Canine squamous cell carcinoma: Electrochemotherapy association with surgery and correlation with overall survival

Squamous cell carcinoma (SCC) is an important malignancy in dogs, due to its incidence and clinical presentation, which can be of locally aggressive single or multiple lesions with a metastatic potential. The objective of this investigation was to evaluate SCC response to treatment, anatomopathological and immunohistochemical characteristics, disease-free interval and overall survival time. 54 dogs with histopathologically diagnosed SCC were included in this study. Their mean age was 9.16 years with a range of 1–14 years. Of the 54 animals in the study, 34 (65.4%) had white skin and white fur coats. There was a significant correlation between fur coat colour and the development of tumours in areas of sun exposure (p = .001). Animals with tumours in areas of the body exposed to the sun had longer overall survival time than animals with tumours in areas not associated with sun exposure (p = .001). Surgery combined with electrochemotherapy (ECT) yielded a survival rate 32% higher than using a surgical approach alone (HR = 0.32, p = .038, IC = 0.11–0.94). ECT, with or without surgery, had an objective response rate of 90.9%. Local lymph node and/or distant site metastasis at diagnosis, or at some point during follow-up, occurred in 34.6% (18/52) of animals. Animals with tumours in sun exposed locations had more aggressive histopathological characteristics but had longer overall survival time. This is probably due to individualised therapeutic treatment with both surgery and ECT.     

Growth and survival of the freshwater clam,Galatea paradoxa (Born 1778) cultured on different substrata at the Volta estuary,Ghana

This study was conducted to assess the optimum clam size and substratum type for the culture of Galatea paradoxa. The experiment was conducted over a 90‐day period at the Volta Estuary, Ghana. Three size classes of G. paradoxa categorized as small (27.6 ± 0.4 mm), medium (36.8 ± 0.4 mm) and large (50.0 ± 0.6 mm), were used to ascertain the effect of a sandy and muddy substratum on growth performance and survival. Growth increased from the small‐sized clams to the large‐sized clams at 2.54, 3.03 and 3.43 g, respectively, over the experimental period. Growth was higher (P < 0.05) in the muddy substratum (3.58 g) compared with sandy (2.41 g). Similarly, the specific growth rate increased from 0.99% day^?1 in the small‐sized clams to 1.36% day^?1 in the large‐sized clams. Survival rates were significantly higher (99.4%) for the large‐sized clams compared with the medium (78.8%) and the small‐size clams (74.1%). There was significant interaction between the size class and the substratum type with a trend towards increasing survival rate from the small to the large size clams in both substrata. The survival rate was lower in the muddy substratum (71.4%) compared with the sandy substratum (96.7%). The yield was significantly higher in the muddy (2.01 kg m^?2 90 day^?1) compared with sandy substratum (1.87 kg m^?2 90 day^?1). The results of this study indicate that the culture of small‐sized clams is best practiced on sandy substratum due to their elevated survival rates, whereas the muddy substratum appears most suitable for the culture of larger clams (>40 mm) because of their relatively higher survivorship and better growth performance.     

Tail Muscle Free Amino Acid Concentration of Pacific White Shrimp,Litopenaeus vannamei,Fed Diets Containing Protein‐bound versus Crystalline Amino Acids

Tail muscle tissue free amino acids (FAA) concentration is reported for subadult (4.3 g) Litopenaeus vannamei fed diets containing only protein‐bound amino acids, crystalline lysine as a supplement to protein‐bound lysine, and only crystalline amino acids (CAA). FAA were determined in shrimp at 0.00, 0.25, 0.5, 1, 2, 4, and 8 h postfeeding. Highest total tissue concentrations of essential amino acids and individual amino acids were typically observed 4 h postfeeding for shrimp fed intact and crystalline lysine‐supplemented feeds. Those shrimp offered diets containing only CAA showed no tissue maxima. Similar uptake patterns for lysine shown by shrimp fed the intact and crystalline lysine‐supplemented diet indicate the potential use of small amounts of crystalline lysine in shrimp feeds.     

Investigations into inhibitor type and mode, simulated gastrointestinal digestion, and cell transport of the angiotensin I-converting enzyme-inhibitory peptides in Pacific hake (Merluccius productus) fillet hydrolysate

Fish protein hydrolysate (FPH) produced by incubation of Pacific hake fillet with 3.00% Protamex at pH 6.5 and 40 degrees C for 125 min demonstrated in vitro ACE-inhibitory activity (IC50 = 165 microg/mL), which was enhanced by ultrafiltration through a 10 kDa molecular weight cutoff membrane (IC50 = 44 microg/mL). However, after simulated gastrointestinal digestion, FPH and ultrafiltrate had similar ACE-inhibitory activity (IC 50 = 90 microg/mL), indicating that FPH peptides act as "pro-drug type" inhibitors and that enrichment by ultrafiltration may be unnecessary. Matrix-assisted laser desorption/ionization-time of flight mass spectrometry confirmed that the molecular weights of major peaks were <1 kDa regardless of ultrafiltration. ACE-inhibitory activities of digested hydrolysates were not significantly affected by preincubation with ACE ( P > 0.05) and exhibited a competitive inhibitory mode. A permeability assay using fully differentiated colorectal adenocarcinoma (Caco-2) cells showed an apical to basolateral transport of peptides that ranged from approximately 2 to 20% after 2 h at 37 degrees C. Pacific hake fillet hydrolysates are a potentially bioavailable source of ACE-inhibitory peptides awaiting further in vivo study.     

Leaf structure and function of sweet cherry tree (Prunus avium L.) cultivars with open and dense canopies

Variations in morpho-anatomy, chemical composition, gas exchange and stem water potential (Ψ_stem) were studied in leaves of Prunus avium L. cultivars with different growth habit: Burlat and Summit (upright, dense canopies), and Van (spreading, open canopy). Light transmittance was highest in Van canopies. Therefore, Van allowed an easier penetration of the visible radiation through the canopy, which improves photosynthetic rates, especially at harvest. Conversely, no differences in Ψ_stem were found among cultivars. Van showed the highest leaf mass per unit area and variations in this parameter were mainly associated with alterations in leaf thickness than in leaf density. Leaves of Summit and Van were thicker than those of Burlat, mainly due to increased palisade and spongy parenchyma thickness. However, Burlat leaves had the highest palisade/spongy ratio. Total chlorophyll concentration per area and the chlorophyll a/b ratio were always higher in Van leaves than in the other two cultivars. In contrast, Van leaves had the lowest concentrations of soluble carbohydrates and starch. In dense canopies of Summit and Burlat, light reduction caused a notable increase in the relative concentration of total phenols. The above results provide useful information about the biological behaviour of sweet cherry cultivars with open and dense canopies in order to help the agronomical decisions regarding the local environmental conditions and the consumers’ preference. In particular, it is important to manipulate factors such as orchard exposure, row orientation, plant spacing, training system and pruning, which are able to reduce shading in the canopy.     

Polymerase Chain Reaction assay for the detection of Kudoa paniformis and Kudoa thyrsites in Pacific Hake (Merluccius productus)

Myoliquefaction of Pacific hake has been attributed to proteolytic action associated with parasitic infection. Among the two infecting species of Kudoa, Kudoa paniformis and Kudoa thyrsites, the former is reported to be more virulent for the "soft flesh" phenomenon in Pacific hake. The objective of this research was to develop a sensitive and specific polymerase chain reaction (PCR) assay to detect infection of hake by K. paniformis. Primers based on specific regions ( approximately 1562 bp) of the small subunit ribosomal DNA of K. paniformis successfully amplified the target DNA segments from both spore and muscle extracted DNA templates. DNA sequencing confirmed the veracity of this method to distinguish parasitic infection by K. paniformis versus K. thyrsites. The established PCR method was applied to investigate Kudoa infection in 44 Pacific hake samples using DNA extracted from muscle and/or spores, and the results were compared to infection evaluated by microscopic examination of extracted spores.     

The production of azadirachtin by in-vitro tissue cultures of neem,Azadirachta indica

Callus produced from leaves of a Ghanaian strain of the neem tree, Azadirachta indica A. Juss has been shown to produce the natural insecticide azadirachtin when grown in a defined medium. The azadirachtin was isolated by standard procedures of solvent partition and column chromatography monitored by supercritical fluid chromatography. Biological activity was monitored with antifeedant tests using the desert locust (Schistocerca gregaria Forsk.). The azadirachtin was identified by chromatography on three independent chromatographic systems (SFC, HPLC & TLC) and two thin-layer colour tests. It has 100% antifeedant activity at < 0.04mg litre^?1. The yield of azadirachtin was 0.0007% based on dry weight of callus.     

Alternative low doses and routes of administering a prostaglandin F2�� analogue to induce luteolysis in Nelore cows

The present study was conducted in order to verify the efficacy of lower doses and alternative routes of a prostaglandin F2α analogue, luprostiol (PGF), for the induction of luteolysis and the precipitation of estrus in nonlactating Nelore cows (Bos taurus indicus). A conventional dose (15 mg) of PGF was compared to doses lower than the conventional dose, which ranges from 10 to 50%, that were administered intramuscularly (IM), intravulvosubmucosally (IVSM), or in the Bai-hui acupuncture site located within the lumbosacral area. The cows were administered PGF 8 day after estrus in the presence of a corpus luteum, and randomly assigned to the following groups: G1 (positive control), 15 mg, IM (n = 23); G2, 7.5 mg, IM (n = 23); G3, 3.75 mg, IM (n = 24); G4, 7.5 mg, IVSM (n = 25); G5, 3.75 mg, Bai-hui acupoint (n = 24); and G6, 1.5 mg, Bai-hui acupoint (n = 25). The results indicated that 50% of a conventional dose of PGF (7.5 mg) resulted in a complete luteal regression (plasma progesterone <1 ng/ml) at Hour 48, and hastened estrus, regardless of whether or not PGF was administered IM or IVSM. Comparatively, 10 or 25% of the conventional dose, even when administered to the Bai-hui acupoint, resulted in an initial reduction in the concentration of progesterone at Hour 24, followed by an increase observed at Hour 48. In conclusion, 25% of a conventional PGF dose administered via the Bai-hui acupoint proved inadequate to induce a complete luteal regression, whereas 50% of a conventional dose administered IM or IVSM was found to be the minimal dose required to induce effectively a complete luteal regression, and to precipitate the onset of estrus in nonlactating Nelore cows.     

Cross comparison of seminal plasma proteins from cattle and buffalo (Bubalus bubalis)

The objective of this study was to evaluate seminal plasma proteins from cattle and buffalo (Bubalus bubalis), to identify differences between related species. Sixteen buffaloes and 16 cattle between 30 and 60 months of age were used. Semen collection was performed by electroejaculation, followed by macroscopic and microscopic subjective analyses. After analysis, the samples were centrifuged at 800 g for 10 min, and the supernatant (seminal plasma) was recentrifuged at 10,000 g for 30 min at 4°C. The total protein concentration was determined by the Bradford method, and the proteins were digested in solution for mass spectrometry (nLC-MS/MS). Multivariate statistical analysis was used to evaluate the proteomics results by non-hierarchical clustering the considering exponentially modified protein abundance index (emPAI). Principal component analysis (PCA) and partial least squares discriminant analysis (PLS-DA) were used for clustering. Proteomics identified 78 proteins, and multivariate analysis showed 4 that were over-expressed in buffaloes (cystatin C, prosaposin, peptide YY and keratin type II cytoskeletal 5) and 9 in cattle (spermadhesin-1, seminal plasma protein PDC-109, ribonuclease 4, metalloproteinase inhibitor 2, acrosin inhibitor 1, seminal ribonuclease, C-type natriuretic peptide, angiogenin-1 and osteopontin). Among the proteins identified in seminal plasma, the C-type natriuretic peptide and metalloproteinase inhibitors were described for the first time in buffaloes. Some protease inhibitors were found over-expressed in buffaloes, and important proteins in seminal plasma of cattle were not identified or were found at lower expression levels in buffaloes, which can contribute to reproductive performance in this species.