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The influence of Modal–cotton (MC) fibre blend ratio and ring frame machine parameters such as front top roller loading and break draft on the blended yarn properties has been studied. Compact MC blended yarn samples of 14.75 tex with three different MC fibre blend ratio has been produced in a LR 6 ring spinning frame fitted with Suessen Compact drafting system. A robust design optimisation to minimise the variations of the output yarn properties such as blended yarn tenacity, yarn unevenness and hairiness caused because of the variations in the material as well as machine setting parameters is achieved through the Taguchi parametric design approach. It is found that the maximum compact MC blended yarn tenacity is 23.76 g/tex, which is influenced very much by MC fibre blend ratio but meagrely by top roller loading and break draft. Similarly, the minimum 9.54 U% and 3.59 hairiness index are achieved with 100:0 and 70:30 MC fibre blend ratio, respectively, at 23-kg top roller loading. Statistical ANOVA analysis is performed on the results and optimum values are obtained within the 95% confidential level through confirmation experiments.

No abstract available. Article truncated after 150 words. A 70-year-old woman with a history of hypertension presented with left-sided weakness, headache, nausea, and vomiting. She denied loss of consciousness or seizure activity. On examination, she had receptive aphasia. Pupils were equal, round and reactive. She had neck pain on flexion. Her left upper extremity was plegic. Computed tomography of the brain showed acute hemorrhage involving the right thalamus, extending into the ventricular system, and a midline mass. She underwent a computed tomography angiogram, which showed a large, complex arteriovenous malformation (AVM) with a dilated branch of the right suprasellar internal carotid artery feeding the AVM, which then drained into the vein of Galen and straight sinus (Figure 1). She was monitored in the intensive care unit without worsening neurological deficit. She was discharged to a rehabilitation facility, having had no intravascular or surgical intervention. AVMs are intracranial vascular anomalies which occur in 0.1% of the population (1). Clinical …

Proton pump inhibitors (PPIs) have been associated with adverse clinical outcomes amongst clopidogrel users after an acute coronary syndrome. Recent pre-clinical results suggest that this risk might extend to subjects without any prior history of cardiovascular disease. We explore this potential risk in the general population via data-mining approaches.Using a novel approach for mining clinical data for pharmacovigilance, we queried over 16 million clinical documents on 2.9 million individuals to examine whether PPI usage was associated with cardiovascular risk in the general population.In multiple data sources, we found gastroesophageal reflux disease (GERD) patients exposed to PPIs to have a 1.16 fold increased association (95% CI 1.09-1.24) with myocardial infarction (MI). Survival analysis in a prospective cohort found a two-fold (HR = 2.00; 95% CI 1.07-3.78; P = 0.031) increase in association with cardiovascular mortality. We found that this association exists regardless of clopidogrel use. We also found that H2 blockers, an alternate treatment for GERD, were not associated with increased cardiovascular risk; had they been in place, such pharmacovigilance algorithms could have flagged this risk as early as the year 2000.Consistent with our pre-clinical findings that PPIs may adversely impact vascular function, our data-mining study supports the association of PPI exposure with risk for MI in the general population. These data provide an example of how a combination of experimental studies and data-mining approaches can be applied to prioritize drug safety signals for further investigation.

Off-label drug use, defined as use of a drug in a manner that deviates from its approved use defined by the drug's FDA label, is problematic because such uses have not been evaluated for safety and efficacy. Studies estimate that 21% of prescriptions are off-label, and only 27% of those have evidence of safety and efficacy. We describe a data-mining approach for systematically identifying off-label usages using features derived from free text clinical notes and features extracted from two databases on known usage (Medi-Span and DrugBank). We trained a highly accurate predictive model that detects novel off-label uses among 1,602 unique drugs and 1,472 unique indications. We validated 403 predicted uses across independent data sources. Finally, we prioritize well-supported novel usages for further investigation on the basis of drug safety and cost.

BACKGROUND: Following traumatic vertebral injuries and resultant spinal cord injury, most patients are doomed to a life either of quadriplegia or paraplegia. Current treatment option is limited to the stabilization of the vertebral fracture along with medications to prevent secondary damage leading to further deterioration and wishful waiting for recovery. In most instances recovery is insignificant. Safety of intrathecal injection of autologous bone marrow stem cells is proven but its efficacy varies between patients (1). Intralesional application has been reported to be more efficacious than intrathecal application (2, 3, 4). We have analyzed our experience in human patients followed up for 3 year period and have found several grey areas in spinal cord injury(5) one of them is to explore the differences between Intrathecal and intralesional application of stem cells with and without scaffolds in the latter technique. Towards achieving this goal we started a pilot study in animals where instead of post-vertebral fixation intrathecal injection, we have performed intralesional application of autologous BMSC along with scaffolds (6). These scaffolds not only help retain the transplanted cells at the site of injury but also allow more neural precursors to grow compared to application without scaffolds (7). This study analyses the data retrospectively to plan further prospective studies with a view to improvise the results. MATERIALS AND METHODS: Study 1 : 100 to 120 ml of Bone marrow was tapped from the right posterior iliac crest under local anesthesia from human spinal injury victims (n=108; 76 males, 32 females) about 3 weeks to 18 months after surgical fixation of the vertebrae. The Level of injury was varied- Cervical (13 patients.) Upper Thorax- T1-T7 (35 patients) Lower thorax T8-T12 (46 patients) Lumbar (2 patients.) Age Group Range: 8 yrs to 55 yrs. The bone marrow mononuclear cells were processed under cGMP SOP’s Class 10000 clean room and class 100 Biosafety hood as reported earlier (1) and were injected intrathecally into the subarachnoid space of the patients below L2 level after endotoxin tests and confirming CD34 status using flow cytometry. Study 2: 20-30ml of Bone marrow was tapped from the right posterior iliac crest under local anesthesia of canine spinal injury victims immediately after the injury and the bone marrow processed as reported earlier (6) were injected intralesionally embedded in thermoreversible hydrogel scaffolds at the site of the injury after endotoxin tests and CD34 analysis using flow cytometry. Both the animals had an Olby score of 1 with no CP reflex, Patellar reflex and deep pain reflex. RESULTS:Study-1Parameters 4-6 months follow-up(1) 12-36 months follow-up(8) Follow up attrition rate: 14.02% 69% Motor power improvement (atleast 2 grade of power post injection) 14.11% 7.69% Motor power improvement resulting in functional recovery 4.70% 3 walk with support, 1 walking without support 5.13% Subjective sensory improvement 16.97% 25.64% Abnormal sensation 0% 0% Autonomic improvement by Urodynamic studies 9.41% 10.26% Study-2: The first canine died due to cystitis on the 7th day after the procedure and the cause of death was confirmed to be unrelated to the procedure by autopsy that revealed co-morbid conditions like cystitis, nephritis and transmissible venereal tumor. Histopathology of the engrafted area revealed sustainability of aggregated stem cells that were transplanted revealing an ideal biocompatibility of the construct prepared with bone marrow mononuclear cells and polymer hydrogel for spinal cord regeneration in dogs. The second canine had complete recovery of hind limb function and coordination after 6 months, Olby score has improved to 14 (normal 15) with normal bowel and bladder control.DISCUSSION: It was hypothesized that intralesional application at time of surgery may be superior to Intrathecal application of bone marrow stem cells simply because the former technique ensured the delivery of a higher proportion of cells in the damaged area. It is possible that the cells injected intrathecally are carried along with the CSF to parts other than that damaged as well. This has been supported by study 2 when it was applied with scaffold. The injury model used in study 2 is a natural traumatic model and is more akin to real life than any other controlled spinal cord model that one could create in the lab. This study 2 which is still in process allows for the animal to live to its entire course enabling us to follow up the neurological recovery of the patient and on its death perform an autopsy to not only determine the cause of death but to also examine the fate of stem cells injected intralesionally. We hope to determine the percentage of stem cells remaining as stem cells and to determine the nature and magnitude of histopathological changes that might have taken place which facilitated /non facilitated the recovery in the study animalsCONCLUSION:It is understood from the study 1, that the factors determining outcome are multiple and includes the age of patients, level of injury, time interval between injury and ABMMC injection, dosage of stem cells injected and all these need to be evaluated in future studies. More studies are necessary to ascertain the efficacy. Safety of both intrathecal and intralesional injection with scaffold have been proven in this studies. Inclusion of larger number of cases with a long term follow up is necessary to know the efficacy of intralesional therapy with scaffolds References1.Abraham S, Manjunath S, Baskar S, Senthil Kumar R, Dedeepiya V, Terunuma H, Ravikumar R, Narayanan R, Selvan R ,Sankaranarayanan S, Jasper J, Parthiban JKBC, Arjundas D and Subramanniyan SR. Autologous Stem Cell Injections for Spinal Cord Injury - A multicentric Study with 6 month follow up of 108 patients 7th Annual Meeting of Japanese Society of Regenerative Medicine, Nagoya, Japan, 13 - 14 March 2008 2.Vaquero J, Zurita M, Oya S, Santos M. Cell therapy using bone marrow stromal cells in chronic paraplegic rats: systemic or local administration? Neurosci Lett. 2006 May.3.de Haro J, Zurita M, Ayllón L, Vaquero J. Detection of 111.In-oxine-labeled bone marrow stromal cells after intravenous or intralesional administration inchronic paraplegic rats. Neurosci Lett. 2005 Mar4.Yano S, Kuroda S, Lee JB, Shichinohe H, Seki T, Ikeda J, Nishimura G, Hida K, Tamura M, Iwasaki Y. In vivo fluorescence tracking of bone marrow stromal cells transplanted into pneumatic injury model of rat spinal cord. J Neurotrauma.2005 Aug5.Ravikumar R, Narayanan S, Baskar S, SenthilNagarajan R, Abraham S. Autologous Stem Cell Injection for Spinal Cord Injury- A Clinical Study from India. PASRM 2007-0046.Justin William B, Ayyappan S, DhanaJayaRao G, Pushkinraj H, Kannan TA , Sureshkumar R, Manjunath S, Murugan P, Srinivasan V, Abraham S. Intralesional Application of Autologous Bone Marrow Stem Cells with Scaffold in Canine for Spinal Cord Injury. PASRM, IV annual symposium , 2009.7.Yasuda H, Kuroda S, Shichinohe H, Kamei S, Kawamura R, Iwasaki Y. Effect of biodegradable fibrin scaffold on survival, migration, and differentiation of transplanted bone marrow stromal cells after cortical injury in rats. J Neurosurg. 2010 ;112(2):336-44. 8.Abraham et al. Stem cell Clinical trials Session. Stem cell Global blue Print Conference (21-22 May2010) Toronto, Canada

BackgroundPeripheral arterial disease (PAD) is a growing problem with few available therapies. Cilostazol is the only FDA-approved medication with a class I indication for intermittent claudication, but carries a black box warning due to concerns for increased cardiovascular mortality. To assess the validity of this black box warning, we employed a novel text-analytics pipeline to quantify the adverse events associated with Cilostazol use in a clinical setting, including patients with congestive heart failure (CHF).Methods and resultsWe analyzed the electronic medical records of 1.8 million subjects from the Stanford clinical data warehouse spanning 18 years using a novel text-mining/statistical analytics pipeline. We identified 232 PAD patients taking Cilostazol and created a control group of 1,160 PAD patients not taking this drug using 1:5 propensity-score matching. Over a mean follow up of 4.2 years, we observed no association between Cilostazol use and any major adverse cardiovascular event including stroke (OR = 1.13, CI [0.82, 1.55]), myocardial infarction (OR = 1.00, CI [0.71, 1.39]), or death (OR = 0.86, CI [0.63, 1.18]). Cilostazol was not associated with an increase in any arrhythmic complication. We also identified a subset of CHF patients who were prescribed Cilostazol despite its black box warning, and found that it did not increase mortality in this high-risk group of patients.ConclusionsThis proof of principle study shows the potential of text-analytics to mine clinical data warehouses to uncover 'natural experiments' such as the use of Cilostazol in CHF patients. We envision this method will have broad applications for examining difficult to test clinical hypotheses and to aid in post-marketing drug safety surveillance. Moreover, our observations argue for a prospective study to examine the validity of a drug safety warning that may be unnecessarily limiting the use of an efficacious therapy.

Ischemic preconditioning is a neuroprotective mechanism whereby a sublethal ischemic exposure is protective against a subsequent lethal ischemic attack. We previously demonstrated that SIRT1, a nuclear localized stress-activated deacetylase, is vital for ischemic preconditioning neuroprotection. However, a recent study demonstrated that SIRT1 can also localize to the mitochondria. Mitochondrial localized SIRT1 may allow for a direct protection of mitochondria following ischemic preconditioning. The objective of this study was to determine whether ischemic preconditioning increases brain mitochondrial SIRT1 protein levels and to determine the role of PKCɛ and HSP90 in targeting SIRT1 to the mitochondria. Here we report that preconditioning rats, with 2 min of global cerebral ischemia, induces a delayed increase in non-synaptic mitochondrial SIRT1 protein levels which was not observed in synaptic mitochondria. This increase in mitochondrial SIRT1 protein was found to occur only in neuronal cells and was mediated by PKCε activation. Inhibition of HSP90, a protein chaperone involved in mitochondrial protein import, prevented preconditioning induced increases in mitochondrial SIRT1 and PKCε protein. Our work provides new insights into a possible direct role of SIRT1 in modulating mitochondrial function under both normal and stress conditions, and to a possible role of mitochondrial SIRT1 in activating preconditioning induced ischemic tolerance.

In order to ensure the that equipment functions well, both equipment users and the equipment maintenance and repair team must be trained. Users must be trained in basic care and maintenance of equipment, and the equipment team must be trained to undertake repairs and more complex maintenance tasks.

BACKGROUND: Vascular endothelial cells play a major role in wound healing and also in growth of the tumors. Angiogenesis can be a target for treating diseases that are due to either poor vascularisation or decreased blood supply as in stroke, ulcers, heart disease, etc or abnormal and increased vasculature like in tumours. Application of specific compounds that may inhibit or induce the creation of new blood vessels in the body may help in the treatment of such diseases (1). Ex vivo generation of blood vessels may offer an excellent alternative to the synthetic valves that are being currently used in cardiology. Micro gravity also referred to, as weightlessness is not essentially zero gravity but rather minimal gravity. According to cell type, micro gravity causes variety of changes in proliferation and differentiation of cells while also affecting the migration of cells and cellular functions (2, 3). Siamwala et al from AUKBC have already studied the effects of microgravity on the microvascular endothelial cells from bovine lung and macrovascular endothelial cells from the bovine pulmonary artery. It was observed that the proliferation and migration of macrovascular endothelial cells were increased in microgravity (4, 5). Nitric oxide production was also studied and observed that microgravity treatment did not change nitric oxide production by microvascular endothelial cells (4)OBJECTIVE: Isolation and Comparison of culture characteristics of Human microvascular endothelium cultured conventionally and in novel nanomaterial scaffold and further study the morphological and molecular characteristics of microvascular endothelial cells under normal gravity against simulated micro gravityMATERIALS AND METHODS: The human Omentum samples were obtained using surgical procedures after informed consent. The microvascular endothelial cells were isolated following the protocol described by Scott et al (6).The isolated cells were seeded in two groups; Group I - Cells in Nano Polymer, Overlaid M199 medium +20%FBS and, Group II - Cells in Conventional M199 medium+20%FBS. The cultures were maintained at 37C with 5%CO2. The medium was changed every 3 days thereafter. The cells were observed regularly for their morphological characteristics and images were documented. At day 7 and day 13 the cells were harvested and the cells were subjected to two assays to confirm the presence of nitric oxide (NO) in samples, which will in turn confirm the presence of endothelial cells. In the first assay the isolated cell suspension was probed with diaminorhodamine-4M (DAR-4FM). In the second assay, the total nitrite level in the isolated cells suspended media was measured using Griess Assay protocol. RESULTS: A 21% increase in number of NO positive cells was observed in the cells cultured on the novel nanopolymer, while a 1.7 fold increase in nitrite production was detected in Group I in comparison to that of Group II.CONCLUSION: Our data suggests the presence of more NO driven nitrite in the Microvascular endothelial cells cultured in the nanopolymer. In the next phase of the study, the cells thus isolated from both the groups will be subjected to simulated microgravity of 10-3 g in a clinostat at the AUKBC Vascular Biology Lab and the morphological and molecular characteristics of Microvascular endothelial cells will be studied to identify whether the effects of microgravity on Microvascular endothelial cells is organ specific. References:1.Jain RK. Antiangiogenic therapy for cancer: current and emerging concepts. Oncology (Williston Park). 20052.Freed LE, Pellis N, Searby N, de Luis J, Preda C, Bordonaro J, Vunjak-Novakovic G. Microgravity cultivation of cells and tissues. Gravit Space Biol Bull. 19993.Dedolph RR, Dipert MH. The physical basis of gravity stimulus nullification by clinostat rotation. Plant Physiol. 19714.Siamwala JH, Majumder S, Tamilarasan KP, Muley A, Reddy SH, Kolluru GK, Sinha S, Chatterjee S. Simulated microgravity promotes nitric oxide-supported angiogenesis via the iNOS-cGMP-PKG pathway in macrovascular endothelial cells.FEBS Lett. 20105.Siamwala JH, Reddy SH, Majumder S, Kolluru GK, Muley A, Sinha S, Chatterjee S. Simulated microgravity perturbs actin polymerization to promote nitric oxide-associated migration in human immortalized Eahy926 cells. Protoplasma. 20106.Scott PA, Bicknell R. The isolation and culture of Microvascular endothelium. J Cell Sci. 1993

BACKGROUND: Recent studies have shown that mesenchymal stem cells isolated from post natal human dental pulp, (Dental pulp stem cells-DPSCs) which is from permanent teeth and SHED (stem cells from human exfoliated deciduous teeth),the Periodontal ligament stem cells (PDLSC) and Stem cells from root Apical papilla(SCAP)have the potential to differentiate into cells of a variety of tissues including heart, muscle, cartilage, bone, nerve, salivary glands, teeth etc(1,2,3,4).This multipotential ability of DPSCs is being researched for clinical application for treating a variety of diseases like myocardial infarction, muscular dystrophy, neuro-degenerative disorders, cartilage replacement, tooth regeneration and for repair of bone defects to mention a few. Moreover, the isolation of stem cells from teeth is minimally invasive, readily accessible and the non immunogenic characteristic of dental stem cells has paved the way for efforts to store the exfoliated deciduous teeth or milk teeth which is usually discarded, for use in the future. In this study we have isolated and expanded in vitro, the cells obtained from human dental pulp. MATERIALS AND METHODS: After obtaining written informed consent, 24 teeth that were extracted for therapeutic or cosmetic reasons from 16 patients were used in this study. The specimens were transported from the clinic to NCRM lab taking 6 to 48 Hrs. For removal of the pulp tissue, the teeth were split obliquely at the Cementoenamel junction and the pulp tissue was isolated using brooches. The extracted pulp tissues were subjected to digestion using Collagenase type-I and type II at 37˚C for 15- 30 minutes. The digested cells were filtered with 70µm filter and centrifuged at 1800 rpm for 10 minutes. The pellet was then suspended in Dulbecco’s modified Eagle’s medium (DMEM)/Ham’s F12 supplemented with 15% fetal bovine serum , 100 U/ml penicillin, 100 µg/ml streptomycin,2 m M L -glutamine, and 2 m M nonessential amino acids (5) .Cell counting was done by Trypan Blue dye exclusion method and the cells were seeded in 6 well culture plates. The plates with cells were incubated at 37˚C with 5% CO2 for varying periods from 14 days-28 days. The cells were observed daily and media change was done every three days. RESULTS: Viable Dental Pulp tissue-cells were obtained after transportation of up to 48 hrs and the in vitro growth of cells was initially slow but colonies were identified from the 10th day onwards. The cells were harvested at different intervals of 14-28 days for each sample based on their growth and subjected to H & E staining .The H & E staining of the cultured cells of all the samples showed positive resultsCONCLUSION: We are able to transport extracted teeth and derive viable dental pulp tissue cells after enzymatic digestion and multiply them in culture after a maximum of 48 hrs after transportation. The cells could be grown in culture with a morphology resembling dental pulp stem cells while in culture expansion and in H&E studies. Further characterization of the cells is necessary to confirm their Stemness. References1.Gronthos S, Mankani M, Brahim J, Robey PG, Shi S. Postnatal human dental pulp stem cells (DPSCs) in vitro and in vivo. Proc Natl Acad Sci U S A. 20002.Nosrat IV, Widenfalk J, Olson L, Nosrat CA. Dental pulp cells produce neurotrophic factors, interact with trigeminal neurons in vitro, and rescue motoneurons after spinal cord injury. Dev Biol. 2001 Oct 3.Iohara K, Zheng L, Ito M, Tomokiyo A, Matsushita K, Nakashima M. Side population cells isolated from porcine dental pulp tissue with self-renewal and multipotency for dentinogenesis, chondrogenesis, adipogenesis, and neurogenesis. Stem Cells. 2006 Nov4.Gandia C, Armiñan A, García-Verdugo JM, Lledó E, Ruiz A, Miñana MD, Sanchez-Torrijos J, Payá R, Mirabet V, Carbonell-Uberos F, Llop M, Montero JA, Sepúlveda P. Human dental pulp stem cells improve left ventricular function, induce angiogenesis, and reduce infarct size in rats with acute myocardial infarction. Stem Cells. 2008 Mar5.Kerkis I, Kerkis A, Dozortsev D, Stukart-Parsons GC, Gomes Massironi SM, Pereira LV, Caplan AI, Cerruti HF. Isolation and characterization of a population of immature dental pulp stem cells expressing OCT-4 and other embryonic stem cell markers. Cells Tissues Organs. 2006

BACKGROUND: The Enteric Nervous system (ENS) is a part of the Peripheral nervous system (PNS) that controls the peristaltic activity of the gut wall which is essential for propulsion of food in the digestive tract. It is composed of a large number of neurons and glial cells, distributed throughout the length of the gut. These ganglion cells develop from the neural crest in the embryo. Failure of complete colonization of the gut by these enteric neural crest cells during early development of life results in absence of ganglia or neurons in a portion of the gut, usually the colon which leads to aperistaltis and severe intestinal obstruction. This is known as Hirschsprung’s disease (HSCR) also known as congenital megacolon. HSCR affects 1 in 4500 newborns (1, 2). It appears either sporadically or has a familial basis and is often associated with other developmental defects. The main forms of treatment of HSCR are surgical resection of the aganglionic segment and pull through of the normal bowel. At present research is aimed at developing Cell based therapies for replacement of ganglion cells or enteric neuronal cells in the aganglionic portion of the gut thus aiming at restoring the function of the gut (1, 3, 5). In this study we have isolated, in vitro expanded and characterized the Enteric Neuronal cells derived from human gut full thickness biopsy samplesMATERIALS AND METHODS: The postnatal gut full thickness biopsy samples of size 2-4 mm were obtained using from 13 patients undergoing gut resection surgery after informed consent. The samples were washed in Phosphate Buffer saline and using forceps, the outer smooth muscle layers along with the myenteric plexus were peeled off from the underlying tissue as strips. The strips were washed in Phosphate Buffer saline (PBS) and treated with 1mg/ml Collagenase/Dispase mixture in PBS for 30-45 min at 37°C. The digested cells were filtered with 70µm filter and the cell suspensions were centrifuged at 1800rpm for 10 mins. The pellet obtained was suspended in DMEM/F12 medium supplemented with penicillin (100U/ml), streptomycin (100 µg/ml), L-glutamine (2 mmol/L), growth factors like bFGF (20ng/ml) and EGF (20ng/ml)(2). Cell counting was done by Trypan Blue dye exclusion method and the cells were seeded in cell culture dishes coated with Fibronectin. The flasks with cells were incubated at 37˚C with 5% CO2 for varying periods from 18 days-28 days. The cells were observed daily and media change was done every 2-3 days. RESULTS: In all the samples, the Neurosphere like bodies (NLBs) were observed in the culture from 10th day onwards which were then subjected to histological and immunohistochemical studies. H&E staining showed positive for neural cells and Immunohistochemistry yielded positive for S-100 ,normally present in cells derived from the neural crest and Neuron Specific Enolase (NSE) a neuronal specific marker.CONCLUSION: We could successfully isolate and expand Human Enteric Neuronal cells from postnatal gut biopsy samples. Further research is warranted to utilize these Enteric Neuronal Cells for Cell based therapies to treat Hirschsprung’s disease. References:1. Marco Metzger, Claire Caldwell, Amanda J. Barlow, Alan J. Burns, and Nikhil Thapar. Enteric nervous system stem cells derived from Human gut mucosa for the Treatment of Aganglionic gut disorders. Gastroenterology. 2009 136:2214-2225.2. Nadege Bondurand, Dipa Natarajan, Nikhil Thapar, Chris Atkins and Vassilis Pachnis. Neuron and glia generating progenitors of the mammalian enteric nervous system isolated from foetal and postnatal gut cultures. Development and disease. 130(25):6387-6400. 3. Ulrich Rauch, Andrea Hänsgen, Cornelia Hagl, Stefan Holland-Cunz, Karl-Herbert Schäfer. Isolation and cultivation of neuronal precursor cells from the developing Human enteric nervous system as a tool for cell therapy in dysganglionosis. Int J Colorectal Dis. 2006 21:554–559.4. Richard M. Lindley, Daniel B. Hawcutt, M. Gwen Connell, David H. Edgar, Simon E. Kennyb. Properties of secondary and tertiary human enteric nervous system neurospheres. Journal of Pediatric Surgery 2009 44:1249–1256. 5. Petra M. Bareiss, Marco Metzger, Kai Sohn, SteVen Rupp, Julia S. Frick, Ingo B. Autenrieth, Florian Lang, Heinz Schwarz, Thomas Skutella, Lothar Just. Organotypical tissue cultures from adult murine colon as an in vitro model of intestinal mucosa. Histochem Cell Biol. 2008 129:795–804.

BACKGROUND: Patients with complete spinal cord injury at cervical level, mostly lead a life with quadriplegia. We report a case of 22 year old male, who became quadriplegic after C4-C5 injury and has improved after five injections of in-vitro expanded autologous bone marrow mononuclear cells (ABMMNC).MATERIALS & METHODS:The patient developed complete quadriplegia following fall from height. MRI suggested C5 burst fracture with retropulsion of C5 vertebrae and cord contusion at C4-C5 and underwent decompression and fixation four days post injury. After surgical fixation, bone marrow was harvested twice and a portion of the MNCs were expanded in-vitro and injected and another portion was cryopreserved which were later expanded and injected through lumbar puncture. The first injection was given 13 days after surgery. The first specimen of BM had CD34 of 0.18% and was injected twice; once immediately after harvesting (CD34:1.57%) and later after cryopreservation for 11 months (CD34: 3.33%). The second specimen had a CD34 of 0.52% and was injected thrice; twice after harvesting with CD34 of 1.28% and 6.23% respectively and later after cryopreservation for four months (CD34: 14.03%). The expansion was done in a GMP compliant clean room using autologous serum and the specimens were subjected to aerobic culture and Endotoxin tests before each administration.RESULTS: Investigations done (i)Post injury, (ii) Post decompression and (iii) one year (iv) One and half year after the injury have shown objective improvements with Light touch score (20-22-40-48), Pin prick score (20-20-32-40), Anal sensations (Nil-Nil-Present), Motor Score (11-11-22-26), Sensory level (C6-C6-C6-C6), Motor level (C5-C5-C6-C6), Neurological level (C5-C6-C6-C6), Spinal cord independence measure (9-12-45-50) and Barthel index (0-0-30-40). Repeat MRI show gliosis at C5. The ASIA score has improved from A to B with no adverse reactions.CONCLUSION: Multiple intrathecal injections of in-vitro expanded ABMMNCs in cervical-spinal cord injury was found to be safe and gradual objective improvements have been observed over a year and a half without any adverse outcome

Background: The adult Bone Marrow Stem Cells (BMSCs) have distinct advantages over the other types of stem cells. They are multipotent, can be stored for upto 10 years and considered to be one of the best sources of hematopoietic and mesenchymal stem cells in an adult body. Genetically inherited diseases such as Retinitis Pigmentosa and Degenerative diseases such as Age Related Macular Degeneration remain unsolved as no definitive treatment is available to repair the damages caused to the RPE and Photoreceptors as of now. In this scenario, the technique of Bone Marrow aspiration & isolation of Mono Nucleated Cells (MNCs) & intra-vitreal injection of a very small volume of MNCs in human retinal disorders has been standardized and is safe and feasible for human studies (Mohanty et al) and autotransplantation of RPEs from periphery to affected area are underpractice(Coffey et al). In this study we report our research work on different approaches to the above diseases using cell therapeuticsStudy 1 Materials & methods: Ciliary Pigment Epithelium was harvested from donor eyes from Aditya Jyot Eye Hospital, Mumbai and was taken to and grown at NCRM lab. The cells were grown in the earlier reported methodology of Brenda et al (Science 2004). Results: The CPE derived Retinal stem cells grew well in the lab. However, the practical difficulties of harvesting the same in patients limited our further steps in this study. Study II:? Materials & methods: Cadaver eye RPE cells were harvested and grown using polymer scaffolds after transporting them over 6 to 12 Hrs. The RPEs were grown on conventional methods and in polymer scaffolds and were subjected to RT-PCR. Results: Human RPEs were able to grow without amniotic membrane and the same was proven by RT-PCR. This would make it possible for the peripheral RPEs taken from patients to be stored and later expanded and used for replacing the diseased cells of the central portion of the retina in future, without having to harvest the RPEs again. Study III: Materials & methods: Bone marrow mono nuclear cells were isolated and transported in cold containers (4-8oC) over a period of 6-12 Hrs and viability was evaluated.Results: The bone marrow mononuclear cells were viable up to 12 Hrs in our methodology with a viability of more than 95% making it possible for cells isolated from Chennai centre to be taken to Mumbai or any other destination within a reach of 12 Hrs for application as reported in earlier studies.Conclusion: The in-vitro expansion of RPEs without Human Amniotic Membrane is expected to open up a new possibility for treating the Retinal Degenerative Diseases. However an animal study is needed before clinical application. Intra vitreal application of Bone Marrow Mono Nuclear cells to treat RP and AMD as reported earlier are considered safe. We plan to undertake treatment and long term follow-up of more numbers of patients with RP and AMD.

Bullous Keratopathy (BK) affects thousands of people in India every year. Though in early stages it is manageable medically, advanced disease warrants either total corneal transplantation or partial thickness transplantation for which a donor-cadaver cornea is necessary. Amano et al have reported the successful treatment of BK in animal models using in-vitro expanded human corneal endothelial precursors; though the rabbits had to be kept facing eye down to allow gravity assisted settling of the cells to the summit of the cornea where the damage had been created. For successful treatment using the above method, a human being has to lie prone with eyes immobilized for 24-36 Hrs. This is extremely discomforting and hence not practical. Corneal endothelium removed from the button and transported at varying temperature conditions for 48Hrs was successfully cultured in NCRM and this was reported earlier. We are working on a suitable scaffold to retain the cells in situ until their attachment to the damaged portion of the corneal endothelium enabling it to heal without the patient having to lie prone. With such capability, we envisage to make a corneal endothelial precursor/stem cell (CES) bank named as CESBANK to make in-vitro expanded CES available for patients with corneal diseases, most commonly Bullous Keratopathy (BK).

In the published paper, the details of the implementation of Newman's P2D model was incomplete. Additionally, the acknowledgement section failed to note the contributions of a collaborator who provided the P2D simulation code used in the original work.

Background - Use of autologous bone marrow stem cell is a newly evolving treatment modality for end stage cardiac failure as reported in the literature. We report our experience with two patients with dilated cardiomyopathy who underwent this treatment after failure of maximal conventional therapy. Methods - A 29 year old Male patient with history of orthopnea and PND, with a diagnosis of dilated cardiomyopathy and echocardiographic evidence of severe LV dysfunction was referred for further treatment. His echo on admission showed EF of 17% and no other abnormal findings except elevated bilirubin levels. He was in NYHA functional class IV. He received intracoronary injection of autologous bone marrow stem cells in January 2009. 254X106 cells were injected with a CD34+ of 0.20%. His clinical condition stabilized and he was discharged home. He received a second injection of 22X106 in vitro expanded stem cells with a CD34+ of 0.72% in Aug 2009. He is now in NYHA class II-III with EF 24%. A 31year old Male patient with history of increasing shortness of breath, severe over the past 3-4 days was admitted for evaluation and treatment. His echo on admission showed EF of 20% and was in NYHA functional class IV. Coronary angiogram was normal and he was stabilized on maximal anti failure measures. He received intracoronary autologous bone marrow stem cell injection of 56X106 with a CD34+ of 0.53% in August 2009. His clinical condition stabilized over the next 10 days and he was discharged home. Conclusions - In our experience of two cases of dilated cardiomyopathy, safety of intracoronary injection of autologous bone marrow stem cells both isolated and in vitro expanded has been proven in both the cases with efficacy proven in one of the cases. Long term follow-up of these two cases and inclusion of more number of similar cases where all available conventional therapies have not resulted in significant improvement for such studies are planned.

A three year old male non-descriptive companion dog was presented to the Small Animal Orthopedic Unit of Madras Veterinary College Teaching Hospital (MVC) with paraplegia of fourth degree neurological deficit of hind limbs due to automobile trauma. Radiographic views were suggestive of dislocation at T8-T9 vertebral segment with fracture of L2 vertebra. Myelography confirmed the signs of abrupt stoppage of the contrast column cranial to dislocated area and was interpretive of transected spinal cord at L2 level. Construct was prepared with bone marrow mononuclear cells (BMMNC) isolated from bone marrow aspirate of femur and the cells were seeded in Thermoreversible Gelatin Polymer (TGP) at the cell processing facility of Nichi-In Centre for Regenerative Medicine (NCRM) as per GMP protocols and was engrafted after hemilaminectomy and durotomy procedures in the MVC. Postoperatively the animal was clinically stable; however the animal died on the 7th day. Autopsy revealed co-morbid conditions like cystitis, nephritis and transmissible venereal tumor. Histopathology of the engrafted area revealed sustainability of aggregated stem cells that were transplanted revealing an ideal biocompatibility of the construct prepared with bone marrow mononuclear cells and polymer hydrogel for spinal cord regeneration in dogs. Further studies in similar cases will have to be undertaken to prove the long term efficacy.

Introduction: Autologous Immune Enhancement therapy (AIET) is one of the lesser used modality of therapy in our country for advanced cancers. AIET uses patient’s own immune cells such as NK cells and T-Lymphocytes to fight against cancer.Aim: To study the effectiveness of AIET in Advanced Carcinoma of pancreas.Case History: A 40 years old male was diagnosed with inoperable locally advanced Carcinoma of the head of pancreas when he was investigated for obstructive jaundice. Palliative bypass surgery was done, since his tumor was inoperable. Following his surgery he received 6 cycles of adjuvant chemotherapy and 3 cycles of AIET.? After completion of chemotherapy and 2 cycles of AIET therapy his tumor marker decreased significantly with decrease in tumor size.? So re-laparotomy was attempted for removal of tumor, which was unsuccessful. After a long gap he received his 3rd cycle of AIET.? He was irregular in receiving his AIET treatment. He survived for 18 months after the diagnosis. Conclusion: The median life span of patients following diagnosis of advanced Carcinoma of pancreas is usually not more than 3-6 months. Longer survival of our patient could be due to AIET, but we need further studies to prove it. There were no adverse effects encountered due to AIET.

Abstract Increased oxidative stress and mitochondrial dysfunction have been identified as common pathophysiological phenomena associated with neurodegenerative disorders such as Alzheimer's disease (AD), Parkinson's disease (PD) and Huntington's disease (HD). As the age-related decline in the production of melatonin may contribute to increased levels of oxidative stress in the elderly, the role of this neuroprotective agent is attracting increasing attention. Melatonin has multiple actions as a regulator of antioxidant and prooxidant enzymes, radical scavenger and antagonist of mitochondrial radical formation. The ability of melatonin and its kynuramine metabolites to interact directly with the electron transport chain by increasing the electron flow and reducing electron leakage are unique features by which melatonin is able to increase the survival of neurons under enhanced oxidative stress. Moreover, antifibrillogenic actions have been demonstrated in vitro, also in the presence of profibrillogenic apoE4 or apoE3, and in vivo, in a transgenic mouse model. Amyloid-β toxicity is antagonized by melatonin and one of its kynuramine metabolites. Cytoskeletal disorganization and protein hyperphosphorylation, as induced in several cell-line models, have been attenuated by melatonin, effects comprising stress kinase downregulation and extending to neurotrophin expression. Various experimental models of AD, PD and HD indicate the usefulness of melatonin in antagonizing disease progression and/or mitigating some of the symptoms. Melatonin secretion has been found to be altered in AD and PD. Attempts to compensate for age- and disease-dependent melatonin deficiency have shown that administration of this compound can improve sleep efficiency in AD and PD and, to some extent, cognitive function in AD patients. Exogenous melatonin has also been reported to alleviate behavioral symptoms such as sundowning. Taken together, these findings suggest that melatonin, its analogues and kynuric metabolites may have potential value in prevention and treatment of AD and other neurodegenerative disorders.

Abstract Aging is associated with a decline in immune function (immunosenescence), a situation known to correlate with increased incidence of cancer, infectious and degenerative diseases. Innate, cellular and humoral immunity all exhibit increased deterioration with age. A decrease in functional competence of individual natural killer (NK) cells is found with advancing age. Macrophages and granulocytes show functional decline in aging as evidenced by their diminished phagocytic activity and impairment of superoxide generation. There is also marked shift in cytokine profile as age advances, e.g., CD3+ and CD4+ cells decline in number whereas CD8+ cells increase in elderly individuals. A decline in organ specific antibodies occurs causing reduced humoral responsiveness. Circulating melatonin decreases with age and in recent years much interest has been focused on its immunomodulatory effect. Melatonin stimulates the production of progenitor cells for granulocytes-macrophages. It also stimulates the production of NK cells and CD4+ cells and inhibits CD8+ cells. The production and release of various cytokines from NK cells and T-helper lymphocytes also are enhanced by melatonin. Melatonin presumably regulates immune function by acting on the immune-opioid network, by affecting G protein-cAMP signal pathway and by regulating intracellular glutathione levels. Melatonin has the potential therapeutic value to enhance immune function in aged individuals and in patients in an immunocompromised state.