Single Cell Deposition and Patterning with a Robotic System

Integrating single-cell manipulation techniques in traditional and emerging biological culture systems is challenging. Microfabricated devices for single cell studies in particular often require cells to be spatially positioned at specific culture sites on the device surface. This paper presents a robotic micromanipulation system for pick-and-place positioning of single cells. By integrating computer vision and motion control algorithms, the system visually tracks a cell in real time and controls multiple positioning devices simultaneously to accurately pick up a single cell, transfer it to a desired substrate, and deposit it at a specified location. A traditional glass micropipette is used, and whole- and partial-cell aspiration techniques are investigated to manipulate single cells. Partially aspirating cells resulted in an operation speed of 15 seconds per cell and a 95% success rate. In contrast, the whole-cell aspiration method required 30 seconds per cell and achieved a success rate of 80%. The broad applicability of this robotic manipulation technique is demonstrated using multiple cell types on traditional substrates and on open-top microfabricated devices, without requiring modifications to device designs. Furthermore, we used this serial deposition process in conjunction with an established parallel cell manipulation technique to improve the efficiency of single cell capture from ∼80% to 100%. Using a robotic micromanipulation system to position single cells on a substrate is demonstrated as an effective stand-alone or bolstering technology for single-cell studies, eliminating some of the drawbacks associated with standard single-cell handling and manipulation techniques

Mineralization of sheep manure and its influence on lettuce production

Diversos resíduos orgânicos são utilizados na agricultura sem o adequado conhecimento da sua dinâmica de mineralização. Avaliou-se a mineralização de esterco de ovinos e sua influência na produção de alface. Utilizou-se o delineamento experimental de blocos ao acaso com três repetições. Foram utilizadas 25 t ha-1 como dose de esterco para cada um dos seguintes tratamentos: 1) esterco de ovinos que se alimentaram de feno de mandioca (PAM); 2) esterco de ovinos que se alimentaram de subproduto de ervilha (ERV); 3) esterco de ovinos que se alimentaram de feno de capim coast-cross (FCC); 4) esterco de ovinos que se alimentaram de subproduto de saccharina (SAC) e 5) solo sem aplicação de esterco (testemunha). Foi determinada semanalmente a respiração basal do solo, utilizada como indicador de mineralização da matéria orgânica. A massa fresca de alface foi avaliada como medida de produção. Os tratamentos ERV, FCC e SAC apresentaram ganhos de massa fresca na ordem de 68, 65 e 62% em relação à testemunha e de 43, 39 e 33% em relação ao PAM, respectivamente. A produção menor promovida pelo PAM, em relação às demais, pode ser explicada pela forma de mineralização da matéria orgânica que apresentou elevada respiração microbiana cinco dias após o transplantio, com acentuado declínio, nas medições subseqüentes, ao longo do ciclo da cultura. Os demais tratamentos apresentaram mineralização sincronizada com conseqüente aumento na produção de massa fresca. ____________________________________________________________________________________________________________ ABSTRACTSeveral organic wastes are used in agriculture with no precise knowledge about the mineralization dynamics of these materials. In this study the sheep manure mineralization and its influence on the lettuce production was evaluated. A randomized block design with three replications was used. Five treatments were studied using 25 t ha-1 as dose of manure: 1) sheep manure obtained from animals fed with cassava straw (PAM); 2) sheep manure obtained from animals fed with residue of pea crop (ERV); 3) sheep manure obtained from animals fed with Coast-Cross hay (FCC), 4) sheep manure obtained from animals fed with saccharin residue (SAC) and 5) soil without application of manure (control). Weekly the basal respiration was determined and used as an indicator of organic matter mineralization. Lettuce fresh mass was evaluated as a measure of production. Treatments ERV, FCC and SAC showed superior weight gains of 68, 65 and 62% compared to the control and 43, 39 and 33% compared to MAP, respectively. Lower production promoted by the MAP in relation to the other treatments can be explained by organic matter mineralization that showed a high microbial respiration five days after transplanting, with marked decline in subsequent measurements during the crop cycle. The other systems showed mineralization synchronized with the production increase of lettuce fresh mass

Genome of the Avirulent Human-Infective Trypanosome—Trypanosoma rangeli

Background: Trypanosoma rangeli is a hemoflagellate protozoan parasite infecting humans and other wild and domestic mammals across Central and South America. It does not cause human disease, but it can be mistaken for the etiologic agent of Chagas disease, Trypanosoma cruzi. We have sequenced the T. rangeli genome to provide new tools for elucidating the distinct and intriguing biology of this species and the key pathways related to interaction with its arthropod and mammalian hosts.  Methodology/Principal Findings: The T. rangeli haploid genome is ,24 Mb in length, and is the smallest and least repetitive trypanosomatid genome sequenced thus far. This parasite genome has shorter subtelomeric sequences compared to those of T. cruzi and T. brucei; displays intraspecific karyotype variability and lacks minichromosomes. Of the predicted 7,613 protein coding sequences, functional annotations could be determined for 2,415, while 5,043 are hypothetical proteins, some with evidence of protein expression. 7,101 genes (93%) are shared with other trypanosomatids that infect humans. An ortholog of the dcl2 gene involved in the T. brucei RNAi pathway was found in T. rangeli, but the RNAi machinery is non-functional since the other genes in this pathway are pseudogenized. T. rangeli is highly susceptible to oxidative stress, a phenotype that may be explained by a smaller number of anti-oxidant defense enzymes and heatshock proteins.  Conclusions/Significance: Phylogenetic comparison of nuclear and mitochondrial genes indicates that T. rangeli and T. cruzi are equidistant from T. brucei. In addition to revealing new aspects of trypanosome co-evolution within the vertebrate and invertebrate hosts, comparative genomic analysis with pathogenic trypanosomatids provides valuable new information that can be further explored with the aim of developing better diagnostic tools and/or therapeutic targets

Survival of atraumatic restorative treatment (ART) sealants and restorations: a meta-analysis

The purpose of this study is to perform a systematic investigation plus meta-analysis into survival of atraumatic restorative treatment (ART) sealants and restorations using high-viscosity glass ionomers and to compare the results with those from the 2005 ART meta-analysis. Until February 2010, four databases were searched. Two hundred four publications were found, and 66 reported on ART restorations or sealant survival. Based on five exclusion criteria, two independent reviewers selected the 29 publications that accounted for the meta-analysis. Confidence intervals (CI) and or standard errors were calculated and the heterogeneity variance of the survival rates was estimated. Location (school/clinic) was an independent variable. The survival rates of single-surface and multiple-surface ART restorations in primary teeth over the first 2 years were 93% (CI, 91–94%) and 62% (CI, 51–73%), respectively; for single-surface ART restorations in permanent teeth over the first 3 and 5 years it was 85% (CI, 77–91%) and 80% (CI, 76–83%), respectively and for multiple-surface ART restorations in permanent teeth over 1 year it was 86% (CI, 59–98%). The mean annual dentine lesion incidence rate, in pits and fissures previously sealed using ART, over the first 3 years was 1%. No location effect and no differences between the 2005 and 2010 survival rates of ART restorations and sealants were observed. The short-term survival rates of single-surface ART restorations in primary and permanent teeth, and the caries-preventive effect of ART sealants were high. Clinical relevance: ART can safely be used in single-surface cavities in both primary and permanent teeth. ART sealants have a high caries preventive effect

IMPACT OF ROUX-EN-Y GASTRIC BYPASS SURGERY (RYGB) ON METABOLIC SYNDROME COMPONENTS AND ON THE USE OF ASSOCIATED DRUGS IN OBESE PATIENTS

ABSTRACT BACKGROUND The prevalence of obesity and metabolic syndrome is increasing worldwide and both behavior modification and drug therapy have low adherence. Gastric bypass has shown effective results in both reducing weight and improving comorbidities. OBJECTIVE To evaluate the impact of Roux-en-Y Gastric Bypass Surgery (RYGB) on both metabolic syndrome components and the use of associated drugs in obese patients. METHODS Historical cohort of patients subjected to Roux-en-Y Gastric Bypass Surgery (RYGB) between January 2007 and March 2014 in a private clinic. The sample consisted of 273 obese class II and III individuals, 86.4% of whom were female, with age ≥20 years, followed up for 2 months after surgery. Sociodemographic, anthropometric, biochemical, clinical, and drug-use data were collected from patients’ medical records. RESULTS Significant differences were found in weight, body mass index and waist circumference, after 60 postoperative days. Components for metabolic syndrome diagnosis (hypertension P=0.001; hyperglycemia P<0.001; hypertriglyceridemia P=0.006) were reduced after 60 days of postoperative, with the exception HDL-c (P=0.083). There was a significant reduction in the use of antihypertensive (P<0.001), hypoglycemic (P=0.013), lipid lowering (P<0.001), and antiobesity (P=0.010) drugs and increased use of gastroprotective drugs, vitamins, and minerals (P<0.001) after 60 postoperative days. CONCLUSION Patients subjected to Roux-en-Y Gastric Bypass Surgery exhibited both weight loss and significant improvement not only in metabolic syndrome components (except for HDL-c) but in the use of drugs associated with obesity and metabolic syndrome