Spatial transcriptomics landscape of lesions from non-communicable inflammatory skin diseases.

Abundant heterogeneous immune cells infiltrate lesions in chronic inflammatory diseases and characterization of these cells is needed to distinguish disease-promoting from bystander immune cells. Here, we investigate the landscape of non-communicable inflammatory skin diseases (ncISD) by spatial transcriptomics resulting in a large repository of 62,000 spatially defined human cutaneous transcriptomes from 31 patients. Despite the expected immune cell infiltration, we observe rather low numbers of pathogenic disease promoting cytokine transcripts (IFNG, IL13 and IL17A), i.e. >125 times less compared to the mean expression of all other genes over lesional skin sections. Nevertheless, cytokine expression is limited to lesional skin and presented in a disease-specific pattern. Leveraging a density-based spatial clustering method, we identify specific responder gene signatures in direct proximity of cytokines, and confirm that detected cytokine transcripts initiate amplification cascades of up to thousands of specific responder transcripts forming localized epidermal clusters. Thus, within the abundant and heterogeneous infiltrates of ncISD, only a low number of cytokine transcripts and their translated proteins promote disease by initiating an inflammatory amplification cascade in their local microenvironment.

Aptamer-based Field-Effect Biosensor for Tenofovir Detection.

During medical treatment it is critical to maintain the circulatory concentration of drugs within their therapeutic range. A novel biosensor is presented in this work to address the lack of a reliable point-of-care drug monitoring system in the market. The biosensor incorporates high selectivity and sensitivity by integrating aptamers as the recognition element and field-effect transistors as the signal transducer. The drug tenofovir was used as a model small molecule. The biointerface of the sensor is a binary self-assembled monolayer of specific thiolated aptamer and 6-mercapto-1-hexanol (MCH), whose ratio was optimized by electrochemical impedance spectroscopy measurements to enhance the sensitivity towards the specific target. Surface plasmon resonance, performed under different buffer conditions, shows optimum specific and little non-specific binding in phosphate buffered saline. The dose-response behavior of the field-effect biosensor presents a linear range between 1 nM and 100 nM of tenofovir and a limit of detection of 1.2 nM. Two non-specific drugs and one non-specific aptamer, tested as stringent control candidates, caused negligible responses. The applications were successfully extended to the detection of the drug in human serum. As demonstrated by impedance measurements, the aptamer-based sensors can be used for real-time drug monitoring.

Dose reduction to normal tissues as compared to the gross tumor by using intensity modulated radiotherapy in thoracic malignancies.

BACKGROUND AND PURPOSE: Intensity modulated radiotherapy (IMRT) is a powerful tool, which might go a long way in reducing radiation doses to critical structures and thereby reduce long term morbidities.The purpose of this paper is to evaluate the impact of IMRT in reducing the dose to the critical normal tissues while maintaining the desired dose to the volume of interest for thoracic malignancies. MATERIALS AND METHODS: During the period January 2002 to March 2004, 12 patients of various sites of malignancies in the thoracic region were treated using physical intensity modulator based IMRT. Plans of these patients treated with IMRT were analyzed using dose volume histograms. RESULTS: An average dose reduction of the mean values by 73% to the heart, 69% to the right lung and 74% to the left lung, with respect to the GTV could be achieved with IMRT.The 2 year disease free survival was 59% and 2 year overall survival was 59%. The average number of IMRT fields used was 6. CONCLUSION: IMRT with inverse planning enabled us to achieve desired dose distribution, due to its ability to provide sharp dose gradients at the junction of tumor and the adjacent critical organs.

Intensity modulated radiotherapy in abdominal malignancies: our experience in reducing the dose to normal structures as compared to the gross tumor.

BACKGROUND AND PURPOSE: A better understanding of appropriate sequencing and use of multimodality approach in the management and subsequent improvement in overall survival mandates a vigil on quality of life issues. Intensity modulated radiotherapy (IMRT) is a powerful tool, which might go a long way in reducing radiation doses to critical structures and thereby reduce long term morbidities. The purpose of this paper is to evaluate the impact of IMRT in reducing the dose to the critical normal tissues while maintaining the desired dose to the volume of interest for abdominal malignancies. MATERIALS AND METHODS: During the period January 2002 to March 2004, 11 patients of various sites of malignancies in the abdominal region were treated using physical intensity modulator based IMRT. Plans of these patients treated with IMRT were analyzed using dose volume histograms. RESULTS: An average dose reduction of the mean values by 50% to the liver, 57% to the right kidney, 56% to the left kidney, 66% to the cord and 27% to the bowel, with respect to the GTV could be achieved with IMRT. The two-year disease free survival was 79% and two-year overall survival was 88%. The average number of IMRT fields used was six. CONCLUSION: IMRT with inverse planning enabled us to achieve desired dose distribution, due to its ability to provide sharp dose gradients at the junction of tumor and the adjacent critical organs.

Magnetic resonance imaging in carcinoma cervix--does it have a prognostic relevance.

PURPOSE: To evaluate the prognostic relevance of tumor size as determined on Magnetic Resonance Imaging (MRI) in cervical cancer. METHODS AND MATERIALS: A total of 70 consecutive patients were included in the study. 15 patients underwent surgery alone (Group A), 27 patients underwent surgery followed by adjuvant radiation (Group B), 14 patients underwent concomitant chemo radiation (Group C), and 14 patients underwent radical radiation alone (Group D). External radiation was delivered followed by intra cavitary brachytherapy. Serial MRI scans were performed in all patients before and after completion of treatment on a 1.0 Tesla MRI scanner. Patients were divided into three groups based upon MR volumes 100 cc. A correlation between MR volume, FIGO stage, disease free survival (DFS) and overall survival (OS) was done. Disease free and overall survivals were calculated using Kaplan Meier survival curves according to stage, MR volume and treatment protocol. RESULTS: In group I (MR volume 100 cc), 57% patients had stage II, 14% had stage III and 29% patients had stage IV disease. The DFS and OS did not achieve a level of statistical significance when evaluated as per protocol [DFS at p = 0.0685 and OS p = 0.3242], however a statistical significance was seen when DFS and OS were evaluated according to MR volumes [DFS, p = 0.0015 and OS, p = 0.0001]. CONCLUSION: In cervical cancer, the volume of disease as assessed on MRI may be a better prognostic indicator than FIGO staging and needs further evaluation.

Genes involved in sister chromatid separation and segregation in the budding yeast Saccharomyces cerevisiae.

Accurate chromosome segregation requires the precise coordination of events during the cell cycle. Replicated sister chromatids are held together while they are properly attached to and aligned by the mitotic spindle at metaphase. At anaphase, the links between sisters must be promptly dissolved to allow the mitotic spindle to rapidly separate them to opposite poles. To isolate genes involved in chromosome behavior during mitosis, we microscopically screened a temperature-sensitive collection of budding yeast mutants that contain a GFP-marked chromosome. Nine LOC (loss of cohesion) complementation groups that do not segregate sister chromatids at anaphase were identified. We cloned the corresponding genes and performed secondary tests to determine their function in chromosome behavior. We determined that three LOC genes, PDS1, ESP1, and YCS4, are required for sister chromatid separation and three other LOC genes, CSE4, IPL1, and SMT3, are required for chromosome segregation. We isolated alleles of two genes involved in splicing, PRP16 and PRP19, which impair alpha-tubulin synthesis thus preventing spindle assembly, as well as an allele of CDC7 that is defective in DNA replication. We also report an initial characterization of phenotypes associated with the SMT3/SUMO gene and the isolation of WSS1, a high-copy smt3 suppressor.

Understanding the broader context: the health of the urban native Canadian.

The average life expectancy for a Native male is 66, and for a non-Native male, 76. It is undisputed that the health of the Native Canadian is poor, yet substantial inequalities remain. One of the reasons these inequalities remain is the shallow, limited understanding of the poor health of the urban Native Canadian which, in turn, leads to quick fixes and temporary solutions. The purpose of the following paper is to give nursing leaders an accurate description of the poor health status of the urban Native Canadian, as well as a description of the perpetuating events leading up to their current situation. Once this is understood, the nursing profession can work with urban Native Canadians towards sustainable, long-term solutions. The following paper provides an overview of the historical oppression of Native Canadians, a discussion on the effects of marginalization and the subsequent adaptation process, an overview of the current health status of the urban Native Canadian and a practical assessment tool. The paper concludes with practical suggestions and directions for the future.

The conserved protein kinase Ipl1 regulates microtubule binding to kinetochores in budding yeast.

Chromosome segregation depends on kinetochores, the structures that mediate chromosome attachment to the mitotic spindle. We isolated mutants in IPL1, which encodes a protein kinase, in a screen for budding yeast mutants that have defects in sister chromatid separation and segregation. Cytological tests show that ipl1 mutants can separate sister chromatids but are defective in chromosome segregation. Kinetochores assembled in extracts from ipl1 mutants show altered binding to microtubules. Ipl1p phosphorylates the kinetochore component Ndc10p in vitro and we propose that Ipl1p regulates kinetochore function via Ndc10p phosphorylation. Ipl1p localizes to the mitotic spindle and its levels are regulated during the cell cycle. This pattern of localization and regulation is similar to that of Ipl1p homologs in higher eukaryotes, such as the human aurora2 protein. Because aurora2 has been implicated in oncogenesis, defects in kinetochore function may contribute to genetic instability in human tumors.

A single-step stent delivery, deployment, and expansion system.

Placement of intracoronary stents now comprises a significant number of the percutaneous coronary revascularization procedures being performed in high-volume interventional laboratories in the United States. Stents have become popular because of the reliable acute results, and decreased restenosis. The most common stent currently in use in the United States is the Palmaz-Schatz balloon-expandable stent (PS stent). This stent delivery system (SDS) usually requires predilatation with a balloon and another dilatation, after stent deployment, with a high-pressure balloon in order to adequately expand the stent. We report on a new hybrid stent-delivery system using the PS stent, the sheath from the SDS, and an NC Bandit balloon. This new hybrid system is safe and significantly decreases the procedure time, fluoroscopy time, and need for additional stents due to angiographic dissection resulting from inexact positioning of the high-pressure balloon within the initially deployed stent, and/due to inadequate placement of the first stent. This may be an ideal delivery system for single stent deployment using currently approved FDA equipment.

Effects of free glycerol contained in intravenous fat emulsion on plasma triglyceride determination.

BACKGROUND: Plasma triglyceride analyses to evaluate the clearance of IV fat emulsion are necessary in patients receiving parenteral nutrition. Enzymatic kits for triglyceride analysis measure the glycerol that is hydrolyzed from triglyceride. This study investigates the effect of fat emulsion free glycerol on plasma triglyceride determination. METHODS: Venous blood from fasting volunteers (n = 10) was drawn into tubes containing EDTA and was centrifuged to separate the plasma. An IV fat emulsion containing 2.25% glycerol was added to plasma samples to create serial dilutions ranging from 0.049% to 0.245% final lipid concentration. Total triglyceride and free glycerol concentrations were determined in each dilution and control sample. The free glycerol concentration was subtracted from the total triglyceride concentration to yield a true triglyceride value. RESULTS: Increasing concentrations of fat emulsion added to donor plasma produced increases in total triglyceride and free glycerol concentrations. The increase in free glycerol concentration produced significant concentration-related differences between total and true triglyceride concentrations. CONCLUSIONS: The total triglyceride level overestimated the true triglyceride concentration due to fat emulsion free glycerol. To assure reliable triglyceride results in patients receiving fat emulsion, blanking for free glycerol should be considered.

Changing profile of haematogenous osteomyelitis in a teaching hospital.

A total of 155 consecutive patients of osteomyelitis were studied clinically, radiologically and bacteriologically. The follow up ranged from 2 1/2 to 4 1/2 yr (average 3.5 yr). Age varied from 8 months to 50 yr. Onset of the disease was acute in 106 and insidious in 49 patients. Of these 116 cases were treated conservatively which include 12 treated by incision and drainage. The rest underwent surgery (saucerization, curettage, sequestrectomy etc.). There was a trend for osteomyelitis to shift from the known incidence in early age to adulthood, acute to insidious onset and infection by Gram positive to Gram negative organisms.