PPAR Agonists Promote the Differentiation of Porcine Bone Marrow Mesenchymal Stem Cells into the Adipogenic and Myogenic Lineages.

PURPOSE: The aim of this work was to evaluate the effect of PPAR agonists on the differentiation and metabolic features of porcine mesenchymal stem cells induced to the adipogenic or myogenic lineages. METHODS: Bone marrow MSCs from neonate pigs were isolated and identified by cell proliferation, cell surface markers or the gene expression of stem cells (CD44, CD90, CD105 or Oct4 and Nanog, respectively). Cells were differentiated into adipose or muscle cells and treated with the PPAR agonists; adipogenic and myogenic differentiation was promoted by adding these compounds. The expression of PPARγ (an adipose marker) and MyoD1 and MyHC (muscle markers), metabolic changes and expression levels of metabolic enzymes involved in glycolysis, lipogenesis, lipolysis and the pentose phosphate pathway were tested by qPCR. RESULTS: MSCs from neonate pigs exhibited high proliferation and were positive for CD44, CD90 and CD105 markers and Oct4 and Nanog expression. The treatment that promoted the highest expression of PPARγ was 50 µM of conjugated linoleic acid (CLA) c9 t11 (6.44 ± 0.69-fold, p ≤ 0.0001) in the adipose differentiation, and upregulation of HX2, ACCAα, ATGL, LPL and G6DP (p ≤ 0.0001) and downregulation of PFK and ACCAß (p ≤ 0.0001) were found. For muscle differentiation, the best treatment was 50 µM of CLA c10 t12 (59.72 ± 4.72-fold, p ≤ 0.0001), and metabolic changes were upregulation of PFK, ACCAß, G6DP, CPT1 and PPARß/δ (p ≤ 0.0001), but no effect was observed with HX2 and ACCAα (p ≥ 0.05). CONCLUSIONS: Our results suggest that differentiated cells exhibit a typical cell lineage metabolism and higher efficiencies both in anabolism and catabolism.

[Utility of topical vancomycin application in the prevention of surgical site infection of the lumbosacral spine]. / Utilidad de la aplicación de vancomicina tópica en la prevención de infección de sitio quirúrgico de la columna lumbosacra.

INTRODUCTION: surgical site infections (SSI) remain a significant cause of morbidity and mortality and one of the most representative causes of nosocomial infections. The use of intrawound vancomycin in lumbar spine surgery is a potential prophylactic measure against SSI; however, evidence regarding its efficacy is contradictory. Our study was designed to research if intrawound vancomycin significantly prevents SSI in lumbar spine surgery. MATERIAL AND METHODS: this is a randomized, double-blinded, controlled clinical trial; 233 patients who underwent lumbar spine surgery, were randomly assigned to a group in which intrawound vancomycin was instilled in the incision before closure (109), or to a control group (114). The main outcome is the presence of SSI; we determined its prevalence and searched for difference between groups for association between SSI and independent variables. RESULTS: global SSI prevalence was 1.8%, in the experimental group was 0.9%, in the control group was 2.6%. There was no significant difference between these values, p = 0.622. The relative risk of SSI in the experimental group was 0.35 (95% CI 0.037-3.30), that of the control group was 2.87 (95% CI 0.30-27.16). The number needed to treat is 58.3. We did not find a significant association between the independent variables studied and the appearance of SSI. CONCLUSIONS: we did not find a significant difference in the prevalence of SSI between groups nor a significant association between SSI and independent variables.

INTRODUCCIÓN: las infecciones postoperatorias del sitio quirúrgico son una importante causa de morbimortalidad y una de las formas más comunes de infecciones nosocomiales. La aplicación de vancomicina al terminar una intervención de columna lumbar es una potencial práctica profiláctica de infecciones del sitio quirúrgico (ISQ). La evidencia que sostiene su uso es controversial. Nuestro estudio investiga si la aplicación de vancomicina disminuye en forma significativa la prevalencia de ISQ. MATERIAL Y MÉTODOS: ensayo clínico aleatorizado, controlado, cegado; 223 pacientes intervenidos de la columna lumbar fueron aleatoriamente asignados a un grupo experimental de 109 pacientes en quienes se colocó vancomicina y a un grupo control de 114 pacientes que no recibió vancomicina. El principal desenlace del estudio es la aparición de ISQ; se estudió la prevalencia de ISQ en ambos grupos y se buscó si existe diferencia significativa. Se analizó la existencia de factores predictores de ISQ. RESULTADOS: la prevalencia global de infección fue 1.8%; en el grupo experimenta 0.09% y en el grupo control 2.6%. No hubo diferencia significativa entre estas cifras, p = 0.622. El riesgo relativo de ISQ en el grupo experimental fue 0.35 (IC95% 0.037-3.30), el del grupo control fue 2.87 (IC95% 0.30-27.16). El número necesario para tratar es 58.3. No encontramos asociación significativa entre las variables independientes estudiadas y la aparición de ISQ. CONCLUSIONES: no encontramos evidencia suficiente de que la aplicación de vancomicina disminuya significativamente la prevalencia de ISQ ni asociación significativa de ISQ con las variables independientes estudiadas.

[Ciprofloxacin vs cefazolin in the prevention of infection in cirrhotic patients with gastrointestinal bleeding]. / Cefazolina vs. Ciprofloxacino en la Profilaxis de Infecciones en Pacientes Cirróticos con Sangrado Digestivo.

AIM: To determine if prophylaxis with cefazolin produces a significant reduction in infections in cirrhotic patients with gastrointestinal bleeding when compared with ciprofloxacin. METHODS: Randomized clinical trial. Patients 18 years or older with diagnosis of cirrhosis, gastrointestinal bleeding and no clinical or laboratory evidence of infection who were admitted to the gastrointestinal bleeding unit of HNERM between July 2008 and July 2010 were included. Patients were allocated to receive either i.v. ciprofloxacin 200 mg bid or i.v. cefazolin 1 gm tid for 7 days. RESULTS: 98 patients were included, 53 in the cefazolin group and 45 in the ciprofloxacin one. Age average was 66 +/- 10 years, 61% were male, 59,2% had ascites. Overall rate of infections was 14,3% (14/98). Rebleeding rate was 8,1% and mortality 4,1%. There were no differences in age, sex, Child Pugh score, ascites, hepatic encephalopathy nor in billirubin, albumin, PT and creatinine levels between the study groups. Infection rate in the cefazolin groups was 11,3% while in the ciprofloxacin one 17,8% (p=0,398).When Child-Pugh A and patients without ascites were excluded of the analysis, the cefalozin group had 22,2% of infections and 26,9% in the ciprofloxacin one (p=0,757). CONCLUSION: there were no differences in infection rates between patients with prophylaxis with cefazolin and ciprofloxacin after an episode of gastrointestinal bleeding.

Beware: Gastrointestinal symptoms can be a manifestation of COVID-19. / Alerta: los síntomas gastrointestinales podrían ser una manifestación de la COVID-19.

BACKGROUND: There is an increasing number of reports on the presentation of gastrointestinal symptoms in cases of COVID-19. AIM: To review the studies reporting gastrointestinal symptoms in COVID-19. RESULTS: Fifteen articles (2,800 patients) were identified. Gastrointestinal symptom frequency varied from 3.0% to 39.6% and included diarrhea (7.5%), náusea (4.5%), anorexia (4.4%), vomiting (1.3%), abdominal pain (0.5%), and belching/reflux (0.3%). Those symptoms can be the first manifestation of COVID-19, but whether they reflect a better or worse prognosis, is controversial. The potential relation of the angiotensin converting enzyme 2 receptor in the digestive tract as an entry route for the virus is discussed. CONCLUSION: Gastrointestinal symptoms may be common in COVID-19, in some cases appearing as the first manifestation, even before fever and respiratory symptoms. Therefore, clinicians and gastroenterologists must be aware of those atypical cases during the current pandemic, as well as of the fecal-oral route and corresponding preventive measures.

Open abdomen management with human acellular dermal matrix in liver transplant recipients.

BACKGROUND: Abdominal wall closure after liver transplantation is not always feasible and may result in increased intra-abdominal pressure along with associated complications. Various temporary closure techniques as well as open wound management have been used to address this complex problem. The aim of this series was to describe an approach to definitive wound closure of the open abdomen in liver transplant patients. METHODS: We performed a retrospective review of all liver transplant patients at our institution from September 2005 to November 2007. The management of the open abdomen in 10 liver transplant patients was reviewed, and a novel approach described to manage these defects. RESULTS: Ten patients with open wounds were closed during the study period using human acellular dermal matrix (HADM). There were 7 men and 3 women of median age 55 years. Average size of HADM was 235 cm(2). The median follow-up is 10 months with no incidence of evisceration or hernia. In 1 patient, the graft failed along the lateral side due to infection; it dislodged during vacuum-assisted closure dressing change in another patient at 5 months after closure. Fascial closure was not possible due to organ edema (n = 3), a large liver (n = 4) or wound infection with dehiscence (n = 3). CONCLUSIONS: HADM can be used for primary wound closure in both clean and contaminated wounds as an alternative to an open abdomen post-liver transplantation.

Belatacept and Eculizumab for Treatment of Calcineurin Inhibitor-induced Thrombotic Microangiopathy After Kidney Transplantation: Case Report.

Thrombotic microangiopathy (TMA) after kidney transplantation is an uncommon and challenging cause of graft dysfunction and is associated with early graft loss. An idiosyncratic endothelial reaction to calcineurin inhibitors (CNIs) has been implicated as a frequent cause of TMA. This reaction is marked by uncontrolled activation of complement and subsequent cellular destruction. Usual therapy consists of withdrawal of the inciting drug and plasmapheresis to minimize levels of circulating complement. Recently, eculizumab, a monoclonal antibody to complement component C5, has been used for the treatment of atypical hemolytic uremic syndrome. Belatacept, an inhibitor of T cell costimulatory protein CTLA-4 has been used in immunosuppression strategies aimed at minimization of CNI. Here we report the first case of treatment of CNI-associated TMA/hemolytic uremic syndrome with withdrawal of tacrolimus and initiation of both belatacept and eculizumab. The case describes a favorable clinical course for both graft and patient, and is accompanied by a review of the literature.

Hepatocellular carcinoma in renal transplant patients.

INTRODUCTION: Chronic liver diseases, especially those related to hepatitis B (HBV) and C viruses (HCV), are a common problem in renal transplant patients. Hepatocellular carcinoma (HCC) is a complication of chronic liver diseases, incidence in the renal transplant cohort is higher than in the general population (1.4% to 4% vs 0.005% to 0.015%). METHODS: We retrospectively evaluated the incidence of HCC, its clinical presentation, the treatments, and the relation to chronic viral hepatitis among the population transplanted at our center between January 1980 and December 1998 and followed to August 2003. RESULTS: During the study period, six recipients among 534 renal transplants displayed HCC (incidence 1.12% of the entire population and 2.29% of patients with chronic viral hepatitis). Among the cohort five were men, and all had chronic viral hepatitis: three HBV, one HCV, and 2, a coinfection. HCC was diagnosed 124.1 (range 45 to 244) months after transplantation. All patients presented with abnormal liver function tests and tumors larger than 5 cm. Four had more than three tumors and three had an alpha-fetoprotein level higher than 400 IU/mL. Three patients received no treatment (survivals 1, 1, and 4 months); two patients, chemoembolization (survival 6 and 12 months); and one, surgical ethanol injections (survival 4 months). The overall survival was 4.5 months. CONCLUSION: HCC in renal transplant recipients is a common complication among patients with chronic viral hepatitis. The outcome was poor because HCC was detected at an advanced stage. Screening strategies for early diagnosis must be prospectively evaluated.

Surgery in elderly patients in Mexico. Portal hypertension surgery as an example.

Although people older than 65 years represent less than 5% of Mexico's registered population, medical care for elderly patients requires a multidisciplinary approach. In our academic university hospital, they are managed by a team of specialists. As an example of this approach, we evaluated the surgical treatment of bleeding portal hypertension in a highly selected elderly population. A retrospective study was done reviewing the files of 25 patients older than 65 years. All had good liver function (Child-Pugh class A and B) and had undergone elective surgery. Sixteen of them were women. The mean age was 68.8 years (age range, 65-76 years), and most had a diagnosis of cirrhosis. All patients were treated with portal blood flow-preserving procedures (selective shunts or Sugiura-Futagawa procedures). The operative mortality was 8%. Eight later deaths were recorded, with a mean follow-up of 25 months (range, 2-110 months). Survival (Kaplan-Meier) was 87% at 12 months, 54% at 60 months, and 45% at 110 months. Two rebleeding incidents were recorded as well as 3 cases of postoperative encephalopathy. We concluded that well-selected elderly patients, undergoing elective surgery with portal blood flow-preserving procedures, have a good postoperative outcome.

Comparative study of 2 variants of a modified esophageal transection in the Sugiura-Futagawa operation.

OBJECTIVE: To compare 2 techniques of esophageal transection in our modification of the Sugiura-Futagawa procedure for the treatment of bleeding portal hypertension in low-risk patients who cannot undergo surgery to have shunts placed. DESIGN: A prospective controlled trial comparing 2 variants of transection (classic, complete section of the anterior muscularis externa and whole mucosa; modified, placement of a circumferential running suture without opening the mucosa). SETTING: Academic university hospital. PATIENTS: Eighty-three low-risk patients (Child-Pugh score A and B) with a history of bleeding portal hypertension were operated on (35 classic, 48 modified transections) between 1989 and 1996. Both groups were comparable. MAIN OUTCOME MEASURES: Postoperative dehiscence of the transection was evaluated as well as fistulization, postoperative stenosis, rebleeding, postoperative endoscopic findings, survival, and mortality. RESULTS: Fistulization was observed in 1 (2%) of the patients in the modified group, and dehiscence in 1 patient (2%). In the classic group, 3 (8%) of the patients had dehiscence (relative risk, 2.6) and 1 (2%) of the patients, fistulization. No differences were observed regarding rebleeding (6 patients [6%] vs 5 patients [7%]), postoperative stenosis (4 patients [8%] vs 5 patients [10%]), postoperative endoscopic findings, survival, and mortality (early and late). CONCLUSION: The modified variant of the transection has a lower frequency of postoperative dehiscence, with the same long-term results.

Isoenzyme patterns of Leishmania isolates from Colombia.

Promastigotes from four cutaneous leishmaniasis cases from Colombia were tested by cellulose acetate electrophoresis using nine enzyme systems. The isoenzyme profiles of the Colombian isolates were indistinguishable from each other and from Panamanian Leishmania braziliensis panamensis controls, but were distinct from an isolate of Leishmania braziliensis guyanensis from Brazil and three isolates from the Leishmania mexicana complex for the enzyme phosphogluconate dehydrogenase.

Diagnosis of cutaneous and mucocutaneous leishmaniasis in Colombia: a comparison of seven methods.

Seven methods of diagnosing leishmaniasis were compared in 177 patients presenting with lesions of the skin (165) or mucosa (12) in Tumaco and Cali, Colombia. The three methods of visualizing amastigotes in tissue samples (histological staining of tissue sections, impression smears of punch biopsies, and smears of dermal scraping from slits in the lesion margins) were less sensitive than the four Leishmania isolation methods (aspiration of lesion border cultured in biphasic media, aspirate inoculated into hamster nasal tissue, culture of punch biopsy macerate, and hamster inoculation of macerate). The aspirate-culture and biopsy-hamster methods employed in this study proved most sensitive of the four methods for the recovery of parasites. The combined overall sensitivity of the 7 methods was 67% for all enrolled patients and 75% for Montenegro skin test-positive patients. The individual sensitivities for the methods for all patients and Montenegro-positive positive, patients, respectively, were: histopathology 14% and 16%, impression smear 19% and 21%, dermal scraping 22% and 26%, aspirate-culture 58% and 64%, aspirate-hamster 38% and 41%, biopsy-culture 50% and 55%, and biopsy-hamster 52% and 57%. All methods were less sensitive in lesions of greater than 6 months duration than in lesions of more recent onset. Mucosal lesions were best diagnosed by the culture or hamster inoculation of a macerated mucosal biopsy. The diagnosis by inoculation of hamsters was achieved within 2 to 12 weeks, a mean of 34.5 days. Promastigotes were seen on Senekjie's medium within 3-8 days.(ABSTRACT TRUNCATED AT 250 WORDS)

Leishmania braziliensis from the Pacific coast region of Colombia: foci of transmission, clinical spectrum and isoenzyme phenotypes.

Tegumentary leishmaniasis is highly prevalent in the Pacific coast region of Colombia. We have identified 90 foci of transmission in this region based on 179 parasitologically diagnosed patients. Human transmission occurred in mangrove forests, secondary growth and intervened tropical rain forest. A parasitological diagnosis, that is, either isolation or visualization of Leishmania was made in 68.6% of suspected cases. Three phenotypically distinguishable groups of L. braziliensis were encountered based on isoenzymes: L. b. panamensis variants (82%), variants of L. b. braziliensis (14.5%), and stocks intermediate between L. b. panamensis and L. b. guyanensis reference strains (3.5%). The L. b. braziliensis variants produced cutaneous disease alone relatively infrequently (12% of classified cutaneous stocks) but were more frequently (38% of all mucosal stocks) isolated from mucosal lesions. Leishmania infection of the mucous membranes caused a wide spectrum of disease, severity being closely related to time of evolution. Both contiguous and metastatic spread to the mucous membranes was supported by the clinical course of 19 mucosal cases.

Cardiovascular reflexes and pudendal evoked responses in chronic haemodialysis patients.

The bulbocavernous reflex (BCR), cortical pudendal evoked responses (CPERs), non-invasive cardiovascular tests and nerve conduction studies were performed in 16 patients on chronic haemodialysis and in a group of normal subjects. BCR and CPERs were more severely affected in patients with impotence. There was a significant correlation between the Valsalva ratio and P1 latency of the CPERs. The BCR and CPERs are alternative techniques for the assessment of impotence in uremic patients.

[Incidence, etiology and prognostic value of persistent significant proteinuria in kidney transplants]. / Incidencia, etiología y pronóstico de la proteinuria persistente significativa en el trasplante renal.

We investigated persistent significant proteinuria (PSP), defined as proteinuria > 1 gr/24 hours on three consecutive months, in renal allograft recipients. The clinical records of 273 patients (288 grafts) were reviewed and 236 grafts (178 live related, 58 cadaver donor) that functioned for at least 4 months (230 patients, 148 men and 82 women) were selected for analysis. The histological diagnoses of 226 grafts and 35 native kidneys were also reviewed. PSP was present in 67 grafts (28.4%); 43 of these grafts were studied histologically (transplant glomerulopathy (TxGPT) 19, idiopathic glomerulopathy (GP) 13, and chronic rejection 11). Patients with an idiopathic GP in the graft usually presented with the nephrotic syndrome (65%); this presentation was infrequent in patients with chronic rejection. The appearance of proteinuria was strongly associated with functional deterioration in grafts with chronic rejection and TxGPT; in grafts with PSP and a histological diagnosis of idiopathic GP, renal function was usually normal. Within grafts with PSP no statistically significant differences in actuarial survival (AS) could be established when the time of appearance or magnitude of PSP, the presence or absence of arterial hypertension, the immunosuppressive regimen, and the histological diagnosis were considered. Contrariwise, the difference in AS was highly significant (p < 0.0001) when grafts with and without PSP were compared. The former had an AS at 5 and 10 years of 74.6% and 55.7%, while in the case of the latter AS was 57.3% and 32.1%, respectively. In conclusion, in the present series 28.4% of grafts that functioned 4 months or more presented PSP. The most frequent glomerular lesion was TxGPT. The presence of PSP was a marker for poorer prognosis, since AS at 5 and 10 years was significantly less in this group.

[Neurological complications of renal transplant]. / Complicaciones neurológicas del trasplante renal.

Advances in surgical procedures and new immunosuppressor therapies have improved the outcome of renal grafts. However, these changes have been accompanied by infectious, neoplastic and neurologic complications. The purpose of this study was to determine the incidence of neurologic complications among 542 patients receiving a renal transplant (from living or cadaveric donors) at CEMIC between 1970 and 1996. Neurologic complications occurred in 43 patients (8%) as follows: 8 meningitis (1.5%), 8 acute confusional syndrome (1.5%), 7 encephalitis (1.3%), 7 cerebrovascular accidents (1.3%), 6 convulsions (1.1%), 3 tumors (0.5%), 3 femoral nerve lesion (0.5%), and 1 epidural lipomatosis (0.1%). Etiologic agents most commonly observed in meningitis were: Cryptococcus neoformans, Listeria monocytogenes and Mycobacterium tuberculosis. Major difficulties arose in the diagnosis of encephalitis. Diagnosis of the above complications required clinical astuteness and repeated bacteriologic, serologic and imaging studies.

Retransplantation of the liver: review of current literature for decision making and technical considerations.

Liver transplantation (LTx) is an established treatment modality for patients with end-stage liver disease, metabolic disorders, and patients with acute liver failure. When a graft fails after primary LTx, retransplantation of the liver (reLTx) is the only potential cure. ReLTx accounts for 7%-10% of all LTx in the United States. Early causes of graft failure for which reLTx may be indicated include primary graft nonfunction and vascular inflow thrombosis. ReLTx in such cases in the early postoperative period is usually straightforward as long as an appropriate secondary allograft is secured in a timely fashion. Late indications may include ischemic cholangiopathy, chronic rejection, and recurrence of the primary liver disease. ReLTx performed in the late period is often more complex and selection criteria are more stringent due to the persistent shortage of organs. The question of whether to retransplant patients with recurrent hepatitis C remains controversial, but these practices are likely to change as the epidemic progresses and new treatments evolve. We also present recent results with reLTx from Yale-New Haven Transplant Center and early results with the use of living donors for reLTx.

Conversion to sirolimus allows preservation of renal function in kidney and kidney-pancreas allograft recipients.

The major causes of graft failure are chronic allograft nephropathy (CAN) and patient mortality. Sirolimus (SRL) is a powerful immunosuppressant with a less nephrotoxic profile as well as a lower incidence of cancer. The aim of this study was to evaluate the impact of conversion to SRL from calcineurin inhibitor (CNI)-based therapy in kidney (KT) and kidney-pancreas (SPK) allograft recipients. We analyzed renal function, allograft and patient survival, and SRL-associated adverse effects in 93 adult patients (86 KT and 7 SPK), who were converted to SRL between January 2001 and November 2008. The main reason for conversion was CAN (76; 9%) and 52 (7%) were receiving tacrolimus. Conversion occurred at a median 26.2 months. There was a significant improvement in creatinine clearance (CCr) at 6 months after conversion (CCr(baseline) 51.4 vs CCr(6m) 60.4 mL/min; P < .0001), without changes at 12 and 24 months. However, proteinuria increased significantly at 6 months compared with the baseline: 150 mg/24 hours (0-453) versus 0 mg/24 hours (range, 0-309), respectively (P < .0001), but did not progress at 12 or 24 months. At the same time we observed more extensive use of angiotensin-converting enzyme inhibitors/angiotensin receptor blockers: 60/5%; 65/3% and 70/2% at 6, 12, and 24 months. There were no changes in blood pressure control. Cholesterol significantly increased at 6 months (218.2 +/- 37 vs. 186.6 +/- 44 mg/dL; P < .0001). Graft and patient survivals at 4 years were 88% and 95%, respectively. Our experience suggested that conversion to SRL constituted a safe alternative with excellent results in patient and graft survival.