorded litter mass daily until the 14th day of lactation. Midway through the experiment (day 8) half of the mothers from each group had fur from the dorsal body surface removed to increase their thermal conductance and facilitate heat dissipation. Our results showed that neither high nor low BMR mouse lines benefited from increasing their thermal conductance at peak lactation. On the contrary, growth of the litters reared by the low BMR females was compromised. Thus, our results do not support the heat dissipation limitation hypothesis.

What determines the basal rate of metabolism?

The basal rate of metabolism (BMR) is the most reported estimate of energy expenditure in endotherms. Its principal determinant is body mass, but BMR also correlates with a variety of behavioral and ecological factors that do not determine basal rate: they are byproducts of the mechanisms that are its determinate. In mammals, mass-independent BMR increases when muscle mass is >40% of total body mass and BMR is then ≥100% of the value expected from body mass. Mammals with muscle masses <30% of body mass have lower BMRs, a diminished capacity to regulate body temperature and often have reduced activity levels. At muscle masses <42% of body mass, birds have body temperatures and basal rates higher than mammals with the same muscle mass. Their high basal rates derive from fast blood flow and increased mitochondrial density in their pectoral muscles. These enhancements also occur in the flight muscles of bats. Oxygen transport to the pectoral muscles of birds is facilitated by an increase in heart mass and hematocrit. This arrangement avoids transporting a large muscle mass to fuel flight, thereby reducing the cost of flight. Pectoral muscle masses <9% of body mass correlate with a flightless condition in kiwis, rails and ducks but some fruit pigeons have BMRs as low as those measured in kiwis, while remaining volant. The mass-independent BMRs of endotherms principally reflect changes of muscle activity and mass. An increase in muscle mass may have contributed to the evolution of endothermy.

Post-operative analysis of pediatric esotropia associated with high hypermetropia.

BACKGROUND: To describe clinical features, evaluation, surgical management and outcomes in children with esotropia associated with high hypermetropia. METHODS: Medical records of healthy children who received strabismus surgery for accommodative esotropia with hypermetropia larger than spherical equivalence of + 4.0 diopters from 2009 to 2015, were reviewed. RESULTS: A total of 47 patients were identified. The average age was 2.9 years old. The average spherical equivalence of cycloplegic refraction was + 6.0 diopters (D). All patients presented with large angle esotropia with spectacle correction. Average age of esotropia onset was 1.3 years. Average time between the onset of esotropia to spectacle correction was 7.2 months. Average duration between onset of constant esotropia to strabismus surgery was 28.1 months. Average duration between spectacle correction to strabismus surgery was 21.8 months. Post-operatively, 74.5% of patients achieved ocular alignment within 10 prism diopters (PD) of orthotropia. Overall, 66.0% patients developed sensory fusion. For patients who achieved surgical success, 71.4% developed sensory fusion, compared at 50.0% for patients who were over- or under-corrected (p = 0.18). For patients who received hyperopic spectacles within 6 months of esotropia onset, 92.3% developed sensory fusion, compared with 54.5% for patients who received hyperopic spectacles at 6 month or later after esotropia onset (p = 0.02). CONCLUSIONS: Strabismus surgery for esotropia with high hypermetropia has high rate of surgical success with low rate of under- or over-correction. There is a trend toward higher rate of sensory fusion for patients with surgical success. Shorter time interval between esotropia onset and receiving hyperopic spectacles is associated with higher rate of sensory fusion development.

A general theory of effect size, and its consequences for defining the benchmark response (BMR) for continuous endpoints.

A general theory on effect size for continuous data predicts a relationship between maximum response and within-group variation of biological parameters, which is empirically confirmed by results from dose-response analyses of 27 different biological parameters. The theory shows how effect sizes observed in distinct biological parameters can be compared and provides a basis for a generic definition of small, intermediate and large effects. While the theory is useful for experimental science in general, it has specific consequences for risk assessment: it solves the current debate on the appropriate metric for the Benchmark response in continuous data. The theory shows that scaling the BMR expressed as a percent change in means to the maximum response (in the way specified) automatically takes "natural variability" into account. Thus, the theory supports the underlying rationale of the BMR 1 SD. For various reasons, it is, however, recommended to use a BMR in terms of a percent change that is scaled to maximum response and/or within group variation (averaged over studies), as a single harmonized approach.

Individual variation in metabolic reaction norms over ambient temperature causes low correlation between basal and standard metabolic rate.

Basal metabolic rate (BMR) is often assumed to be indicative of the energy turnover at ambient temperatures (Ta) below the thermoneutral zone (SMR), but this assumption has remained largely untested. Using a new statistical approach, we quantified the consistency in nocturnal metabolic rate across a temperature range in zebra finches (N=3213 measurements on 407 individuals) living permanently in eight outdoor aviaries. Foraging conditions were either benign or harsh, and body mass and mass-adjusted BMR (BMRm) and SMR (SMRm) were lower in individuals living in a harsh foraging environment. The correlation between SMRm at different Ta was high (r=0.91), independent of foraging environment, showing that individuals are consistently ranked according to their SMRm However, the correlations between BMRm and SMRm were always lower (average: r=0.29; range: 0

Contaminants and energy expenditure in an Arctic seabird: Organochlorine pesticides and perfluoroalkyl substances are associated with metabolic rate in a contrasted manner.

Basal metabolic rate (BMR), the minimal energetic cost of living in endotherms, is known to be influenced by thyroid hormones (THs) which are known to stimulate in vitro oxygen consumption of tissues in birds and mammals. Several environmental contaminants may act on energy expenditure through their thyroid hormone-disrupting properties. However, the effect of contaminants on BMR is still poorly documented for wildlife. Here, we investigated the relationships between three groups of contaminants (organochlorines (OCs), perfluoroalkyl substances (PFASs), and mercury) with metabolic rate (MR), considered here as a proxy of BMR and also with circulating total THs (thyroxine (TT4) and triiodothyronine (TT3)) in Arctic breeding adult black-legged kittiwakes (Rissa tridactyla) from Svalbard, during the chick rearing period. Our results indicate a negative relationship between the sum of all detected chlordanes (∑CHLs) and MR in both sexes whereas perfluorotridecanoate (PFTrA) and MR were positively related in females only. MR was not associated with mercury. Additionally, levels of TT3 were negatively related to ∑CHLs but not to PFTrA. The findings from the present study indicate that some OCs (in both sexes) and some PFASs (only in females) could disrupt fine adjustment of BMR during reproduction in adult kittiwakes. Importantly, highly lipophilic OCs and highly proteinophilic PFASs appear, at least in females, to have the ability to disrupt the metabolic rate in an opposite way. Therefore, our study highlights the need for ecotoxicological studies to include a large variety of contaminants which can act in an antagonistic manner.

Antibody-Based Protective Immunity against Helminth Infections: Antibody Phage Display Derived Antibodies against BmR1 Antigen.

Helminth parasite infections are significantly impacting global health, with more than two billion infections worldwide with a high morbidity rate. The complex life cycle of the nematodes has made host immune response studies against these parasites extremely difficult. In this study, we utilized two phage antibody libraries; the immune and naïve library were used to identify single chain fragment variable (scFv) clones against a specific filarial antigen (BmR1). The V-gene analysis of isolated scFv clones will help shed light on preferential VDJ gene segment usage against the filarial BmR1 antigen in healthy and infected states. The immune library showed the usage of both lambda and kappa light chains. However, the naïve library showed preferential use of the lambda family with different amino acid distributions. The binding characteristics of the scFv clones identified from this work were analyzed by immunoassay and immunoaffinity pull down of BmR1. The work highlights the antibody gene usage pattern of a naïve and immune antibody library against the same antigen as well as the robust nature of the enriched antibodies for downstream applications.

Heat dissipation does not suppress an immune response in laboratory mice divergently selected for basal metabolic rate (BMR).

The capacity for heat dissipation is considered to be one of the most important constraints on rates of energy expenditure in mammals. To date, the significance of this constraint has been tested exclusively under peak metabolic demands, such as during lactation. Here, we used a different set of metabolic stressors, which do not induce maximum energy expenditures and yet are likely to expose the potential constraining effect of heat dissipation. We compared the physiological responses of mice divergently selected for high (H-BMR) and low basal metabolic rate (L-BMR) to simultaneous exposure to the keyhole limpet haemocyanin (KLH) antigen and high ambient temperature (Ta). At 34°C (and at 23°C, used as a control), KLH challenge resulted in a transient increase in core body temperature (Tb) in mice of both line types (by approximately 0.4°C). Warm exposure did not produce line-type-dependent differences in Tb (which was consistently higher by ca. 0.6°C in H-BMR mice across both Ta values), nor did it result in the suppression of antibody synthesis. These findings were also supported by the lack of between-line-type differences in the mass of the thymus, spleen or lymph nodes. Warm exposure induced the downsizing of heat-generating internal organs (small intestine, liver and kidneys) and an increase in intrascapular brown adipose tissue mass. However, these changes were similar in scope in both line types. Mounting a humoral immune response in selected mice was therefore not affected by ambient temperature. Thus, a combined metabolic challenge of high Ta and an immune response did not appreciably compromise the capacity to dissipate heat, even in the H-BMR mice.

Transgenic characterization of two testis-specific promoters in the silkworm, Bombyx mori.

Sex-specific regulatory elements are key components for developing insect genetic sexing systems. The current insect genetic sexing system mainly uses a female-specific modification system whereas little success was reported on male-specific genetic modification. In the silkworm Bombyx mori, a lepidopteran model insect with economic importance, a transgene-based, female-specific lethality system has been established based on sex-specific alternative splicing factors and a female-specific promoter BmVgp (vitellogenin promoter) has been identified. However, no male-specific regulatory elements have yet been identified. Here we report the transgenic identification of two promoters that drive reporter gene expression in a testis-specific manner in B. mori. Putative promoter sequences from the B. mori Radial spoke head 1 gene (BmR1) and beta-tubulin 4 gene (Bmß4) were introduced using piggybac-based germline transformation. In transgenic silkworms, expression of the reporter gene enhanced green fluorescent protein (EGFP) directed by either BmR1 promoter (BmR1p) or Bmß4p showed precisely testis-specific manners from the larval to adult stage. Furthermore, EGFP expression of these two transgenic lines showed different localization in the testis, indicating that BmR1p or Bmß4p might be used as distinct regulatory elements in directing testis-specific gene expression. Identification of these testis-specific promoters not only contributes to a better understanding of testis-specific gene function in insects, but also has potential applications in sterile insect techniques for pest management.