Effect of silage from ryegrass intercropped with winter or common vetch for grazing dairy cows in small-scale dairy systems in Mexico

The objective was to determine the effect of including silages of annual ryegrass (Lolium multiflorum) intercropped with winter vetch (Vicia villosa) (ARG-VV) or with common vetch (Vicia sativa) (ARG-VS) compared with maize silage (MS) on milk yield and milk composition of dairy cows grazing cultivated perennial ryegrass–white clover pastures with supplemented concentrate during the dry season. Six Holstein dairy cows with a mean yield of 19.0 kg/cow/day at the beginning of the experiment were randomly assigned to a 3 × 3 repeated Latin square. Treatments were: 8 h/day intensive grazing, 3.6 kg of dry matter (DM) per cow per day of concentrate plus MS, and ARG-VV or ARG-VS ad libitum at a stocking rate of 3.0 cows/ha for three experimental periods of 3 weeks each. Milk yield (MY) and milk composition, live weight and body condition score as well as silage and concentrate intakes were recorded during the third week of each experimental period, and pasture intake was estimated indirectly from utilised metabolisable energy. Economic analysis was obtained by preparing partial budgets. There were no statistical differences (P > 0.10) in MY, milk fat or protein content nor for live weight, but there was significant difference (P < 0.10) in body condition score. There were non-statistical differences in silage DM intake (P < 0.11); however, significant differences (P < 0.10) were obtained for estimated grazed herbage intake whilst no differences for total DM intake. Slightly higher economic returns (10%) were obtained with ARG-VS over MS, and this was 7% higher than ARG-VV. It is concluded that ARG-VS could be an option for complementing grazing for small-scale dairy production systems in the dry season as it is comparable to MS in animal performance and slightly better in economic terms.     

Venous hepatic segmentation in dogs (Canis lupus familiaris—L. 1758)

The external shape of the liver is varied and determines specific vascular arrangements. This morphological relationship is important to establish hepatic segmentation in different species submitted to surgeries that aim to preserve a larger area of liver parenchyma. After observing 60 livers injected with Neoprene Latex and three plastic moulds obtained by corrosion, eight hepatic venous segments were identified, drained by six hepatic veins agrouped into segmental veins, which drained one sector (segments I, VI, VII and VIII) and intersegmental veins, which drained more than one sector (segments II/III and IV/V). They were described as follows: left intersegmental vein, formed by a segmental vein from the papillary process (segment I), two to three lateral left segmental veins that drained the segment II, and one to five left paramedian segmental veins that drained the segment III; sagittal intersegmental vein, formed by the confluence between segmental vein of the quadrate lobe (segment IV) and the medial right paramedian segmental vein, which derived from the segment V; lateral right paramedian vein drained the dorsocranial sector of the segment VI; the lateral right segmental vein, formed by one to four vessels that drained segment VII, and the segmental vein of the caudate process, which drained the segment VIII. Understanding the number and disposition of the hepatic veins in lobate livers is essential to reduce bleeding risks in surgeries. The nomenclature based on segmentation analogy of non-lobate liver could be less confusing and, therefore, be more useful in the surgical approaches of lobate livers.     

Transforming fusions of FGFR and TACC genes in human glioblastoma

The brain tumor glioblastoma multiforme (GBM) is among the most lethal forms of human cancer. Here, we report that a small subset of GBMs (3.1%; 3 of 97 tumors examined) harbors oncogenic chromosomal translocations that fuse in-frame the tyrosine kinase coding domains of fibroblast growth factor receptor (FGFR) genes (FGFR1 or FGFR3) to the transforming acidic coiled-coil (TACC) coding domains of TACC1 or TACC3, respectively. The FGFR-TACC fusion protein displays oncogenic activity when introduced into astrocytes or stereotactically transduced in the mouse brain. The fusion protein, which localizes to mitotic spindle poles, has constitutive kinase activity and induces mitotic and chromosomal segregation defects and triggers aneuploidy. Inhibition of FGFR kinase corrects the aneuploidy, and oral administration of an FGFR inhibitor prolongs survival of mice harboring intracranial FGFR3-TACC3-initiated glioma. FGFR-TACC fusions could potentially identify a subset of GBM patients who would benefit from targeted FGFR kinase inhibition.     

Intra-city Differences in Cardiac Expression of Inflammatory Genes and Inflammasomes in Young Urbanites: A Pilot Study

Southwest Mexico City (SWMC) air pollution is characterized by high concentrations of ozone and particulate matter < 10 μm (PM(10)) containing lipopolysaccharides while in the North PM(2.5) is high. These intra-city differences are likely accounting for higher CD14 and IL-1β in SWMC v NMC mice myocardial expression. This pilot study was designed to investigate whether similar intra-city differences exist in the levels of myocardial inflammatory genes in young people. Inflammatory mediator genes and inflammasome arrays were measured in right and left autopsy ventricles of 6 southwest/15 north (18.5 ± 2.6 years) MC residents after fatal sudden accidental deaths. There was a significant S v N right ventricle up-regulation of IL-1β (p=0.008), TNF-α (p=0.001), IL-10 (p=0.001), and CD14 (p=0.002), and a left ventricle difference in TNF-α (p=0.007), and IL-10 (p=0.02). SW right ventricles had significant up-regulation of NLRC1, NLRP3 and of 29/84 inflammasome genes, including NOD factors and caspases. There was significant degranulation of mast cells both in myocardium and epicardial nerve fibers. Differential expression of key inflammatory myocardial genes and inflammasomes are influenced by the location of residence. Myocardial inflammation and inflammasome activation in young hearts is a plausible pathway of heart injury in urbanites and adverse effects on the cardiovascular system are expected.     

Chemical restraint of African mole‐rats (Fukomys sp.) with a combination of ketamine and xylazine

ObjectiveTo establish an accurate anaesthetic dose for chemical restraint of African mole-rats using ketamine and xylazine.Study designProspective nonrandomized laboratory study.AnimalsSixteen adult Ansell’s mole-rats (Fukomys anselli) and eight giant mole-rats (F. mechowii).MethodsFukomys anselli of different ages, sexes and reproductive status were systematically anaesthetized starting with an intramuscular injection of ketamine (2.5 mg kg⁻¹) and increasing the doses in steps of 0.5 mg kg⁻¹ until loss of the righting reflex (LRR) was observed. Xylazine was added to a constant dose of ketamine, starting at 0.5 mg kg⁻¹ that was increased by 0.5 mg kg⁻¹ in further trials. Once an effective combination was established and evaluated in F. anselli, it was also tested in F. mechowii. Heart and respiratory rates and rectal temperatures were measured during anaesthesia. anova for repeated measures and Student’s t-test were used to compare means.ResultsChemical restraint was accomplished at a dose of 6 mg kg⁻¹ ketamine combined with 2.5 mg kg⁻¹ xylazine. LRR lasted on average mean 56 ± SD 19 minutes (F. anselli) and 140 ± 41 minutes (F. mechowii). Loss of pedal withdrawal reflex (LPR) lasted for 20 ± 15 minutes (F. anselli) and for 29 ± 2 minutes (F. mechowii), respectively. All animals recovered satisfactorily. Heart and respiratory rates were stable during anaesthesia, but rectal temperature fell significantly in F. mechowii after losing the righting reflex (LRR) from T1 (32.6 ± 0.6 °C) to T3 (30.4 ± 0.9 °C).Conclusions and Clinical relevanceAfrican mole-rats (Bathyergidae) live in closed burrow systems under particular conditions (hypercapnia, hypoxia, stable temperature, humidity, darkness) and show several physiological adaptations. Injectable anaesthetics in the dose rates used in other rodents are not appropriate for use in these subterranean species. Here, a reliable protocol for chemical restraint is provided.     

Detection of the intranuclear microsporidian Enterospora nucleophila in gilthead sea bream by in situ hybridization

Enterospora nucleophila is an intranuclear microsporidian responsible for emaciative microsporidiosis of gilthead sea bream (GSB). Its minute size and cryptic nature make it easily misdiagnosed. An in situ hybridization (ISH) technique based on antisense oligonucleotide probes specific for the parasite was developed and used in clinically infected GSB in combination with calcofluor white stain (CW) and other histopathological techniques. The ISH method was found to label very conspicuously the cells containing parasite stages, with the signal concentrating in merogonial and sporogonial plasmodia within the infected cell nuclei. Comparison with CW demonstrated limited ISH signal in cells containing mature spores, which was attributed mostly to the scarcity of probe targets present in these stages. Although spores were detected in other organs of the digestive system as well as in the peripheral blood, proliferative stages or parasite reservoirs were not found in this work outside the intestines. The study demonstrated a frequent disassociation between the presence of abundant spores and the intensity of the infections as determined by the parasite activity. The ISH allows confirmatory diagnosis of GSB microsporidiosis and estimation of infection intensity and will be a valuable tool for a more precise determination of parasite dissemination pathways and pathogeny mechanisms.     

Sysmex XT‐2000iV scattergram analysis in a cat with basophilia

A 13‐year‐old female Domestic Shorthair cat was presented to the Veterinary Teaching Hospital of the University of Milan for an interscapular mass suspected to be a mesenchymal malignant tumor. A preoperative CBC performed with Sysmex XT‐2000iV showed leukocytosis with neutrophilia and eosinophilia. The Sysmex WBC/DIFF scattergram showed an additional, well‐separated cluster of events between the neutrophil, eosinophil, and lymphocyte clusters. Blood smear evaluation revealed the presence of a significant number of basophils; thus, it was hypothesized that the additional cluster could represent the basophilic population. A second CBC, 24 days later, showed the same pattern on the WBC/DIFF scattergram in the absence of leukocytosis and neutrophilia. After surgical excision of the mass, a definitive diagnosis of feline injection site sarcoma was made. To the author's knowledge, there are no previous reports about the identification of feline basophils in the WBC/DIFF scattergram of Sysmex XT‐2000iV.     

A transgenic durum wheat line that is free of marker genes and expresses 1Dy10

As currently practiced, genetic engineering of monocots requires the use of selective agents, such as herbicides and antibiotics, and marker genes for resistance to favor the multiplication of the initially transformed cells. In the present paper we have used “minimal gene cassettes” and positive selection to generate transgenic durum wheat lines free of herbicide and antibiotic resistance marker genes. Two biolistic transformation experiments were carried out using three “minimal gene cassettes” consisting of linear DNA fragments each excised from the source plasmids. The targeted trait genes were two bread wheat sequences encoding the Dx5 and Dy10 high-molecular-weight (HMW) glutenin subunits, which have been associated with superior bread-making quality and which are absent from durum wheats. The positive selectable marker was the Escherichia coli phosphomannose isomerase (pmi) gene, whose product catalyzes the reversible interconversion of mannose-6-phosphate and fructose-6-phosphate, allowing plant cells to utilize mannose as a carbon source. PCR assays of genomic DNA from regenerated plants identified 15 T₀ plants that contained the pmi marker gene for an overall transformation efficiency of 1.5%, which is similar to biolistic transformation efficiencies of durum wheat with intact circular plasmids. Line TC-52, which initially contained pmi, non-expressed 1Dx5, and expressed 1Dy10 HMW glutenin subunit transgenes, was further investigated. PCR was used to follow inheritance of the pmi marker gene and 1Dx5 from the T₁ to T₃ generations. Transgene expression was monitored by the chlorophenol-red assay for pmi and SDS-PAGE of seed proteins for 1Dy10. From these analyses, we observed that the 1Dy10, 1Dx5 and pmi transgenes were not linked, allowing us in the T₃ generation to identify 1Dy10 transgenic segregants that contained no marker or silent 1Dx5 transgenes. Homozygotes containing and expressing only the 1Dy10 transgene were identified in the T₄ generation. These experiments show that it is possible to combine biolistic transformation by minimal gene cassettes with genetic segregation to make marker-free transgenic wheat plants with new traits.