Animal-derived Products Research Articles

Unrestricted somatic stem cells, as a novel feeder layer: Ex vivo culture of human limbal stem cells.

Ex vivo culture of limbal stem cells (LSCs) is a current promising approach for reconstruction of the ocular surface. In this context, 3T3 feeder layer cells (mouse embryo fibroblast) are generally utilized to maintain and expand LSCs. The aim of this study is to develop a novel culture method (animal-derived products free) to expand LSCs, using umbilical cord derived human unrestricted somatic stem cells (hUSSCs) instead of 3T3 cell with an emphasis on maintaining of the Stemness in LSCs. Using flow-cytometer, isolated hUSSCs were characterized for CD105, CD90, CD166, CD34, CD45, CD31 cell surface markers and their differentiation capability into adipogenic as well as osteogenic lineages were evaluated. In addition to colony-forming efficiency (CFE), epithelial lineage differentiation and karyotyping, LSC properties were evaluated for ABCG2, ΔNP63-α, CK19, CK3, and CK12 mRNA and protein expressions using quantitative RT-PCR (qRT-PCR) and immunocytochemistry, when these cells were co-cultured with hUSSCs (in comparison with 3T3 feeder layer). LSCs, co-cultured with hUSSCs, showed normal karyotype (46, XX), while they could efficiently form colony (86 ± 3) and display up-regulation of the genes associated with stemness and down-regulation of corneal epithelial differentiation genes. Consistent with 3T3 feeder cells, hUSSCs with spindle-shaped morphology and quick splitting up properties had ability to preserve the stem like-cell phenotype of LSCs. These findings were confirmed by qRT-PCR and flow-cytometer. Findings of present study suggest hUSSCs as a promising alternative method for 3T3 feeder layer cells, to preserve growth and stemness of LSCs ex vivo culture.

Novel method for the determination of average molecular weight of natural polymers based on 2D DOSY NMR and chemometrics: Example of heparin

Apart from the characterization of impurities, the full characterization of heparin and low molecular weight heparin (LMWH) also requires the determination of average molecular weight, which is closely related to the pharmaceutical properties of anticoagulant drugs.To determine average molecular weight of these animal-derived polymer products, partial least squares regression (PLS) was utilized for modelling of diffused-ordered spectroscopy NMR data (DOSY) of a representative set of heparin (n=32) and LMWH (n=30) samples. The same sets of samples were measured by gel permeation chromatography (GPC) to obtain reference data. The application of PLS to the data led to calibration models with root mean square error of prediction of 498Da and 179Da for heparin and LMWH, respectively. The average coefficients of variation (CVs) did not exceed 2.1% excluding sample preparation (by successive measuring one solution, n=5) and 2.5% including sample preparation (by preparing and analyzing separate samples, n=5). An advantage of the method is that the sample after standard 1D NMR characterization can be used for the molecular weight determination without further manipulation. The accuracy of multivariate models is better than the previous results for other matrices employing internal standards. Therefore, DOSY experiment is recommended to be employed for the calculation of molecular weight of heparin products as a complementary measurement to standard 1D NMR quality control. The method can be easily transferred to other matrices as well.

Occurrence of pyrrolizidine alkaloids in animal- and plant-derived food: results of a survey across Europe

ABSTRACTPyrrolizidine alkaloids (PAs) are secondary metabolites of plant families such as Asteraceae or Boraginaceae and are suspected to be genotoxic carcinogens. Recent investigations revealed their frequent occurrence in honey and particularly in tea. To obtain a comprehensive overview of the PA content in animal- and plant-derived food from the European market, and to provide a basis for future risk analysis, a total of 1105 samples were collected in 2014 and 2015. These comprised milk and milk products, eggs, meat and meat products, (herbal) teas, and (herbal) food supplements collected in supermarkets, retail shops, and via the internet. PAs were detected in a large proportion of plant-derived foods: 91% of the (herbal) teas and 60% of the food supplements contained at least one individual PA. All types of (herbal) teas investigated were found to contain PAs, with a mean concentration of 460 µg kg−1 dry tea (corresponding to 6.13 µg L−1 in [herbal] tea infusion). The highest mean concentrations were found in rooibos tea (599 µg kg−1 dry tea, 7.99 µg L−1 tea infusion) and the lowest in camomile tea (274 µg kg−1 dry tea, 3.65 µg L−1 tea infusion). Occurrence of PAs in food supplements was found to be highly variable, but in comparable ranges as for (herbal) tea. The highest concentrations were present in supplements containing plant material from known PA-producing plants. In contrast, only 2% of the animal-derived products, in particular 6% of milk samples and 1% of egg samples, contained PAs. Determined levels in milk were relatively low, ranged between 0.05 and 0.17 µg L−1 and only trace amounts of 0.10–0.12 µg kg−1 were found in eggs. No PAs were detected in the other animal-derived products.

Top-Down Inhibition of BMP Signaling Enables Robust Induction of hPSCs Into Neural Crest in Fully Defined, Xeno-free Conditions.

SummaryDefects in neural crest development have been implicated in many human disorders, but information about human neural crest formation mostly depends on extrapolation from model organisms. Human pluripotent stem cells (hPSCs) can be differentiated into in vitro counterparts of the neural crest, and some of the signals known to induce neural crest formation in vivo are required during this process. However, the protocols in current use tend to produce variable results, and there is no consensus as to the precise signals required for optimal neural crest differentiation. Using a fully defined culture system, we have now found that the efficient differentiation of hPSCs to neural crest depends on precise levels of BMP signaling, which are vulnerable to fluctuations in endogenous BMP production. We present a method that controls for this phenomenon and could be applied to other systems where endogenous signaling can also affect the outcome of differentiation protocols.

Bithionol residue analysis in animal-derived food products by an effective and rugged extraction method coupled with liquid chromatography-tandem mass spectrometry.

Herein, we developed a simple analytical procedure for the quantitation of bithionol residues in animal-derived food products such as porcine muscle, eggs, milk, eel, flatfish, and shrimp using a modified quick, easy, cheap, effective, rugged, and safe (QuEChERS) extraction method coupled with liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI+/MS-MS). Samples were extracted with 0.1% solution of formic acid in acetonitrile and the extract was purified using a C18 sorbent. Separation was performed on a Waters XBridge™ C18 reversed-phase analytical column using 0.1% solution of formic acid/acetonitrile as the mobile phase. Six-point matrix-matched calibration indicated good linearity, with the calculated coefficients of determination (R2) being≥0.9813. Intra- and inter-day recoveries (determined at spiking levels equivalent to 1×and 2×the limit of quantitation (0.25μg/kg)) ranged between 80.0 and 94.0%, with the corresponding relative standard deviations (RSDs) being≤8.2%. The developed experimental protocol was applied to different samples purchased from local markets in Seoul, which were tested negative for bithionol residues. In conclusion, the proposed method proved to be versatile and precise, being ideally suited for the routine detection of bithionol residues in animal-derived food products with various protein and fat contents.

Umbilical Cord Blood Platelet Lysate as Serum Substitute in Expansion of Human Mesenchymal Stem Cells.

The diverse clinical applications for human mesenchymal stem cells (hMSCs) in cellular therapy and regenerative medicine warrant increased focus on developing adequate culture supplements devoid of animal-derived products. In the present study, we have investigated the feasibility of umbilical cord blood-platelet lysate (UCB-PL) as a standard substitute for fetal bovine serum (FBS) and human peripheral blood-PL (PB-PL). In this experimental study, platelet concentrates (PC) from UCB and human PB donors were frozen, melted, and sterilized to obtain PL. Quality control included platelet cell counts, sterility testing (viral and microbial), total protein concentrations, growth factor levels, and PL stability. The effects of UCB-PL and PB-PL on hMSCs proliferation and differentiation into osteocytes, chondrocytes, and adipocytes were studied and the results compared with FBS. UCB-PL contained high levels of protein content, platelet-derived growth factor- AB (PDGF-AB), and transforming growth factor (TGF) compared to PB-PL. All growth factors were stable for at least nine months post-storage at -70˚C. hMSCs proliferation enhanced following treatment with UCB-PL. With all three supplements, hMSCs could differentiate into all three lineages. PB-PL and UCB-PL both were potent in hMSCs proliferation. However, PB promoted osteoblastic differentiation and UCB-PL induced chondrogenic differentiation. Because of availability, ease of use and feasible standardization of UCB-PL, we have suggested that UCB-PL be used as an alternative to FBS and PB-PL for the cultivation and expansion of hMSCs in cellular therapy.

Environmental impacts of food consumption by dogs and cats.

In the US, there are more than 163 million dogs and cats that consume, as a significant portion of their diet, animal products and therefore potentially constitute a considerable dietary footprint. Here, the energy and animal-derived product consumption of these pets in the US is evaluated for the first time, as are the environmental impacts from the animal products fed to them, including feces production. In the US, dogs and cats consume about 19% ± 2% of the amount of dietary energy that humans do (203 ± 15 PJ yr-1 vs. 1051 ± 9 PJ yr-1) and 33% ± 9% of the animal-derived energy (67 ± 17 PJ yr-1 vs. 206 ± 2 PJ yr-1). They produce about 30% ± 13%, by mass, as much feces as Americans (5.1 ± Tg yr-1 vs. 17.2 Tg yr-1), and through their diet, constitute about 25–30% of the environmental impacts from animal production in terms of the use of land, water, fossil fuel, phosphate, and biocides. Dog and cat animal product consumption is responsible for release of up to 64 ± 16 million tons CO2-equivalent methane and nitrous oxide, two powerful greenhouse gasses (GHGs). Americans are the largest pet owners in the world, but the tradition of pet ownership in the US has considerable costs. As pet ownership increases in some developing countries, especially China, and trends continue in pet food toward higher content and quality of meat, globally, pet ownership will compound the environmental impacts of human dietary choices. Reducing the rate of dog and cat ownership, perhaps in favor of other pets that offer similar health and emotional benefits would considerably reduce these impacts. Simultaneous industry-wide efforts to reduce overfeeding, reduce waste, and find alternative sources of protein will also reduce these impacts.

Development and application of a population physiologically based pharmacokinetic model for penicillin G in swine and cattle for food safety assessment

Penicillin G is a widely used antimicrobial in food-producing animals, and one of the most predominant drug residues in animal-derived food products. Due to reduced sensitivity of bacteria to penicillin, extralabel use of penicillin G is common, which may lead to violative residues in edible tissues and cause adverse reactions in consumers. This study aimed to develop a physiologically based pharmacokinetic (PBPK) model to predict drug residues in edible tissues and estimate extended withdrawal intervals for penicillin G in swine and cattle. A flow-limited PBPK model was developed with data from Food Animal Residue Avoidance Databank using Berkeley Madonna. The model predicted observed drug concentrations in edible tissues, including liver, muscle, and kidney for penicillin G both in swine and cattle well, including data not used in model calibration. For extralabel use (5× and 10× label dose) of penicillin G, Monte Carlo sampling technique was applied to predict times needed for tissue concentrations to fall below established tolerances for the 99th percentile of the population. This model provides a useful tool to predict tissue residues of penicillin G in swine and cattle to aid food safety assessment, and also provide a framework for extrapolation to other food animal species.

Efficacy of Humanized Mesenchymal Stem Cell Cultures for Bone Tissue Engineering: A Systematic Review with a Focus on Platelet Derivatives.

Fetal bovine serum (FBS) is the most commonly used supplement for ex vivo expansion of human mesenchymal stem cells (hMSCs) for bone tissue engineering applications. However, from a clinical standpoint, it is important to substitute animal-derived products according to current good manufacturing practice (cGMP) guidelines. Humanized alternatives to FBS include three categories of products: human serum (HS), human platelet derivatives (HPDs)-including platelet lysate (PL) or platelet releasate (PR), produced by freeze/thawing or chemical activation of platelet concentrates, respectively, and chemically defined media (serum-free) (CDM). In this systematic literature review, the in vitro and in vivo osteogenic potential of hMSCs expanded in humanized (HS-, HPD-, or CDM-supplemented) media versus hMSCs expanded in FBS-supplemented media, was compared. In addition, PL and PR were compared in terms of their growth factor (GF)/cytokine-content and cell-culture efficacy. When using either 10-20% autologous or pooled HS, 3-10% pooled HPDs or CDM supplemented with GFs, in comparison with 10-20% FBS, a majority of studies reported similar or superior in vitro proliferation and osteogenic differentiation, and in vivo bone formation in ectopic or orthotopic rodent models. Moreover, a trend for higher GF content was observed in PL versus PR, although evidence for cell culture efficacy is limited. In summary, humanized supplements seem at least equally effective as FBS for hMSC expansion and osteogenic differentiation. Although pooled HPDs appear to be the most favorable supplement for large-scale hMSC expansion, further efforts are needed to standardize the preparation and composition of these products in compliance with cGMP standards.

Detection of Mycobacteria by Culture and DNA-Based Methods in Animal-Derived Food Products Purchased at Spanish Supermarkets.

Mycobacteria include obligate and opportunistic pathogens that cause significant human and animal disease. The burden of tuberculosis has been largely reduced in developed territories but remains a huge problem worldwide. The significance of nontuberculous mycobacteria is growing considerably, especially in developed regions with higher life expectancy and more therapy-related immunosuppressed individuals. Due to their robustness mycobacteria can contaminate animal products by direct transmission from infected individuals or by environmental contamination during processing. The situation at market level is poorly known. Most studies analyzing commercially available foods are limited to a small or local scale and mainly focused on a particular mycobacterial species. There is a need to investigate if animal products that have passed the established controls to be for sale at main supermarkets could represent a route of contact with any mycobacteria. Thus, our goal was to study the prevalence of mycobacteria in these foods to assess if this could represent a source of human exposure. Five stores from the main supermarket chains in Spain were selected. 138 dairy and 119 meat products were purchased. All were processed using culture and multiplex real-time PCR methods. Additional molecular methods were used to specifically identify any positive result. Mycobacterium avium subsp. hominissuis (2), M. avium subsp. avium (1), and M. fortuitum (1) were isolated from powdered infant formula and ground beef, chicken sausage, and mortadella cold cut, respectively. Mycobacterial DNA (M. avium, M. tuberculosis complex and other nontuberculous mycobacteria) was detected in 15% of dairy products and 2% of meat products. These results show that the prevalence of viable mycobacteria in foods of animal origin obtained at the supermarket was not substantial although a considerable proportion of them contained mycobacterial DNA. Contact with mycobacteria through this route could be ensured over time. Further investigation is necessary to determine the real impact of foodborne mycobacterial exposure on human health and identify critical points in the food production system to enable setting up more stringent control measures.

Residual detection of buparvaquone, nystatin, and etomidate in animal-derived food products in a single chromatographic run using liquid chromatography-tandem mass spectrometry

A reliable and highly sensitive screening method based on liquid chromatography coupled withtriple-quadrupoleelectrospray tandemmass spectrometry (LC-MS/MS) analysis has been developed for the detection and quantification of three veterinary drugs, including buparvaquone, nystatin, and etomidate impurity B CRS. The tested drugs were extracted from samples of porcine muscle, pasteurized whole milk, and eggs using 10mM ammonium formate in acetonitrile followed by liquid-liquid purification with n-hexane. Chromatographic separation was achieved on a Phenomenex Luna C18 analytical column using 0.1% formic acid in ultrapure water (A) and acetonitrile (B) as mobile phases. All the matrix-matched calibration curves were linear (R2≥0.9756) over the concentration levels of the drugs tested. Recovery at two spiking levels (equivalent to the limit of quantification (LOQ)=5ng/g and 2×LOQ) ranged from 72.88% to 92.59% with intra- and inter-day precisions <17%, except for porcine muscle spiked with 5ng/g nystatin (RSD=25.15%). Samples collected from markets located in Seoul, Republic of Korea, tested negative for all the drugs analyzed. In summary, this method is suitable for screening and quantifying the selected drugs in a single chromatographic run and with high selectivity in animal-derived food products meant for human consumption.

Eco-friendly management of Sitophilus oryzae and Rhyzoper thadominica in stored wheat at Pantnagar, Uttarakhand

he laboratory experiments were conducted to study the efficacy of some indigenous products such as plant products (chopped neem leaves ajwain seed powder, garlic capsules, saw dust), animal derivedproducts ( cow dung cake powder, cow urine) and inert materials ( ash powder, sand, talcum powder, salt) @ 2g/100g of wheat grains against rice weevil, Sitophilus oryzae (Linn.) and lesser grain borer, Rhyzoper thadominica (Fab.) on stored wheat by undertaking various parameters viz., percent adult emergence, percent seed damage, per cent weight loss and per cent germination after six months of storage during April to October 2015. After 180 days of treatments, the minimum number of adults weevils of S. oryzae( 4.00, 6.00,7.67, 9.00, 9.67 and 10.33) were emerged in wheat grains treated with ash powder, sand, cow dung powder, talcum powder, ajwain seed powder and neem leaf powder, respectively with minimum grain damage (7.33 %, 8.67 %,11.67 % and 12.00 %) was observed on ash powder, sand, ajwain seed powder and talcum powder, respectively, whereas minimum weight loss (2.67 %, 3.67 %, 5.67 % and 6.67 %) was recorded on ash powder, sand, cow dung powder, talcum powder and ajwain seed powder, respectively. Similarly, the minimum number of adult beetles of R. dominica were emerged (2.00,2.33, 6.67,7.00 and 8.33) in ash power, sand, talcum powder, cow dung cake powder and ajwain seed powder, respectively with minimum grain damage (4.00 %,8.00 %. 8.67 % and 12.67 % and weight loss (1.33 %, 4.00 %, 7.33 % and 11.67 %) in sand, ash powder, talcum powder and cow dung cake treated wheat grains, respectively. It has been clearly observed that seed germination was not affected adversely in any treatment during six months of storage. The present study clearly revealed that these naturally occurring indigenous products could be used to manage the storage insect pests in wheat.

Occurrence, Toxicity, and Analysis of Major Mycotoxins in Food.

Mycotoxins are toxic secondary metabolites produced by certain filamentous fungi (molds). These low molecular weight compounds (usually less than 1000 Daltons) are naturally occurring and practically unavoidable. They can enter our food chain either directly from plant-based food components contaminated with mycotoxins or by indirect contamination from the growth of toxigenic fungi on food. Mycotoxins can accumulate in maturing corn, cereals, soybeans, sorghum, peanuts, and other food and feed crops in the field and in grain during transportation. Consumption of mycotoxin-contaminated food or feed can cause acute or chronic toxicity in human and animals. In addition to concerns over adverse effects from direct consumption of mycotoxin-contaminated foods and feeds, there is also public health concern over the potential ingestion of animal-derived food products, such as meat, milk, or eggs, containing residues or metabolites of mycotoxins. Members of three fungal genera, Aspergillus, Fusarium, and Penicillium, are the major mycotoxin producers. While over 300 mycotoxins have been identified, six (aflatoxins, trichothecenes, zearalenone, fumonisins, ochratoxins, and patulin) are regularly found in food, posing unpredictable and ongoing food safety problems worldwide. This review summarizes the toxicity of the six mycotoxins, foods commonly contaminated by one or more of them, and the current methods for detection and analysis of these mycotoxins.

Detection of ovine milk adulteration using taqman real-time pcr assay

Food safety, quality and composition have become the subjects of increasing public concern. To prevent fraud and enhance quality assurance, credible analysis of dairy products is crucial. Bovine milk is more widely available and cheaper than milk of sheep and goat. Bovine milk is also processed in large quantities to produce a range of dairy produce. DNA-based methods have proven to be more reliable, because of the stability of DNA under the conditions of high temperature, high pressure, and chemical treatment used during the processing of some food products. The commercial InnuDETECT cheese assay based on the principle TaqMan real-time PCR systems have been tested for the identification and quantification of bovine DNA in ovine milk samples. DNA was extracted using the InnuPREP DNA Mini Kit and quantified by the QuantiFluor dsDNA system. The assay showed good linearity, with correlation coefficient of R2 = 0.983 and efficiency of 86%. The internal control amplified fragment from different mammalian species (cow, sheep and goat), with similar CT values. Detection of bovine DNA in milk mixtures was achieved even in samples containing 0.5% of bovine milk. The InnuDETECT cheese assay has been successfully used to measure bovine DNA in ovine milk, and will prove useful for bovine species identification and quantitative authentication of animal-derived products.

A Review of Research-Grade Human Induced Pluripotent Stem Cells Qualification and Biobanking Processes.

Human induced pluripotent stem cell (hiPSC) biobanks are invaluable resources for basic and clinical research, since they provide a sustainable supply of accessible cell lines that meet high quality and safety standards. hiPSCs are particularly useful for understanding disease mechanisms, creating cell models for drug development, and generating novel clinical therapies. For clinical applications and drug discovery, it is fundamental that the acquired pluripotent cell lines never touch animal-derived products nor xenogeneic reagents (Good Manufacturing Practice-grade); whereas for research grade, it is sufficient to operate under Good Laboratory Practice conditions. However, regardless of the end use, it is important that every step in the whole process, starting from the original cells throughout expansion and manipulation, must be performed and recorded rigorously. Here, we describe our biobanking management system that is applied specifically to human pluripotent stem cells.

Mutilating Procedures, Management Practices, and Housing Conditions That May Affect the Welfare of Farm Animals: Implications for Welfare Research.

Simple summaryIntensive farming systems are confronted with a number of animal welfare issues such as injuries from horns in cattle and feather pecking in poultry. To solve these problems, mutilating procedures, such as dehorning in cattle and goats and beak trimming in laying hens, are applied routinely. These and other procedures such as early maternal separation, overcrowding, and barren housing conditions impair animal welfare. Scientific underpinning of the efficacy of these interventions and management practices is poor. We advocate that all stakeholders, in particular animal scientists and veterinarians, take the lead in evaluating common, putative mutilating and welfare reducing procedures and management practices to develop better, scientifically supported alternatives, focused on adaptation of the environment to the animals, to ensure uncompromised animal welfare.A number of mutilating procedures, such as dehorning in cattle and goats and beak trimming in laying hens, are common in farm animal husbandry systems in an attempt to prevent or solve problems, such as injuries from horns or feather pecking. These procedures and other practices, such as early maternal separation, overcrowding, and barren housing conditions, raise concerns about animal welfare. Efforts to ensure or improve animal welfare involve adapting the animal to its environment, i.e., by selective breeding (e.g., by selecting “robust” animals) adapting the environment to the animal (e.g., by developing social housing systems in which aggressive encounters are reduced to a minimum), or both. We propose adapting the environment to the animals by improving management practices and housing conditions, and by abandoning mutilating procedures. This approach requires the active involvement of all stakeholders: veterinarians and animal scientists, the industrial farming sector, the food processing and supply chain, and consumers of animal-derived products. Although scientific evidence about the welfare effects of current practices in farming such as mutilating procedures, management practices, and housing conditions is steadily growing, the gain in knowledge needs a boost through more scientific research. Considering the huge number of animals whose welfare is affected, all possible effort must be made to improve their welfare as quickly as possible in order to ban welfare-compromising procedures and practices as soon as possible.

A medieval physician: Amirdovlat Amasiatsi (1420-1495).

We aimed to acquaint the reader with a medieval physician, Amirdovlat Amasiatsi, who lived and practiced in the 15th century Anatolia. Amirdovlat wrote several books on medicine mainly focusing on phytotherapy and pharmacology using medicinal plants, animal-derived products and minerals. All his works were written in Middle Armenian, spoken Armenian language of the time. In his writings, Amirdovlat described unique recipes that represent a portrayal of medical knowledge and practice at the time in Anatolia where he lived and worked. He discussed the physical and therapeutic properties as well as geographic distributions of various plants and minerals, using different languages, mainly Turkish, Greek, Arabic, French and Persian. Amirdovlat's works not only enhanced our understanding of Armenian medical practices but also provided great deal of information on those of Ancient Greco-Roman as well as Islamic world, demonstrating close relationship of Armenian medicine with Greco-Roman and Islamic medicine. Amirdovlat accomplished this by amalgamating the past and contemporary practices of his time. In this regard, Amirdovlat's works, in particular "Useless for the Ignorant", are very unique playing a significant role in preserving traditions and heritage of different cultures.

Mycobacterium avium subsp. paratuberculosis and associated risk factors for inflammatory bowel disease in Iranian patients

BackgroundInflammatory bowel disease (IBD) is described as a relapsing condition with high morbidity and uncertain complex pathogenesis. The association of Mycobacterium avium ssp. paratuberculosis (MAP) with Crohn’s disease (CD) in human has been debated for decades, however there is no confirmed data to verify such relations in Iran. The aim of this study was to investigate risk factors and a possible role of MAP in Iranian patients with CD.MethodsAnti-MAP antibodies were detected in serum of IBD patients and subjects without IBD (nIBD) through ELISA; MAP DNA and viable MAP cells were identified in patients’ biopsies through nested PCR and direct culture methods, respectively. Principal component analysis (PCA) was used to investigate the risk factors in relation to IBD and MAP infection.ResultsPositivity for IS900 PCR was detected in 64% (n = 18) of CD, 33% (n = 10) of ulcerative colitis (UC) and 9.7% (n = 6) of nIBD samples. Live MAP cells were isolated from biopsies of 2 CD patients only. Among 28 patients with CD, 46% (n = 13) and 39% (n = 11) were positive for antibodies against MAP3865c133–141 and MAP3865c125–133 peptides, respectively, whereas much lower seroreactivity was detected in UC subjects accounting for 3% (n = 1) for MAP3865c133–141 and 16.7% (n = 5) for MAP3865c125–133. A high immune reactivity to MAP epitopes among CD patients was positively correlated with consumption of fast food meals and IBD familiarity. For both CD and UC, breastfeeding period and consumption of fruit/vegetables presented negative correlation with the presence of anti-MAP antibodies.ConclusionsThis study provided evidences that high prevalence of MAP DNA and anti-MAP antibodies in CD patients is significantly associated with the development of CD. Despite the role of several factors contributing to IBD, the presence of MAP DNA and anti-MAP antibodies in Iranian CD patients highlights a possible transmission of MAP from animal-derived products to humans.

Exploiting leech saliva to treat osteoarthritis: A provocative perspective

Plant and animal-derived products are crucial components in complementary and alternative medicine. Although modern medicine has provided numerous innovations and advancements, these often fail to reveal new and dependable, inexpensive treatments nor real cures that are relatively free of adverse side effects. We present evidence that hirudotherapy, which utilizes leeches, improves certain diseases, including osteoarthritis. Osteoarthritis, a disease in joints, could benefit from use of medicinal peptides found in leech saliva, components of its immune system.

Improvement ofDe NovoCholesterol Biosynthesis Efficiently Promotes the Production of Human Immunodeficiency Virus Type 1-Derived Lentiviral Vectors

The use of lentiviral vectors (LVs) for gene transfer in research, technological, or clinical applications requires the production of large amounts of vector. Mass production of clinical-grade LVs remains a challenge and limits certain perspectives for therapeutic use. Some improvements in LV production protocols have been possible by acting on multiple steps of the production process. The addition of animal-derived cholesterol to the culture medium of producer cells is known to increase the infectivity of LVs. To avoid the use of this animal-derived product in clinical settings, an alternative approach is to increase de novo the production of cholesterol by overexpressing a crucial cholesterogenic enzyme, namely, 3-hydroxy-3-methylglutaryl-coenzyme A reductase (HMGCR). This project evaluates the impact of such an approach on the production, infectivity, and stability of LVs. We demonstrated that the overexpression of human HMGCR isoform 1 (hHMGCR1) in LV producer cells efficiently increased de novo cholesterol biosynthesis and enhanced by 2- to 3-fold the physical and infectious titers of LVs. We also observed that LVs produced in hHMGCR1-overexpressing cells were comparable in stability to LVs produced under classical conditions and were capable of transducing human CD34+ hematopoietic stem/progenitor cells efficiently. Interestingly, we also showed that LV production in the absence of fetal calf serum (FCS) but under hHMGCR1-overexpressing conditions allowed a viral production yield comparable to that achieved under classical conditions in high FCS content, leading the way to the establishment of new LV production protocols on adherent cells without serum.