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The advancement of human retinal pigment epithelial cell (hRPE) replacement therapy is partly dependent on optimization of cell culture, cell preservation, and storage medium. This study was undertaken to search for a suitable storage temperature and storage medium for hRPE. hRPE monolayer sheets were cultured under standard conditions at 37°C and then randomized for storage at six temperatures (4, 16, 20, 24, 28, and 37°C) for 7 days. After revealing a suitable storage temperature, hRPE sheets were subsequently stored with and without the silk protein sericin added to the storage medium. Live/dead assay, light microscopy, pH, and phenotypic expression of various proteins were used to assess cell cultures stored at different temperatures. After 7 days of storage, hRPE morphology was best preserved at 4°C. Addition of sericin to the storage medium maintained the characteristic morphology of the preserved cells, and improved pigmentation and levels of pigmentation-related proteins in the cultured hRPE sheets following a 7-day storage period at 4°C.
B-cell non-Hodgkin lymphoma (B-cell NHL) is the second commonest malignancy in the stomach. We determined the distribution of Helicobacter pylori outer membrane protein Q (HopQ) allelic type, cytotoxin-associated gene (cag)-pathogenicity activity island (cag-PAI) and vacuolation activating cytotoxin A (vacA) genes, respectively, in patients with B-cell NHL. We also compared them with their distribution in non-ulcer dyspepsia (NUD). H. pylori was cultured from gastric biopsy tissue obtained at endoscopy. Polymerase chain reaction was performed. Of 170 patients enrolled, 114 (63%) had NUD and 56 (37%) had B-cell NHL. HopQ type 1 was positive in 66 (58%) in NUD compared with 46 (82%) (P = 0·002) in B-cell NHL; HopQ type 2 was positive in 93 (82%) with NUD compared with 56 (100%) (P < 0·001) in B-cell NHL. Multiple HopQ types were present in 46 (40%) in NUD compared with 46 (82%) (P < 0·001) in B-cell NHL. CagA was positive in 48 (42%) in NUD vs. 50 (89%) (P < 0·001) in B-cell NHL; cagT was positive in 35 (31%) in NUD vs. 45 (80%) (P < 0·001) in B-cell NHL; left end of the cagA gene (LEC)1 was positive in 23 (20%) in NUD vs. 43 (77%) (P < 0·001) in B-cell NHL. VacAs1am1 positive in B-cell NHL in 48 (86%) (P < 0·001) vs. 50 (44%) in NUD, while s1am2 was positive in 20 (17%) in NUD vs. 46 (82%) (P < 0·001) in B-cell NHL. H. pylori strains with multiple HopQ allelic types, truncated cag-PAI evidenced by expression of cagA, cagT and cag LEC with virulent vacAs1 alleles are associated with B-cell NHL development.
L-carnitine (B-hydroxy-Y-N-trimethyl aminobutyrate) is a water-soluble product, found in animals, plants and microorganisms. Its synthesis takes place from two important amino acids, lysine and methionine. Essentially, L-carnitine has an intermediary role in metabolism. It promotes energy metabolism for cell and regulates co-enzyme A concentration in cystosol and mitochondria, which are important in glucose and lipid metabolism. L-carnitine was first isolated from the chicken embryo in a significant amount, but is absent in the egg. Under normal physiological conditions, the endogenously synthesised levels are sufficient for normal growth and functions. Its requirements are increased in stressful conditions and during metabolic and physiological higher demands, such as growth and laying periods. L-carnitine increases energy production, fat metabolism and improves immune status in birds, which is energy demanding. In this review, several aspects of the beneficial effects of dietary supplementation of L-carnitine on poultry health and production are briefly summarised.
Mycotoxins are secondary fungal metabolites of structurally diverse groups occurring worldwide as contaminants of cereals such as wheat, corn, maize, peanut, and barley, which comprise the main ingredients of poultry diets. Milk thistle (Silybum marianum) has wide range of beneficial properties, including data showing its positive effects in protecting the liver of birds exposed to aflatoxins. Trials have shown that in broilers affected by aflatoxicosis, which adversely affected feed intake, weight gain, feed efficiency, serum biochemistry and immune status, supplementation of diets with silymarin reduced the detrimental effects, thereby improving the productivity and health. Although promising results have been reported, established doses and further research on the standardisation of this herb will increase its uses as alternative to synthetic drugs. The following review discusses several important aspects of the beneficial properties of milk thistle on poultry growth performance in experimentally induced aflatoxicosis.
The present study was conducted to appraise the ontogenic radio-sensitivity of a serious tropical pest, Spodoptera litura (Fabr.). The molecular responses pertaining to the phenoloxidase (PO) pathway and an anti-oxidant defense mechanism were evaluated in order to understand its implication in pest control at pre-harvest and post-harvest intervals. Irradiation exhibited an inverse relationship with age with respect to impact on developmental and transcriptional responses. Transcript abundance of PO cascade enzymes, prophenoloxidase (slppo-2), its activating enzyme (slppae-1) and free-radical scavenging enzymes, superoxide dismutase (slsod) and catalase (slcat) was evaluated upon gamma irradiation alone and the dual-stress of radiation plus microbial challenge. The slppo-2, slppae-1, slsod and slcat transcripts were significantly up-regulated in F1 L6 larvae (6th-instar) resulting from 100 Gy sub-sterilized male adults and unirradiated female moths. The extent of upregulation was relatively higher in comparison with L6 survivors (6th-instar larvae) developed from irradiated neonates (L1) treated with 100 Gy. Upon Photorhabdus challenge, the transcripts were down-regulated in irradiated L1 suggesting increased larval susceptibility to bacterial infections. Radioresistance increased with the age of the insect, and molecular responses (transcript abundance) of insect defense mechanism were less influenced when older age (F1 progeny) were irradiated. These findings will help to optimize the gamma dose to be employed in inherited sterility technique for (pre-harvest) pest suppression and (post-harvest) phytosanitation and quarantine, and suggest compatible integration of biorational tactics including nuclear technology.
Based upon its economical perspective, the phenomenon of forced moulting in the poultry industry has become a common practice to increase the productivity and reproductive life span of birds. Different feed supplements, including vitamins, minerals, probiotics and prebiotics have been extensively used by poultry farmers for many years. In the last decade, researchers have reported advantageous effects of these supplements in improving different health biomarkers of post moult poultry birds. Therefore, including with these supplements in post moult feed is believed to exhibit better results than moulting alone. The current review is aimed at highlighting the empirical data available on the importance of various feed supplements that are considered favourable in ameliorating the health status of moulted poultry birds.
Phenotype of cultured ocular epithelial transplants has been shown to affect clinical success rates following transplantation to the cornea. The purpose of this study was to evaluate the relationship between cell nucleus morphometry and phenotype in three types of cultured epithelial cells. This study provides knowledge for the development of a non-invasive method of determining the phenotype of cultured epithelium before transplantation. Cultured human conjunctival epithelial cells (HCjE), human epidermal keratinocytes (HEK), and human retinal pigment epithelial cells (HRPE) were analyzed by quantitative immunofluorescence. Assessments of nucleus morphometry and nucleus-to-cytoplasm ratio (N/C ratio) were performed using ImageJ. Spearman’s correlation coefficient was employed for statistical analysis. Levels of the proliferation marker PCNA in HCjE, HEK, and HRPE correlated positively with nuclear area. Nuclear area correlated significantly with levels of the undifferentiated cell marker ABCG2 in HCjE. Bmi1 levels, but not p63α levels, correlated significantly with nuclear area in HEK. The N/C ratio did not correlate significantly with any of the immunomarkers in HCjE (ABCG2, CK7, and PCNA) and HRPE (PCNA). In HEK, however, the N/C ratio was negatively correlated with levels of the undifferentiated cell marker CK14 and positively correlated with Bmi1 expression. The size of the nuclear area correlated positively with proliferation markers in all three epithelia. Morphometric indicators of phenotype in cultured epithelia can be identified using ImageJ. Conversely, the N/C ratio did not show a uniform relationship with phenotype in HCjE, HEK, or HRPE. N/C ratio therefore, may not be a useful morphometric marker for in vitro assessment of phenotype in these three epithelia.
We studied the prevalence of Helicobacter pylori virulence markers, e.g. cytotoxin associated gene (cagA), cagA promoter, vacuolating associated cytotoxin A (vacA) alleles induced by contact with epithelium (iceA type), and outer membrane protein Q (hopQ) in expatriates and compared them with those in local residents. Gastric biopsies were obtained at endoscopy for culture, histology and PCR for virulence marker and hopQ. Of 309 patients, 236 (76%) were males with a mean age of 45 years. A total of 102 patients were expatriates. hopQ type 1 was present in 98 (47%) local residents compared to 88 (86%) expatriates (P < 0·001), while hopQ type 2 was present in 176 (85%) local residents, compared to 60 (59%) expatriates (P < 0·001). H. pylori virulence marker cagA was positive in 97 (47%) local residents compared to 86 (84%) expatriates (P < 0·001) while cagA-P was positive in 72 (35%) local residents compared to 87 (85%) expatriates (P < 0·001). iceA type 1 was positive in 157 (76%) local residents compared to 45 (44%) expatriates (P < 0·001), while iceA type 2 was positive in 81 (39%) local residents compared to 86 (84%) expatriates (P < 0·001). Distribution of H. pylori cagA, cagA promoter, iceA and hopQ type in local residents and expatriates was different. H. pylori virulence markers were associated with severe pathology in expatriates.
Delineating site-specific management zones within fields can be helpful in addressing spatial variability effects for adopting precision farming practices. A 3-year (2008/09 to 2010/11) field study was conducted at the Postgraduate Agricultural Research Station, University of Agriculture, Faisalabad, Pakistan, to identify the most important soil and landscape attributes influencing wheat grain yield, which can be used for delineating management zones. A total of 48 soil samples were collected from the top 300 mm of soil in 8-ha experimental field divided into regular grids of 24 × 67 m prior to sowing wheat. Soil and landscape attributes such as elevation, % of sand, silt and clay by volume, soil electrical conductivity (EC), pH, soil nitrogen (N) and soil phosphorus (P) were included in the analysis. Artificial neural network (ANN) analysis showed that % sand, % clay, elevation, soil N and soil EC were important variables for delineating management zones. Different management zone schemes ranging from three to six were developed and evaluated based on performance indicators using Management Zone Analyst (MZA V0·1) software. The fuzziness performance index (FPI) and normalized classification entropy NCE indices showed minimum values for a four management zone scheme, indicating its appropriateness for the experimental field. The coefficient of variation values of soil and landscape attributes decreased for each management zone within the four management zone scheme compared to the entire field, which showed improved homogeneity. The evaluation of the four management zone scheme using normalized wheat grain yield data showed distinct means for each management zone, verifying spatial variability effects and the need for its management. The results indicated that the approach based on ANN and MZA software analysis can be helpful in delineating management zones within the field, to promote precision farming practices effectively.
Poultry red mite (Dermanyssus gallinae), the haematophagous pest of egg laying hens, is an important problem in poultry production in many parts of the world. Control has typically relied on synthetic acaricides in spite of advancements in immunological, biotechnological and genetic approaches. However, the repeated long term use of these compounds has resulted in the development of drug resistant populations of poultry mites, therefore, the availability of effective acaricides is diminishing rapidly. Due to this, there is need for the continuous availability of new chemical acaricides to replace the older types, however the development and registration of new acaricides is a long expensive process, therefore, preserving and maintaining the effectiveness of available acaricides is essential.
This review has two parts; the first is to provide the basic understanding of acaricide resistance in D. gallinae while the second part provides more detailed knowledge about the management of resistance to preserve the efficacy of available acaricides.
Fusarium wilt caused by Fusarium oxysporum, Schlecht. emend. Snyd. & Hans. f. sp. ciceri is prevalent in most chickpea-growing countries and is a major devastating disease. Host plant resistance is the most practical method of disease management. Indigenous chickpea germplasm reveals a heterogeneous genetic make-up and the response of resistance to wilt is an unexplored potential source for disease resistance. There are 70 indigenous germplasm lines selected on the basis of their agronomic performance and diverse areas of collections in the country. Of these, four accessions had a highly resistant score of 1 and six had a score of 3 using a 1–9 rating scale, indicating their level of resistance to Fusarium wilt (race 4). Other germplasm accessions of chickpea were found to be moderately resistant to highly susceptible disease reaction. Likewise, the same set of germplasm was also screened for Meloidogyne incognita (race 1) using pot culture under controlled condition. Only one accession was found to be resistant to this pest. These resistant gene sources can be utilised effectively for race-specific chickpea wilt and root-knot resistance breeding programmes.
Poultry researchers and nutritionists are looking for viable alternative feed additives since conventional supplements have been criticised for their potential negative impact on the food chain. Among the currently available poultry feed additives, natural herbs and plants have been widely advocated due to their reported widespread beneficial effects. Garlic (Allium sativum) is one such potential feed supplement which has recently been reported as having a wide range of beneficial effects on the production performance and physiological biochemistry of broilers and laying hens. Notable beneficial effects have been seen on growth, feed efficiency, egg production and quality, as well as stimulation of immune system and lowering blood cholesterol levels in poultry birds. The results reported vary from author to author probably due to variations in the dose of the product fed, the duration of feeding and processing techniques employed.
Two reviews are available in published literature relating to the effects of ascorbic acid on poultry performance. The first review was written by Pardue and Thaxton in 1986 who reviewed the role of ascorbic acid in alleviating the negative effect of heat stress in poultry birds. The second review was presented by Whitehead and Keller in 2003, who detailed the general effects of ascorbic acid in poultry birds, not taking into consideration ‘heat stress’. The first review is deficient in the literature published after 1986, while the second review does not describe the effect of ascorbic acid in heat stressed birds. The present review describes the past and present knowledge of ascorbic acid in alleviating heat stress in poultry birds with new aspects. Heat stress is associated with compromised performance and productivity through a decline in feed intake, nutrient utilisation, growth rate, egg production and quality, feed efficiency and immunity. Heat stress is also characterised by a reduced antioxidant status in birds, resulting in increased oxidative stress. Moreover, male fertility is decreased when birds are exposed to heat stress. Supplementation of 250 mg of ascorbic acid per kg of feed has been found to be optimum to improve feed intake, body weight gain, feed efficiency, egg production and quality, nutrient digestibility, immune response and antioxidant status in poultry birds. This work compiles past and present information about the role of ascorbic acid in heat-stressed poultry.
Due to the potentially undesirable effects of antibiotics as growth promoters in poultry production, researchers are looking for viable alternative to limit or replace their use. One such class of comparable alternative is natural source of herbs and medicinal plants. In the last decade, these alternatives have been increasingly used in broiler, layer and Japanese quail diets. Reports have variously claimed that medicinal plants, used as either the whole plant, their leaves or flowers, can enhance poultry performance. From the available literature, it can be concluded that thyme (Thymus vulgaris) belongs to such class of medicinal plant and may be an effective alternative to antibiotics in poultry production. In this review, its effects on different parameters of production performance in poultry are briefly discussed.
In the last decade, there has been growing interest in the use of natural herbs and medicinal plants as feed additives in poultry diets to maximise their potential output. Ginger is one such potential rhizome with a wide range of medicinal effects. In broilers and layers, this plant has been used in different forms, doses and durations. In this review, documented effects of ginger in poultry feed on feed intake and feed conversion ratio, growth and weight gain, carcass yield, egg production and quality, antioxidants and blood biochemistry, with their possible mechanisms of action, are discussed.
We describe the first reported case of a malignant glomus tumour of the larynx.
Case report and review of the world literature concerning malignant glomus tumours.
A 37-year-old man presented with progressive hoarseness and dyspnoea. A smooth, right-sided laryngeal mass was found on flexible nasolaryngoscopy. Initial biopsy of this lesion was reported as a true ‘glomus tumour’, which is a benign lesion. Wide local excision was performed. The final histology of this specimen showed it to be a ‘malignant glomus tumour’. Immunohistochemistry was positive for smooth muscle actin, collagen type IV, vimentin and cluster of differentiation 34 glycoprotein. Total laryngectomy was then performed as a definitive curative procedure.
Glomus tumour is rarely found in visceral locations. This tumour and its malignant counterpart are rare differential diagnoses of laryngeal masses.
Polyurethanes (PU) have been widely used as biomaterial in recent years, while thrombus may still occur when contacting with blood especially for extended period of time. Poly(ethylene glycol) (PEG) and phosphorylcholine (PC)-based polymers are commonly employed for surface modification to create protein repellent surfaces. PC-based polymers have been investigated as biomimetic materials because PC is the major component in the outer layer of cell membranes. In this study, the biomimetic copolymer brush of PEG-b-poly(2-methacryloyloxyethyl phosphorylcholine) on PU surfaces was synthesized via atom transfer radical polymerization (ATRP) with a surface initiator. The flexible PEG chain was 200 g·mol-1, while the poly(2-methacryloyloxyethyl phosphorylcholine) (poly(MPC)) chain length was controlled by the ratio of monomer to sacrificial initiator in solution. The topology of the modified surfaces was characterized by the phase image of atomic force microscopy (AFM) to study the synergy effect between PEG chains and poly(MPC) chains. The unmodified and modified surfaces were characterized by Fourier transform infrared (FTIR), X-ray photoelectron spectroscopy (XPS), water contact angle and platelet adhesion. The results demonstrated that efficient grafting of PEG-b-poly(MPC) brushes on the surfaces was achieved. The PU surfaces modified with PEG and phosphorylcholine zwitterionic brushes showed effective resistance to platelet adhesion and high hemocompatibility in vitro. These PEG and PC-grafted PU materials might be potentially applied in blood-contacting materials or devices due to their good mechanical and hemocompatible properties.
There is growing interest in developing natural alternatives to antibiotic growth promoters in order to maintain both birds’ performance and health. In the last decade, Turmeric has been extensively used in poultry diets. Turmeric is a natural herb of the ginger family, Zingiberaceae. Wide range medicinal properties of this plant have been advocated. In poultry feed, Turmeric has been extensively used in different concentrations, dosages and durations. In this review, the beneficial effects of this plant on growth, weight gain, feed conversion ratio, ameliorative effect on liver health, immunomodulatory and antioxidative effects are reviewed.