9 research outputs found
Onset mechanism of a female patient with Dent disease 2
No full textBackground Approximately 15% of patients with Dent disease have pathogenic variants in theOCRLgene on Xq25-26, a condition that is referred to as Dent disease 2 (Dent-2). Dent-2 patients sometimes show mild extrarenal features of Lowe syndrome, such as mild mental retardation, suggesting that Dent-2 represents a mild form of Lowe syndrome. To date, eight female patients with Lowe syndrome have been reported, but no female Dent-2 patients have been reported. Methods In this study, we performed genetic testing of the first female Dent-2 patient to detect the presence of anOCRLvariant. Aberrant splicing was demonstrated by in vivo, in vitro, and in silico assays, and skewed X-chromosome inactivation (XCI) in our patient and asymptomatic mothers of three Lowe patients with the heterozygousOCRLvariant was evaluated by HUMARA assays using genomic DNA and RNA expression analysis. Results Our patient had anOCRLheterozygous intronic variant of c.1603-3G > C in intron 15 that led to a 169-bp insertion in exon 16, yielding the truncating mutation r.1602_1603ins (169) (p.Val535Glyfs*6) in exon 16. HUMARA assays of leukocytes obtained from this patient demonstrated incompletely skewed XCI (not extremely skewed). On the other hand, the asymptomatic mothers of 3 Lowe patients demonstrated random XCI. These results may lead to our patient's Dent-2 phenotype. Conclusions This is the first report of a female patient clinically and genetically diagnosed with Dent-2 caused by anOCRLheterozygous splicing site variant and skewed XCI. Skewed XCI may be one of the factors associated with phenotypic diversity in female patients with Lowe syndrome and Dent-2
The clinical utility of an index of global oxygenation for guiding red blood cell transfusion in cardiac surgery
No full textScrutinization of 2D and mixed convection flow of generalized Newtonian fluid with nanoparticles and magnetic field
No full textIn this research paper we focuses on presenting the local non-similar solutions for two-dimensional steady and mixed convective flow of electrically conducting Carreau nano-fluid under the influence of Brownian motion and thermophoresis effects. This research paper also present the new mass flux boundary condition of nanoparticles. The governing partial differential equations are converted into the ordinary differential equations by using the local non-similar transformations named as Sparrow-Quack-Boerner local non-similar method. A numerical approach is used to investigate the local nonsimilar solutions of the entire problem. Outcomes for the skin friction, rate of heat and mass transfer have been obtained and discussed for parametric variations of the buoyancy parameter ξ, Magnetic parameter M, Weissenberg number We, viscosity ratio parameter β^{∗}, Brownian motion parameter N_{b}, thermophoresis parameter N_{t}, Prandtl number Pr and Schmidt number Sc. From the obtained results a considerable increase in the skin friction coefficient is observed with the increase in buoyancy parameter ξ. Also ξ shows the relative growth for the momentum and concentration profiles.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author
Role of Cerebral Oximetry in Reducing Postoperative End-Organ Dysfunction After Major Non-Cardiac Surgery: A Randomised Controlled Trial
No full textBackground/Objectives: An increasing number of older individuals require general anaesthesia for major non-cardiac surgery, with 20% displaying postoperative complications. Regional cerebral oxygen saturation (rSO2) correlates with the gold standard of mixed venous oxygen saturation, indicating global perfusion. We hypothesised that rSO2-based anaesthesia reduces organ dysfunction and morbidity after major non-cardiac surgery. Methods: In Singapore and Toronto, we conducted a prospective, double-blind, randomised controlled trial in elderly patients undergoing major non-cardiac surgery, after obtaining research ethics board permission and informed consent. This RCT followed the CONSORT guidelines. Patients received bilateral cerebral oximetry sensors, and the control group received standard care. In the intervention group, an algorithm restored rSO2 if it dropped 10% below baseline for >15 s by adjusting cerebral perfusion pressure, inspired oxygen concentration, end-tidal carbon dioxide, depth of anaesthesia, haemoglobin, and cardiac index. Postoperative complications and outcomes were noted. Categorical data were analysed using Chi-square or Fisher’s exact tests and continuous data using a t-test or a Mann–Whitney U test. The study was powered for 394 patients, but due to the COVID-19 pandemic and funding constraints, this study was terminated at 101 patients. Results: Of 101 patients, 49 were randomised to the control and 52 to the intervention group. A total of 31 (63%) patients in the control group and 30 (58%) in the interventional exhibited bilateral cerebral desaturation. Time of cumulative cerebral desaturation was longer in the control group (23 ± 48 min vs. 9 ± 15 min, respectively, p = 0.01). A total of 142 algorithm-based treatments were employed, restoring rSO2 in 29 (86%) patients. Both groups displayed equal postoperative outcomes. Conclusions: In major non-cardiac surgery, cerebral desaturation is prevalent in over 85% of patients. Although algorithm-guided therapy restored rSO2 in the majority of patients, it did not result in reduced postoperative morbidity
Notes about the Khazar Bricks, Blocks and Fortresses
No full textСтаттю присвячено критичному аналізові публікацій Г. Є. Афанасьєва про переважання візантійських мір довжини в розмірах хозарських цегли, блоків та фортець. Автор "Нотаток" в принципі підтримує тезу про вплив Візантії на виробництво цегли і блоків в Хозарському каганаті, але "теоретико-методологічний підхід" Г. Є. Афанасьєва до проблеми вважає неприйнятним. Автор не знаходить візантійського впливу на хозарську фортифікацію, за винятком фортець Саркел, Правобережна Цимлянська та Хумара.Статтю присвячено критичному аналізові публікацій Г. Є. Афанасьєва про переважання візантійських мір довжини в розмірах хозарських цегли, блоків та фортець. Автор "Нотаток" в принципі підтримує тезу про вплив Візантії на виробництво цегли і блоків в Хозарському каганаті, але "теоретико-методологічний підхід" Г. Є. Афанасьєва до проблеми вважає неприйнятним. Автор не знаходить візантійського впливу на хозарську фортифікацію, за винятком фортець Саркел, Правобережна Цимлянська та Хумара.This article is dedicated to the critical analysis of G. Ye. Afanasiev’s publications about predominating of the Byzantine linear measures in the sizes of the Khazar bricks, blocks and fortresses. The author of “Notes” in principle supports a thesis about influence of Byzantium on the production of bricks and blocks in the Khazar kaganate. However he considers that “theoretical-methological approach” of G. Ye. Afanasiev to the problem is unacceptable. Author does not find Byzantine influence on Khazar fortification, except for the fortresses of Sarkel, Right-Bank Tsymlyanskaya and Humara
Notes about the Khazar Bricks, Blocks and Fortresses
No full textСтаттю присвячено критичному аналізові публікацій Г. Є. Афанасьєва про переважання візантійських мір довжини в розмірах хозарських цегли, блоків та фортець. Автор "Нотаток" в принципі підтримує тезу про вплив Візантії на виробництво цегли і блоків в Хозарському каганаті, але "теоретико-методологічний підхід" Г. Є. Афанасьєва до проблеми вважає неприйнятним. Автор не знаходить візантійського впливу на хозарську фортифікацію, за винятком фортець Саркел, Правобережна Цимлянська та Хумара.Статтю присвячено критичному аналізові публікацій Г. Є. Афанасьєва про переважання візантійських мір довжини в розмірах хозарських цегли, блоків та фортець. Автор "Нотаток" в принципі підтримує тезу про вплив Візантії на виробництво цегли і блоків в Хозарському каганаті, але "теоретико-методологічний підхід" Г. Є. Афанасьєва до проблеми вважає неприйнятним. Автор не знаходить візантійського впливу на хозарську фортифікацію, за винятком фортець Саркел, Правобережна Цимлянська та Хумара.This article is dedicated to the critical analysis of G. Ye. Afanasiev’s publications about predominating of the Byzantine linear measures in the sizes of the Khazar bricks, blocks and fortresses. The author of “Notes” in principle supports a thesis about influence of Byzantium on the production of bricks and blocks in the Khazar kaganate. However he considers that “theoretical-methological approach” of G. Ye. Afanasiev to the problem is unacceptable. Author does not find Byzantine influence on Khazar fortification, except for the fortresses of Sarkel, Right-Bank Tsymlyanskaya and Humara
Clonality - X Chromosome Inactivation Assay
No full textAuthor: Molecular Profiling Initiative, NCI
This method was successful in our lab using prostate tissue and for our specific objectives. Investigators must be aware that they will need to tailor the following protocol for their own research objectives and tissue under study.
Investigators can utilize X chromosome inactivation (methylation) to determine the clonality status of a tumor or premalignant lesion in females. The technique is based on a methylation-sensitive restriction enzyme and analysis of a polymorphic locus on the X chromosome. Clonal cell populations will show "loss" of the non-methylated allele after restriction digest. The assay can be performed on DNA recovered from microdissected samples. Both frozen tissue and fixed-embedded archival tissue can be utilized.
### Materials
1. DNA sample (see [Processing of Microdissected Tissue - DNA-based Analysis](http://cgap-mf.nih.gov/Protocols/Processing/DNA.html))
- Proteinase K (Sigma)
- Proteinase K buffer (0.05 M Tris-HCL, 0.001 M EDTA, 1% Tween 20, 0.1 mg/ml proteinase K, pH 8.0)
- Phenol:chloroform:isoamyl alcohol (1:1:1, v:v:v) (Gibco)
- 3 M Na Acetate, pH 5.2 (Life Technologies)
- Isopropanol
- Glycogen, 10 mg/ml (GenHunter)
- 70% ethanol
- Buffer 1 (50 mM Tris-HCl, pH 8.0, 10 mM MgCl2, 50 mM NaCl)
- Hha I restriction enzyme (Life Technologies)
- REact 2 Buffer (Life Technologies)
- 10X PCR buffer (100 mM Tris-HCL pH 8.3, 500 mM KCL, 15 mM MgCl2, 0,01 % w/v gelatin, autoclaved) (Perkin Elmer)
- dNTP, each 10 mM (Perkin Elmer)
- 7-deaza dGTP, 10 mM (Boehringer Mannheim)
- DMSO, minimum 99.5%, GC (Sigma)
- Forward and reverse primers, each 20 µM (Human Androgen Receptor Gene (HUMARA) CAG trinucleotide repeat microsatellite primers are described in Allen et al)
- AmpliTaq Gold, 5 U/ml (Perkin Elmer)
- a 32P-dCTP, 20 µCi/µl (NEN)
- Gel-Mix 6 (Life Technologies)
- 10X TBE (1 M Tris, 0.9 M Boric Acid, and 0.01 M EDTA) (Life Technologies)
- Loading dye (95% formamide, 20 mM EDTA, 0.05% bromophenol blue, 0.05% xylene cyanol)
### Methods
*TIP: Investigators must be especially careful when using this methodology to analyze archival tissue specimens. Formalin-fixation in particular results in DNA that is difficult to amplify and often produces inconsistent PCR results, including artifactual allelic loss and poor amplification of large products. If this technique is utilized for analysis of archival samples, we highly recommend that replicate experiments (multiple independent dissections, triplicate PCR reactions, etc.) be used to verify results*.
**A: LCM and Proteinase K Treatment**
1. Obtain microdissected cells using the [LCM procedure](http://cgap-mf.nih.gov/Protocols/Microdissection/LCM.html).
- *TIP: The number of cells needed to successfully perform the assay varies depending on the quality and processing conditions of the tissue samples. One thousand cells is recommended as a good starting point*.
- Suspend approximately 1000 microdissected cells in 20 µl proteinase K buffer.
- Incubate overnight at 37°C.
**B: DNA Purification**
1. Lyse the cells overnight at 42°C, in 50 µl phenol:chloroform:isoamyl alcohol.
- Boil 10 min at 95°C.
- Vortex 1 min.
- Centrifuge 20 min at 14,000 rpm at RT.
- Pipet upper phase into new tube and discard lower phase.
- Add 5 µl 3M NaAc.
- Add 250 µl isopropanol.
- Add 2 µl glycogen.
- Vortex briefly.
- Place on dry ice for 30 min.
- Centrifuge 14,000 rpm for 30 min.
- Add 300 µl 70% ethanol.
- Vortex briefly.
- Centrifuge 2 min at 14,000 rpm at RT.
- Discard supernatant.
- Dry pellet at RT for 5 min, or 37°C for 2 min.
- Resuspend the pellet in 20 µl buffer 1.
- Incubate at 37°C for 3 min to completely dissolve the pellet.
**C: DNA Digestion**
1. Pipet 8 µl of the resuspended DNA into a reaction tube.
- Add 1 µl REact buffer 2.
- Add 1 µl Hha I.
- Incubate overnight at 37°C.
- Incubate at 90°C for 10 min.
- Use 2 µl of this digested DNA for PCR (see below).
- Use 2 µl of the non-digested DNA from B18 above as a negative control for PCR.
- *TIP: Parallel analysis of control DNA that is known to be from a monoclonal population is recommended to verify the efficiency of the restriction digest. DNA recovered from a tissue specimen that was processed similar to the tissue sample under study is ideal*.
**D: PCR**
1. Set up the following PCR reaction:
2.Cycles are as follows:
**E: Gel Electrophoresis**
1. Prepare gel consisting of 6% acrylamide as in [LOH protocol](http://cgap-mf.nih.gov/Protocols/DNARNAProteomicAnalysis/DNA/DNAProtocols/LOH.html).
- Add 4 µl dye to 20 µl PCR product.
- Denature the samples for 5 min at 94°C.
- Load 3 µl onto gel.
- Electrophorese at 1800 volts for 1 to 2 hours.
- *TIP: In clonality analysis, the PCR products are ~200 bp. Therefore do not overload the gel with sample and run the gel for a longer period of time to sharpen the bands and increase separation*.
- Transfer the gel to 3 mm Whatman paper and dry. Perform an autoradiograph with Kodak BIOMAX film as described in [LOH protocol](http://cgap-mf.nih.gov/Protocols/DNARNAProteomicAnalysis/DNA/DNAProtocols/LOH.html).
- *TIP: Try a short exposure (1 hour) time first and only re-expose depending on the results*.
### Results
A sample is considered to be composed of a monoclonal cell population when two alleles are recognized in the undigested DNA sample and complete absence of one allele is seen in the digested DNA sample.
- *TIP: DNA that is recovered from microdissected samples and "semi-purified" using a one-step proteinase K buffer strategy will sometimes produce "non-specific" PCR products in addition to the true alleles. Normal-cell DNA from archival specimens from control subjects serves as a good comparator and can assist in interpretation of allele patterns*.
### References
1. Allen RC, Zoghbi HY, Moseley AB, Rosenblatt HM, Belmont JW. [Methylation of HpaII and HhaI sites near the polymorphic CAG repeat in the human androgen-receptor gene correlates with X chromosome inactivation.](http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?db=m&form;=6&uid;=93098238&Dopt;=r) *Am J Hum Genet* 51:1229-39, 1992 .
- Enomoto T, Fujita M, Inoue M, Tanzawa O, Nomura T, Shroyer K. [Analysis of clonality by amplification of short tandem repeats. Carcinomas of the female reproductive tract.](http://www.ncbi.nlm.nih.gov/htbin-post/Entrez/query?db=m&form;=6&uid;=95171001&Dopt;=r) *Diagnostic Molecular Pathology* 3:292-7, 1994
Protocol for a phase 3, randomised, active-control study of four-factor prothrombin complex concentrate versus frozen plasma in bleeding adult cardiac surgery patients requiring coagulation factor replacement: the LEX-211 (FARES-II) trial
No full textIntroduction Reduced thrombin generation is an important component of post cardiopulmonary bypass (CPB) coagulopathy. To replenish coagulation factors and enhance thrombin generation in bleeding surgical patients, frozen plasma (FP) and four-factor prothrombin complex concentrate (4F-PCC) are used. However, the efficacy–safety balance of 4F-PCC relative to FP in cardiac surgery is unconfirmed.Methods and analysis LEX-211 (FARES-II) is an active-control, randomised, phase 3 study comparing two coagulation factor replacement therapies in bleeding adult cardiac surgical patients at 12 hospitals in Canada and the USA. The primary objective is to determine whether 4F-PCC (Octaplex/Balfaxar, Octapharma) is clinically non-inferior to FP for haemostatic effectiveness. Inclusion criteria are any index (elective or non-elective) cardiac surgery employing CPB and coagulation factor replacement with 4F-PCC or FP ordered in the operating room for bleeding management. Patients will be randomised to receive 1500 or 2000 international units of 4F-PCC or 3 or 4 units of FP, depending on body weight. The primary endpoint of haemostatic treatment response is ‘effective’ if no additional haemostatic intervention is required from 60 min to 24 hours after the first initiation of 4F-PCC or FP; or ‘ineffective’ if any other haemostatic intervention (including a second dose of study drug) is required. An estimated 410 evaluable patients will be required to demonstrate non-inferiority (one-sided α of 0.025, power ≥90%, non-inferiority margin 0.10). Secondary outcomes include transfusions, bleeding-related clinical endpoints, coagulation parameters and safety.Ethics and dissemination The trial has been approved by the institutional review boards of all participating centres. Trial completion is anticipated at the end of 2024, and results will be disseminated via publications in peer-reviewed journals and conference presentations in 2025. The results will advance our understanding of coagulation management in bleeding surgical patients, potentially reducing the need for allogeneic blood products and improving outcomes in surgical patients.Trial registration number NCT05523297
Vasopressor use after noncardiac surgery: an international observational study
No full textBackground: Hypotension after major noncardiac surgery is associated with increased morbidity, mortality, and costs, and is often treated with postoperative vasopressor infusions. The frequency of administration in the postoperative period is unknown. Methods: This international prospective cohort study was conducted between October 2020 and October 2023. At each hospital, adults undergoing noncardiac surgery were enrolled into two cohorts: all consecutive patients for 1 week (Cohort A) and an additional sample of up to 30 consecutive patients administered postoperative vasopressor infusions within 1 yr (Cohort B). The primary outcome (Cohort A) was the incidence of postoperative vasopressor infusions, defined as any continuous infusion of vasopressors. Secondary outcomes included in-hospital mortality, organ dysfunction, length of hospital stay, and complications associated with postoperative vasopressor infusions (both cohorts). Results: In total, 25 675 participants were enrolled from 228 hospitals across 42 countries. In Cohort A, 770/19 768 (3.9%) participants received postoperative vasopressor infusions, with vasopressor use ranging between 0% and 18% across hospitals (median odds ratio: 2.30 [credible interval 1.96–2.73]). This variability did not alter after adjustment for case-mix and procedural characteristics. For both cohorts, postoperative vasopressor infusions were associated with higher (15.5%) in-hospital mortality, higher rates of organ failure, and longer hospital stay. Conclusions: Administration of postoperative vasopressors after noncardiac surgery varied across hospitals and was associated with worse outcomes. Variable practice across hospitals could not be explained by differences in case-mix. Clinical trial registration: https://clinicaltrials.gov/study/NCT03805230, ESAIC tracking ID: ESAIC_CTN_SQUEEZE
