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Introduction HIV-1-infected patients develop haematological disorders such as cytopenias. One possible explanation is the inhibition of haematopoiesis at the level of differentiation of CD34+ haematopoietic progenitor stem cells. Based on our previous studies, we hypothesised that there may be viral encoded, or host cellular factors which participate in the process of inhibition of haematopoiesis. Materials and methods Virus-depleted media from infected CD4+ T cells was prepared by filtration and added to CD34+ cell differentiation semisolid medium. We have also used the virus-depleted media to isolate host/viral factors including miRNA. Isolated miRNAs were screened for their haematopoietic inhibitory function using the miRNA mining approach. Results Addition of virus-depleted media caused a 40% inhibition of differentiation of CD34+ cells into myeloid and erythroid colony formation. Real-time RT-PCR showed miR-15a and miR-24 from both pIndie-C1 and pNL4.3 HIV-1-infected cells showed a significant differential expression when compared to control media. Conclusion In this study, we have identified two miRNAs, miR-15a and miR-24 secreted from purified HIV-1-infected CD4+ T cells that inhibited CD34+ haematopoietic progenitor stem cell differentiation into myeloid and erythroid colonies in vitro.

Stromal vascular fraction (SVF) can easily be obtained from a mini-lipoaspirate procedure of fat tissue and platelet rich plasma (PRP) can be obtained from peripheral blood. We evaluated the safety and preliminary efficacy of administering SVF and PRP intra-articularly into patients with osteoarthritis grade 1 and 2. A total of ten patients underwent a local tumescent liposuction procedure to remove approximately 100 ml of fat tissue from the abdomen. SVF was isolated using an enzyme digestion and resuspended in PRP for intra-articular injection in the knee. The Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score and six-minute walk distance (6MWD) were used to evaluate clinical effects and included measure of patient’s subjective assessment of pain, joint mobility, and physical disability. WOMAC score, 6MWD and laboratory tests were repeated at 3 and 6 months and 1, 1.5 and 2 years. XRAY and MRI were completed at 1 year. The average total WOMAC score was 64 at baseline and significantly reduced to 52 at 3 months, 46 at 6 months, 42 at 1 year, 38 at 1.5 years, and 41 at 2 years. Patients walked an average of 1310 feet at baseline and demonstrated a statistically significant improvement at 3 and 6 months and 1, 1.5, and 2 years post treatment. Cartilage thickness as determined by MRI improved by at least 0.2 mm in six patients, was unchanged in two patients and decreased by at least 0.2 mm in two patients. Overall, all of the patients were pleased with the treatment results. They reported a reduction in pain levels, especially after 3 months. More importantly, the procedure demonstrated a strong safety profile with no severe adverse events or complications reported. Trial registration NCT03089762; Name of registry: http://www.clinicaltrials.gov

Background Stromal vascular fraction (SVF) can easily be obtained from a mini-lipoaspirate procedure of fat tissue and platelet rich plasma (PRP) can be obtained from peripheral blood. We evaluated the safety and preliminary efficacy of administering SVF and PRP intra-articularly into patients with osteoarthritis grade 1 and 2. Methods A total of ten patients underwent a local tumescent liposuction procedure to remove approximately 100 ml of fat tissue from the abdomen. SVF was isolated using an enzyme digestion and resuspended in PRP for intra-articular injection in the knee. The Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) score and six-minute walk distance (6MWD) were used to evaluate clinical effects and included measure of patient’s subjective assessment of pain, joint mobility, and physical disability. WOMAC score, 6MWD and laboratory tests were repeated at 3 and 6 months and 1, 1.5 and 2 years. XRAY and MRI were completed at 1 year. Results The average total WOMAC score was 64 at baseline and significantly reduced to 52 at 3 months, 46 at 6 months, 42 at 1 year, 38 at 1.5 years, and 41 at 2 years. Patients walked an average of 1310 feet at baseline and demonstrated a statistically significant improvement at 3 and 6 months and 1, 1.5, and 2 years post treatment. Cartilage thickness as determined by MRI improved by at least 0.2 mm in six patients, was unchanged in two patients and decreased by at least 0.2 mm in two patients. Conclusions Overall, all of the patients were pleased with the treatment results. They reported a reduction in pain levels, especially after 3 months. More importantly, the procedure demonstrated a strong safety profile with no severe adverse events or complications reported. Trial registration NCT03089762; Name of registry: http://www.clinicaltrials.gov

Spinal cord injury is a traumatic neurological condition which makes the patient disable. Its management still remains challenging but advancements in the regenerative medicine have changed the approach of treating this serious debilitating condition of the central nervous system. Cell based therapies can restore function in spinal cord injury by replacing the lost neural tissue. These therapies also rejuvenate the existing intact neurons by facilitating remyelination and by repairing and reducing progressive tissue damage and scarring.Autologous bone marrow stem cells were collected from the patients. 5 ml of the processed sample was injected back into the patients via lumbar puncture at L1/L2 level. The bone marrow harvesting and administration was repeated every 4 weeks 3 times (12 weeks).Significant improvements were noticed following the injections into the patients with the duration of injury less than 6 months. ASIA grade improvements were observed in 6 out of 10 patients. VTC and walking, at least with the support, was restored in eight patients. Bladder control and sexual functions improved in three and five patients respectively. Eight patients exhibited decreased spasticity.We believe that autologous bone marrow stem cells contributed towards the neuroplaticity and/or paracrine effect due to which we observed the considerable improvements in the conditions of the patients.This preliminary proof of patient improvement reinforces the potential of autologous bone marrow stem cell treatment in the patients suffering from Spinal Cord Injury. Although the results are encouraging further studies are needed to substantiate the claims.

Human mesenchymal stem cells from bone marrow (hMSCs) have broad therapeutic potential. These cells can be are readily isolated from bone marrow by their property to adhere to tissue culture treated culture wares. However, the proliferation rates and other properties of the cells gradually change during expansion. This study aims to validate the protocol of isolation and differentiation of hMSCs from bone marrow for therapeutic applications.Sixty ml of bone marrow was extracted from 5 patients and MSCs were isolated. These were characterized by Flow Cytometry, CFU assay and were differentiated into bone, fat cells and neurocytes.The cells were having healthy morphology. These were positive for the markers CD105, CD90 and CD73 and negative for CD45, CD34 and HLA-DR. The cells could differentiate into fat, bone and neural cells.MSCs from the bone marrow were isolated and differentiated. These cells were morphologically healthy and passed CFU assay. The cells exhibited differentiation potential into bone, fat and neural tissue. These cells can be used in therapeutic applications.

The regeneration patterns are innate, inherited and evolutionarily conserved mechanisms. In every individual there are certain cells and signaling networks which work together to proliferate the desired tissue lineages to replace the dead, lost and injured counter parts. This homeostasis mechanism keeps functioning of the organ system intact. There are some tissues such as skin, gut, blood, respiratory tract, uterine endometrium, testis must perpetually renew the majority of cells. As the aging advances the turnover potentials decreases under normal circumstances, some of these respond inefficiently to regenerative pressures (eg: brain and heart) while other respond quite well. Therefore creating an optimized micro environment using external means through non-invasive or invasive procedures preferably minimally by utilizing appropriately subjected stem cells/stem cell secretome to induce regeneration at the target sites where it does not take place spontaneously.Stem cell biology is one of the most attractive areas of biomedical research, as emerges for the execution of biotechnology towards the regenerative medicine continues to expand. The presumed potential of the stem cell populations with clonogenic capabilities are harnessed for the therapeutic applications. Advancements in the research technology and the idea of inducing innate regeneration by stem cell based approaches can generate the potential cure for many degenerative disorders, age related disabilities and accidental tissue damages.

Stem cell therapy is a promising treatment for cerebral palsy, which refers to a category of brain diseases that are associated with chronic motor disability in children. Autologous bone marrow stem cells may be a better cell source and have been studied for the treatment of cerebral palsy because of their functions in tissue repair and the regulation of immunological processes.To assess autologous marrow stem cells as a novel treatment for patients with moderate-to-severe cerebral palsy, a total of 10 cerebral palsy patients were enrolled in this clinical study with 24 months follow-up. A total of 10 cerebral palsy patients received autologous bone marrow cells transplantation (4.5 × 10We recorded the gross motor function measurement scores, manual ability function measurement score, and adverse events up to 24 months post-treatment. The gross motor function measurement scores were significantly higher at month 6 post-treatment compared with the baseline scores and were stable up to 24 months follow-up. The increase in manual ability and communication function measurement scores at 6 months were not significant when compared to the baseline score. All the 10 patients survived and none of the patients experienced any serious adverse events or complications.Our results indicated that bone marrow derived MNCs are safe and effective for the treatment of motor deficits related to cerebral palsy. Further randomized clinical trials are necessary to establish the efficacy of this procedure.

Avascular Necrosis (AVN) of hip is a devastating condition seen in younger individuals. It is the ischemic death of the constituents of the bone cartilage of the hip. The femoral head (FH) is the most common site for AVN. It results from interruption of the normal blood flow to the FH that fits into the hip socket. Earlier studies using autologous bone marrow stem cell concentrate injections have shown encouraging results with average success rates. The current study was designed to improve significantly the cartilage regeneration and clinical outcome.Total of 48 patients underwent autologous bone marrow stem cell and activated platelet-rich plasma derived growth factor concentrate (PRP-GFC) therapy for early and advanced stages AVN of femoral head in a single multi-specialty center. The total treatment was divided into three phases. In the phase I, all the clinical diagnostic measurements such as magnetic resonance imaging (MRI), computed tomography (CT) etc. with respect to the AVN patients and bone marrow aspiration from posterior iliac spine from the patients were carried out. In the phase II, isolation of stem cells and preparation from the patients were performed. Subsequently, in phase III, the stem cells and PRP- GFCs were transplanted in the enrolled patients.Ninety three percent of the enrolled AVN patients showed marked enhancement in the hip bone joint space (more than 3mm) after combined stem cells and PRP-GFC treatment as evidenced by comparison of the pre- and post-treatment MRI data thus indicative of regeneration of cartilage. The treated patients showed significant improvement in their motor function, cartilage regrowth (3 to 10mm), and high satisfaction in the two-year follow-up.Combination of stem cell and PRP-GFC therapy has shown promising cartilage regeneration in 45 out of 48 patients of AVN. This study clearly demonstrates the safety and efficacy of this treatment. Larger numbers of patients need to be evaluated to better understand the efficacy of the combined stem cell and PRP-GFC therapy on AVN patients.

Autism is a neural disorder presenting in the early developmental period, usually in the first 2 years of life. It is characterized by persistent deficits in social communication and social interaction, restricted and repetitive patterns of behavior, interests, or activities, and causes clinically significant impairment in social, occupational, or other important areas of functioning.Cellular therapy is an advanced approach to treat disorders where current therapies do not offer a cure or efficient symptom relief. Herein we present the data of ten autistic patients who were enrolled in this study utilizing stem cells. All patients were scored on ISAA and WeeFIM scales before and at 3, 6, 12 or 24 months following the injection of stem cells. The ISAA scores of the patients improved with the injection, and no adverse effects were noted. We report promising results in this small pilot study using autologous Bone Marrow Aspirate Concentrate (BMAC) injected intrathecally into these ten enrolled autistic patients.

The objective is to investigate the safety and clinical efficacy of Autologous Platelet Rich Plasma Concentrated Spray (Keratogrow®), for hair loss. Autologous -PRP spray, prepared from a small volume of blood, was applied on the selected patients' scalps at least twice daily. Three months treatments were given for each patient. The effectiveness of the medication was measured by changes in hair regrowth after 3 months determined by physical exam and digital photography. At the end of the 3 cycles of treatment, the patients presented clinical improvement in the mean number of hairs, with a mean increase of hairs in the target area, and a mean increase in total hair density compared with baseline values.

Platelet-rich plasma (PRP) has shown remarkable beneficial effects without any major adverse reactions in the treatment of androgenic alopecia .The growth factors in activated autologous PRP induces the proliferation of dermal papilla cells.To investigate the clinical efficacy of Platelet Rich Plasma prepared using Merisis One Step Gel Separation Technology in treatment of androgenic alopecia.Five patients were given autologous PRP injections on the affected area of alopecia over a period of three months at interval of two - three weeks and results were assessed.Three months after the treatment, the patients presented clinical improvement in the hair counts, hair thickness, hair root strength and overall alopecia.PRP appears to be a cheap, effective and promising therapy for androgenic alopecia with no major adverse effects.

Late stage detection of COVID-19 disease pathogenesis induced by SARS-CoV-2 infection may prove to be fatal to the infected patients with the severity increasing, if inhibition of differentiation of hematopoietic endothelial

One of Nature's gifts to mankind is mesenchymal stem cells (MSC's). They are multipotent in nature and are present literally in every tissue. Since, they possess certain characteristics of stem cells such as self-renewal and differentiation they are known to be one of the key players in normal tissue homeostasis. This novel function of mesenchymal stem cells has been explored by scientists in the field of regenerative medicine. This review gives an insight of the various sources of mesenchymal stem cells available for tissue engineering with regard to tendon and ligament and the mechanism involved during regeneration.

Key modalities of integrative medicine known to rejuvenate the mind and body are meditation, yoga, and controlled diet. It has been shown previously that intensive or prolonged mind and body therapies (MBT) may have beneficial effects on the well-being of healthy people and in patients. Telomerase activity and levels of peripheral blood adult pluripotent stem cells (PB-APSC) are reliable markers of long-term well-being that are known to decrease with age. The objective of this study is to understand the effect of our MBT program on telomerase activity and stem cells in blood collected from the participants.Here, we have investigated the effects of an intensive three weeks MBT retreat on telomerase activity and the peripheral blood stem cells in participants before and after the MBT. A total of 108 people were enrolled in the study; 38 men and 70 women (aged 18-90) randomly assigned for the study.Telomerase activity was greater in retreat participants at the end of the MBT retreat. About 45% of people showed more than one-fold increase of telomerase activity after our MBT program. Furthermore, about 27% of people showed more pronounced fold increase (2-fold) in telomerase activity after the MBT. In addition, a substantial percentage of people (about 90%) exhibited increased stem cell counts after the MBT.The data suggest increased telomerase activity and stem cells count in peripheral blood from MBT retreat participants that may lead to increased longevity and better quality of life at latter age.

Methanol ingestion leads to severe damage to visual pathways and permanent loss of vision. Current treatment is aimed at removal of methanol from system and prevention of generation of toxic metabolites along with symptomatic management of patient. Autologous bone marrow mononuclear stem cells (MNC) can be used to rejuvenate the damaged retinal cells and restoration of vision.Five patients suffering from methanol induced complete blindness within three months of insult and no known comorbidities during the past 6 months were enrolled to receive autologous bone marrow derived mononuclear cell fraction on compassionate grounds. The visual acuity and visual evoked responses (VER) were done at the time of enrollment and during follow-up visits.Visual acuity of these patients at the time of enrollment: no perception of light. Improvement in visual acuity was recorded by 7 days which reached maximum at 3 weeks after treatment in three patients and three months in two patients. The patients had acuity of 6/9, finger counting and reading with magnifying glasses with no subsequent improvement till 2 years of follow-up. Visual Evoked Responses demonstrated improvements following treatment. No adverse reactions were noticed during follow-up.Treatment with Autologous Bone marrow derived MNC offers a new line of management in patients with loss of vision following methanol ingestion. The efficacy and safety of this line of management needs to be evaluated in controlled clinical trials.

HIV-1 V3 loop clones of virus isolates derived from patients suffering from thrombocytopenia were used for infection of the human thymus/liver conjoint hematopoietic organ that developed in the severe combined immunodeficient mouse (SCID-hu Thy/Liv). The V3 loop clones showed a significantly greater degree of inhibition of megakaryopoiesis than myelopoiesis and erythropoiesis of the human CD34+ progenitor cells, in vivo. Inhibition of megakaryopoiesis occurs through reduction in c-Mpl expression and consequent decrease in STAT5 activation. Therefore HIV-1 V3 loop sequences of thrombocytopenic patients exhibit preferential inhibition of megakaryocyte lineage-specific differentiation of CD34+ progenitor cells, thus reflecting the patients' clinical condition.

To report bone marrow derived stem cell application in a patient with bilateral visual loss due to accidental ingestion of methanol.A 22-year-old male patient with bilateral visual loss at discharge after treatment for methanol ingestion was treated with retrobulbar and intrathecal autologus bone marrow derived stem cells in an outpatient setting. The procedure was followed by Vit B12 Injection 500 mcg alternate days and oral vitamins B1, B2, B6, in therapeutic dose for 3 months.The patient demonstrated favorable outcome three days after the treatment. Visual acuity improved to 20/20 in both eyes by third week.Treatment with retrobulbar and intrathecal autologous bone marrow derived stem cell in an adult male patient with bilateral visual loss due to accidental ingestion of methanol was effective. Larger studies are warranted to explore the real impact of this procedure.

Safer and greater efficacy biologically occurring 3-O-sulfogalactosylceramide, sulfatide ( +/-) poietins for containment of HIV replication and rescue from hematopoietic cell depletion in HIV-1 Infection is our objective. We also discuss novel biomarkers detection with the intention to later design new array of fluorescence molecule (and their protein complexes) labeling methods of monoclonal antibodies using modeling through bioinformatics.

Hematopoietic stem cells which are present in bone marrow, cord blood and liver, terminally differentiate into multiple lineages of blood cell types and thus have applications in bone marrow transplantation. The determination of alleviation of cytopenias in HIV infection in vivo [severe combined immunodeficient transplanted with human fetal thymus and liver tissues, SCID-hu] has been utilized through assay of inhibition of hematopoietic stem cell colony formation ex vivo [1]. Cytopenias or hematopoietic inhibition (decreased blood cells formation) is a common occurrence in HIV infected patients both due to the virus but also caused in synergistic manner by the antiretroviral drugs (AZT or Zidovudine, ddI, certain protease inhibitors) [2]. Experimental observations for the indirect inhibition of the hematopoietic CD34+ stem cell colony forming activity by HIV infected T-cells were previously proposed or reported by us elsewhere in 1990’sand also bysome other investigators, from other institutions [1 and references therein]. The potential biologically occurring Sulfatide (3-O-sulfogalactosylceramide), which is non-specific to only

Live attenuated HIV vaccines offer a means to introduce exogenous sequences into the viral genome to target the virus elimination in vivo. Foreign genes inserted into the nef region of HIV-1 NL4-3 were found to be rapidly deleted following virus infection and/or replication, in a size dependent manner, in the human fetal Thymus/Liver implants of severe combined immunodeficient mouse (SCID-hu) model. When the murine heat stable antigen (HSA) of 283 bp was substituted into HIV-1 nef region, the viral loads in vivo were comparable to the negative control nef attenuated HIV-1, and the reporter HSA gene was not deleted upon infection. However, the murine Thy1.2 gene (505 bp) substituted into the nef attenuated HIV-1, upon infection and replication, deleted 441 bp in vitro and 437 bp in vivo, of the inserted Thy1.2 gene. When the enhanced green fluorescence protein (eGFP) gene (720 bp) was substituted for nef, virus replication was aborted in vivo in the Thy/Liv implants, as seen by the background levels of viral loads, comparable to mock infected implants, and the eGFP gene was deleted. When the herpes simplex virus thymidine kinase gene, HSV-TK (1.15 kbp), or HSA gene, was substituted into the viral vpr gene, TK but not HSA gene was deleted, upon infection in vitro. Moreover, NL-TKI reporter virus with both intact nef and vpr genes shows deletion of TK gene both in vitro and in vivo. Excision of foreign genes occurred within the exogenous segments but not in the viral own regions. These results suggest that larger "suicide" genes introduced via HIV-1 can be deleted upon infection. However, smaller size nucleotide sequences or genes (approximately 300 bp) inserted in place of viral nef or vpr gene may be used to target the virus or its components, for attack and elimination in vivo, and thus have implications for the development of live attenuated HIV vaccines.

We have since the 1970's embarked on the development of biologically derived fluorophore-protein complexes that will find applications in the communicable and non-communicable disease etiology processes and their cures. We have since then become largely successful in these endeavors along with interspersed contributions also from investigators who have generally restricted to working in confined disciplines. Their encompassment with our works as this investigator has traversed his definitely chosen and not merely a circumstantial, coincidental, or accidental step-wise multi-disciplinary scientific path from biophysics to regenerative medicine spanning these lines of investigations for last four decades have finally yielded the much necessitated disease related applied biological interventions for human benefits. Taking a cue from our early investigations and findings on which we call attention to the identification and characterization of the use of the primary light-emitting lumazine precursor of riboflavin which is 6,7-dimethyl-8-ribityl lumazine-protein complex which we had derived from the bioluminescence bacterium (Photobacterium phosphoreum) wherein it functions as a naturally occurring fluorescence light emitter (LumP). These in vivo phenomena have been a precursor to the subsequent developments in vitro. This in vivo to in vitro investigation path of ours has been also comprised among others of binding of photosynthetic light-harvesting marine dinoflagellate algae (Glenodinium sp.) derived peridinin-chlorophyll a-protein (PerCP) complex-labeled monoclonal antibodies useful in the development of flow cytometry. These fluorescence labeled antibodies bound antigens which include those of communicable infectious diseases (HIV/AIDS - env-gp160, gag-p24), non-communicable but also potential hereditary and malignant disorders (Cancer/Tumor Markers - Melan-A/Mart-1 of melanoma), normal immune response cells (Human/Mouse/species cellular MHC/TCR/CD45/CD33/CD56/CD19/CD41), and of types of stem cells (CD34/CD38/c-Mpl/Oct4/Neuropilin-1/SOX17). Such antigens have been analyzed by us and other investigators by fluorescence-activated cell sorting (FACS - cell surface and intracellular binding), confocal fluorescence microscopy, or/and immunohistochemistry, to determine qualitative and quantitative antigen expression levels and their mechanistic implications. We have followed stem cell differentiation patterns and signaling mechanisms through marker antigen-antibody binding wherein the antibodies are labeled with covalently linked fluorophore-protein complexes or fluorescence emitting chromophores. These complexes among others also have included PerCP. We are also now in the process of developing flow cytometry applications of additional visible light emitting chromophore-protein complexes through industrial collaborations. The United States Navy has long been known for interest in the estimation of changes in illumination intensity in and under the oceans to track movements of enemy ships and other naval vessels.

Chemotherapy drugs and radiotherapy are highly toxic and both damage adjacent healthy cells. Side effects may be acute (occurring within few weeks after therapy), intermediate or late (occurring months or years after the therapy). Some important side effects of chemotherapy are: nausea, vomiting, diarrhea, mucositis, alopecia, constipation etc; whereas radiation therapy though administered locally, can produce systemic side effects such as fatigue, anorexia, nausea, vomiting, alteration in the taste, sleep disturbance, headache, anemia, dry skin, constipation etc. Late complications of these therapies also include pharyngitis, esophagitis, laryngitis, persistent dysphagia, fatigue, hepatotoxicity, infertility and cognitive deficits. These arrays of side effects have a devastating effect on the quality of life of cancer survivors. Due to the inadequacy of most of the radio-protectors and chemo-protectors in controlling the side effects of conventional cancer therapy the complementary and alternative medicines have attracted the view of researchers and medical practitioners more recently. This review aims at providing a comprehensive management protocol of above mentioned chemo-radiotherapy induced side effects based on Ayurveda, which is an ancient system of traditional medicine practiced in Indian peninsula since 5000 BC. When the major side effects of chemo-radiotherapy are looked through an ayurvedic perspective, it appears that they are the manifestations of aggravated pitta dosha, especially under the group of disorders called Raktapitta (haemorrhage) or Raktadushti (vascular inflammation). Based on comprehensive review of ancient vedic literature and modern scientific evidences, ayurveda based interventions are put forth. This manuscript should help clinicians and people suffering from cancer to combat serious chemo-radiotherapy related side effects through simple but effective home-based ayurveda remedies. The remedies described are commonly available and safe. These simple ayurveda based solutions may act as an important adjuvant to chemo-radiotherapy and enhance the quality of life of cancer patients.

Hematopoietic homeostasis is maintained throughout the lifetime of an individual through self-renewal of hematopoietic stem cells. Defects in the self - renewal and differentiation lead to hematopoietic insufficiency and development of malignancies. Leukemic stem cells (LSCs), which are considered to originate from hematopoietic stem or progenitor cells, not only adopt the regulatory machinery operating in normal HSCs but establish their own mechanisms against apoptosis and senescence. Hematopoietic malignancies are of Lymphoid origin with CLL and ALL and myeloid malignancies with AML and CML and in addition there are disease of the plasma multiple myelomas. One of the major therapeutic strategies for hematological malignancies is hematopoietic stem cell transplantation along with combination of chemotherapy. The review gives an insight of different hematological malignancies, the mechanism and therapeutic strategies available at present.

With the cementing of the cancer stem cell (CSC) concept, cancer biology and cancer drug discovery have attained a new avenue to target cancer. Studying the hierarchy of tumor tissue organization and how to inhibit the cell that resides at the very top of this hierarchy has opened up a new branch of tumor biology and given the opportunity to develop novel cancer-targeting strategies. With the discovery of CSCs in majority of cancer indications there seems to be a universal applicability of the concept. However, the CSC field is still at an early fledgling state and a lot more needs to be done in terms of understanding their emergence, maintenance, role in metastasis and their function in shaping the tumor architecture. CSCs are considered to be responsible for tumor initiation, metastasis and resistance to conventional radio and chemotherapy. Therefore, different approaches to targeting these tumorigenic and rare cells are urgently needed in order to improve the efficacy of anti-cancer therapy. We outline here the cancer stem cell concept and its relevance as well as biotherapeutic approaches to CSC targeting, including oncolytic viruses, monoclonal antibodies, cytokines and cytotoxic T lymphocytes.

Human immunodeficiency virus (HIV)-infected individuals exhibit a variety of hematopoietic dysfunctions. The SCID-hu mouse (severe combined immunodeficient mouse transplanted with human fetal thymus and liver tissues) can be used to model the loss of human hematopoietic precursor cell function following HIV infection and has a distinct advantage in that data can be obtained in the absence of confounding factors often seen in infected humans. In this study, we establish that HIV type 1 (HIV-1) bearing a reporter gene inserted into the viral vpr gene is highly aggressive in depleting human myeloid and erythroid colony-forming precursor activity in vivo. Human CD34 + progenitor cells can be efficiently recovered from infected implants yet do not express the viral reporter gene, despite severe functional defects. Our results indicate that HIV-1 infection alone leads to hematopoietic inhibition in vivo; however, this effect is due to indirect mechanisms rather than to direct infection of CD34 + cells in vivo.

Recent advances in the field of regenerative medicines manifested the unique properties of stem cells including the ability of self-renewal and differentiation to make them available for their replacement in tissue injury. Mesenchymal stem cells (MSCs) are important tools in treating immune disorders and in tissue repair due to their multipotency, immunosuppressive properties, and production of cytokines or growth factors. MSC-mediated therapy is a fast-growing field that has proven safe and effective in the treatment of various degenerative diseases and tissue injuries. Generation of induced pluripotent stem (iPS) cells holds a great promise for regenerative medicine and other aspects of clinical applications. The mechanisms governing multipotency in MSCs are not well understood. This review mainly throws light on the biology of MSCs, including their efficiency in treating several diseases and also the progress of the use of iPSC-derived MSC widely in the clinic.

Cancer stem cells (CSCs) are stem-like tumor populations that are reported to contribute towards tumor growth, maintenance and recurrence after therapy. Hypoxia increases CSC fraction and promotes acquisition of a stem-cell-like state. Cancer stem cells are critically dependant on the hypoxia-inducible factor-1 (HIF-1) for survival, self-renewal, tumor growth and maintenance of their undifferentiated phenotype. Recent researches show that stage of differentiation of the tumor cells is predictive of their susceptibility to natural killer cell (NK) cell mediated cytotoxicity and cancer stem cells are significant targets of NK cell cytotoxicity. Studies also show that reversion of tumor cells to a less-differentiated phenotype can be achieved by blocking NFκB. Yoga therapy (yogic lifestyle modifications encompassing physical postures, breathing practices, relaxation techniques and meditations) is known to modulate neural, endocrine and immune functions at the cellular level through influencing cell cycle control, aging, oxidative stress, apoptosis and several pathways of stress signaling molecules. Yoga therapy has also been shown to enhance natural killer cell activity and modulate stress and DNA damage in breast cancer patients receiving radiotherapy. Recent study found that brief daily yogic meditation may reverse the pattern of increased NFκB-related transcription of pro-inflammatory cytokines in leukocytes. Thus, yoga therapy has the potential to reduce cancer stem cell survival, self -renewal and tumor growth by modifying the tumor micro-environment through various mechanisms such as; 1) reducing HIF-1 activity by enhanced oxygenation, 2) promoting NK cell activity directly (or indirectly through down regulating NFκB expression), thereby enhancing NK cell mediated CSC lysis, and 3) by minimizing the aberrant expressions or activities of various hormones, cytokines, chemokines and tumor signaling pathways. Yoga therapy may have a synergistic effect with conventional modalities of treatment in preventing cancer progression and recurrences.

Complementary and alternative therapies (CAM) are gaining popularity amongst patients as add on to conventional medicine. Yoga stands third amongst all CAM that is being used by cancer patients today. Different schools of yoga use different sets of practices, with some using a more physical approach and many using meditation and/or breathing. All these modules are developed based on the needs of the patient. This paper is an attempt to provide the basis for a comprehensive need based integrative yoga module for cancer patients at different stages of treatment and follow up. In this paper, the holistic modules of the integrated approach of yoga therapy for cancer (IAYTC) have been developed based on the patient needs, as per the observations by the clinicians and the caregivers. Authors have attempted to systematically create holistic modules of IAYTC for various stages of the disease and treatment. These modules have been used in randomized trials to evaluate its efficacy and have shown to be effective as add-on to conventional management of cancer. Thus, the objective of this effort was to present the theoretical basis and validate the need based holistic yoga modules for cancer patients.Literature from traditional texts including Vedas, Ayurveda, Upanishads, Bhagavat Gita, Yoga Vasishtha etc. and their commentaries were looked into for references of cancer and therapeutic directives. Present day scientific literature was also explored with regards to defining cancer, its etiopathology and its management. Results of studies done using CAM therapies were also looked at, for salient findings. Focused group discussions (FGD) amongst researchers, experienced gurus, and medical professionals involved in research and clinical cancer practice were carried out with the objectives of determining needs of the patient and yoga practices that could prove efficient. A list of needs at different stages of conventional therapies (surgery, chemotherapy and radiation therapy) was listed and yoga modules were developed accordingly. Considering the needs, expected side effects, the energy levels and the psychological states of the participants, eight modules evolved.The results of the six steps for developing the validated module are reported. Step 1: Literature review from traditional yoga and ayurveda texts on etiopathogenesis and management of cancer (arbuda), and the recent literature on cancer stem cells and immunology of cancer. Step 2: Focused group discussions and deliberations to compile the needs of patients based on the expected side effects, energy levels and the psychological state of the patient as observed by the caregivers and the clinicians. Step 3: Content validation through consensus by the experts for the eight modules of IAYTC that could be used as complimentary to conventional management of cancer at different stages during and after the diagnosis was created. Step 4: Field testing for safety and feasibility of the modules through pilot studies. Step 5: Compilation of the results of efficacy trials through RCTs and step 6: A review of our studies on mechanisms to offer evidence for action of IAYTC on psycho-neuro-immunological pathways in cancer.The evidence from the traditional knowledge and recent scientific studies validates eight modules of integrated approach of yoga therapy for cancer that can be used safely and effectively as complimentary during all conventional cancer therapies.

The two neighboring southwestern states of India, Karnataka and Maharashtra, have high incidence of HIV/AIDS and are among the six most high prevalence HIV infected states. In Karnataka state, the northern districts of Bagalkot, Belgaum and Bijapur (the three Bs) and in Maharashtra state, the southern districts of Sangli, Satara, and Solapur (the three Ss) are the areas with the highest incidence of HIV/AIDS. We have evaluated the incidence of maternal to child transmission (MTCT) of HIV-1 infection in Belgaum District which is more than 500 kilometers distance by road from the campus in greater Bangalore (Karnataka State). We have obtained the prenatal CD4 counts of HIV infected pregnant mothers. We have also screened the HIV infected children in two orphanages (rehabilitation centres for HIV infected children) in Belgaum District. The clinical conditions of these infected children were assessed for their CD4 counts, anti-retroviral therapy (ART) intake status, outpatient illnesses and body composition. We have observed that there is an influence of the age factor on the CD4 counts of the HIV infected children. Further, in view of the role of our recently found involvement of sulfatide, 3-O- galactosylceramide, in inhibition of HIV-1 replication and enhancement of hematopoiesis which is otherwise inhibited due to such infection, we have discussed the possible role of sulfatides that biologically occur in the fetal adnexa (placentatrophoblasts /amnion/chorion-umbilical cord), in containing HIV infection as a potential safer alternative to the ART regimens currently approved to be clinically practiced. Lastly, we have discussed the complementary and alternative medicine (CAM) therapies such as evidence based yoga and ayurveda as add-on to ART in potential elimination of MTCT of HIV infection. Out of a total of 150 children delivered by HIV infected mothers, 13 children were found to be positive as determined by the dried blood smear (DBS) for virological testing, giving an incidence of about 8.66% in the Belgaum district during the last two years, in spite of the prescription of currently available ART regimens. All the 13 HIV-transmitting mothers had normal vaginal deliveries. Though 12% of the total 150 deliveries required lower segment caesarean section (LSCS), none among them resulted in MTCT of HIV. Comparison of the prenatal CD4 counts between transmitting and non-transmitting mothers did not show significant differences (p=0.25) thus suggesting indirectly that HIV-1 proviral loads (undetermined / unavailable) need not necessarily determine the fate of incidence of vertical transmission. The mean age of 44 HIV infected children (14 females, 30 males) that were screened in two orphanages was 10.8±3.1 years. Out of these 44 children, 27 were taking ART (61.36%) with mean duration of consumption being 2.8±2.28 years. Fifty percent (n=22) of the children were suffering from at least one outpatient illness, out of which 13 were taking ART. Their mean basal metabolic rate (BMR), body mass index (BMI), muscle mass, fat mass and fat % were 795.45±106.9, 14.55±1.9 kg/m(2), 9.54±3.4 kg, 3.69±2.24 kg and 15.04±7.8% respectively. Comparison between the children taking ART (on-ART, n=27) and those not taking ART (non-ART, n= 17) showed that though there was no significant difference in the average age of the two groups, on-ART children had significantly higher BMR (p=0.05), and muscle mass (p=0.004), than non-ART. The CD4 counts, BMI, fat mass and fat percentage did not show significant statistical differences between the two groups. The CD4 counts of the children (both on-ART and non-ART) of age 8 years and below (n=12) were found to be significantly higher (p=0.04) than those of age 14 and above (n=10). All the children in age group of 14 years and above (n=10) except one child were on ART, whereas 7 out of 12 children in age group of 8 years and below were on-ART. In one of the rehabilitation centers called Aadhar, among non-ART children, a significant correlation was observed between the age of the child and CD4 counts (measured separately in the months of June 2011 and December 2011). Both the CD4 counts measured in June 2011 (n=6; r=-0.82, p= 0.04) as well as in December 2011 (n=6; r=-0.97, p=0.001) showed a significant decline as the age progressed. Also, at the same center, among on-ART children, the CD4 counts in June 2011 (n=7) and December 2011 (n=8) were significantly different between the children in the age group of 8 below years, and those in the age group of 14 years and above (p= 0.005). As HIV infected children grow in age, they may lose maternal derived immunity as shown by the decrease in CD4 counts, irrespective of their ART status. It is to be expected from these results that the conferred maternal immunity (possibly primarily humoral and secondarily cytotoxic immune responses) to the virus acquired at child birth taper off and eventually overcome by the generation of mutant HIV strains in the children, as the life spans of the infected children progress. We have discussed safer therapeutic interventions whose efficacy on HIV/AIDS may be synergistic to or even substitute the existing treatment strategies. Some of such interventions may even be customized to help eliminate MTCT. Further, these virus infected pregnant mother patient blood / serum samples could prove useful in the vaccine development against HIV infection.

New combination drug treatment(s) now available to patients with HIV-1 infection allows them to live longer lives with good quality of life although they suffer from the incurable HIV-1 infection. In a previous study we found that sulfatide was efficient in lowering HIV-1 viral loads in SCID mice engrafted with human fetal liver/thymus tissues (SCID-hu). Current antiviral treatments carry an increased risk of other complications like cardiovascular disease and diabetes after long-term use. There is a need for new potent safe pharmaceutical agents. Endogenous sulfatide is a mixture of -isoforms, i.e. sulfatide molecules with different long-chain bases and fatty acid chain lengths and saturation. Sulfatide isoforms may have different physicochemical properties i.e, they are of different potency at different target cells. Other investigators have shown that incubation of cultured cells with sulfatide incorporated into the plasma membrane inhibited HIV-1 entry into the cells thereby inhibiting intracellular HIV-1 replication. We have shown that CD1d dependent stimulation by sulfatide may activate pDC antigen expressing cells that produce type I inteferons. Type I inteferons are known to reduce HIV-1 replication. This could provide a second likely explanation (after the inhibition of virus entry) for the more efficient lowering of HIV-1 viral loads in sulfatide versus AZT treated mice. This review aims to show the efficiency of sulfatide in reducing HIV-1 viral loads as compared to conventional HAART treatment. We also discuss the risks of HAART treatment and propose a clinical alternative of sulfatide in HIV-1 infection.

Although microglia are the only cells found to be productively infected in the central nervous system of acquired immunodeficiency disease syndrome (AIDS) patients, there is extensive white and gray matter disease nonetheless. This neuropathogenesis is believed to be due to indirect mechanisms other than infection with human immunodeficiency virus 1 (HIV-1). Cytokines and toxic small molecules have been implicated in the clinical and histopathological findings in CNS AIDS. Previously, we have demonstrated in rodent glial cultures the presence of biologically active epitopes of gp120 and gp41 that are capable of inducing interleukin 1 and tumor necrosis factor alpha. In this study, we map the HIV-1 envelope epitopes that induce nitric oxide, inducible nitric oxide synthase, interleukin 1, and tumor necrosis factor alpha in human glial cultures. Epitopes in the carboxy terminus of gp120 and the amino terminus of gp41 induce these proinflammatory entities. In addition, we compare HIV-1 infection and pathology in glial cells derived from human brain taken at different states of maturation (fetal, neonatal, and adult brain) in an effort to address some of the clinical and histological differences seen in vivo. This study demonstrates that, in the absence of virus infection and even in the absence of distinct viral tropism, human glia respond like rodent glia to non-CD4-binding epitopes of gp120/gp41 with cytokine and nitric oxide production. Differences among fetal, neonatal, and adult glial cells' infectivity and cytokine production indicate that, in addition to functional differences of glia at different stages of development, cofactors in vitro and in vivo may also be critical in facilitating the biological responses of these cells to HIV-1.

The pathology in central nervous system (CNS) AIDS suggests that direct infection with HIV-1 is not required for changes in glia and neurons. Induction of a variety of pathological responses in vitro in rodent brain cultures also suggests that CD4 is not the receptor for HIV-1 in the brain, given that human and rodent CD4 are not homologous. This implies that the epitopes on HIV-1 which bind glia and activate them are novel, non-CD4-binding domains. We have therefore mapped the envelope (env) regions required for production in rat glial cultures of interleukin-1 (IL-1) and tumor necrosis factor alpha (TNFα) which we hypothesize are important in CNS AIDS. Serially truncated deletion mutants from the gp120/gp41 carboxy terminus representing folded, glycosylated recombinant env proteins were expressed in HeLa cells via a vaccinia virus vector. These proteins, linear gp120/gp41 peptides, as well as polyclonal and monoclonal antibodies reactive to defined regions of gp120/gp41 were used to map the epitopes involved in production of IL-1 and TNFα. Compared to HeLa cell and wild-type vaccinia virus controls, the vaccinia recombinant env protein gp160 containing cleaved gp120 and gp41 induced both IL-1 and TNFa. If gp160 was not cleaved into gp120 and gp41, IL-1 but not TNFa induction was reduced. Peak production of TNFa by gp120/gp41 was at 4 h while IL-1 production was still significantly elevated at 44 h at the highest concentrations of env protein. Using the truncation deletions, the V3 loop of gp120 appeared to be critical for IL-1. Glycosylation and folding of V3 is probably important in IL-1 induction since a V3 peptide was not as active. While removal of glycosylated, folded V4 and C4 regions had no effect on IL-1, linear peptides in the region from the V4 loop to the C4 domain were strong inducers of IL-1. Non-glycosylated, linear V4 loop peptide induced more IL-1 than the V4 in protein generated in HeLa cells, suggesting that glycosylation and/or conformational structures sequester V4 inducer epitopes. Using the truncation deletions, the carboxy terminus region (V4-C5) of gp120 as well as gp41 were shown to be critical for TNFa production. Peptides representing linear epitopes in the V3 loop, C5 domain of gp120, and the ectodomain of gp41 were all strong inducers of TNFα; a protein representing almost the entire gp41 was the strongest inducer of TNFα. This suggests both conformational and linear determinants induce TNFα. Glycosylated, folded VI-3 and C1-3, exclusive of other gp120 domains, did not induce TNFα production. These data were further confirmed by antibody inhibition of env regions. Cytokine induction by env proteins was transcriptionally regulated, since messenger RNA levels were increased as determined by Northern blot analysis. In summary, virus infection is not required and gp160 epitopes, other than the classical CD4 binding domains, are involved in induction of IL-1 and TNFa in glial cell cultures. These data suggest that HIV-1 env proteins in their native form, as well as those that are partially denatured (linearized or non-glycosylated) could induce two proinflammatory cytokines which are possibly important in CNS AIDS.

This article is expected to contribute towards understanding the therapeutic benefits of specific yoga modules on the inhibition of replication and enhancement to normal levels of hematopoiesis in HIV-1 infected subjects. More unique could be the effects of yoga on the indirect effects of HIV-1 induced hematopoietic inhibition of the CD34+ progenitor stem cells, via the CD4+ T lymphocytes. Such indirect effects may be caused by host cellular factors. Yoga practices may also improve the self renewal capacity (a step that precedes commitment of CD34+ progenitor cells to terminal differentiation), via STAT5 gene regulation. This may eliminate the need for constitutive STAT5 gene expression through gene therapy. In this article recent research and ancient Indian literature are reviewed to devise yoga modules for the potential treatment of hematopoietic inhibition in HIV-1 infection. The possible mechanisms through which hematopoietic inhibition may occur in HIV-1 infected patients are first described followed by the role of stress in the progression of HIV where probable involvement of psycho-neuro-immunological axis (PNI) is highlighted. Yoga therapy is introduced and its effectiveness in terms of evidence in relevant area is reviewed. Further, the basic principles of Integrated Approach of Yoga Therapy [IAYT] are described and depending on the potential mechanisms through which yoga therapy may act, both modern scientific research and ancient "scriptural" evidence are provided at all the five levels of existence (body, life force, emotional, intellectual and bliss). This will enable to design comprehensive yoga modules that may intervene in this indirect inhibition of haematopoiesis in HIV-1 infected individuals and potentially restore normal levels of haematopoiesis.

The unusually high 88% one-year cadaver kidney graft survival rate in patients with IgA nephropathy (IgAN) prompted us to investigate the influence of IgA anti-HLA class I antibodies on subsequent graft survival. We found that patients with various original diseases with IgA antibodies to the HLA molecule had high 91% one-year graft survival compared with 58% one-year survival for those who did not have preformed IgA antibodies against the HLA molecule prior to transplantation (P < 0.0005). The IgA antibodies were detected by reaction with class I HLA molecules isolated by capture with monoclonal antibody and detected with an enzyme-linked immunosorbent assay. In contrast, IgG antibodies to the HLA molecule resulted in a lower one-year graft survival rate (74%) than in those patients without IgG antibodies (87%) (p = 0.08). IgA antibodies to the HLA molecule, when present, tended to react at a high frequency on a random lymphocyte panel. These findings suggest that sensitization resulting in IgA anti-HLA antibodies may counteract the deleterious effect of an IgG antibody response in clinical kidney transplantation.

The pathogenesis of HIV-1 infection is a complex process in which Natural Killer (NK) and Natural Killer T (NKT) cells play an important role. NKT cells express markers for NK cells and a TCR of the conventional T cells and recognize lipid antigens presented by the non-polymorphic CD1 molecule. CD1d-restricted type I NKT cells express an invariant TCR and can recognize αGalCer, whereas a major subset of type II NKT expressing diverse TCR can recognize a self-glycolipid, sulfatide. It has been shown that CD4+ type I NKT cells are infected by HIV-1 and decreased in HIV-1-infected individuals. However, their exact role in HIV-1 infection as well as the biology and function of the type II NKT cell subset in HIV-1 infection and disease progression are not known. Our earlier studies have shown that activation of CD1d-restricted type II NKT cells by sulfatide and their interactions with plasmacytoid dendritic (pDC) and myeloid dendritic (mDC) cells result in anergy induction in type I NKT cells in several models. Here we used SCID-Hu (Thy/Liv) animals, co-implanted with human fetal liver and thymus, and found that these implants contain both type I and type II NKT cells, CD161+CD3+ NKT cells, NK cells and dendritic cells during HIV-infection. We found that the administration of sulfatide (bi-weekly, 20 μg/animal, i.p.) in SCID-Hu animals inhibits HIV-1 replication more efficiently than treatment with the nucleoside analog reverse transcriptase inhibitor, AZT. Virus replication was lowered significantly up to 4-8 weeks post infection. Furthermore sulfatide administration also resulted in significant retention of hematopoeisis that is lost during HIV-1 infection. Advantageously, sulfatide administration itself was not associated with anemia or bone marrow suppression, that are severe side effects of HAART. Since the CD1d-mediated immune pathway is highly conserved between rodents and humans, sulfatide treatment may represent a novel HLA-independent approach for intervention of HIV-1 pathogenesis.

Paraoxanase-2 (PON2) activity was increased upon HIV-1 infection of the CD34+CD4+ hematopoietic cell line TF-1. Thymocytes derived from the human fetal conjoint thymus/liver hematopoietic organ of SCID-hu mice also exhibited an increase in PON2 activity. Additionally, a remarkable increase of PON2 mRNA expression was also observed in both TF-1 and thymocytes following HIV-1 infection. The phosphorylation of STAT5 was decreased in TF-1 cells upon HIV-1 infection. Interestingly, phosphorylation of STAT5 does not occur in GM-CSF "starved" TF-1 cells; however, PON2 protein, activity and mRNA expression are increased under these conditions, similar to HIV-1 infection. We conclude that PON2 is induced in HIV-1 infection through a mechanism that may involve STAT5 inactivation.

Tumors contain a minority population of cancer stem cells (CSC) that maintain the tumor. In marked contrast to the CSC, the tumor cells have either no capacity or a markedly diminished capacity, to form new tumors. Therefore, to treat cancer effectively, the CSC must be eliminated. The tumor will rapidly recur if the therapy eliminates cancer cells but spares a significant population of the cancer stem cells. We report here for the first time the utilization of the SCID-hu Thy/Liv chimeric small animal model system for induction of human cancer stem cells and their early detection. This model system allows long-term systemic human T-cell reconstitution in vivo, and also provides both human antigen-presenting cells (APCs) and effector cells for induction of anti-tumor immune responses. We were able to generate human hematopoietic cancer stem cells (HCSC) using the SCID-hu Thy/Liv system, and confirmed the expression of both the CD34 marker and two human tumor antigens in these cells. Importantly, we observed an enhanced expression of several embryonic stem cell markers in the HCSC, as well as morphological appearance typical for undifferentiated cells, suggesting the acquisition of highly malignant aggressive properties. Therefore, these findings are an important step in our understanding of the mechanisms involved in the early formation of the cancer stem cells, and in demonstrating the stem cell origin of cancer.

Cytopenias arising from hematopoietic abnormalities are a severe common complication contributing to early mortality in HIV/AIDS patients. The proto-oncogene c-mpl, identified as the thrombopoietin receptor is involved in multilineage differentiation of CD34+ hematopoietic progenitor cells. We have introduced the c-mpl gene into CD34+ cells via transduction of the lentivirus p156RRLsinPPTmPGK-CMPL-PRE. The lentiviral construct expresses c-mpl on approximately 90% of purified CD34+ cells. These transduced cells have then been reconstituted into human fetal thymus/liver implants in severe combined immunodeficient mice (SCID-hu Thy/Liv). The c-mpl expression on transduced CD34+ cells is not susceptible to downregulation due to the effects of HIV-1 infection. Reconstituted CD34+ cells transduced with control lentivirus, p156RRLsinPPTmPGK-EGFP-PRE, express EGFP at90%. Reconstituted c-mpl expressing SCID-hu implants show almost maximum rescue (approximately 90%) of myelopoiesis, erythropoiesis and megakaryopoiesis, during HIV-1 infection in vivo, at 6 weeks post-infection. We also show that the differentiated multi-lineage progeny colonies and thymocytes in mice reconstituted with the c-mpl transduced CD34+ cells, carry the HLA Class I loci phenotypes of these donor cells, in the implants of the recipient SCID-hu animals. We propose a gene therapeutic strategy, with c-mpl as the major genetic component, to address the morbidity and mortality resulting from cytopenias in HIV infected patients.