Prevalence and epidemiological characteristics of methicillin-resistant Staphylococcus aureus isolated from cattle in Bangalore India as a part of the One Health approach.

In India, limited studies are available on the epidemiological aspects of methicillin-resistant Staphylococcus aureus (MRSA) infections in both animal and human settings. Herein, we investigated the prevalence, antimicrobial resistance profile and molecular characteristics of MRSA isolates recovered from cattle using the One Health approach. Out of 66 mecA-positive staphylococci, species-specific multiplex PCR detected 24 % (n=16) of isolates as MRSA. Maximum antibiotic resistance was seen against cloxacillin (94 %, n=15) and least for enrofloxacin and cephalothin (each 13 %, n=2). Overall, 13 % (n=2) of MRSA isolates were multidrug-resistant. Molecular characterization by SCCmec typing identified 88 % (n=14) of MRSA isolates as type V. Twelve isolates (75 %) belonged to novel spa-type t17242, of which 67 % (n=8) belonged to agr type I. MLST analysis revealed ST 1687 (50 %, n=8) as the most predominant sequence type. Circulation of different MRSA clones among the cattle populace offers a risk of transmission to humans through direct contact, food chain or environmental contamination. Thus, continuous monitoring of MRSA strains is imperative for early diagnosis and for establishing effective treatment strategies to restrain the disease burden caused by MRSA infections.

Novel pneumococcal capsule type 33E results from the inactivation of glycosyltransferase WciE in vaccine type 33F.

The polysaccharide (PS) capsule is essential for immune evasion and virulence of Streptococcus pneumoniae. Existing pneumococcal vaccines are designed to elicit anticapsule antibodies; however, the effectiveness of these vaccines is being challenged by the emergence of new capsule types or variants. Herein, we characterize a newly discovered capsule type, 33E, that appears to have repeatedly emerged from vaccine type 33F via an inactivation mutation in the capsule glycosyltransferase gene, wciE. Structural analysis demonstrated that 33E and 33F share an identical repeat unit backbone [→5)-ß-D-Galf2Ac-(1→3)-ß-D-Galp-(1→3)-α-D-Galp-(1→3)-ß-D-Galf-(1→3)-ß-D-Glcp-(1→], except that a galactose (α-D-Galp) branch is present in 33F but not in 33E. Though the two capsule types were indistinguishable using conventional typing methods, the monoclonal antibody Hyp33FM1 selectively bound 33F but not 33E pneumococci. Further, we confirmed that wciE encodes a glycosyltransferase that catalyzes the addition of the branching α-D-Galp and that its inactivation in 33F strains results in the expression of the 33E capsule type. Though 33F and 33E share a structural and antigenic similarity, our pilot study suggested that immunization with a 23-valent pneumococcal PS vaccine containing 33F PS did not significantly elicit cross-opsonic antibodies to 33E. New conjugate vaccines that target capsule type 33F may not necessarily protect against 33E. Therefore, studies of new conjugate vaccines require knowledge of the newly identified capsule type 33E and reliable pneumococcal typing methods capable of distinguishing it from 33F.

Estimation of baseline IgG antibody levels to 23 pneumococcal vaccine-type capsular polysaccharides in healthy vaccine naïve Indian adults.

Since immunological responses to pneumococcal vaccines are assessed by a fold-increase in antibody levels relative to pre-immunization levels, it is therefore critical to determine baseline antibody levels to establish putative threshold as a measure of normal response. Herein, for the first time, we measured baseline IgG antibody levels in 108 healthy unvaccinated Indian adults using WHO-recommended ELISA. Median baseline IgG concentration ranged between 0.54 µg/mL to 12.35 µg/mL. Highest levels of baseline capsule polysaccharide (cPS)-specific IgG were found against types 14, 19A, and 33F. Whereas, lowest baseline IgG levels were observed against types 3, 4, and 5. Overall, ∼79% of study population had median baseline IgG levels ≥1.3 µg/mL against 74% of cPS's. Substantial baseline antibody levels in unvaccinated adults were observed. The study would be critical in bridging gaps in baseline immunogenicity data and may offer a valuable foundation for evaluating immune response of Indian adults to pneumococcal vaccination.

Clinical Profile, Management, and Outcome of Visceral Artery Pseudoaneurysms: 5-Year Experience in a Tertiary Care Hospital.

Visceral artery pseudoaneurysms are potentially lethal lesions and tend to rupture in a high proportion of cases, thereby warranting an immediate and active intervention. We present our experience of splanchnic visceral artery pseudoaneurysms in a university hospital over a 5-year time interval with emphasis on etiology, clinical presentation, management (endovascular/surgical), and final outcome. This was a retrospective study in which we searched our image database for pseudoaneurysms of visceral arteries over a period of 5 years. The clinical and operative details were retrieved from the medical record section of our hospital. The lesions were analyzed for the vessel of origin, size, etiology, clinical features, mode of treatment, and outcome. Twenty-seven patients with pseudoaneurysms were encountered. Pancreatitis (8) was the most common cause, followed by previous surgery (7) and trauma (6). Fifteen were managed by the interventional radiology (IR) team, 6 by surgery, and in 6 no intervention was done. Technical and clinical success was achieved in all patients in the IR group with few minor complications. Surgery and no intervention carry a high mortality in such a setting (66 and 50%, respectively). Visceral pseudoaneurysms are potentially fatal lesions, commonly encountered after trauma, pancreatitis, surgeries, and interventional procedures. These lesions are easily salvageable by minimally invasive interventional techniques (endovascular embolotherapy), and surgeries carry a lot of morbidity and mortality in such cases and a prolonged hospital stay.

Discovery and Characterization of Pneumococcal Serogroup 36 Capsule Subtypes, Serotypes 36A and 36B.

Streptococcus pneumoniae can produce a wide breadth of antigenically diverse capsule types, a fact that poses a looming threat to the success of vaccines that target pneumococcal polysaccharide (PS) capsule. Yet, many pneumococcal capsule types remain undiscovered and/or uncharacterized. Prior sequence analysis of pneumococcal capsule synthesis (cps) loci suggested the existence of capsule subtypes among isolates identified as "serotype 36" according to conventional capsule typing methods. We discovered these subtypes represent two antigenically similar but distinguishable pneumococcal capsule serotypes, 36A and 36B. Biochemical analysis of their capsule PS structure reveals that both have the shared repeat unit backbone [→5)-α-d-Galf-(1→1)-d-Rib-ol-(5→P→6)-ß-d-ManpNAc-(1→4)-ß-d-Glcp-(1→] with two branching structures. Both serotypes have a ß-d-Galp branch to Ribitol. Serotypes 36A and 36B differ by the presence of a α-d-Glcp-(1→3)-ß-d-ManpNAc or α-d-Galp-(1→3)-ß-d-ManpNAc branch, respectively. Comparison of the phylogenetically distant serogroup 9 and 36 cps loci, which all encode this distinguishing glycosidic bond, revealed that the incorporation of Glcp (in types 9N and 36A) versus Galp (in types 9A, 9V, 9L, and 36B) is associated with the identity of four amino acids in the cps-encoded glycosyltransferase WcjA. Identifying functional determinants of cps-encoded enzymes and their impact on capsule PS structure is key to improving the resolution and reliability of sequencing-based capsule typing methods and discovering novel capsule variants indistinguishable by conventional serotyping methods.

Heat distribution and the condition of hypothermia in the multi-layered human head: A mathematical model.

The conduction, perfusion and metabolic heat generation based partial differential equation has been used to study the heat transfer in human head. The main objective of this study is to predict the temperature distribution at the multi-layered human head that results in hypothermic condition. The temperature profiles have been estimated at the interface points of brain, skull and scalp with respect to various parameters including atmospheric temperature, arterial temperature and metabolic heat generation. The variational finite element method and analytical method based on Laplace transform has been employed to establish the solution of the formulated model, and the resulting outcomes are illustrated graphically. Under cold exposure, the blood capillaries around scalp exchange core heat with the external cold environment and experience lowering in the tissue temperature of the blood in the scalp. It is reflected in the graphical view of the model that the prolonged exposure to cold transmits its effect into the deep brain capillaries, wherein the temperature gradually lowers down below the normal body temperature that results hypothermia and hence abnormal body homoeostasis.

The effectiveness of image-guided percutaneous catheter drainage in the management of acute pancreatitis-associated pancreatic collections.

PURPOSE: Acute pancreatitis is commonly complicated by the development of pancreatic collections (PCs). Symptomatic PCs warrant drainage, and the available options include percutaneous, endoscopic, and open surgical approaches. The study aimed to assess the therapeutic effectiveness and safety of image guided percutaneous catheter drainage (PCD) in the management of acute pancreatitis related PCs. MATERIAL AND METHODS: This was a single-centre prospective study covering a 4-year study period. Acute pancreatitisrelated PCs complicated by secondary infection or those producing symptoms due to pressure effect on surrounding structures were enrolled and underwent ultrasound or computed tomography (CT)-guided PCD. The patients were followed to assess the success of PCD (defined as clinical, radiological improvement, and the avoidance of surgery) and any PCD-related complications. RESULTS: The study included 60 patients (60% males) with a mean age of 43.1 ± 21.2 years. PCD recorded a success rate of 80% (16/20) for acute peripancreatic fluid collections (APFC) and pancreatic pseudocysts (PPs), 75% (12/16) for walled-off necrosis (WON), and 50% (12/24) for acute necrotic collections (ANCs). Post-PCD surgery (necrosectomy ± distal pancreatectomy) was needed in 50% of ANC and 25% of WON. Only 20% of APFCs/PPs patients required surgical/endoscopic treatment post-PCD. Minor procedure-related complications were seen in 4 (6.6%) patients. CONCLUSION: PCD is an effective, safe, and minimally invasive therapeutic modality with a good success rate in the management of infected/symptomatic PCs.

The spectrum of imaging findings in pulmonary hydatid disease and the additive value of T2-weighted magnetic resonance imaging in its diagnosis.

PURPOSE: To describe the spectrum of imaging findings in pulmonary echinococcosis and to study the additive value of T2-weighted magnetic resonance imaging (MRI) in the characterisation of pulmonary hydatid disease. MATERIAL AND METHODS: This was a descriptive, prospective study conducted for a period of 3 years from December 2016 to November 2019. Patients suspected of having pulmonary echinococcosis (n = 110) on preliminary chest radiography were examined with chest computed tomography (CT). Among them 41 cases were additionally examined with T2-weighted MRI of thorax. Final diagnosis was based on surgery or histopathology. RESULTS: Of the 110 patients enrolled for the study 15 were lost to attrition, and among the final cohort of 95 patients CT correctly diagnosed 68/84 (80.9%) as hydatid cyst, whereas 16/84 (19.1%) received an erroneous alternate diagnosis on CT. Based on the classical findings of hyperintense pulmonary cystic lesion with T2-weighted hypointense rim or detached internal T2-weighted hypointense membrane, a correct diagnosis of hydatid cyst was possible in 30 patients whereas a correct alternate diagnosis was made in 8 cases. T2-weighted MRI was found to have sensitivity of 96.7%, specificity of 80%, positive predictive value (PPV) of 93.7% and negative predictive value (NPV) of 88.9% with an overall diagnostic accuracy of 92.6%. Using the McNemar test, MRI was found to be diagnostically superior to CT (p = 0.019). CONCLUSIONS: Most of the pulmonary hydatid cysts can be diagnosed on CT; however, sometimes the findings may be indeterminate or atypical, leading to a diagnostic dilemma. MRI, owing to its ability to demonstrate hypointense endocyst, can act as a useful adjunct to correctly diagnose hydatid cyst or suggest an alternative diagnosis.

Evaluation of various patient-, lesion-, and procedure-related factors on the occurrence of pneumothorax as a complication of CT-guided percutaneous transthoracic needle biopsy.

PURPOSE: To assess the influence of various patient-, lesion-, and procedure-related variables on the occurrence of pneumothorax as a complication of CT-guided percutaneous transthoracic needle biopsy. MATERIAL AND METHODS: In a total of 208 patients, 215 lung/mediastinal lesions (seven patients were biopsied twice) were sampled under CT guidance using coaxial biopsy set via percutaneous transthoracic approach. Incidence of post procedure pneumothorax was seen and the influence of various patient-, lesion-, and procedure-related variables on the frequency of pneumothorax with special emphasis on procedural factors like dwell time and needle-pleural angle was analysed. RESULTS: Pneumothorax occurred in 25.12% (54/215) of patients. Increased incidence of pneumothorax had a statistically significant correlation with age of the patient (p = 0.0020), size (p = 0.0044) and depth (p = 0.0001) of the lesion, and needle-pleural angle (p = 0.0200). Gender of the patient (p = 0.7761), emphysema (p = 0.2724), site of the lesion (p = 0.9320), needle gauge (p = 0.7250), patient position (p = 0.9839), and dwell time (p = 0.9330) had no significant impact on the pneumothorax rate. CONCLUSIONS: This study demonstrated a significant effect of the age of the patient, size and depth of the lesion, and needle-pleural angle on the incidence of post-procedural pneumothorax. Emphysema as such had no effect on pneumothorax rate, but once pneumothorax occurred, emphysematous patients were more likely to be symptomatic, necessitating chest tube placement. Gender of the patient, site of the lesion, patient position during the procedure, and dwell time had no statistically significant relation with the frequency of post-procedural pneumothorax. Surprisingly, needle gauge had no significant effect on pneumothorax frequency, but due to the small sample size, non-randomisation, and bias in needle size selection as per lesion size, further studies are required to fully elucidate the causal relationship between needle size and post-procedural pneumothorax rate. The needle should be as perpendicular as possible to the pleura (needle-pleural angle close to 90°), to minimise the possibility of pneumothorax after percutaneous transthoracic needle biopsy.

Evaluation of intracranial tuberculomas using diffusion-weighted imaging (DWI), magnetic resonance spectroscopy (MRS) and susceptibility weighted imaging (SWI).

OBJECTIVE:: The present study was aimed to evaluate patients of suspected intracranial tuberculomas with diffusion-weighted imaging (DWI), magnetic resonance spectroscopy (MRS) and susceptibility-weighted imaging (SWI). METHODS:: The present study evaluated 116 patients known or suspected of having central nervous system tuberculosis with advanced MRI techniques comprising of DWI, MRS and SWI in addition to the conventional MRI. RESULTS:: Apparent diffusion coefficient value of tuberculomas was not significantly different (p > 0.05) from apparent diffusion coefficient value of metastatic lesions and high-grade gliomas. MRS revealed that NAA/Cr and NAA/Cho ratios of tuberculomas were not significantly different (p > 0.05) from that of malignant brain lesions. However, Cho/Cr ratio of tuberculomas (1.36 ± 0.41) was significantly lower from that of malignant brain lesions (2.63 ± 0.99). SWI revealed a complete and regular hypointense peripheral ring in 42 cases of tuberculomas (58%) and in none of the malignant brain lesions. CONCLUSION:: DWI offers no clear advantage in differentiating tuberculomas from metastasis and gliomas. Tuberculomas may be differentiated from metastases and gliomas by their unique metabolite pattern on MRS. Presence of a complete and regular peripheral hypointense ring in SWI favors the diagnosis of tuberculomas. ADVANCES IN KNOWLEDGE:: The results from the present study suggest promising role of SWI in the discrimination of tuberculomas from metastatic brain lesions and gliomas with the presence of a complete and regular peripheral hypointense ring favoring the diagnosis of tuberculomas.

A comparison of imaging guided double percutaneous aspiration injection and surgery in the treatment of cystic echinococcosis of liver.

OBJECTIVE: To compare the results and complications of treatment by double percutaneous aspiration injection (DPAI) in cystic echinococcosis (CE) of the liver with those of surgery. To the best of our knowledge, such a study has not been carried out till date. METHODS: From November 2012 to November 2015, 43 patients were randomly allocated to DPAI group (n = 22) and surgery group (n = 21). After the intervention, patients were evaluated monthly for 3 months, then at the sixth month and 6 monthly thereafter. RESULTS: Average hospital stay was 2.38 days in DPAI group and 8.23 days in the surgery group. Response to DPAI was categorized as successful in 95.3% (n = 20) patients and incomplete in 4.7% (n = 1) patients. Response to surgery was characterized as successful in 85.7% (n = 18) patients and incomplete in 4.7% (n = 1) patients, and recurrence was seen in 9.5% (n = 2) patients. Using a 10% margin for non-inferiority, treatment response in the DPAI group was non-inferior to that of the surgery group. In the DPAI group, 19 patients had no complications, minor complications were seen in 4.7% (n = 1) patients and a major complication was seen in 4.7% (n = 1) patients. In the surgery group, no complications were seen in 13 patients, major complications were seen in 28.57% (n = 6) patients and minor complications were seen in 9.5% (n = 2) patients. CONCLUSION: Over a follow-up period of 3 years, DPAI is non-inferior to surgery in the treatment of CE of the liver, while there is a statistically significant difference in the hospital stay and occurrence of complications. Advances in knowledge: DPAI offers advantages such as a short hospital stay, minimal invasiveness and morbidity, while being non-inferior to surgery. Total Immunoglobulin G antibody titres have limited utility in follow-up of patients treated.

Impact of centralization of pancreaticoduodenectomy coupled with fast track recovery protocol: a comparative study from India.

BACKGROUND: Fast track strategy in the management of patients undergoing intra-abdominal surgery of various types has emerged as a landmark approach to reduce surgical stress and accelerate recovery. This study was to evaluate the effect of fast track strategy on patients subjected to pancreaticoduodenectomy (PD) from an individual unit during transit from low to a high volume center. METHODS: A total of 142 PD patients who had been subjected to fast track strategy between June 2008 and September 2012 were compared with 46 patients who had received conventional surgery between January 2006 and May 2008. Comparative analysis was made of postoperative complications, postoperative recovery, length of hospital stay and patient readmission requirement. RESULTS: The patients subjected to fast track strategy had a faster recovery and a shorter hospital stay than those who were treated conventionally (7.8 vs 12.1 days). The intraoperative events like operative blood loss (417.9+/-83.8 vs 997.4+/-151.8 mL, P<0.001), blood transfused (a median of 0 vs 1 unit, P<0.001) and operative time taken (125 vs 245 minutes, P<0.001) were significantly lower in the fast track group. The frequency of pancreatic fistula (4.9% vs 13.0%) and delayed gastric emptying (7.0% vs 17.4%) was also significantly reduced with fast track treatment. Nevertheless, the readmission rate (11.3% vs 6.5%) was found relatively higher within the fast track group. However, increased readmission rates in this study seem to be independent of fast track protocol. CONCLUSIONS: This preliminary analysis suggests that the fast track approach might be beneficial to the well-being of the patients after PD, for it accelerates the immediate clinical recovery of patients and significantly shortens their length of hospital stay.

Computational Identification of Tumor Anatomic Location Associated with Survival in 2 Large Cohorts of Human Primary Glioblastomas.

BACKGROUND AND PURPOSE: Tumor location has been shown to be a significant prognostic factor in patients with glioblastoma. The purpose of this study was to characterize glioblastoma lesions by identifying MR imaging voxel-based tumor location features that are associated with tumor molecular profiles, patient characteristics, and clinical outcomes. MATERIALS AND METHODS: Preoperative T1 anatomic MR images of 384 patients with glioblastomas were obtained from 2 independent cohorts (n = 253 from the Stanford University Medical Center for training and n = 131 from The Cancer Genome Atlas for validation). An automated computational image-analysis pipeline was developed to determine the anatomic locations of tumor in each patient. Voxel-based differences in tumor location between good (overall survival of >17 months) and poor (overall survival of <11 months) survival groups identified in the training cohort were used to classify patients in The Cancer Genome Atlas cohort into 2 brain-location groups, for which clinical features, messenger RNA expression, and copy number changes were compared to elucidate the biologic basis of tumors located in different brain regions. RESULTS: Tumors in the right occipitotemporal periventricular white matter were significantly associated with poor survival in both training and test cohorts (both, log-rank P < .05) and had larger tumor volume compared with tumors in other locations. Tumors in the right periatrial location were associated with hypoxia pathway enrichment and PDGFRA amplification, making them potential targets for subgroup-specific therapies. CONCLUSIONS: Voxel-based location in glioblastoma is associated with patient outcome and may have a potential role for guiding personalized treatment.

Standardisation and evaluation of a quantitative multiplex real-time PCR assay for the rapid identification of Streptococcus pneumoniae.

Rapid diagnosis of Streptococcus pneumoniae can play a significant role in decreasing morbidity and mortality of infection. The accurate diagnosis of pneumococcal disease is hampered by the difficulties in growing the isolates from clinical specimens and also by misidentification. Molecular methods have gained popularity as they offer improvement in the detection of causative pathogens with speed and ease. The present study aims at validating and standardising the use of 4 oligonucleotide primer-probe sets (pneumolysin [ply], autolysin [lytA], pneumococcal surface adhesion A [psaA] and Spn9802 [DNA fragment]) in a single-reaction mixture for the detection and discrimination of S. pneumoniae. Here, we validate a quantitative multiplex real-time PCR (qmPCR) assay with a panel consisting of 43 S. pneumoniae and 29 non-pneumococcal isolates, 20 culture positive, 26 culture negative and 30 spiked serum samples. A standard curve was obtained using S. pneumoniae ATCC 49619 strain and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) gene was used as an endogenous internal control. The experiment showed high sensitivity with lower limit of detection equivalent to 4 genome copies/µl. The efficiency of the reaction was 100% for ply, lytA, Spn9802 and 97% for psaA. The test showed sensitivity and specificity of 100% with culture isolates and serum specimens. This study demonstrates that qmPCR analysis of sera using 4 oligonucleotide primers appears to be an appropriate method for the genotypic identification of S. pneumoniae infection.

Casein kinase 2α regulates glioblastoma brain tumor-initiating cell growth through the ß-catenin pathway.

Glioblastoma (GBM) is the most common and fatal primary brain tumor in humans, and it is essential that new and better therapies are developed to treat this disease. Previous research suggests that casein kinase 2 (CK2) may be a promising therapeutic target for GBMs. CK2 has enhanced expression or activity in numerous cancers, including GBM, and it has been demonstrated that inhibitors of CK2 regressed tumor growth in GBM xenograft mouse models. Our studies demonstrate that the CK2 subunit, CK2α, is overexpressed in and has an important role in regulating brain tumor-initiating cells (BTIC) in GBM. Initial studies showed that two GBM cell lines (U87-MG and U138) transduced with CK2α had enhanced proliferation and anchorage-independent growth. Inhibition of CKα using siRNA or small-molecule inhibitors (TBBz, CX-4945) reduced cell growth, decreased tumor size, and increased survival rates in GBM xenograft mouse models. We also verified that inhibition of CK2α decreased the activity of a well-known GBM-initiating cell regulator, ß-catenin. Loss of CK2α decreased two ß-catenin-regulated genes that are involved in GBM-initiating cell growth, OCT4 and NANOG. To determine the importance of CK2α in GBM stem cell maintenance, we reduced CK2α activity in primary GBM samples and tumor spheres derived from GBM patients. We discovered that loss of CK2α activity reduced the sphere-forming capacity of BTIC and decreased numerous GBM stem cell markers, including CD133, CD90, CD49f and A2B5. Our study suggests that CK2α is involved in GBM tumorigenesis by maintaining BTIC through the regulation of ß-catenin.

Effect of co-transplantation of mesenchymal stem cells and hematopoietic stem cells as compared to hematopoietic stem cell transplantation alone in renal transplantation to achieve donor hypo-responsiveness.

INTRODUCTION: We evaluated donor hypo-responsiveness in renal allograft recipients to donor adipose tissue-derived mesenchymal stem cell (h-AD-MSC) +hematopoietic stem cell transplantation (HSCT) vs. HSCT alone. METHODS: Patients were divided into 2 demographically equal groups (n = 100) A and B subjected to equal non-myeloablative conditioning of target-specific irradiation, anti-T + B cell antibodies and cyclophosphamide with HSCT. Group A was administered h-AD-MSC additionally. Transplantation was performed following favorable cross-matching. Cyclosporine, 3 mg/kg BW/day + prednisone, 20 mg/day were immunosuppressants for first 3 months, cyclosporine was replaced by azathioprine subsequently and prednisone lowered to 5-10 mg/day. Peripheral blood chimerism (PBC) was studied using fluorescent in situ hybridization technique at 3/18 months post transplant. Biopsy was performed for graft dysfunction and reported as per Banff criteria,'05. RESULTS: Mean nucleated HSC counts (n × 10(8)/kgBW) was 7.32 with mean CD34+ yield 0.09% in group A; and 6.98 and 0.40% in group B, respectively; CD45-/90+ was 13.49% in former. Over 18 months post transplant, former had mean serum creatinine (SCr), 1.59 mg%, 12% acute rejection (AR) episodes, 3% patient, 1% patient +graft loss; latter had mean SCr 1.49 mg%, 18% AR episodes, 1% patient, 6% graft and 8% patient +graft losses. PBC was higher (4%) in former than later (1.8%). CONCLUSION: Combined h-AD-MSC +HSCT under non-myeloablative conditioning was safe, more effective than HSCT alone to achieve donor hypo-responsiveness with adequate stable graft function and reduced rejection episodes.