Genetic diversity and pectinolytic activity of epiphytic yeasts from grape carposphere.

The genetic diversity of epiphytic yeasts from grape carposphere is susceptible to environmental variations that determine the predominant carposphere microbiota. Understanding the diversity of yeasts that inhabit grape carposphere in different environments and their pectinolytic activity is a way to understand the biotechnological potential that surrounds us and help improve winemaking. Therefore, this study aimed to evaluate the pectinolytic activity and characterize the genetic diversity of isolated epiphytic yeasts from grape carposphere. Grapes of the Bordeaux cultivar were collected from different regions of Paraná and Rio Grande do Sul States, in Brazil, and the yeasts were isolated from these grape carpospheres. Monosporic isolates were morphologically and genetically characterized on potato dextrose agar medium and by PCR-RFLP and rep-PCR (BOX-PCR) in the ITS1-5.8S-ITS2 region of rDNA. The index of pectinolytic activity of isolates was also evaluated estimating the ratio between the halo diameter of enzymatic degradation and the diameter of the colony when the isolates were grown in cultivation medium containing 10 g/L pectin, 5 g/L yeast extract, 15 g/L agar, 0.12% (w/v) Congo red, and pH 6.2. We observed that the grape carposphere is an environment with a great genetic diversity of epiphytic yeasts of the following genera: Cryptococcus (31.25%), Pichia (25.0%), Candida (25.0%), Dekkera (12.5%), and Saccharomyces (6.25%). The PCR-RFLP technique allowed analyzing existing polymorphism among individuals of a population based on a more restrict and evolutionarily preserved region, mostly utilized to differentiate isolates at the genus level. Approximately 33% of yeast isolates presented pectinolytic activity with potential biotechnological for wine and fruit juice production. This great genetic variability found indicated that it is a potential reservoir of genes to be applied in viniculture improvement programs.

Characterization of the inaA gene and expression of ice nucleation phenotype in Pantoea ananatis isolates from Maize White Spot disease.

Maize White Spot (MWS), a foliar disease caused by Pantoea ananatis, could cause up to 60% yield loss. Some strains of P. ananatis harboring the ice nucleation gene inaA catalyze the formation of ice nuclei, causing tissue damage at temperatures slightly below freezing. Little is known about the relationship between the presence of the ina gene in this maize pathogen and its expression during the phenomenon of ice nucleus formation. Here, we attempted to verify the presence of the inaA gene and the expression of phenotype in vitro. The identity of the isolates and the presence of the inaA gene were determined by P. ananatis species-specific primers. The expression of the inaA gene was assessed in vitro by the visualization of ice-crystal formation in water at subzero temperatures. A total of ninety P. ananatis isolates from MWS lesions were characterized. The presence of the inaA gene was confirmed by gel electrophoresis of the 350-400-bp PCR products. The inaA primers did not lead to DNA fragment amplification in three isolates. The ice nucleation phenotype was expressed in 83.34% of the isolates carrying the inaA gene. Our study showed that the ice nucleation in P. ananatis isolated from MWS lesions was dependent on the presence of a functional ina gene in the genome. We also found evidence indicating that some P. ananatis strains have a mutated form of the inaA gene, producing a non-functional ice nucleation protein. This is the first report on inaA gene characterization in P. ananatis isolates from Maize White Spot.

Genomic variability of Pantoea ananatis in maize white spot lesions assessed by AFLP markers.

Measures to control maize white spot (MWS) caused by Pantoea ananatis are preferentially based on resistant cultivars. A lack of knowledge on the genetic variability of pathogens could interfere with the development and utilization of controlling strategies in this pathosystem. The main goals of this study were to investigate the genetic variability of 90 P. ananatis isolates from three different eco-geographical regions of Brazil by amplified fragment length polymorphism (AFLP), and to determine the presence of a universal P. ananatis plasmid in isolates from tropical Brazil. Analysis of genetic similarity by AFLP allowed us to categorize the 90 isolates into two groups. However, no correlation between the collecting sites and genetic groupings was observed. The polymorphism percentage found in P. ananatis ranged between 24.64 and 92.46%, and genetic diversity was calculated to be 0.07-0.09. The analysis of molecular variance showed that 99.18% of genetic variability was within the populations, providing evidence that evolutionary forces were acting on these populations. All P. ananatis isolates showed the P. ananatis universal plasmid (280 or 352 kb). This is the first report on the presence of a universal P. ananatis plasmid from MWS lesions in the tropical area.

Photoprotective and Antimutagenic Activity of Agaricus subrufescens Basidiocarp Extracts.

The photoprotective and antimutagenic activity of opened and closed basidiocarps of Agaricus subrufescens (=A. blazei; =A. brasiliensis) obtained by different extraction methods were evaluated on Aspergillus nidulans conidia submitted to ultraviolet (UV) light. The aqueous extracts were obtained by three extraction methods: maceration, infusion, and decoction, at two different extraction times. The extracts of A. subrufescens did not present toxicity for A. nidulans conidia. A suspension of A. nidulans conidia was submitted to extracts before and after the exposure to UV light. All basidiocarp extracts, regardless of the extraction method or development stage, protected A. nidulans conidia against the damaging effects of the mutagenic agent. The antimutagenic and photoprotective activity was strengthened with extracts obtained by 168-h maceration, followed by 24-h maceration and 60-min infusion and, at last, by 30-min infusion. Although the extracts presented protector effect as well as recoverer effect to the action of UV light, the preventive effect was more evident. Differences in the biological activity in function of the different development stages were detected with greater antimutagenic and photoprotective activity for the opened basidiocarps. However, the extraction method is the most important factor to be considered when compared to the basidiocarp development stage to obtain better antimutagenic and photoprotective activity of A. subrufescens basidiocarps.

Screening of basidiomycetes in submerged cultivation based on antioxidant activity.

Submerged cultivation of medicinal basidiomycetes is a reproducible and efficient method of producing mycelia and metabolites. The antioxidant activity indicates its medicinal properties and is an important tool for basidiomycete screening. In this study, we analyzed the production of mycelial biomass and exopolysaccharides and the antioxidant activity of basidiomycete strains in submerged cultivation. Twenty-five strains were used for submerged cultivation in extract malt medium, and the production of mycelial biomass and exopolysaccharides was evaluated. Antioxidant activity was determined using the 1,1-diphenyl-2-picrylhydrazyl method. Among the 25 evaluated strains, Lentinus crinitus produced the highest biomass, reaching 1190 ± 52 mg·L(-1)day(-1); Agaricus subrufescens strains had the highest exopolysaccharide production from 18.96 ± 0.15 to 20.97 ± 2.10 mg L(-1)·day(-1). Additionally, A. subrufescens showed the highest total antioxidant activity, reinforcing the therapeutic potential of this basidiomycete. No significant correlation was found between mycelial biomass or exopolysaccharide production and antioxidant activity; however, the results depended on each species and the strains of the same species. We found large variations in the production of mycelial biomass and exopolysaccharides and in antioxidant activity among different species and among strains of the same species. Thus, evaluating the total antioxidant activity is an important tool for identifying strains with biotechnological potential.

Serum levels and mesenteric fat tissue expression of adiponectin and leptin in patients with Crohn's disease.

Crohn's disease (CD) is characterized by inflammation and an aetiology that is still unknown. Hypertrophy of mesenteric fat is a reflection of disease activity, as this fat covers the entire length of the affected area. Adipocytes synthesize leptin and adiponectin, adipocytokines responsible for pro- and anti-inflammatory effects. Therefore, we evaluated serum levels of adiponectin and leptin, as well as mesenteral expression of adiponectin in active CD and those in remission. Sixteen patients with ileocaecal CD followed at the Outpatient Clinic, Coloproctology Unit of University of Campinas Clinical Hospital, participated in the study. Analysis of serum adiponectin and leptin by enzyme-linked immunosorbent assay was performed in patients with active CD (ACD group), remission CD (RCD group) and in six healthy controls. Ten patients with active ileocaecal CD (FCD group) and eight patients with non-inflammatory disease selected for surgery were also studied. The specimens were snap-frozen and the expression of adiponectin was determined by immunoblot of protein extracts. Serum C-reactive protein levels were higher in the ACD group when compared to the others and no difference of body mass index was observed between the groups. Serum adiponectin was lower in the ACD group when compared to control, but no differences were seen when comparing the ACD and RCD groups. Mesenteric adiponectin expression was lower in the FCD group when compared to the FC group. Serum leptin was similar in all groups. The lower levels of serum and mesenteric adiponectin in active CD suggest a defective regulation of anti-inflammatory pathways in CD pathogenesis.

Influence of particle size on the SARS-CoV-2 spike protein detection using IgG-capped gold nanoparticles and dynamic light scattering.

Due to the unprecedented and ongoing nature of the coronavirus outbreak, the development of rapid immunoassays to detect severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and its highly contagious variants is an important and challenging task. Here, we report the development of polyclonal antibody-functionalized spherical gold nanoparticle biosensors as well as the influence of the nanoparticle sizes on the immunoassay response to detect the SARS-CoV-2 spike protein by dynamic light scattering. By monitoring the increment in the hydrodynamic diameter (ΔDH) by dynamic light scattering measurements in the antigen-antibody interaction, SARS-CoV-2 S-protein can be detected in only 5 min. The larger the nanoparticles, the larger ΔDH in the presence of spike protein. From adsorption isotherm, the calculated binding constant (K D ) was 83 nM and the estimated limit of detection was 13 ng/mL (30 pM). The biosensor was stable up to 90 days at 4 °C. Therefore, the biosensor developed in this work could be potentially applied as a fast and sensible immunoassay to detect SARS-CoV-2 infection in patient samples.

Outcome of carpal tunnel release surgery in patients with and without Linburg-Comstock anomaly.

Anatomical variations may entail increased volume within the carpal tunnel and consequently the development of carpal tunnel syndrome (CTS) symptoms. The clinical importance of anomalous connection between the flexor pollicis longus (FPL) and flexor digitorum profundus (FDP) tendons was described by Linburg and Comstock. The objective of the present study was to compare postoperative outcomes of carpal tunnel release surgery in patients with and without Linburg-Comstock anomaly (LCA). Hands of CTS patients with and without LCA were evaluated for finger sensitivity, grip, tip, key and tripod pinch strength, pain intensity and Boston Carpal Tunnel Questionnaire score. Patients were evaluated preoperatively and 1, 3 and 6 months after surgery. Forty-six hands in 36 CTS patients with LCA and 50 hands in 38 patients without LCA were included. On all study parameters, no significant difference was found between CTS patients with versus without LCA. As outcomes are similar in both cases, LCA resection is not useful in patients with LCA and CTS. Preoperative diagnosis is unnecessary and screening for LCA is non-contributive for CTS evaluation.

Linburg-Comstock anomaly: A comparison of the prevalence in women with and without clinical carpal tunnel syndrome.

We aimed to compare the prevalence of the Linburg-Comstock anomaly in women with and without a clinical diagnosis of carpal tunnel syndrome. The prevalence of the Linburg-Comstock anomaly was evaluated in 400 hands from 200 women over 40 years of age who were diagnosed clinically with carpal tunnel syndrome (CTS), designated as the CTS group. The volunteer group consisted of 400 hands from 200 healthy women over 40 years of age. The women from both groups were asked to carry out the clinical flexion and pain tests described by Linburg and Comstock (1979) as a basis for the clinical diagnosis. CTS patient ages ranged from 40 to 90 (mean 55.8) years, while volunteer group ages ranged from 40 to 93 (mean 55) years. The flexion test was positive in 305 (76%) hands in the CTS group and 242 (60%) hands in the volunteer group. The pain test was positive in 261 (65%) hands in the CTS group and 108 (27%) hands in the volunteer group. Both tests were positive in 244 (61%) hands in the CTS group and 98 (24%) hands in the volunteer group. All these differences were statistically significant. Based on clinical examination using the flexion and pain tests, the prevalence of Linburg-Comstock anomaly was statistically higher in the group of women with carpal tunnel syndrome than in healthy volunteers.

Stability and surgical complications of tooth-borne and bone-borne appliances in surgical assisted rapid maxillary expansion: a systematic review.

The objective of this systematic review was to evaluate the stability and complications of tooth-borne (TB), bone-borne (BB) and hybrid (TB-BB) appliances in surgically assisted rapid maxillary expansion (SARME). Database searches were conducted (PubMed, Embase, Cochrane Library and SciELO), as well as a grey literature search (Google Scholar) and hand searches of reference lists. Forty-six articles were included after study selection (κ=0.854). After eligibility assessment, 16 articles and one article from the grey literature were processed (κ=0.866) and six articles were selected by hand searching, for a total of 23 articles included. Regarding stability, TB appliances showed width relapse rates ranging from 4 to 35% in canines, from 1 to 37% in premolars and from 0.2 to 49.5% in molars. In BB appliances, width relapse rates were 1.7-21% in canines, 1.5% in premolars and 4.6-11.5% in molars. In hybrid appliances, the width relapse rate was 14% in premolars, with a 1.8% overexpansion reported in the molar region. In TB and BB appliances, skeletal relapse rates were similar on the nasal floor (11-53% and 41.6%, respectively) and at the level of the maxilla (18% and 16%, respectively). The most common complications were bone resorption in TB appliances (18.14%) and appliance-related complications in BB appliances (17.9%). The risk of bias was high in 19 studies, medium in three studies and low in one study. The TB and BB appliances used in SARME were considered to have a high long-term stability. BB appliances appeared to have fewer relapses than TB appliances due to a more parallel distribution of forces exerted. However, relapse appears to be highly influenced by postorthodontic treatments, where arch-form coordination is achieved in the consolidation period with the purpose of overexpansion correction, alignment and final vertical adjustments. Further randomised controlled trials with long-term data and large sample sizes are needed to support evidence-based clinical decision-making and to allow meta-analytic studies of stability outcomes regarding the type of anchorage in SARME.

Chronic exercise reduces platelet activation in hypertension: upregulation of the L-arginine-nitric oxide pathway.

Nitric oxide (NO) inhibits platelet function and plays a key role in the regulation of cardiovascular homeostasis. Essential hypertension is characterized by an increased risk of thrombus formation, and by an inhibition of intraplatelet NO bioactivity. We have previously shown that membrane transport of L-arginine is a rate-limiting step for platelet-derived NO synthesis. This study examined the effects of exercise on the platelet L-arginine-NO pathway and aggregation and systemic inflammation markers in 13 sedentary hypertensive patients subjected to 60 min of training activity (exercise group), predominantly aerobic, three times a week for a period of 12 weeks. Six sedentary hypertensive patients participated in the control group. After 12 weeks, L-arginine transport was significantly increased and associated with increased platelet NO synthase activity and cGMP levels and reduced platelet aggregation. Moreover, exercise training reduced plasma concentrations of fibrinogen and C-reactive protein and blood pressure. The control group did not change their previous intraplatelet L-arginine-NO results and systemic inflammatory markers levels. Thus, exercise training reduces inflammatory responses, restores NO synthesis in platelets and thereby contributes to the beneficial effects of exercise in hypertension. The present study adds exercise as a new tool to reduce morbidity and mortality associated with platelet activation in hypertension.

Role of nitrites in the genesis of adenocarcinoma associated with Barrett's esophagus.

BACKGROUND: Barrett's esophagus (BE) is one of the complications of gastroesophageal reflux disease (GERD) and a premalignant condition. It consists of a process of replacement of the squamous epithelium of the esophagus by intestinal columnar epithelium containing goblet cells, known as specialized intestinal metaplasia with goblet cells, and several factors have been related to its pathogenesis. The objective of this study was to evaluate an experimental model of duodenogastroesophageal reflux and the effect of ingestion of sodium nitrite solution on the genesis of adenocarcinoma associated with Barrett's esophagus. MATERIALS AND METHODS: Sixty male Wistar rats were divided into four groups. Twenty were not submitted to surgery and served as controls (10 animals ingesting only water and 10 ingesting water plus a solution of sodium nitrite), while the remaining 40 animals were submitted to side-to-side duodenogastroesophageal anastomosis (20 animals ingesting only water and 20 ingesting water plus the sodium nitrite solution). The Vienna classification for dysplasia and adenocarcinoma was used in the analysis of results. RESULTS: After 42 weeks of observation, Barrett's esophagus was found in 26.3% (5/19) of the animals submitted to surgery that had not ingested nitrites compared to 72.3% (13/18) of the animals in the group submitted to surgery and given nitrites. Six cases of adenocarcinoma (33.3%) were also found in this latter group. Barrett's esophagus was not found in any of the animals that were not submitted to surgery. Categories 2, 3 and 5 of the Vienna classification were only found in the animals submitted to surgery that also received sodium nitrite (66.7%). CONCLUSION: The ingestion of sodium nitrite associated with duodenogastroesophageal reflux plays an important role in the genesis of adenocarcinoma associated with Barrett's esophagus.

Hierarchy of surgical stability in orthognathic surgery: overview of systematic reviews.

The purpose was to perform an overview of systematic reviews in order to create a hierarchical scale of stability in orthognathic surgery with the aid of the highest level of scientific evidence. The systematic search was conducted in the PubMed, Embase, and Cochrane Library databases. The grey literature was investigated in Google Scholar and a manual search was done of the references lists of included studies. Fifteen studies were included in the final sample, of which eight were systematic reviews and seven were meta-analyses. These were assessed for methodological quality using the AMSTAR 2 tool and all were considered to be of medium to high methodological quality. The clinical studies included in the 15 reviews and meta-analyses were classified by the review authors as having a moderate to high potential for risk of bias. The hierarchical pyramid of stability in orthognathic surgery was established, with two surgical procedures considered highly unstable: (1) maxillary expansion with semi-rigid internal fixation evaluated at the dental level in the posterior region, and (2) clockwise rotation of the mandible with rigid internal fixation of bicortical screws in the sagittal direction.

Extragastrointestinal stromal tumor and liver transplantation: case report and review.

The occurrence of de novo malignant neoplasias has been shown in postransplant patients under imunosuppression. It is the second leading cause of late death in liver transplant recipients. The greatest incidence is seen in cancers associated with chronic infection by human papilloma virus, skin cancers, oropharyngeal, and gastrointestinal (GI) malignancies. GI stromal tumors (GISTs) are the most common mesenchymal tumors of the GI tract. Rare cases are identified outside the GI tract are collectively known as extragastrointestinal stromal tumors (EGISTs). We present an EGIST case in a liver transplantation patient. A 64-year-old man underwent liver transplantation because of cirrhosis (hepatitis B virus and alcoholism) and hepatocellular carcinoma. Histopathologic findings revealed 2 trabecular hepatocellular carcinomas: a 3.5-cm-diameter lesion located at segment VIII and another 2-cm one at segment V. Seven months later, he noticed a hardened, mobile, painless, 3-cm subcutaneous nodule in the perineum localized in the right lateral quadrant 2 cm distant from the anus. A surgical resection with 1 cm margin yielded a histopathology report of a 5.0 x 3.0 cm spindle cell stromal tumor. The immunohistochemical profile was compatible with a GIST, with 5 mitosis per 50 high-powered fields. This tumor is extremely rare after liver transplantation but has shown a good outcome up to now.

Increased bone contact to a calcium-incorporated oxidized commercially pure titanium implant: an in-vivo study in rabbits.

The aim of this study was to evaluate the bone response to an oxidized titanium implant (Ox) and a calcium-incorporated oxidized titanium implant (Ca). A blasted titanium implant (Bl) was used as control. The implants were topographically characterized using an optical interferometer and placed: one in each distal femoral metaphysis and two in each proximal tibial metaphysis in rabbits. The rabbits were killed 12 weeks after implant insertion, and the implants and their surrounding tissues were removed en bloc for histomorphometrical evaluations. Topographical evaluation revealed three different surfaces: average height deviation (S(a), microm) values for Ca:Ox:Bl implants were 0.3:0.6:0.9, developed surface area ratios (%) 17:44:31, number of summits per microm(2) 208:136:118, and core fluid retention index values 1.33:1.33:1.38. The mean percentages of bone contact to the implants placed in the tibia (Ca:Ox:Bl) were 47:30:34 and to the implants placed in the femur (Ca:Ox) 32:20. The mean percentages of surrounding bone area for the implants placed in the tibia were 40:47:37 and for the implants placed in the femur 43:46. A significant increase in bone contact was found for smooth (S(a) <0.5 microm) but more densely peaked calcium-incorporated oxidized implants when compared to slightly rougher (S(a)=0.5-1.0 microm) oxidized or blasted implants.

Histiocytic Ulcerative Colitis in an American Staffordshire Terrier.

A 9-year-old female American Staffordshire terrier was presented to a veterinary hospital with diarrhoea, severe prostration, hypothermia, dehydration and anaemia. The dog died 6 days after the first consultation. At necropsy examination the serosa of the large intestine showed a granular appearance and the mucosa was thickened, ulcerated and red, with prominent folding. Histological examination revealed marked inflammatory infiltration of macrophages into the mucosa and submucosa of the large intestine. These cells stained positively by the periodic acid-Schiff reaction. Immunohistochemistry showed marked presence of intracytoplasmic Escherichia coli in the macrophages. Bacteriological examination of intestinal sections yielded E. coli growth and the isolate displayed atypical characteristics when compared with strains associated with previously published cases of histiocytic ulcerative colitis (HUC). The molecular characterization showed that the isolate harboured none of the genes associated with enterotoxigenic E. coli strains and harboured only a limited number of genes associated with extra-intestinal pathotypes. Adherent and invasive E. coli are unlikely to have been involved in the pathogenesis of HUC in the present case. HUC is a rare disease with a predisposition for boxer dogs; however, sporadic occurrence in other breeds may occur. This is the first reported case of HUC in an American Staffordshire terrier.

Stability and surgical complications in segmental Le Fort I osteotomy: a systematic review.

This systematic review was conducted to evaluate the stability and surgical complications of segmental Le Fort I osteotomy. The search was divided into a main search (PubMed, Embase, and Cochrane Library), grey literature search (Google Scholar), and manual search. Twenty-three studies were included: 14 evaluating stability as the outcome and nine evaluating surgical complications. The level of agreement between the authors was considered excellent (κ=0.893 for study selection and κ=0.853 for study eligibility). The segmental Le Fort I osteotomy provides stable outcomes in the sagittal plane, is less stable dentally than skeletally in the transverse plane, and provides little stability in the posterior segment after downward movement. The most frequent complications are oral fistula (six studies) and damage to the adjacent teeth (five studies), but the most prevalent complication is postoperative infection (32.62%). Four studies evaluating stability as the outcome showed a medium potential risk of bias, whereas all studies addressing surgical complications showed a high potential risk of bias. The segmental Le Fort I osteotomy should not be excluded from the technical armamentarium in orthognathic surgery. On the contrary, the literature consulted suggests it to be a useful tool for the three-dimensional surgical correction of maxillary malposition.

Polyethylene glycol-mediated fusion between primary mouse mesenchymal stem cells and mouse fibroblasts generates hybrid cells with increased proliferation and altered differentiation.

Bone marrow-derived mesenchymal stem cells (MSCs) can differentiate into different cell lineages with the appropriate stimulation in vitro. Transplantation of MSCs in human and other animal models was found to repair tissues through the fusion of transplanted MSCs with indigenous cells. We have generated mouseâmouse hybrid cell lines in vitro by polyethylene glycol-mediated fusion of primary mouse MSCs with mouse fibroblasts to investigate the characteristics of hybrid cells, including their potentials for proliferation and differentiation. Similar to the parental MSCs, hybrid cells are positive for the cell-surface markers CD29, CD44, CD49e, and Sca-1, and negative for Gr-1, CD11b, CD13, CD18, CD31, CD43, CD45, CD49d, CD90.2, CD445R/B220, and CD117 markers. The hybrid cells also produce a high level of tissue nonspecific alkaline phosphatase compared to the parental cells. Conditioned medium of hybrid cells contain biologically active factors that are capable of stimulating proliferation of other cells. Although the parental MSCs can differentiate into adipogenic and osteogenic lineages, hybrid cells held disparate differentiation capacity. Hybrid cell lines in general have increased proliferative capacity than the primary MSCs. Our study demonstrates that proliferative hybrid cell lines can be generated in vitro by induced fusion of both immortal and primary somatic cells with primary MSCs.

Mesenchymal stem cells: isolation, in vitro expansion and characterization.

Mesenchymal stem cells (MSC), one type of adult stem cell, are easy to isolate, culture, and manipulate in ex vivo culture. These cells have great plasticity and the potential for therapeutic applications, but their properties are poorly understood. MSCs can be found in bone marrow and in many other tissues, and these cells are generally identified through a combination of poorly defined physical, phenotypic, and functional properties; consequently, multiple names have been given to these cell populations. Murine MSCs have been directly applied to a wide range of murine models of diseases, where they can act as therapeutic agents per se, or as vehicles for the delivery of therapeutic genes. In addition to their systemic engraftment capabilities, MSCs show great potential for the replacement of damaged tissues such as bone, cartilage, tendon, and ligament. Their pharmacological importance is related to four points: MSCs secrete biologically important molecules, express specific receptors, can be genetically manipulated, and are susceptible to molecules that modify their natural behavior. Due to their low frequency and the lack of knowledge on cell surface markers and their location of origin, most information concerning MSCs is derived from in vitro studies. The search for the identity of the mesenchymal stem cell has depended mainly on three culture systems: the CFU-F assay, the analysis of bone marrow stroma, and the cultivation of mesenchymal stem cell lines. Other cell populations, more or less related to the MSC, have also been described. Isolation and culture conditions used to expand these cells rely on the ability of MSCs, although variable, to adhere to plastic surfaces. Whether these conditions selectively favor the expansion of different bone marrow precursors or cause similar cell populations to acquire different phenotypes is not clear. The cell populations could also represent different points of a hierarchy or a continuum of differentiation. These issues reinforce the urgent need for a more comprehensive view of the mesenchymal stem cell identity and characteristics.