Effects of Annealing and Concentration of Calcium Salts on Thermal and Rheological Properties of Maize Starch During an Ecological Nixtamalization Process

The objective of the present work was to study the effect of annealing and concentration of Ca(OH)₂ (lime) and calcium salts on the thermal and rheological properties of maize starch during an ecological nixtamalization process. Thermal and rheological properties of maize starch changed during the ecological nixtamalization process because of three main causes: the annealing phenomenon, type of calcium salt, and calcium salt concentration. In all treatments thermal properties (T_o, T_p, and T_f) of nixtamal starch increased owing to the annealing process, whereas the type of salt or lime increased thermal properties and decreased pasting properties in this order: CaCl₂ > CaSO₄ > Ca(OH)₂ ≈ CaCO₃. This behavior was because of the dissociation of each salt or lime in water. Anions (OH⁻) can penetrate much more easily into the starch granule and start the gelatinization process by rupturing hydrogen bonds. Additionally, amylose‐lipid complexes were formed during the nixtamalization processes, as indicated by an increasing peak at 4.5 Å in X‐ray diffraction patterns.     

Clinical application of multidetector computed tomography and magnetic resonance imaging for evaluation of cranial nerves in horses in comparison with high resolution imaging standards

Although horses are affected by cranial nerve disease, our understanding of these structures' imaging anatomy is limited, and the optimal modality for imaging of each of these nerves is unclear. The aim of this study was to describe the imaging appearance of the equine cranial nerves on high‐resolution 1.5T magnetic resonance imaging (MRI) and computed tomography (CT) scans of a cadaver head, and with these as standards, examine the utility of MRI and CT performed in clinical cases. High‐resolution MRI and CT images were prospectively acquired of the head of a normal Thoroughbred gelding following euthanasia. Ten clinical cases undergoing high‐field MRI under general anaesthesia and 10 clinical cases undergoing CT in the standing horse under sedation were retrospectively evaluated by three reviewers to assess cranial nerve visibility. On high‐resolution, thin‐slice, MRI scans of the normal cadaver head, each of the 12 cranial nerves and their topographic location could be appreciated. On high‐resolution cadaver CT, cranial nerves II, V and VII were clearly visible, but others were less easily identified; osseous structures were clearly visualised. Clinical MRI and CT allowed for variable visualisation of the cranial nerves, dependent on the sequence and the orientation of scan planes. High‐field MRI allowed excellent visualisation of equine cranial nerves, whereas CT allowed for more detailed visualisation of the osseous canals and foramina. In live horses, the ability to identify all 12 nerves is challenging with either MRI or CT; however, high‐field MRI enables better visualisation of the nerve bundles than CT.     

Early Pregnancy Detection of Iraqi Riverine Buffalo (Bubalus bubalis) Using the BioPRYN Enzyme‐Linked Immunosorbent Assay for PSPB and the Progesterone Assay

This study was undertaken to detect pregnancy in Iraqi riverine buffalo (Bubalus bubalis) using three different methods (rectal palpation, plasma progesterone concentration and detection of the presence of pregnancy‐specific protein B (PSPB) with the BioPRYN^® enzyme‐linked immunosorbent assay (ELISA) test. The aim of the study was to identify the most sensitive, early and accurate method for detecting pregnancy. Twenty‐two female riverine buffalo that were 6.0 ± 0.93 years old were used. Four blood samples per buffalo were taken via jugular venipuncture at days 22–24, 32–34, 42–44 and 58–61 post‐mating (PM) to measure the progesterone concentration (ng/ml) and to detect the presence of plasma PSPB. The rectal palpation method was employed to evaluate all buffalo on days 42–44 and 58–61 PM. The BioPRYN^® test differed (p < 0.01) from the other tests with earlier accuracy for detecting pregnant and non‐pregnant buffalo. Eighty‐eight percent of pregnant and 76.9% of non‐pregnant buffalo were distinguished early (days 22–24 PM) using BioPRYN^® and plasma PSPB‐ELISA level (2.09 ± 0.12 ng/ml) in relation to 66.7% and 53.9% detected using the progesterone assay at similar days (4.30 ± 0.40 ng/ml). In conclusion, these results described, for the first time, the early and accurate pregnancy detection of water riverine buffalo using BioPRYN^® technology and provided the plasma levels of PSPB using an ELISA test. These findings will improve the reproductive and productive efficiency of Iraqi riverine buffalo by adapting the recent management and reproductive strategies in Iraq and in the world.     

Ventricular Arrhythmias during Race Recovery in Standardbred Racehorses and Associations with Autonomic Activity

Background: A lack of information on normal heart rhythm at maximal effort hampers investigation of poor performance and sudden death in Standardbred racing. Hypothesis/Objective: To characterize rhythm variations during scheduled racing in clinically normal Standardbred horses. Animals: Two hundred and eighty‐eight Standardbred horses competing in 40 scheduled races at a Southern Ontario racetrack. Methods: Observational study, convenience sampling: Heart rhythm was monitored by ECG from harnessing to postrace recovery and assessed visually and by examining heart rate intervals. Rhythm variations were used as response variables in multivariate analysis of race records detailing signalment, race, and race outcome. Results: Monitoring involved 345 individual horse or race events. Occasional, isolated premature cycles, only, occurred during the race. Postrace, sudden cardiac slowing (punctuated deceleration [PD]) appeared in 42 events (12.2%). Only premature ventricular complexes were exhibited in 40 events postrace (11.6%), whereas 55 (15.9%) exhibited complex ventricular arrhythmias (CVA) including torsades‐like polymorphic ventricular tachycardia, 34.5% of these being closely associated with PD (odds ratio = 8). Predispositions to CVA were found for horses parked at the 1/2 mile (odds ratio = 3), and trotters breaking in the stretch (odds ratio = 38). Horses spontaneously reverted to sinus rhythm and no sudden death events were encountered. Conclusions and Clinical Importance: Arrhythmias occur frequently in racing Standardbreds during cardiac deceleration, often associated with sudden, rapid increases in vagal tone. Circumstances imposing unusual demand and racing at the trot appear to predispose. Findings provide insight into possible mechanisms of sudden death.     

Genome‐Wide Identification of MicroRNAs Responsive to High Temperature in Rice (Oryza sativa) by High‐Throughput Deep Sequencing

MicroRNAs (miRNAs) are known to play important roles in plant growth and stress response. Heat stress is a severe abiotic stresses by adversely affecting plant growth and yield. To identify heat‐responsive miRNAs at the genome‐wide level in rice (Oryza sativa), we constructed two small RNA libraries from young panicles treated or not with heat conditions. Ion torrent sequencing of the two libraries identified 294 known miRNAs and 539 novel miRNAs. Differential expression analysis showed that 26 miRNAs were downregulated and 21 miRNAs were upregulated in response to heat stress. Among them, five heat‐responsive miRNAs, including miR162b, miR529a‐p5, PC‐5P‐62245‐9, miR171b and miR169n, were validated by quantitative real‐time polymerase chain reaction. A total of 44 target genes of the differentially expressed miRNAs were predicted. These target genes are most significantly overrepresented in the cell growth process. The results demonstrated that rice miRNAs play critical roles in the heat stress response. This study opens up a new avenue for understanding the regulatory mechanisms of miRNAs involvement in the heat stress response in rice.     

Effect of catheter size and injection rate of contrast agent on enhancement and image quality for triple‐phase helical computed tomography of the liver in small dogs

Rapid contrast injection is recommended for triple‐phase helical computed tomography (CT) of the liver. However, a large‐gauge catheter is needed for faster contrast injection and this is not practical for small breed dogs or cats. The purpose of this crossover group study was to evaluate applicability of a lower injection rate with a small‐gauge (G) catheter for triple‐phase hepatic CT in small dogs. Triple‐phase CT images were acquired for six beagle dogs using three protocols: an injection rate of 1.5 ml/s with a 24 G catheter, 3.0 ml/s with a 22 G catheter, and 4.5 ml/s with a 20 G catheter. Enhancement of the aorta, portal vein, and hepatic parenchyma was measured in each phase (arterial, portal, and delayed) and image quality was scored subjectively by two observers. Injection duration, time to scan delay, and time to peak enhancement were also recorded. Contrast injection duration decreased with a higher injection rate (n = 6, P ≤ 0.01), but time to peak enhancement and time to scan delay were not significantly affected by injection rates and catheter sizes. Contrast injection rate did not significantly affect aortic, portal, and hepatic enhancement. In addition, separation between each phase and quality of images was subjectively scored as good regardless of injection rate. Findings from the current study supported using an injection rate of 1.5 ml/s with a catheter size of 24 G for triple‐phase hepatic CT in small dogs (weight < 12 kg).     

Effects of five cryoprotectants on proliferation and differentiation‐related gene expression of frozen‐thawed bovine calf testicular tissue

The cryopreservation of testicular tissue is a potential method for preserving male fertility. However, the effect of cryopreservation on bovine calf testicular tissue is scarce. This study investigated the effect of different cryoprotectants on bovine calf testicular tissue at the molecular level. Testicular tissue from ten immature bovine calves (6 months) was collected after slaughter and cryopreserved in an extender containing different concentrations of the following five cryopreservation solutions (CP): bovine serum albumin (BSA) with 5% dimethyl sulfoxide (DMSO), trehalose with 5% DMSO, DMSO and glycerol and ethylene glycol (EG). After 7‐day cryopreservation, the expression levels of three spermatogonial stem cell (SSC)‐related genes, octamer‐4 (OCT4), KIT ligand (MGF/SCF) and kit oncogene (C‐KIT), were investigated by quantitative PCR (qPCR). The cell viability was highest for the tissues preserved with 30 mg/ml BSA (77.82% ± 1.22) and 40 mg/ml trehalose (74.23% ± 1.16) compared with other groups (p < 0.05), and the level of expression of the three genes was highest with 30 mg/ml BSA (p < 0.05). Compared with other CPs, the 30 mg/ml BSA and 40 mg/ml trehalose have the better cryopreserve protection. The 30 mg/ml BSA is the most viable media for the cryopreservation of testicular tissue from cattle.     

Characterization of an Alginate Lyase,FlAlyA, from Flavobacterium sp. Strain UMI-01 and Its Expression in Escherichia coli

A major alginate lyase, FlAlyA, was purified from the periplasmic fraction of an alginate-assimilating bacterium, Flavobacterium sp. strain UMI-01. FlAlyA showed a single band of ~30 kDa on SDS-PAGE and exhibited the optimal temperature and pH at 55 °C and pH 7.7, respectively. Analyses for substrate preference and reaction products indicated that FlAlyA was an endolytic poly(mannuronate) lyase (EC 4.2.2.3). A gene fragment encoding the amino-acid sequence of 288 residues for FlAlyA was amplified by inverse PCR. The N-terminal region of 21 residues except for the initiation Met in the deduced sequence was predicted as the signal peptide and the following region of six residues was regarded as propeptide, while the C-terminal region of 260 residues was regarded as the polysaccharide-lyase-family-7-type catalytic domain. The entire coding region for FlAlyA was subjected to the pCold I—Escherichia coli BL21(DE3) expression system and ~eight times higher yield of recombinant FlAlyA (recFlAlyA) than that of native FlAlyA was achieved. The recFlAlyA recovered in the periplasmic fraction of E. coli had lost the signal peptide region along with the N-terminal 3 residues of propeptide region. This suggested that the signal peptide of FlAlyA could function in part in E. coli.     

Purification and characterization of two anionic trypsins from the hepatopancreas of carp

SUMMARY: Two trypsins, designated as trypsin A and trypsin B, have been purified from the hepatopancreas of carp. The purification procedures consisted of ammonium sulfate fractionation, and chromatographies on DEAE-Sephacel, Ultrogel AcA54 and Q-Sepharose. Trypsin A was purified to homogeneity with the molecular mass of approximately 28 kDa, while trypsin B gave two close bands of 28.5 kDa and 28 kDa on sodium dodecylsulfate polyacrylamide gel electrophoresis both under reducing and non-reducing conditions. On native-PAGE, both trypsin A and trypsin B showed a single band. Trypsin A and trypsin B revealed optimum temperature of 40°C and 45°C, respectively, and shared the same optimum pH 9.0 using Boc-Phe-Ser-Arg-MCA as substrate. Both enzymes were effectively inhibited by trypsin inhibitors and their susceptibilities were similar. The NH₂-terminal amino acid sequences of trypsin A and trypsin B were determined to 37th and 40th amino acid residue, respectively. Their sequences were very homologous, but not identical to that of a trypsin-type serine proteinase from carp muscle and these of other trypsins. Immunoblotting test using the antibody raised against trypsin A cross-reacted with trypsin B positively.