Tenogenic induction of equine mesenchymal stem cells by means of growth factors and low-level laser technology

Tendons regenerate poorly due to a dense extracellular matrix and low cellularity. Cellular therapies aim to improve tendon repair using mesenchymal stem cells and tenocytes; however, a current limitation is the low proliferative potential of tenocytes in cases of severe trauma. The purpose of this study was to develop a method useful in veterinary medicine to improve the differentiation of Peripheral Blood equine mesenchymal stem cells (PB-MSCs) into tenocytes. PB-MSCs were used to study the effects of the addition of some growth factors (GFs) as TGFβ3 (transforming growth factor), EGF2 (Epidermal growth factor), bFGF2 (Fibroblast growth factor) and IGF-1 (insulin-like growth factor) in presence or without Low Level Laser Technology (LLLT) on the mRNA expression levels of genes important in the tenogenic induction as Early Growth Response Protein-1 (EGR1), Tenascin (TNC) and Decorin (DCN). The singular addition of GFs did not show any influence on the mRNA expression of tenogenic genes whereas the specific combinations that arrested cell proliferation in favour of differentiation were the following: bFGF2 + TGFβ3 and bFGF2 + TGFβ3 + LLLT. Indeed, the supplement of bFGF2 and TGFβ3 significantly upregulated the expression of Early Growth Response Protein-1 and Decorin, while the use of LLLT induced a significant increase of Tenascin C levels. In conclusion, the present study might furnish significant suggestions for developing an efficient approach for tenocyte induction since the external administration of bFGF2 and TGFβ3, along with LLLT, influences the differentiation of PB-MSCs towards the tenogenic fate.     

Generation of Salmonella ghost cells expressing fimbrial antigens of enterotoxigenic Escherichia coli and evaluation of their antigenicity in a murine model

Salmonella Typhimurium ghost cells expressing K88ab, K88ac, K99, and FasA fimbriae of enterotoxigenic Escherichia coli (ETEC) in their envelopes were constructed. The genes encoding the fimbriae were individually cloned into an expression plasmid, pMMP81, carrying the asd gene, which was subsequently electroporated into the Δasd S. Typhimurium mutant. Plasmid pJHLP99, carrying the phiX174 lysis gene E, was also subsequently electroporated into the Salmonella mutant. The presence of the individual fimbriae on the ghost cells was examined by Western blot analysis. Forty BALB/c mice were equally divided into 2 groups of 20 mice each. Group A mice were intramuscularly vaccinated with a mixture of the 4 ghost cells expressing the individual fimbriae. The group B mice were inoculated with sterile phosphate-buffered saline as a control. The antigen-specific serum IgG concentrations were significantly higher in group A than in group B from week 2 until week 6 after inoculation. In addition, the antigen-specific IgA concentrations in fecal samples were significantly higher in group A than in group B at week 2 after inoculation. A large difference between the groups in the number of antigen-specific IgA-secreting cells in the small intestine was observed by immunohistochemical study. Also, the splenic lymphocyte proliferative responses were significantly greater in group A than in the control mice. These results suggest that vaccination with our Salmonella ghost cells can induce both humoral and cell-mediated immune responses and that the increased number of antigen-specific IgA-secreting cells in the small intestine may be correlated with the elevated fecal IgA immune response.     

Loser cows in Danish dairy herds: definition, prevalence and consequences

During the last few years, many Danish dairy farmers have expressed increasing concerns regarding a group of cows, which we have chosen to term ‘loser cows’. Until now, a loser cow has not been described scientifically. We defined a loser cow on the basis of a clinical examination of the cow. A total of 15,151 clinical examinations were made on 6,451 individual cows from 39 randomly selected, large Danish dairy herds with loose-housing systems using a clinical protocol. Scores for the clinical signs lameness, body condition, hock lesions, other cutaneous lesions, vaginal discharge, condition of hair coat and general condition were converted into a loser cow score. Cows with a loser cow score of 8 or more were classified as loser cows. The overall prevalence of loser cows was 2.15%, 4.50% and 2.98% during the first, second and third round of herd visits, respectively.

The associations between the loser cow state and milk production, mortality, morbidity, culling and workload for the farmer were evaluated using data from herd visits and from the Danish Cattle Database and a number of different statistical techniques. It was concluded that the loser cow state has significant negative consequences for both the farmer and the cow. On average, loser cows yielded 0.61 to 2.24 kg energy corrected milk less per day than non-loser cows depending on parity. Hazard ratio for death or euthanasia was 5.69 for loser cows compared to non-loser cows. Incidence rate ratio for disease treatments was 0.69 for non-loser cows compared to loser cows. Loser cows were often culled in an ‘unfavourable’ way and generally caused extra workload for the farmer.

A simplified version of the loser cow score was evaluated and is recommended for future research and use in practice.     

Tumour necrosis factor stimulated gene 6 intrinsically regulates PD-L1 expressions in breast cancer cells,leading to modulation of tumour microenvironment

Recent studies have shown that tumour cells express tumour necrosis factor-inducible gene 6 (TSG-6) and its protein, which is known to play a key role in regulating excessive immune responses and proliferation and growth of mesenchymal stem cells (MSCs). It has not been confirmed whether the inhibition of TSG-6 for tumour cells can suppress tumour cell growth and regulate the activation of immune cells in the tumour microenvironment (TME). TSG-6-specific small interfering RNA was transfected into canine and human breast cancer cells (CIPp, CIPm and BT-20). TSG-6-down-regulated (siTSG-6) cells showed decreased cell proliferation, migration, and invasion abilities. Decreased mRNA expressions of NF-κB, STAT3 and Sox2, confirming that TSG-6 is an upper factor governing tumour growth and metastasis. Notably, siTSG-6 cells showed significantly decreased expression levels of CD44 and PD-L1. Direct and indirect co-culture of canine peripheral blood mononuclear cells (cPBMCs) and the siTSG-6 cells showed significant activation in M1 type macrophages and cytotoxic T cells. They also showed a tendency to decrease in the expression of CTLA-4 and increase in the expression of PD-1. In conclusion, this study suggests that the down-regulation of TSG-6 in breast cancer cells could not only suppress tumour growth and metastasis, and but also regulate TME. Since modulation of immune checkpoint proteins occurs in both tumour cells and immune cells, inhibiting TSG-6 and its protein within the TME could be novel therapeutic target for anticancer treatment.     

Irrigated land retirement

Land retirement is ceasing irrigation withthe goal of reducing load, in general, ofdissolved constituents and, in particular,of trace elements, present in subsurfacedrainage generated from irrigated lands. Retirement is achieved through a process ofgoal setting, strategy development anddetermining effects, developing landselection criteria, implementation, andmonitoring. In this study, effects of landretirement are evaluated using hydrologic,soil and economic models as well as resultsfrom a field demonstration study. From themodeling and field monitoring, a process isdeveloped to meet the goals of a landretirement program in the San JoaquinValley of California.Potential negative effects listed for landretirement included loss of agriculturalproductivity, perhaps permanently, and lossof revenue to surrounding communities. Uncertainties included those associatedwith reuse of retired lands as wildlifehabitat, with retired-land maintenanceincluding dust control, with potentialpreservation of retired lands in reservefor future re-introduction to irrigated ordry-land agriculture, and withinstitutional changes concerning repaymentof federal and state water contracts. Benefits would accrue from economic returnto the landowner from the sale of property,the sale or lease of irrigation watersupply, the reduced cost of handlingdrainage, and allocation of freed-up waterto beneficial uses, and the reduced risk ofselenium exposure to fish and wildlife.A recommended sequential approach to selectand manage retired land is to identifyprimary objectives; formulate and implementarea-specific land retirement scenarios;measure biologic, hydrologic, soils andeconomic consequences in the short term andthe long term and manage and monitorretired lands based on dynamic biologic,hydrologic and soil conditions.     

Introduction of Irrigation Service Fee (ISF) in Indonesia: Institutional development in action for resources management

The inability to properly maintain irrigation systems over time forced the Government of Indonesia (GOI) to seek cost recovery from water users through introduction of an irrigation service fee. The plan is to introduce this fee in all of the technical irrigation systems of Indonesia, covering about 4 million hectares, over a 12 year period. Design and Introduction of this service fee in 4 pilot areas of the 4 major rice producing provinces (West, Central and East Java, and South Sulawesi) during 1989–1991 has shown that users are willing to pay. Results of first ISF collection from over 11,000 farmers in Central and East Java in pilot systems was an encouraging 95%. However that acceptance to pay by users depends on their structured and systematic involvement in defining systems needs. They need to understand a differentiation in payment if service levels are different. They appreciate use of collected funds in the system where collected. They understand the organization of water users associations but this requires their active role in fee determination and collection. They understand the establishment of federations of these associations. ISF requires also active involvement of Local Government officials as facilitators and intermediaries between service receivers (the water users) and service providers (personnel of the Public Works irrigation department) in the introduction period. This article describes the experience of the first 21 months of the ISF project, the concept developed, the principles used as basis for ISF, the introduction at the field level, issues related to acceptance by users and by the institutions involved, and the first results.Abbreviations/Acronyms DGWRD Directorate General of Water Resources Development (Ministry of Public Works) - EOM Efficient Operation and Maintenance - Gabungan group of WUAs - IPAIR Indonesian abbreviation for ISF - ISF Irrigation Service Fee - Juru gate keeper - PBB land tax on irrigated lands - PPL agricultural field extension agent - P3A Indonesian abbreviation for Water Users Associations - PU Indonesian abbreviation for Public Works - SM Special Maintenance - WUA Water Users Associations