Key amino acids residues enhance the ability of CpcR to activate cry gene expression in Bacillus thuringiensis.

Typical Bacillus thuringiensis (Bt) produces one or more parasporal crystals composed of insecticidal Cry proteins during the sporulation, and the parasporal crystals and spores are produced from the same cell. Strain Bt LM1212 is different from typical Bt strains in that its crystals and spores are produced in different cells. Previous studies have found that the cell differentiation process of Bt LM1212 is related to the transcription factor CpcR which activates the cry-gene promoters. In addition, CpcR could activate the Bt LM1212 cry35-like gene promoter (P35) when introduced in the heterologous HD73- strain. It was shown that P35 was only activated in non-sporulating cells. In this study, the peptidic sequences of CpcR homologous proteins found in other strains of the Bacillus cereus group were used as references to identify two key amino acid sites for CpcR activity. The function of these amino acids was investigated by measuring P35 activation by CpcR in strain HD73-. These results will lay a foundation for the optimization of the insecticidal protein expression system in non-sporulating cells.

Construction of Bi-Enzyme Self-Assembly Clusters Based on SpyCatcher/SpyTag for the Efficient Biosynthesis of (R)-Ethyl 2-hydroxy-4-phenylbutyrate.

Cascade reactions catalyzed by multi-enzyme systems are important in science and industry and can be used to synthesize drugs and nutrients. In this study, two types of macromolecules of bi-enzyme self-assembly clusters (BESCs) consisting of carbonyl reductase (CpCR) and glucose dehydrogenase (GDH) were examined. Stereoselective CpCR and GDH were successfully fused with SpyCatcher and SpyTag, respectively, to obtain four enzyme modules, namely: SpyCatcher-CpCR, SpyCatcher-GDH, SpyTag-CpCR, and SpyTag-GDH, which were covalently coupled in vitro to form two types of hydrogel-like BESCs: CpCR-SpyCatcher-SpyTag-GDH and GDH-SpyCatcher-SpyTag-CpCR. CpCR-SpyCatcher-SpyTag-GDH showed a better activity and efficiently converted ethyl 2-oxo-4-phenylbutyrate (OPBE) to ethyl(R)2-hydroxy-4-phenylbutanoate ((R)-HPBE), while regenerating NADPH. At 30 °C and pH 7, the conversion rate of OPBE with CpCR-SpyCatcher-SpyTag-GDH as a catalyst reached 99.9%, with the ee% of (R)-HPBE reaching above 99.9%. This conversion rate was 2.4 times higher than that obtained with the free bi-enzyme. The pH tolerance and temperature stability of the BESCs were also improved compared with those of the free enzymes. In conclusion, bi-enzyme assemblies were docked using SpyCatcher/SpyTag to produce BESCs with a special structure and excellent catalytic activity, improving the catalytic efficiency of the enzyme.

Detection and Identification of the Causal Agents of Dothistroma Needle Blight.

Dothistroma needle blight (DNB) is one of the most damaging foliage diseases of pine in plantations and natural forests worldwide and is caused by two closely related fungi: Dothistroma septosporum and D. pini, which are virtually impossible to differentiate from each other based on morphology. Although diagnosis of DNB based on symptoms is relatively reliable in the later stages of the disease when fruit bodies (conidiomata) are formed, for diagnosis in the early stages, as well as identification of the causal agent at species level, molecular methods are required. In addition, reliable and sensitive diagnostics before sporulation is a prerequisite for early detection to minimize accidental introductions of disease through movement of infected plant materials, especially seedlings. While amplification and sequencing of the ITS region of the rDNA alone is not reliable to differentiate the two species, conventional PCR (cPCR) using species-specific primers or mating type-specific primers and quantitative PCR (qPCR) are widely used and accepted molecular methods to identify and differentiate the DNB pathogens, either from cultures or directly from needles.

ACO/ARO/AIO-21 - Capecitabine-based chemoradiotherapy in combination with the IL-1 receptor antagonist anakinra for rectal cancer Patients: A phase I trial of the German rectal cancer study group.

Purpose: Recent advances in the treatment algorithm of locally advanced rectal cancer (LARC) have significantly improved complete response (CR) rates and disease-free survival (DFS), but therapy resistance, with its substantial impact on outcomes and survival, remains a major challenge. Our group has recently unraveled a critical role of interleukin-1α (IL-1α) signaling in activating inflammatory cancer-associated fibroblasts (iCAFs) and mediating radiation-induced senescence, extracellular matrix (ECM) accumulation, and ultimately therapy resistance. We here summarize the recently initiated ACO/ARO/AIO-21 phase I trial, testing the IL-1 receptor antagonist (IL-1 RA) anakinra in combination with fluoropyrimidine-based chemoradiotherapy (CRT) for advanced rectal cancer. Methods/Design: The ACO/ARO/AIO-21 is an investigator-driven, prospective, open-labeled phase I drug-repurposing trial assessing the maximum tolerated dose (MTD) of capecitabine administered concurrently to standard preoperative radiotherapy (45 Gy in 25 fractions followed by 9 Gy boost in 5 fractions) in combination with fixed doses of the IL1-RA anakinra (100 mg, days -10 to 30). Capecitabine will be administered using a 3 + 3 dose-escalation design (500 mg/m2 bid; 650 mg/m2 bid; 825 mg/m2 bid, respectively) from day 1 to day 30. Response assessment including digital rectal examination (DRE), endoscopy and pelvic magnetic resonance imaging (MRI) is scheduled 10 weeks after completion of CRT. For patients achieving clinical complete response (cCR), primary non-operative management is provided. In case of non-cCR immediate total mesorectal excision (TME) will be performed. Primary endpoint of this phase I trial is the MTD of capecitabine. Discussion: Based on extensive preclinical research, the ACO/ARO/AIO-21 phase I trial will assess whether the IL-1RA anakinra can be safely combined with fluoropyrimidine-based CRT in rectal cancer. It will further explore the potential of IL-1 inhibition to overcome therapy resistance and improve response rates. A comprehensive translational research program will expand our understanding from a clinical perspective and may help translate the results into a randomized phase II trial.

Rapid Detection of Equine Piroplasms Using Multiplex PCR and First Genetic Characterization of Theileria haneyi in Egypt.

Equine Piroplasmosis (EP) is an infectious disease caused by the hemoprotozoan parasites Theileria equi, Babesia caballi, and the recently identified species T. haneyi. Hereby, we used a multiplex PCR (mPCR) targeting the 18S rRNA gene of T. equi and B. caballi for the simultaneous detection of EP in Egyptian equids and examined the presence of T. haneyi infections in Egypt. Blood samples from 155 equids (79 horses and 76 donkeys) collected from different governorates of Egypt were examined by mPCR and PCR targeting T. hayeni. The mPCR method revealed a prevalence of T. equi of 20.3% in horses and of 13.1% in donkeys and a prevalence of B. caballi of 1.2% in horses. B. caballi was not detected in donkeys in the current study. The mPCR method also detected coinfections with both species (2.5% and 1.3% in horses and donkeys, respectively). Additionally, we report the presence of T. haneyi in Egypt for the first time in 53.1% of the horse and 38.1% of the donkey tested samples. Coinfection with T. haneyi and T. equi was found in 13.5% of the samples, while infection with the three EP species was found in 1.9% of the samples.

Resuscitation outcomes of a wireless ECG telemonitoring system for cardiovascular ward patients experiencing in-hospital cardiac arrest.

BACKGROUND/PURPOSE: In-hospital cardiac arrest is a serious issue for hospitalized patients. The documented initial rhythm and detected medical events have been reported to influence the survival of cardiopulmonary resuscitation. This study aimed to identify the effect of continuous real-time electrocardiogram (ECG) monitoring on the prognosis of resuscitated patients in a general cardiac ward. METHODS: We conducted this retrospective study using medical records of hospitalized patients in a cardiovascular ward who experienced an in-hospital cardiac arrest and received cardiopulmonary resuscitation from February 2015 to December 2018. The patients who were considered to be at high risk of cardiac events such as ventricular arrhythmia would receive continuous ECG monitoring. A wireless ECG telemonitoring system was introduced to replace traditional bedside ECG monitors. The outcome measures were the initial success of resuscitation, 24-h survival after resuscitation, and survival to discharge. RESULTS: We enrolled 115 patients with a cardiac arrest during hospitalization, of whom 73 (63%) patients received wireless ECG telemonitoring. Patients receiving continuous ECG monitoring were associated with higher opportunities of initial success of resuscitation and 24-h survival after resuscitation (67.1% vs. 40.5%, p = 0.005; and 49.3% vs. 26.2%, p = 0.015, respectively) when comparing to the non-monitoring group; but no significant difference in survival to discharge (21.9% vs. 16.7%, p = 0.498) was observed. With adjustment of the covariates, the monitoring group was associated with a higher likelihood to reach the initial success of resuscitation (odds ratios [ORs], 3.21; 95% confidence interval [CI], 1.03-9.98). However, the effect of monitoring on 24-h survival and survival to discharge was close to null after adjusting for covariates. CONCLUSION: A wireless ECG telemonitoring system were beneficial to the initial success of resuscitation for patients at high risk of cardiovascular events suffering an in-hospital cardiac arrest; but had less impact on 24-h survival and survival to discharge.

Serological Evidence of West Nile Virus in Wild Birds in Bangladesh.

West Nile Virus (WNV) is a vector-borne zoonotic disease maintained in a sylvatic cycle involving mosquito vectors and birds. To detect WNV and other flavivirus infections in wild resident and migratory birds, we tested 184 samples from 19 identified species within nine families collected during 2012-2016 from four districts in Bangladesh. We tested serum samples for the immunoglobulin G (IgG) antibody against WNV using competitive Enzyme-Linked Immunosorbent Assay (c-ELISA), whereas tracheal and cloacal swabs were subjected to consensus Polymerase Chain Reaction (c-PCR) for the detection of the flavivirus RNA. Overall, we detected 11.9% (n = 22; 95% CI: 0.07-0.16) samples were seropositive, including 15.9% in the migratory wild birds and 10.7% in the resident wild birds. The migratory wild Tufted duck showed 28.5% seropositivity, whereas the resident wild house crows showed 12.5% seropositivity. None of the swab samples was positive for flavivirus RNA infection (0%, n = 184; 95% CI: 0-0.019). These study findings recommend continued surveillance for early detection and to better understand the epidemiology of WNV and other flavivirus circulation in both birds and mosquitoes in Bangladesh.

Free convective PCR: From principle study to commercial applications-A critical review.

Polymerase chain reaction (PCR) is an extremely important tool for molecular diagnosis, as it can specifically amplify nucleic acid templates for sensitive detection. As another division of PCR, free convective PCR was invented in 2001, which can be performed in a capillary tube pseudo-isothermally within a significantly short time. Convective PCR thermal cycling is implemented by inducing thermal convection inside the capillary tube, which stratifies the reaction into spatially separate and stable melting, annealing, and extension zones created by the temperature gradient. Convective PCR is a promising tool that can be used for nucleic acid diagnosis as a point-of-care test (POCT) due to the significantly simplified heating strategy, reduced cost, and shortened detection time without sacrificing sensitivity and accuracy. Here, we review the history of free convective PCR from its invention to development and its commercial applications.

Large-scale study for the photocatalytic degradation of paracetamol using Fe2O3/TiO2 nanocomposite catalyst and CPC reactor under natural sunlight radiations.

Heterogeneous photocatalysis is a promising advanced oxidation process for the degradation of emerging contaminants. In this regard, Hematite (α-Fe2O3) doped TiO2 nanocomposite catalyst was synthesized via sol-gel method. The catalyst was prepared in large quantities (225 g) comparatively with other studies and characterized by X-ray diffraction (XRD), Field emission scanning electron microscopy (FESEM), Energy-dispersive X-ray (EDX), and nitrogen gas physisorption studies. The bandgap of the synthesized catalyst was determined using UV-vis diffused reflectance spectroscopy (DRS), and the point of zero charge (PZC) was identified by measuring the zeta potential (ζ-potential) of the nanoparticles. A large-scale study was conducted using a modified Compound Parabolic Collector Reactor (CPCR) for the degradation of paracetamol under natural sunlight irradiations. The operating parameters including the initial concentration of paracetamol, initial pH of the solution, and catalyst loading were optimized using face-centered central composite design (FCCD) based on response surface method (RSM) to obtain the maximum degradation efficiency of paracetamol. •The simplified and direct sol-gel method described helps in the synthesis of a novel nanocomposite catalyst (Fe2O3/TiO2) in large quantities while maintaining good characteristics compared to other methods.•The described treatment method using the modified CPCR will allow the degradation of paracetamol in a more sustainable and green manner.•Optimizing the operating parameters that have a significant influence on the degradation of paracetamol will contribute towards higher degradation rates.

Validation of [125I]CPCR4.3 as an investigative tool for the sensitive and specific detection of hCXCR4 and mCXCR4 expression in vitro and in vivo.

BACKGROUND: The development and clinical translation of [68Ga] Pentixafor has substantially promoted the relevance of non-invasive PET imaging of CXCR4 expression in a broad spectrum of diseases, including cancer and inflammation. Its pronounced selectivity for the human receptor (hCXCR4), however, precludes the use of [68Ga] Pentixafor for imaging receptor expression and dynamics in CXCR4-related diseases in endogenous mouse models. To overcome this restriction, [125I]CPCR4.3, a structurally related pentapeptide ligand, has been evaluated as a preclinical tool for efficient in vitro and in vivo targeting of hCXCR4 and mCXCR4. RESULTS: Compared to the reference [68Ga] Pentixafor, [125I]CPCR4.3 showed 2.4- to 11-fold increased specific binding to human cancer cell lines with different hCXCR4 expression levels (Jurkat, Daudi, HT-29, SH-5YSY, MCF-7, LNCaP) as well as strong and highly specific binding to mCXCR4 expressing cells (mCXCR4-transfected CHO cells, Eµ-myc 1080, 4 T1), which was not detectable for [68Ga]Pentixafor. This is the consequence of the equally high affinity of iodo-CPCR4 to hCXCR4 and mCXCR4 (IC50 = 5.4 ± 1.5 and 4.9 ± 1.7 nM, respectively) as opposed to [natGa] Pentixafor (hCXCR4: 42.4 ± 11.6 nM, mCXCR4: > 1000 nM). Additionally, [125I]CPCR4.3 showed enhanced tracer internalization (factor of 1.5-2 compared to the reference). In vivo biodistribution studies in immunocompetent Black Six and immunocompromised CD-1 nude mice showed predominant hepatobiliary excretion of [125I]CPCR4.3 (logP = 0.51), leading to high activity levels in liver and intestines. However, [125I]CPCR4.3 also showed high and specific accumulation in organs with endogenous mCXCR4 expression (spleen, lung, adrenals), even at low receptor expression levels. CONCLUSIONS: Due to its excellent hCXCR4 and mCXCR4 targeting efficiency, both in vitro and in vivo, [125I]CPCR4.3 represents a sensitive and reliable tool for the species-independent quantification of CXCR4 expression. Its suboptimal clearance properties will certainly restrict its use for in vivo imaging applications using 123I (for SPECT) or 124I (for PET), but due to its high and specific accumulation in mCXCR4 expressing tissues, [125I]CPCR4.3 holds promise as a powerful preclinical tool for the investigation and quantification of CXCR4 involvement and kinetics in various murine disease models via, e.g., biodistribution and autoradiography studies.

Effect of tris(2-carboxyethyl)phosphine and tertiary butyl alcohol on the performance of convection polymerase chain reaction.

Rapid and on-site DNA-based molecular detection has become increasingly important for sensitive, specific, and timely detection and treatment of various diseases. To prepare and store biomolecule-containing reagents stably, reducing agents are used during protein preparation, and freeze-drying technology has been applied to the protein reagents. Some of the additives used during these processes may affect subsequent processes such as polymerase chain reaction (PCR). In this study, we evaluated the impact of TCEP, a reducing agent, and TBA, a freeze-drying medium, on the performance of convection PCR (cPCR) using a battery-operable PCR device. Singleplex cPCR detection of a 249 bp amplicon from human genomic DNA suggested that approximately 82% of performance was achieved in the presence of 0.1 mM TCEP and 1% TBA. The limit of detection and the minimum number of cycles at which amplicons began to appear was a little lower (~ 82% efficiency) or higher (20 vs 15 cycles), respectively, in the chemical-treated group than in the control group. With larger amplicons of 500 bp, the chemical-treated group revealed approximately 78% of performance and amplicons started to appear at 20 cycles of cPCR in both groups. Similar results were obtained with multiplex cPCR amplification.

Performance of 2 Polymerization Protocols Targeting Cloned Toxoplasma Parasites.

BACKGROUND: Toxoplasma gondii is a common parasitic infection of humans. Infection is usually mild. Serious complications can occur in pregnant and immunocompromised patients. AIM: The present study aims to investigate the performance of 2 different PCR protocols; real-time quantitative molecular assays (qPCR) and conventional molecular assays (cPCR), using 2 different sets of primers and by using cloned purified Toxoplasma genomic substances to be evaluated as reference samples. METHODS: The target DNA was provided in 8 different quantities. RESULTS: Amplification failure was reported only with the cPCR in samples of low concentrations using both primer sets. Quantitative PCR detected the 8 different dilutions of the purified Toxoplasma gondii using the 2 sets of primers while cPCR was sensitive to detect only 6 different dilutions. CONCLUSION: Generally real-time quantitative molecular assays, is easy to use method compared to conventional PCR assay and produces more reliable results within only one hour time but still the possible application of qPCRs in routine diagnosis necessitates analysis of a large number of clinical samples in further studies to make the proper choice.

Multifarious Ga-68 Labeled PET Radiopharmaceuticals in Imaging Various Malignancies.

The gamut of gallium labeled radiopharmaceuticals contributes to augmented variety in molecular imaging approach for in vivo identification of tumor characteristics. The spectrum ranges from somatostatin receptor based target-specific imaging agents, to those used for tumor imaging based on specific receptor types extending into ones used for therapeutic monitoring. The versatility of gallium chemistry provides the needed advantage for imaging which is further exploited in clinical practice influences the specificity of tumor imaging. Ga-68 has revealed applicability in labeling compounds from nanoparticles to micro as well as macromolecules. We in this image, present variety of frequently and infrequently used gallium labeled radiopharmaceuticals, for imaging diverse malignancies other than conventional established tracers.

HDP2: a ribosomal DNA (NTS-ETS) sequence as a target for species-specific molecular diagnosis of intestinal taeniasis in humans.

BACKGROUND: Taenia solium, T. asiatica and T. saginata tapeworms cause human taeniasis and are the origin of porcine and bovine cysticercosis. Furthermore, T. solium eggs can cause human cysticercosis, with neurocysticercosis being the most serious form of the disease. These helminth infections are neglected tropical diseases and are endemic in several countries in the Americas, Asia and Africa. As a result of globalization, migration in particular, the infections have been extending to non-endemic territories. Species-specific diagnosis of taeniasis is subject to drawbacks that could be resolved using molecular approaches. In the present study, conventional and real-time amplification protocols (cPCR and qPCR) based on the T. saginata HDP2 sequence were applied in the differential diagnosis of taeniasis (T. saginata, T. solium) in both fecal samples and proglottids expelled by patients. The HDP2 homolog in T. solium was cloned and characterized. RESULTS: Semi-nested cPCR and qPCR (Sn-HDP2 cPCR and Sn-HDP2 qPCR) amplified T. saginata and T. solium DNA, with an analytical sensitivity of 40 and 400 fg, respectively, and identically in both protocols. Eighteen taeniasis patients were diagnosed directly with T. saginata or T. solium, either from proglottids or fecal samples with/without eggs (detected using microscopy), based on the optimized Sn-HDP2 qPCR. After cloning, the T. solium HDP2 homolog sequence was confirmed to be a ribosomal sequence. The HDP2 fragment corresponded to a non-transcribed sequence/external transcribed repeat (NTS/ETS) of ribosomal DNA. Compared with the T. saginata HDP2 homolog, the T solium HDP2 sequence lacked the first 900 nt at the 5' end and showed nucleotide substitutions and small deletions. CONCLUSIONS: Sn-HDP2 cPCR and Sn-HDP2 qPCR were set up for the diagnosis of human taeniasis, using proglottids and fecal samples from affected patients. The new Sn-HDP2 qPCR protocol was the best option, as it directly differentiated T. saginata from T. solium. The diagnosis of an imported T. solium-taeniasis case and nine European T. saginata cases was relevant. Finally, the cloning and sequencing of the T. solium HDP2 fragment confirmed that HDP2 was part of a ribosomal unit.

Exploration of teaching practice of foreign body airway obstruction first aid / 中华医学教育探索杂志

In order to optimize the teaching process, improve the teaching quality and cultivate of emergency nursing talents with comprehensive quality, we systematically designed the teaching process of foreign body airway obstruction first aid. From the design ideas and curriculum innovation, we focused on the use of a variety of teaching methods, and we combined it with modern information technology, which demonstrated the practicability and operability of first aid skill. This teaching practice has proved that the systematically teaching reform design is helpful to improve the learning efficiency of nursing students and cultivate the comprehensive quality of students' emergency response ability.

Correlation between Success Rates of Cardiopulmonary Cerebral Resuscitation and the Educational Level of the Team Leader; A Cross-Sectional Study.

OBJECTIVES: To determine the correlation between the success rates of the cardiopulmonary cerebral resuscitation (CPCR) and the team's leader education and skill level in Shiraz, southern Iran. METHOD: This cross-sectional study was conducted during a 6-month period from October 2007 to March 2008 in Nemazee hospital of Shiraz. We included all the patients who underwent CPCR due to cardiopulmonary arrest in emergency room of Nemazee hospital during the study period. We recorded the rates of return of spontaneous circulation (ROSC) and discharge rate (DR) of all the patients. The correlation between these two parameters and the team leader's education and skill level was evaluated. RESULTS: Overall we included total number 600 patients among whom there were 349 men (58.1%) and 251(41.8%) women with mean age of 58.9±42.6.  We found that 270 (45.1%) patients had ROSC, while 330 (54.9%) patients died. Overall 18 (6.6%) patients were discharged   from hospital (3% of all participants). We found that the ROSC was significantly higher in those with specialist leader (anesthesiologist or pediatrician) when compared to those in whom CPCR was conducted by technicians (55.2% vs. 30.7%; p=0.001). CONCLUSION: Conducting CPCR by persons with higher medical degrees resulted in higher rate of ROSC but not in more discharge rate. Inspite of the fact that the rate of ROSC following CPCR was closely analogous to that of developed countries, discharge rate was lower. This indicates that in our region, much more attention needs to be paid to post-resuscitation care and organizing training programs and to cover more resuscitation by CPCR team, conducted by the specialists.

Establishment of a universal fluorescent multiplex cPCR method for detection of copy number variations in mice / 中国实验动物学报

Objective To establish a high throughput general multiple competitive polymerase chain reaction ( cPCR) detecting method of copy number variations ( CNVs) for the population of chromosome 1 substitution strains from wild mice.Method The selected 14 loci, including 11 CNVs on chromosome 1 and internal control loci on other three chromosmes (Chr 7, Chr 19 and Chr X), were detected based on the universal fluorescent primer multiple competitive pol-ymerase chain reaction.All specific cloned plasmids were constructed as competitors.Results Altogether 11 CNVs were designed in one panel, and the copy of Chr X accurately reflects the gender.Conclusions A rapid and high-throughput fluorescent multiplex cPCR assay is established which can be used for detection of copy number variations on chromosome 1 in mice.

The Effects of Video Programs of Cardiopulmonary Cerebral Resuscitation Education

PURPOSE: The aim of this study was to identify the effect of video programs of cardiopulmonary cerebral resuscitation (CPCR) education of cardiopulmonary cerebral resuscitation of nurses. METHODS: The subjects of the study were 64 nurses working in a university hospital. Nurse's CPCR performance have been measured four times (pre-test, post-test at immediately, 3 months and 6 months after intervention). Data were collected from February to August 2013. RESULTS: There were significant differences in knowledge, attitude, self-efficacy, and performance between groups by measure time. And there were significant interactions in knowledge, self-efficacy, and performance between groups, within groups, except for the attitude. The video programs of CPCR interventions appear to be effective in the improvement of knowledge, self-efficacy, and performance, as compared to the control group. CONCLUSION: The video programs of CPCR education was an effective intervention to improve and retain the level of knowledge, attitude, self-efficacy and performance. And the video program of CPCR education have an advantage of self-learning effect for nurses with shift work. Therefore video programs of CPCR education will be utilized for continuing nurse's education.

Depleted TGF-ß1 levels in end stage renal disease patients from North India.

End stage renal disease is a clinical state that extends from chronic renal failure and is marked by an irreversible loss of renal function. TGF-ß1 mediated renal fibrosis is a common pathology implicated in this form of kidney disease. In this study circulating protein and mRNA levels of TGF-ß1 cytokine were investigated among ESRD patients and respective controls from North India. Physician diagnosed 192 ESRD patients, on hemodialysis, and 130 normal controls participated in the present study. TGF-ß1 circulating levels were measured by ELISA and its expression was quantified using competitive-PCR. Mean TGF-ß1 protein levels were 2.7-fold lower in ESRD patients as compared to normal controls (p<0.001). Additionally, TGF-ß1 mRNA transcripts of this cytokine were also significantly lower in the diseased population compared to controls (p<0.001). These results imply that TGF-ß1 has not played its anticipated pro-fibrotic role and anti-inflammatory function in the studied population.

A real-time convective PCR machine in a capillary tube instrumented with a CCD-based fluorometer.

This research reports the design, analysis, integration, and test of a prototype of a real-time convective polymerase chain reaction (RT-cPCR) machine that uses a color charged coupled device (CCD) for detecting the emission of fluorescence intensity from an RT-cPCR mix in a microliter volume glass capillary. Because of its simple mechanism, DNA amplification involves employing the cPCR technique with no need for thermocycling control. The flow pattern and temperature distribution can greatly affect the cPCR process in the capillary tube, a computational fluid dynamics (CFD) simulation was conducted in this study for the first time to estimate the required period of an RT-cPCR cycle. This study also tested the PCR mix containing hepatitis B virus (HBV) plasmid samples by using SYBR Green I fluorescence labeling dye to assess the prototype performance. The measured results from the image-processing scheme indicate that the RT-cPCR prototype with a CCD-based fluorometer can achieve similar DNA quantification reproducibility compared to commercial machines, even when the initial DNA concentration in the test PCR mix is reduced to 10 copies/µL.