High water content silk protein-based hydrogels with tunable elasticity fabricated via a Ru(II) mediated photochemical cross-linking method

Silk is very promising in the field of biomaterials as a natural biomacromolecule. Silk protein can be made into various forms of materials, including hydrogels. However, silk protein-based hydrogels have not attracted much attention due to its weak mechanical properties. Here, we report high water content silk protein-based hydrogels with tunable elasticity which were fabricated through Ru(II) mediated photochemically cross-linking tyrosine residues in regenerated silk protein. The regenerated silk protein was characterized by Fourier transform infrared spectroscopy (FTIR). The gelation kinetics of the silk protein was studied by rheology measurements. The compressive mechanical properties of the silk protein-based hydrogels was investigated using compressive tests and dynamic mechanical analysis (DMA). Compressive modulus of the hydrogels reached 349±64 MPa at 15 % strain. The fabricated silk protein-based hydrogels were also characterized by Scanning electron microscopy (SEM), revealing an interconnected porous network structure, typical of hydrogels, with an average pore size of approximately 130 μm. Finally, biocompatibility of the silk protein-based hydrogels was demonstrated through cell culture studies using a human fibroblast cell line, HFL1. The reported silk protein-based hydrogels represent a promising candidate for biomaterial applications.     

Suspected ivermectin toxicosis in a miniature mule foal causing blindness

A 9-week-old miniature mule foal presented to the Veterinary Medical Teaching Hospital for acute blindness, ataxia, and depression following an overdose of an over-the-counter ivermectin-based de-worming medication. Ophthalmic examination and electrodiagnostic evaluation eliminated outer retinal abnormalities as the primary cause of the bilateral blindness, implicating instead a central neurologic effect of the drug. With symptomatic and supportive care, the foal recovered fully and regained its vision.     

Abnormal ocular pigment deposition and glaucoma in the dog

The combined occurrence of ocular pigment deposition and glaucoma has been described in Cairn Terriers. Recently, this condition was also observed in two other breeds: the Boxer (two cases) and the Labrador Retriever (one case). Six dogs were referred to the Ophthalmology section of the Department of Clinical Sciences of Companion Animals and to a private referral clinic because of glaucoma or blindness in one or both eyes. In five cases ophthalmic examination showed pigment depositions in the sclera around the entire circumference of the perilimbal zone. Eight enucleated eyes (four eyes of two Cairn Terriers, three eyes of two Boxers and one eye of a Labrador Retriever) were examined microscopically. All eyes showed the same findings: an extensive infiltration of large melanin-containing cells with an eccentric nucleus, located in the iris, ciliary body, retina, choroids and sclera. Transmission electron microscopy of two of the examined eyes revealed that the morphology of most of these cells was consistent with melanophages. While reports in the veterinary literature concerning this condition are limited the cells concerned have been described to be melanocytes. Further research is needed to conclusively identify the cell type. As described in the present report, the histologic and transmission electron microscopic findings suggest a different etiology of the ocular pigment deposition and glaucoma compared with the pigment dispersal syndrome in humans.     

Risk factors associated with occurrence of nematodes in free range pigs in Busia District,Kenya

Nematode infections are a serious constraint to pig production, especially where free range pig keeping is practiced. This study investigated the epidemiology of nematodes in free range pigs in Busia District, Kenya. Three hundred and six pigs from 135 farms were sampled for faeces that were analysed for nematode eggs per gram (EPG) of faeces using the McMaster technique. The nematode eggs were also identified to genus and species based on morphology. A questionnaire on risk factors was also administered to the pig owners. The overall prevalence and mean nematode EPG were 84.2% and 2,355, respectively. The nematode eggs were identified as those belonging to Oesophagostomum spp. (75%), Strongyloides ransomi (37%), Ascaris suum (18%), Metastrongylus spp. (11%), Trichuris suis (7%) and Physocephalus sexalatus (3%). The prevalence of nematodes was positively correlated (p < 0.05) with the amount of rainfall in the division of the pigs' origin (all nematodes except S. ransomi). The prevalence of nematodes was also associated with the age of the pigs. A lower burden of nematodes was associated (p < 0.05) with a history of deworming (A. suum) and the provision of night housing (S. ransomi and Metastrongylus spp.). In conclusion, this study has provided information on nematode infections and the associated risk factors for free range pigs in Busia District, which can be used when implementing integrated control measures.     

In vitro inhibition of the activation of Pro-matrix Metalloproteinase 1 (Pro-MMP-1) and Pro-matrix metalloproteinase 9 (Pro-MMP-9) by rice and soybean Bowman-Birk inhibitors

The in vitro inhibitory activity of the rice Bowman-Birk inhibitor (rBBI) or soybean Bowman-Birk inhibitor (sBBI) against trypsin-catalyzed activation of pro-matrix metalloproteinase 1 or 9 (pro-MMP-1 or pro-MMP-9), respectively, was investigated using electrophoresis with silver staining, heparin-enhanced zymography, biotinylated gelatin, Biotrak assay, and fluorescence quenched substrate hydrolysis. rBBI at concentrations of 0.08-0.352 mg/mL dose-dependently inhibited the in vitro activation of 45 microg/mL pro-MMP-1 by trypsin. Heparin-enhanced zymography analysis of pro-MMP-1, trypsin-activated MMP-1, and a mixture of pro-MMP-1-trypsin-rBBI showed clear zones associated with trypsin-activated MMP-1 and the absence of clear zones in lanes containing pro-MMP-1 or a mixture of pro-MMP-1, trypsin, and rBBI. The results of the Biotrak assay also indicated that rBBI dose-dependently suppressed the activation of pro-MMP-1 by trypsin. sBBI dose-dependently inhibited the activation of 100 microg/mL of pro-MMP-9 by trypsin. Biotinylated gelatin assays demonstrated that pro-MMP-9 or pro-MMP-9 in the presence of trypsin and BBI did not hydrolyze gelatin, whereas p-aminophenylmercury acetate (APMA)-activated MMP-9 and trypsin-activated MMP-9 caused significant hydrolysis of gelatin. Quenched fluorescence substrate hydrolysis for total MMP activity showed that pro-MMP-1 or pro-MMP-9 did not hydrolyze the substrate Mca-Pro-Leu-Gly-Leu-Dpa-Ala-Arg-NH2; active MMP-1 or MMP-9 hydrolyzed the substrate, but lower substrate hydrolysis was obtained when pro-MMP-1 or pro-MMP-9 was incubated with trypsin in the presence of increasing concentrations of rBBI. The results are discussed in light of the role of MMP-1 and MMP-9 in the process of angiogenesis and the potential of rBBI or sBBI as a functional food ingredient.     

Multilocus sequence analysis supports a low genetic diversity among ‘<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma australasia’ related strains infecting vegetable crops and periwinkle in Egypt

‘Candidatus Phytoplasma australasia’ causes important damages to the Egyptian vegetable crop production. A prerequisite for controlling the different diseases it causes to eggplant, tomato and squash, is to trace its propagation pathways. To allow the differentiation of ‘Ca. P. australasia’ strains, a multilocus sequence analysis protocol was developed. Four conserved phytoplama genes namely tuf, secY, dnaK and dppA, were selected among the CDS of a ‘Ca. P. aurantifolia’ genome draft. The corresponding genes were PCR amplified from tomato, eggplant and squash collected in 2010 from the governorates Sharkia, Elmynia and Beni sueif, as well as from Catharanthus roseus periwinkles collected in 2013 from Kafrelsheikh governorate. Sequence comparisons showed no diversity among the Egyptian isolates of ‘Ca. P. australasia’ that also constitute a distinct cluster within the 16SrII-D taxonomic subgroup. This low diversity supports a common epidemiology for the different diseases affecting vegetable crops and periwinkle in Egypt and suggests that future investigations on insect vector should focus on polyphagous leafhoppers.     

Dynamics of natural bovine herpesvirus-1 (BoHV-1) infection in a dairy herd

In this study, natural cycling of BoHV-1 infection was investigated in two groups of dairy cattle containing 2120 head. Group 1 comprised 127 animals and they were monitored for BoHV-1 infection virologically and serologically in six consecutive sampling periods. It consisted of naive heifers between 6 and 8 months of age, while in group 2, age, sex and the BoHV-1 serostatus of the animals were disregarded. The animals in group 1 were found to have seroconverted at the second sampling. Results of the serological study showed slight antibody response after natural BoHV-1 infection in the herd and neutralizing titres fell below protective levels in the 6–8 months after the peak. During the 2-year study period, one recurrence was detected after primary infection. Virus isolation studies revealed a cytopathic effect indicative of BoHV-1 in two nasal swabs taken during the fifth sampling period from animals with mild upper respiratory tract symptoms. As the study was carried out under natural conditions, it is not known whether the viruses isolated were from recurrences or re-infections. Data from cross-neutralization tests with herd isolates showed higher antibody response than those with the reference virus. The dynamics of BoHV-1 in both groups were found to be statistically similar.     

Analysis of the Mycosporine-Like Amino Acid (MAA) Pattern of the Salt Marsh Red Alga Bostrychia scorpioides

This study presents the validation of a high-performance liquid chromatography diode array detector (HPLC-DAD) method for the determination of different mycosporine-like amino acids (MAAs) in the red alga Bostrychia scorpioides. The investigated MAAs, named bostrychines, have only been found in this specific species so far. The developed HPLC-DAD method was successfully applied for the quantification of the major MAAs in Bostrychia scorpioides extracts, collected from four different countries in Europe showing only minor differences between the investigated samples. In the past, several Bostrychia spp. have been reported to include cryptic species, and in some cases such as B. calliptera, B. simpliciuscula, and B. moritziana, the polyphyly was supported by differences in their MAA composition. The uniformity in the MAA composition of the investigated B. scorpioides samples is in agreement with the reported monophyly of this Bostrychia sp.