Comparison of Cost and Potency of Human Mesenchymal Stromal Cell Conditioned Medium Derived from 2- and 3-Dimensional Cultures.

Mesenchymal stromal cell (MSC)-derived products, such as trophic factors (MTFs), have anti-inflammatory properties that make them attractive for cell-free treatment. Three-dimensional (3D) culture can enhance these properties, and large-scale expansion using a bioreactor can reduce manufacturing costs. Three lots of MTFs were obtained from umbilical cord MSCs produced by either monolayer culture (Monol MTF) or using a 3D microcarrier in a spinner flask dynamic system (Bioreactor MTF). The resulting MTFs were tested and compared using anti-inflammatory potency assays in two different systems: (1) a phytohemagglutinin-activated peripheral blood mononuclear cell (PBMNC) system and (2) a lipopolysaccharide (LPS)-activated macrophage system. Cytokine expression by macrophages was measured via RT-PCR. The production costs of hypothetical units of anti-inflammatory effects were calculated using the percentage of TNF-α inhibition by MTF exposure. Bioreactor MTFs had a higher inhibitory effect on TNF (p < 0.01) than monolayer MTFs (p < 0.05). The anti-inflammatory effect of Bioreactor MTFs on IL-1ß, TNF-α, IL-8, IL-6, and MIP-1 was significantly higher than that of monolayer MTFs. The production cost of 1% inhibition of TNF-α was 11-40% higher using monolayer culture compared to bioreactor-derived MTFs. A 3D dynamic culture was, therefore, able to produce high-quality MTFs, with robust anti-inflammatory properties, more efficiently than monolayer static systems.

Assessment of Novel Curcumin Derivatives as Potent Inhibitors of Inflammation and Amyloid-ß Aggregation in Alzheimer's Disease.

Alzheimer's disease (AD) is the most common neurodegenerative disorder affecting the elderly population worldwide. Brain inflammation plays a key role in the progression of AD. Deposition of senile plaques in the brain stimulates an inflammatory response with the overexpression of pro-inflammatory mediators, such as the neuroinflammatory cytokine. interleukin-6. Curcumin has been revealed to be a potential agent for treating AD following different neuroprotective mechanisms, such as inhibition of aggregation and decrease in brain inflammation. We synthesized new curcumin derivatives with the aim of providing good anti-aggregation capacity but also improved anti-inflammatory activity. Nine curcumin derivatives were synthesized by etherification and esterification of the aromatic region. From these derivatives, compound 8 exhibited an anti-inflammatory effect similar to curcumin, while compounds 3, 4, and 10 were more potent. Moreover, when the anti-aggregation activity is considered, compounds 3, 4, 5, 6, and 10 showed biological activity in vitro. Compound 4 exhibited a strong anti-aggregation effect higher than curcumin. Monofunctionalized curcumin derivatives showed better bioactivity than difunctionalized compounds. Moreover, the presence of bulky groups in the chemical structure of curcumin derivatives decreased bioactivity.

α-Synuclein Misfolding Versus Aggregation Relevance to Parkinson's Disease: Critical Assessment and Modeling.

α-Synuclein, an abundant and conserved presynaptic brain protein, is implicated as a critical factor in Parkinson's disease (PD). The aggregation of α-synuclein is believed to be a critical event in the disease process. α-Synuclein is characterized by a remarkable conformational plasticity, adopting different conformations depending on the environment. Therefore, it is classified as an "intrinsically disordered protein." Recently, a debate has challenged the view on the intrinsically disordered behavior of α-synuclein in the cell. It has been proposed that α-synuclein is a stable tetramer with a low propensity for aggregation; however, its destabilization leads to protein misfolding and its aggregation kinetics. In our critical analysis, we discussed about major issues: (i) why α-synuclein conformational behavior does not fit into the normal secondary structural characteristics of proteins, (ii) potential amino acids involved in the complexity of misfolding in α-synuclein that leads to aggregation, and (iii) the role of metals in misfolding and aggregation. To evaluate the above critical issues, we developed bioinformatics models related to secondary and tertiary conformations, Ramachandran plot, free energy change, intrinsic disordered prediction, solvent accessibility, and FoldIndex pattern. To the best of our knowledge, this is a novel critical assessment to understand the misfolding biology of synuclein and its relevance to Parkinson's disease.

Assessment of serum macro and trace element homeostasis and the complexity of inter-element relations in bipolar mood disorders.

BACKGROUND: Bipolar disorders are complex neuropsychiatric in nature and are clinically classified as Type I, Type II, and Type V. The etiological factors include environmental-genetic inter-relations. Trace metals play a significant role in neurological disorders. There is very limited information on the role of macro and trace elements in bipolar disorders. METHODS: Trace elements namely Na, K, S, Ca, Mg, P, Cu, Fe, Zn, Mn and Al were analyzed in serum samples of 3 bipolar types: bipolar I, bipolar II and bipolar V with a control group using inductively coupled plasma-atomic emission spectrometry (ICP-AES). The patients were assessed as per the standard diagnostic criteria and classified into the bipolar type I, II hypomanic, II depressives and V. RESULTS: In bipolar I (mania), Na, K, P, Cu, Al and Mn were increased significantly (p<0.001). In bipolar II hypomania, Na, S, Al and Mn were increased significantly (p<0.02), while in bipolar II depression, Na, K, Cu and Al were increased (p<0.001). In bipolar V, Na, Mg, P, Cu, and Al were increased significantly (p<0.002), though S (p<0.00001), Fe (p<0.002) and Zn (p<0.004) were decreased in all 3 bipolar groups. CONCLUSIONS: There is a disturbance in the charge distribution and element-element interdependency in bipolar serum when compared to controls. These results suggest that there is a definite imbalance in macro and trace element homeostasis as evidenced by element inter-relationships in serum samples of bipolar groups when compared to controls.

Rheumatic fever and rheumatic heart disease: primary prevention is the cost effective option.

OBJECTIVE: To measure the economic output/input ratios for the various options of prevention of rheumatic fever/rheumatic heart disease (RF/RHD) and check the viability of primary prevention vis-à-vis secondary and tertiary preventions. METHODS: Cost accounting of the various prevention options was calculated for each variable as available in literature. Actual data as obtainable for the financial year ending March 2006 were computed for the Pondicherry population. Both direct and indirect costs (including community/social costs) were worked out using mostly primary data and wherever necessary, secondary data. Certain scientific assumptions were used where exact data was not available. RESULTS: Primary prevention is the definite viable economic option (1:1.56) compared to secondary (1: 1.07) and tertiary (1: 0.12) preventions. In fact, the current stress on only secondary and tertiary preventions is found to be economically unviable. CONCLUSION: It is postulated that primary prevention as a practical policy in tackling RF and RHD can be recommended.

Crop damage and livestock depredation by wildlife: a case study from Nanda Devi Biosphere Reserve, India.

The success of conserving biological resources in any Biosphere Reserve or protected area depends on the extent of support and positive attitudes and perceptions of local people have towards such establishments. Ignoring the dependence of the local people for their subsistence needs on resources of such areas leads to conflicts between protected area managers and the local inhabitants. Crop yield losses and livestock depredation were serious problems observed in most buffer zone villages of Nanda Devi Biosphere Reserve. In the present study 10 villages situated in the buffer zone of Nanada Devi Biosphere Reserve (1612 km2 area) in Chamoli district of Uttaranchal, India were studied during 1996-97 using a questionnaire survey of each household (419 = households; 2253 = total population in 1991; 273 ha = cultivated area). Estimates of crop yield losses were made using paired plots technique in four representative villages for each crop species. The magnitude of crop yield losses varied significantly with the distance of agricultural field from forest boundary. The total crop yield losses were high for wheat and potato in all the villages. The spatial distribution of total crop yield losses in any village indicated that they were highest in the area near to forest and least in the area near to village for all crops. Losses from areas near to forest contributed to more than 50% of total losses for each crop in all villages. However, in Lata, Peng and Tolma villages, the losses are high for kidney bean and chemmi (local variety of kidney bean) which varied between 18.5% to 30% of total losses in those villages. Potato alone represents 43.6% of total crop yield loss due to wildlife in Dronagiri village in monetary terms. Among the crops, the monetary value of yield losses are least for amaranth and highest for kidney bean. The projected total value of crop yield losses due to wildlife damage for buffer zone villages located in Garhwal Himalaya is about Rs. 538,620 (US$ 15,389). Besides food grains, horticultural crops i.e. apple, also suffered maximum damage. Major wildlife agents responsible for crop damage were wild boar, bear, porcupine, monkey, musk deer and partridge (chokor). Monkey and wild boar alone accounted for about 50% to 60% of total crop damage in the study villages. Goat and sheep are the major livestock killed by leopard. The total value of livestock losses at prevailing market rates is about Rs. 1,024,520 (US$ 29,272) in the study villages. Due to existing conservation policies and laxity in implementation of preventive measures, the problems for local inhabitants are increasing. Potential solutions discussed emphasize the need to undertake suitable and appropriate protective measures to minimize the crop losses. Change in cropping and crop composition, particularly cultivation of medicinal plants (high value low volume crops), were also suggested. Besides, fair and quick disbursement of compensation for crop loss and livestock killing need to be adopted. Local people of the buffer zone area already have a negative attitude towards park/reserve establishment due to socio-political changes inducing major economic losses and this attitude may lead to clashes and confrontations if proper ameliorative measures are not taken immediately.

Urinary hydatid antigen detection by coagglutination, a cost-effective and rapid test for diagnosis of cystic echinococcosis in a rural or field setting.

We describe here coagglutination (Co-A), a rapid slide agglutination test for the detection of hydatid antigen in the urine for the diagnosis of cystic echinococcosis (CE). Paired urine and serum samples were collected from 16 patients with surgically confirmed CE, 10 patients with ultrasound-proven CE, 14 patients with clinically diagnosed CE, 24 patients with various parasitic diseases other than CE, and 25 healthy control subjects. Co-A detected excreted hydatid antigen in the concentrated urine of 7 of 16 (43.75%) surgically confirmed cases, 6 of 10 (60%) ultrasound-proven cases, and 8 of 14 (57.14%) clinically diagnosed cases of CE. A false-positive reaction was observed with 12.50% of control urine specimens from patients with parasitic diseases other than CE and 12% of urine samples from healthy controls. The circulating antigen was detected in the serum in 13 of 16 (81.25%) surgically confirmed cases, 6 of 10 (60%) ultrasound-proven cases, and 13 of 14 (92.86%) clinically diagnosed cases of CE. False-positive reactions were observed with three sera (12.5%) from controls with other parasitic diseases. The low sensitivity of Co-A for detection of antigen in the urine of a patient whose serum was positive for the antigen is possibly due to low levels of antigen in the urine. Unlike the collection of blood for serum, which is an invasive procedure and also requires technical expertise and disposable syringes, urine can be collected easily and frequently without causing any inconvenience to the patient. Urine as a clinical specimen alternative to serum would be immensely useful in the diagnosis of CE, particularly in a rural or field setting. In such situations as well as in poorly equipped laboratories, Co-A has the potential to be used as a simple, rapid, and economical slide agglutination test for detection of urinary hydatid antigen in the diagnosis of CE.

A field test for the assessment of faecal contamination of potable water.

A simple, sensitive, rapid, inexpensive paper strip impregnated with Salmonellal E. coli medium (SEM) was formulated, and placed in a test tube. When potable water of 10 ml was added to the test tube it detected the faecal contamination of water samples within 16-48 h when incubated at room temperature from 20 to 35 degrees C. The positive results were indicated when the medium turned black (hydrogen sulfide production) for the presence of Salmonella sp. and/or the formation of a red ring (free indole from tryptophan) when a few drops of Kovac's reagent was added for the presence of coliform bacteria (E. coli). More than 600 water samples were tested with the new test (SEM) and results showed 99% agreement with that of the standard most probable number (MPN) coliform test and also proved highly successful in the field when it was employed to detect both Salmonella and E. coli. Different water testing laboratories involved in a water quality monitoring programme and governmental agencies evaluated the test media and reported that the test was user friendly, reliable and simple to perform in the field and will be especially useful for screening of both urban and rural water supplies for routine monitoring of bacteriological contamination.

Nutrition: an essential element of primary health care.

PIP: In India, major social discrepancies linger despite major advances since 1947 with the result of overeating by the rich and undernutrition of the poor. Anemia affected 65% of pregnant women in Hyderabad hospitals, and in Calcutta it hovered around 88.5-90/1% for males and 96.5-96.7% for females aged 15-44. Malnutrition accounted for 15% of hospitalizations often because of infant diarrhea. Parasitic infestations leading to anemia and subsequent malnutrition also resulted in premature births and low birth weight. Weaning practices including its late initiation and prolongation of breast feeding for up to 12 months (or 18 months in rural areas) led to growth retardation. About 25,000 children become blind annually because of vitamin A deficiency, and 55% of women also suffer from it. Almost 120-170 million people suffer from endemic goiter. Socio economic and socio cultural factors are also associated with nutritional problems. Some Brahmin communities in South India avoid garlic and onions because they are considered taboo foods. The Tridosha Theory holds the there are heat-producing foods, cooling foods, and gas-producing foods that have to be avoided. Some of these are brown sugar, eggs, tomatoes, oranges, cereals, and vegetables. In West Bengal they believe that milk and fish can cause leprosy. The population nearing 800 million is another factor, as smaller families have better nutrition. Education by nurses should include the use of exhibits, songs, and plays to teach women proper nutrition. Nurses also have to change their attitudes to avoid commercial foods, identify harmful practices, and promote nutritional education. The project of the National Institute of Nutrition involving 23 villages combated vitamin A deficiency successfully by laying stress on available vegetables and fruits. If nurses participate if such primary health care programs the objectives of Health for All by 2000 will be achieved.^ieng