Care, management, and use of ferrets in biomedical research.

The ferret (Mustela putorius furo) is a small domesticated species of the family Mustelidae within the order Carnivora. The present article reviews and discusses the current state of knowledge about housing, care, breeding, and biomedical uses of ferrets. The management and breeding procedures of ferrets resemble those used for other carnivores. Understanding its behavior helps in the use of environmental enrichment and social housing, which promote behaviors typical of the species. Ferrets have been used in research since the beginning of the twentieth century. It is a suitable non-rodent model in biomedical research because of its hardy nature, social behavior, diet and other habits, small size, and thus the requirement of a relatively low amount of test compounds and early sexual maturity compared with dogs and non-human primates. Ferrets and humans have numerous similar anatomical, metabolic, and physiological characteristics, including the endocrine, respiratory, auditory, gastrointestinal, and immunological systems. It is one of the emerging animal models used in studies such as influenza and other infectious respiratory diseases, cystic fibrosis, lung cancer, cardiac research, gastrointestinal disorders, neuroscience, and toxicological studies. Ferrets are vulnerable to many human pathogenic organisms, like severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), because air transmission of this virus between them has been observed in the laboratory. Ferrets draw the attention of the medical community compared to rodents because they occupy a distinct niche in biomedical studies, although they possess a small representation in laboratory research.

Assessing trait emotional intelligence and its relationship with stress and health behaviour in the education sector: An empirical study from Uttarakhand, India.

BACKGROUND: Emotional intelligence of teachers can affect their mental and physical health as well their performance in school. Both emotional intelligence and health behavior can have an impact on stress. The majority of Indian studies have examined only one type of teachers, have used indigenous scales which are not internationally valid, and have not studied health behaviour. The role of age and gender on emotional intelligence is also a debatable subject which requires larger studies The present study was undertaken to evaluate the trait emotional intelligence, stress and health behaviour of teachers and to determine their inter-relationship and to assess the role of demographic and professional attributes on emotional intelligence. METHODS: Teachers from different schools, colleges and professional institutes situated in Dehradun and nearby towns in the state of Uttarakhand, India were evaluated by internationally valid tools for the three parameters. RESULTS: Emotional Intelligence of teachers has no relation with age, gender, educational qualification, level of teaching or type of institute. It has a negative correlation with stress and a positive correlation with health behaviour. Further, health behaviour is inversely related to stress. CONCLUSIONS: Assessment of emotional intelligence and health behaviour of teachers should be a part of their routine evaluation and training so that specific interventions to reduce stress and to improve their overall health and performance can be appropriately planned.

Salivary cell-free HSD17B1 and HSPA1A transcripts as potential biomarkers for estrus identification in buffaloes (Bubalus bubalis).

Estrus detection is a major problem in buffaloes because of the poor expression of estrus signs leading to low reproductive efficiency. Salivary transcripts analysis is a promising tool to identify biomarkers; therefore, the present study was carried out to evaluate their potential as estrus biomarkers. The levels of HSD17B1, INHBA, HSPA1A, TES transcripts were compared in saliva during estrous cycle stages [early proestrus (day -2, EP), late proestrus (day-1, LP), estrus (E), metestrus (ME) and diestrus (DE)] of cyclic heifers (n = 8) and pluriparous (n = 8) buffaloes by employing quantitative real-time polymerase chain reaction (qRT-PCR). The levels of HSD17B1 (EP/DE 1.46-2.43 fold, LP/DE 2.49-3.06 fold; E/DE 7.21-11.9-fold p < 0.01; ME/D 1.0-1.16 fold) and HSPA1A (EP/DE 0.93-2.39 fold, LP/DE 2.68-3.23 fold; E/DE 8.52-15.18 fold p < 0.01; ME/D 0.86-1.01 fold) were significantly altered during the estrus than other estrous cycle stages in both cyclic heifers and pluriparous buffaloes. Receiver operating characteristic curve analysis revealed the ability of salivary HSD17B1 (AUC 0.96; p < 0.001) and HSPA1A (AUC 0.99; p < 0.01) to differentiate E from other stages of the estrous cycle. Significantly higher levels of HSD17B1 and HSPA1A transcripts in saliva during the estrus phase suggest their biomarkers potential for estrus detection in buffaloes.

Research on emotional intelligence among Indian teachers: A Systematic Review and meta-analysis of its correlation with health parameters and impact of gender.

Background: Emotional intelligence is the self-perception related to identification and regulation of emotions. Several studies have been done among Indian teachers evaluating emotional intelligence in relation to demographic, professional and various psychological parameters, but the variety of scales, teacher types, and conflicting results makes it difficult to draw any meaningful conclusions from this heterogeneous data.The present work aims to synthesize the available data by both qualitative and quantitative analysis and is the first such attempt to include only Indian studies in this field. The main objectives were to determine the correlation of emotional intelligence with teachers' health parameters and to study the gender difference in emotional intelligence. Methods: After a thorough literature search in Google, Google scholar, Scopus, Web of science and Pubmed, fifty-five Indian studies were selected which empirically examined teachers' emotional intelligence, either alone or in association with another parameter evaluating teachers' psychological health and performance. After qualitative assessment of major findings, quantitative analysis was performed. Three separate meta-analysis were carried out. The first one with fifteen effect sizes among 3291 participants evaluated correlation with personal health parameters. The second with nineteen effect sizes in 4165 participants evaluated correlation with professional health parameters. The third with twenty-six studies involving 6005 participants assessed effect of gender. Results: The results show that almost all studies have used a trait measure, teachers' emotional intelligence is positively correlated with both personal and professional health parameters and gender has no effect on emotional intelligence. Conclusion: Major limitations are a very high degree of heterogeneity of the data, incomplete description of the scales, inadequate randomization and small sample sizes in many studies. The results indicate the importance of emotional intelligence in both personal and professional life of teachers and no effect of gender preparing a solid base for future research.

Comparative Proteome Profiling of Saliva Between Estrus and Non-Estrus Stages by Employing Label-Free Quantitation (LFQ) and Tandem Mass Tag (TMT)-LC-MS/MS Analysis: An Approach for Estrus Biomarker Identification in Bubalus bubalis.

Accurate determination of estrus is essentially required for efficient reproduction management of farm animals. Buffalo is a shy breeder and does not manifest overt signs of estrus that make estrus detection difficult resulting in a poor conception rate. Therefore, identifying estrus biomarkers in easily accessible biofluid such as saliva is of utmost interest. In the current study, we generated saliva proteome profiles during proestrus (PE), estrus (E), metestrus (ME), and diestrus (DE) stages of the buffalo estrous cycle using both label-free quantitation (LFQ) and labeled (TMT) quantitation and mass spectrometry analysis. A total of 520 proteins were identified as DEPs in LFQ; among these, 59 and four proteins were upregulated (FC ≥ 1.5) and downregulated (FC ≤ 0.5) during E vs. PE, ME, and DE comparisons, respectively. Similarly, TMT-LC-MS/MS analysis identified 369 DEPs; among these, 74 and 73 proteins were upregulated and downregulated during E vs. PE, ME, and DE stages, respectively. Functional annotations of GO terms showed enrichment of glycolysis, pyruvate metabolism, endopeptidase inhibitor activity, salivary secretion, innate immune response, calcium ion binding, oocyte meiosis, and estrogen signaling. Over-expression of SERPINB1, HSPA1A, VMO1, SDF4, LCN1, OBP, and ENO3 proteins during estrus was further confirmed by Western blotting. This is the first comprehensive report on differential proteome analysis of buffalo saliva between estrus and non-estrus stages. This study generated an important panel of candidate proteins that may be considered buffalo estrus biomarkers which can be applied in the development of a diagnostic kit for estrus detection in buffalo.

Pulse Oximetry Waveform Represents an Earlier Ventricular Contraction in Relation to the Intraarterial Blood Pressure Tracing and Electrocardiogram on Multichannel Monitors: A Case Series.

Multichannel patient monitors apparently display the electrocardiogram, intra-arterial blood pressure, and pulse oximetry waveforms in synchrony during sinus cardiac rhythm. We recorded 20 events of a premature cardiac contraction on multichannel monitors from 10 patients. Interestingly, during premature cardiac contraction, the low-amplitude arterial trace was aligned with the premature electrocardiogram, but the low-amplitude pulse oximetry plethysmograph was aligned with the next normal electrocardiogram and arterial tracing. In conclusion, the pulse oximetry plethysmograph tracing is offset by 1 ventricular depolarization on the electrocardiogram (QRS) and the arterial tracing on multichannel monitors.

Transcriptome analysis of buffalo granulosa cells in three dimensional culture systems.

Hanging drop (HD) three-dimensional (3D) culture model for buffalo granulosa cells (GC) was reported to mimic the preovulatory stage of ovarian follicles in our previous study. To further verify its reliability, the present study attempted a comparative transcriptome profile of buffalo GC freshly isolated from ovarian follicles (<8 mm diameter) (FC) and their cultures in normal culture dish (ND or 2D), polyHEMA coated dish (PH) and HD culture systems (3D). Out of 223 significantly (-log2 fold change: >3; p < .0005; false discovery rate [FDR]: <0.1) differentially expressed genes (SDEGs) among different culture systems, 137 were found unannotated, and 94, 29, and 66 were exclusively expressed in FC, PH, and HD, respectively. However, on eliminating the fixed points of p values and FDR from the entire raw data, only 11 genes related to long noncoding RNA, 12 genes related to luteinization, and 3 genes related to follicular maturation were exclusively expressed in FC, PH, and HD culture systems, respectively. The quantitative real time-PCR validation and the next generation sequencing data had more than 90% correlation. Bioinformatics analyses of the exclusively expressed SDEG revealed that the freshly aspirated GCs were a true representative of GCs from small follicles (<8 mm diameter), the GC spheroids under PH maintained mitochondrial function, and those cultured in HD system for 6 days simulated the inflammatory milieu required for ovulation. Therefore, the comparative transcriptome profile also reinforced that HD culture system is better in vitro culture method than the other methods analyzed in this study for buffalo GC.

Effect of Vedic music on steroidogenic gene expression in 3D-cultured buffalo granulosa cell spheroids model system, a pilot study.

Music is known for reducing stress, anxiety and depression, improving cognitive performance, and enhancing oestrogen levels. However, its effect on non-auditory mammalian cell system and the molecular events leading to higher oestrogen levels is less explored. Therefore, the present study targeted to know the direct effects of a peaceful Vedic music on 3D cultured buffalo granulosa cell spheroids. The spheroids were daily exposed to the Mahamrityunjaya mantra, a kind of Vedic chants, for 1.5 hr for 6 days. After 6 days, the music effect was analysed by the expression analysis of steroidogenic (CYP19A1, STAR and HSD17ß1) and proliferative marker (PCNA) genes. Interestingly, the CYP19A1 gene expression was significantly upregulated by 3.464 ± 0.15 folds in the music exposed spheroids than the non-exposed spheroids. However, the expression of other steroidogenic and proliferative genes was unaltered. These observations provided a transcriptional clue for higher estradiol levels by the music and a scope to use Vedic chants for increasing the CYP19A1 expression to help tackle some pathophysiological conditions.

Loop-Mediated Isothermal Amplification (LAMP) for Detection of Ureaplasma diversum from Cervico-vaginal Swab of Buffaloes.

The main plan of the current study was to develop a rapid, robust, and field-applicable loop-mediated isothermal amplification (LAMP) assay for the detection of Ureaplasma diversum. A strain-specific 16S rRNA gene of Ureaplasma diversum was used for detection which was cloned, sequenced, and characterized earlier. LAMP results were visualized within 90 min with the naked eye. Cervico-vaginal swabs of 50 buffaloes were randomly collected from Livestock Research Center of NDRI as per the Institute Animal ethics guidelines. Out of 50 cervico-vaginal swab samples collected randomly, 34 were found positive with LAMP while 16 samples were negative. Conventional PCR results showed the same result. Therefore, the accuracy of the developed LAMP was about 100%. The developed LAMP assay can also be used to screen the animals for Ureaplasma diversum infection in cervico-vaginal swab. However, further study is needed to assess sensitivity and accuracy towards their detection and their relationship in disease diagnosis.

Effect of Dioxins in Milk on the 3D Cultured Primary Buffalo Hepatocyte Model System.

Cow and human milk have been reported to contain dioxins ranging from 0.023 to 26.46 and 0.88 to 19 pg/g of fat, respectively. However, the toxic effects of the dioxins in the milk in this range of concentrations were not explored. Therefore, considering the outbred livestock tissues as better models than inbred laboratory animals, the present study targeted to study the effect of dioxins present in the milk on three-dimensionally (3D) cultured buffalo primary hepatocyte spheroids. The spheroids were treated with a model dioxin, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD), directly and also through milk fat at different concentrations (i.e, 0.02-20 pg/mL) for 24 h. Among the liver-cell-specific (ALB, HNF4α, and AFP) genes, a similar ALB and upregulated HNF4α expression at all treatments indicated the functional and transcriptionally active hepatocyte spheroids. Supportingly, no significant difference in the antiapoptotic gene expression between the treatments of milk fat and milk fat containing dioxins indicated the survivability of the spheroids during dioxin treatments. Among the selected TCDD responsive (CYP1A1, CYP1A2, AHR, CYP1B1, and TIPARP) genes, a nonsignificant increasing trend of the CYP1A1 expression was observed from 0.2 to 10 pg/mL of TCDD concentration through milk fat. This pattern was similar to the reported insensitive response of human primary hepatocytes toward dioxins than that of rat primary hepatocytes. This may indicate that the buffalo hepatocyte spheroids could be better models than rats for TCDD hepatotoxic studies. Further, TCDD in the milk in the range of 0.02-20 pg/mL concentration may not be very hepatotoxic.

Over-expression of mammaglobin-B in canine mammary tumors.

BACKGROUND: Mammaglobin, a member of secretoglobin family has been recognized as a breast cancer associated protein. Though the exact function of the protein is not fully known, its expression has been reported to be upregulated in human breast cancer.We focused on studying the expression of mammaglobin-B gene and protein in canine mammary tumor (CMT) tissue. Expression of mammaglobin-B mRNA and protein were assessed by quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC), respectively. RESULTS: High levels of mammaglobin-B mRNA expression (6.663 ± 0.841times) was observed in CMT as compared to age and breed matched healthy controls. Further, expression of mammaglobin-B protein was detected in paraffin-embedded mammary tumor tissues from the same subjects by IHC. Mammaglobin-B protein was overexpressed only in 6.67% of healthy mammary glands while, a high level of its expression was scored in 76.7% of the CMT subjects. Moreover, no significant differences in terms of IHC score and qRT-PCR score with respect to CMT histotypes or tumor grades were observed, indicating that mammaglobin-B over-expression occurred irrespective of CMT types or grades. CONCLUSION: Overall, significantly increased expression of mammaglobin-B protein was found in CMTs with respect to healthy mammary glands, which positively correlates to its transcript. These findings suggest that overexpression of mammaglobin-B is associated with tumors of canine mammary glands.

Three-dimensional culture of buffalo granulosa cells in hanging drop mimics the preovulatory follicle stage.

Granulosa cell (GC) culture models mimicking the intrafollicular environment are limited. Such models have a great potential in reproductive toxicity studies. The buffalo, a monovulatory species like humans, could be a better model than polyovulatory rodents. Therefore, we targeted the development and characterization of three-dimensional (3D) culture systems for buffalo GCs. The GCs from small ovarian follicles (SF) maintained the CYP19 gene expression for 144 hr in a 2D culture system. Hence, GCs from SF were cultured directly in 3D using hanging drop and Poly-([2-hydroxyethyl methacrylate]) (polyHEMA) methods in the DMEM media containing 1 ng/ml FSH and 10 ng/ml IGF-1 for 144 hr. The expression profile of nine GC-specific transcripts; CYP19, TNFAIP6, AMH, PTI, NR4A1, FSHR, RUNX, LHR, and COX2/PTGS2; revealed that 3D-spheroids developed in hanging drop method maintained the GC phenotype of preovulatory follicles. Therefore, hanging drop method is a best method for culturing GCs to mimic the intrafollicular environment.

Reverse transcription loop-mediated isothermal amplification (RT-LAMP), a light for mammalian transcript analysis in low-input laboratories.

Transcript analysis is usually performed by costly, time-consuming, and expertise intensive methods, like real time-PCR, microarray, etc. However, they are not much feasible in low-input laboratories. Therefore, we implemented the reverse transcription loop-mediated isothermal amplification (RT-LAMP) as a means of mammalian transcript analysis. Particularly, RT-LAMP was developed for buffalo aromatase cytochrome P450 (CYP19) transcript, to study its expression in 3D-cultured buffalo granulosa cells, which were exposed to lipopolysaccharide (LPS). The CYP19-RT-LAMP assay rapidly identified the LPS-induced downregulation of the CYP19 gene within 30 min at 63°C in a water bath. The assay was visualized via unaided eye by observing the change in turbidity and fluorescence, which were decreased by increasing the LPS exposure time to granulosa cells. Overall, the developed CYP19-RT-LAMP assay provided a hope on the application of RT-LAMP for mammalian transcript analysis in low-input laboratories.

Development and characterization of 2-dimensional culture for buffalo intestinal cells.

Small intestinal epithelial cells (IEC) play a major role in the absorption of nutrients and toxins. Due to the similarity of genome-wide single copy protein orthologues between cattle and human, establishment of ruminant's primary small IEC culture could be a valuable tool for toxicity studies. Therefore, the current study focused on the development and characterization of buffalo IEC culture, as cattle slaughter is banned in India. The buffalo jejunum fragments were washed consecutively several times in saline, warm phosphate buffered saline (PBS), PBS with 5 mM dithiothreitol, digesting solution and 2% sorbitol in PBS. The cells were cultured on 17 µg/cm2 collagen coated plates and transwell plates with serum (2% Fetal bovine serum (FBS) and 10% FBS) and serum-free culture conditions. The cells were differentiated into typical epithelial cobblestone morphology from day 5 onwards in 50% successful cultures. The cultured IEC were characterized by gene expression of epithelial cell markers, cytokeratin and vimentin, and enterocyte markers like villin, zonula occluden (ZO1), fatty acid binding protein 2 (FABP2) and small intestinal peptidase (IP). Based on the morphology and gene expression profile, 10% FBS has been recommended for culturing primary buffalo IEC on collagen coated plates for 10 days. However, 50% of the successful cultures could not show epithelial phenotype on 10% FBS culture conditions even on collagen coated plates. Interestingly, undifferentiated IEC showed an increasing expression of FABP2, IP and ZO1 transcripts compared to differentiated intestinal cells with 10% FBS on collagen plates. Therefore, future studies are needed to understand the role of FABP2, IP and ZO1 in differentiation of buffalo IEC.

Development of recombinant matrix metalloproteinase-3 based sandwich ELISA for sero-diagnosis of canine mammary carcinomas.

Matrix metalloproteinase-3 is invariably upregulated in cancerous condition. So we aimed to determine serum level of MMP-3 in canine mammary tumors. The gene was expressed in E. coli system as ~43kDa recombinant protein, which was refolded, purified, and confirmed. Hyperimmune serum was raised against the expressed protein in rabbits and mice to standardize sandwich ELISA. ROC analysis revealed largest area under the curve of 0.998 with sensitivity (100%) and specificity (95%) for a cut-off value of 0.363 with respect to histopathological staining. The finding of the present study indicates that MMP-3 can act as a potential molecular marker for serodiagnosis of canine mammary carcinomas.

Asynchrony Between Ventilator Flow and Pressure Waveforms and the Capnograph on Dräger Anesthesia Workstations: A Case Report.

Modern anesthesia workstations display capnography, flow-time, and pressure-time waveforms in real time. We observed that at certain ventilator settings (10 breaths/min) on Dräger workstations, the expiratory phase of the capnograph overlaps both the inspiratory and the expiratory phases of ventilation. This discrepancy disappears at respiratory rates of 16 breaths/min. This synchronous respiratory monitoring display at respiratory rates 16 breaths/min is not physiologically correct, because it implies a synchronization of waveforms that is not actually present. This again becomes asynchronous once the respiratory rate is increased to >18 breaths/min. Such an artifact may not affect the patient's safety in most cases but may mislead clinicians when synchrony between flow/pressure and capnography is needed for diagnostic purposes. We wish to share this discrepancy with clinicians and notify the manufacturer so that potential solutions may be found.

Mammaglobin as a diagnostic serum marker of complex canine mammary carcinomas.

Mammaglobin is a glycoprotein exhibiting homology to uteroglobin gene family. Although the biological function of the protein is not yet known it has been reported to act as marker for breast cancer in women. This study reports the expression of mammaglobin gene in canine mammary tumor condition. The gene was cloned, sequenced and heterologously expressed in Escherichia coli host system as 12 kDa (approx.) recombinant fusion protein. The expressed protein was further purified to homogeneity and confirmed by western blotting. Hyperimmune sera were raised against the expressed protein in rabbits and mice to standardize sandwich ELISA for relative quantification of circulating protein in the sera of dogs with mammary tumors. Based on receiver-operating characteristics analysis, the test was found to be 90% sensitive and 95% specific for a cut-off value of 0.177 with respect to histopathological staining in diagnosing canine mammary tumors and the protein level was not elevated in other diseased conditions. These findings indicate that it can act as a novel molecular marker for detecting mammary tumors in canines.

Dilated ascending aorta is associated with the difficulty in correct placement of pulmonary artery catheter.

The present case report highlights that a tense mega-sized aortic root and ascending aorta can mechanically resist the passage of fully inflated (1.5 ml air) balloon to wedge-trace position in the pulmonary artery. Any attempt to push the catheter rather predisposed its recoiling and rebutting into the right ventricle and the cardiac arrhythmia. Inflating continuous cardiac output catheter balloon with lesser volume of air (1 ml) is suggested to overcome this problem.

Pharmacodynamic evaluation of augmentation effect of isoflurane on mivacurium.

This study evaluated the augmentation effect of isoflurane (ISO) given before or after the mivacurium (MIV) injection. Consented 33 adults (18-58 years), ASA I patients of both sexes were randomly assigned into three groups. In group 1 (no ISO) patients--IV propofol (0.5-1 mg/kg) for induction and (25-50 mg) aliquots every 2-5 minutes, N2O (60%) in O2 by mask, was followed by IV MIV (0.04 mg/kg). In group 2 (ISO before MIV) patients-IV propofol as in group 1 and ISO in N2O (60%) and O2 to achiever end tidal level 1% for 10 minutes was followed by same dose of IV MIV. In group 3 (ISO after MIV) patients-after propofol and IV MIV as above, ISO in N2O (60%) and O2 was given to get end tidal level 1% for 10 minutes. All patients were breathing spontaneously using tight fitting facemask and respiration was assisted to keep ETCO2 (35-40 mmHg), SaO2 100%. To monitor MIV effect, ulnar nerve was stimulated at wrist using supramaximal double burst stimuli (DBS). Adductor pollicis response was recorded on myograph-2000 (Biometer, Denmark). Twitch amplitude (D1) and tetanic fade DBSr (D2/D1) were calculated for each stimulus and recorded. Peak MIV effect (D1 suppression by approximately equal to 90% and abolished D2) was significantly (p < 0.05) more in group 2 patients (ISO before MIV) than in group 1 (control) or group 3 (ISO after MIV) patients. ISO given prior of MIV administration significantly augmented the effect of MIV. When ISO was started after MIV injection probably the quick hydrolysis of MIV limited the augmentation effect to tetanic fade only as compared to the patients, who did not get ISO at any point of study. Prior administration of ISO causes intense MIV block at adductor pollicis.