Intact Membranes Research Articles

Modifications of the respiratory chain of Bacillus licheniformis as an alkalophilic and cyanide-degrading microorganism.

Bacillus licheniformis can use cyanide as a nitrogen source for its growth. However, it can also carry out aerobic respiration in the presence of this compound, a classic inhibitor of mammalian cytochrome c oxidase, indicating that B. licheniformis has a branched respiratory chain with various terminal oxidases. Here, we studied the modifications in the respiratory chain of B. licheniformis when cells were cultured in Nutrient Broth, an alkaline medium with ammonium, or an alkaline medium with cyanide. Then, we measured oxygen consumption in intact cells and membranes, enzyme activities, carried out 1D and 2D-BN-PAGE, followed by mass spectrometry analysis of BN-PAGE bands associated with NADH, NADPH, and succinate dehydrogenase activities. We found that cell growth was favored in a nutrient medium than in an alkaline medium with cyanide. In parallel, respiratory activity progressively decreased in cells cultured in the rich medium, alkaline medium with ammonium, and the lowest activity was in the cells growing in the alkaline medium with cyanide. B. licheniformis membranes contain NADH, NADPH, and succinate dehydrogenases, and the proteomic analysis detected the nitrate reductase and the bc, caa3, aa3, and bd complexes. The succinate dehydrogenase migrated with a molecular mass of 375kDa, indicating its association with the nitrate reductase (115kDa + 241kDa, respectively). The NADH dehydrogenase of B. licheniformis forms aggregates of different molecular mass.

Influence of Schneiderian membrane perforation during maxillary sinus floor augmentation with lateral approach on dental implant survival rates: a retrospective study in a university setting.

The primary objective of this study was to compare implant survival rates (ISR) in patients undergoing maxillary sinus floor augmentation (MSFA) with lateral approach with and without membrane perforation. Secondary objectives were to establish the percentage of perforations in these procedures and to evaluate the influence of perforation size on ISR. This retrospective study included patients requiring MSFA with lateral approach. Cases were assigned to two groups according to the occurrence or not of perforation. The exact size of each perforation was registered. Descriptive statistics and associations between the groups were calculated. This study analyzed data from 90 MSFA in 72 patients. Membrane perforation occurred in 24.44%. A total of 170 implants were placed; 72.35% were placed under intact membranes and 27.65% on repaired membranes. The overall ISR in MSFA procedures with intact membranes was 98.37%, and 93.62% in perforated membranes. No statistically significant relationship between groups was observed although the odds of implant failure increased by 4.125 times when perforation occurred. In turn, no statistically significant relationship was observed between perforation size and ISR. Implants inserted below repaired membranes had a lower ISR compared with implants inserted below intact membranes although the difference was not statistically significant. Moreover, no significant statistical correlation was observed between perforation size and ISR. Membrane perforation does not have a significant influence on subsequent implant survival rates. Knowledge of the exact size of the membrane perforation is essential for deciding on the right treatment plan.

In vitro photocurrents from spinach thylakoids following Mn depletion and Mn-cluster reconstitution

Biohybrid devices that generate an electrical signal under the influence of light due to photochemical reactions in photosynthetic pigment-protein complexes have many prospects. On the one hand, the oxygen-evolving complex of photosystem II allows the use of ubiquitous water as a source of electrons for photoinduced electron transfer in such devices; on the other hand, it is the most vulnerable part of the photosynthetic apparatus. From the perspective of sustainable operation of bio-based hybrid devices, it is helpful to analyze how removing or modifying the Mn cluster will affect the performance of the bio-hybrid device. This work analyzed photocurrent generation in a liquid three-electrode solar cell based on manganese-depleted and reactivated thylakoid membranes. Membranes lacking Mn could not produce any significant photocurrent until manganese chloride was added. After adding MnCl2, the cell could produce current when exposed to light. This current was about a few percent from cells with intact thylakoid membranes. However, the photoactivation procedure made it possible to restore up to 75 % of the photocurrent of cells based on intact thylakoid membranes. The main objective of this work is to answer the question about the possibility of photocurrent generation in a biohybrid system based on thylakoid membranes using artificial analogs of the native oxygen-evolving complex. Photoactivation with manganese chloride is the simplest way to obtain preparations devoid of the native Mn cluster, but capable of oxidizing water.

Choosing The Right Method For Labour Induction: Misoprostol Alone Versus Misoprostol With Foleys Catheter

Background The aim of the retrospective study is to compare effectiveness, safety of oral misoprostol and Foley’s catheter for labour induction with misoprostol. The efficacy is measured by induction to delivery interval, the total number of normal vaginal spontaneous delivery within 24 hours without any complications. The two methods of induction were compared. As there is no ideal method of induction of labour, several methods and a combination method with Foley’s catheter and misoprostol was used to find the advantage of both methods. Materials and Methods: This retrospective study was conducted among the 400 patients of which 200 patients who had oral misoprostol alone and the rest 200 patients who had Foley’s catheter with misoprostol over the period of four months from 2024 May to August. Inclusion criteria consisted of singleton pregnancy greater than or equal to 37 weeks with intact membrane, cephalic presentation,bishops score less than or equal to 4. The exclusion criteria comprised of previous LSCS or uterine surgery, rupture of membrane. Results: The results were analyzed. The comparison of the maternal effects and also the neonatal outcome were studied in each group. We came to a conclusion that oral misoprfostol alone with Foley’s catheter is a better method for labour induction. Conclusion: The synergetic action of Foley’s catheter and misoprostol is remarkable. It is economical and preferable in developing countries.

Choosing The Right Method For Labour Induction: Misoprostol Alone Versus Misoprostol With Foleys Catheter

Background The aim of the retrospective study is to compare effectiveness, safety of oral misoprostol and Foley’s catheter for labour induction with misoprostol. The efficacy is measured by induction to delivery interval, the total number of normal vaginal spontaneous delivery within 24 hours without any complications. The two methods of induction were compared. As there is no ideal method of induction of labour, several methods and a combination method with Foley’s catheter and misoprostol was used to find the advantage of both methods. Materials and Methods: This retrospective study was conducted among the 400 patients of which 200 patients who had oral misoprostol alone and the rest 200 patients who had Foley’s catheter with misoprostol over the period of four months from 2024 May to August. Inclusion criteria consisted of singleton pregnancy greater than or equal to 37 weeks with intact membrane, cephalic presentation,bishops score less than or equal to 4. The exclusion criteria comprised of previous LSCS or uterine surgery, rupture of membrane. Results: The results were analyzed. The comparison of the maternal effects and also the neonatal outcome were studied in each group. We came to a conclusion that oral misoprfostol alone with Foley’s catheter is a better method for labour induction. Conclusion: The synergetic action of Foley’s catheter and misoprostol is remarkable. It is economical and preferable in developing countries.

Mass Spectrometry Probe Combined with Machine Learning to Capture the Relationship between Metabolites and Mitochondrial Complex Activity at the Whole-Cell Level.

Mitochondrial complex activity controls a multitude of physiological processes by regulating the cellular metabolism. Current methods for evaluating mitochondrial complex activity mainly focus on single metabolic reactions within mitochondria. These methods often require fresh samples in large quantities for mitochondria purification or intact mitochondrial membranes for real-time monitoring. Confronting these limitations, we shifted the analytical perspective toward interactive metabolic networks at the whole-cell level to reflect mitochondrial complex activity. To this end, we compiled a panel of mitochondrial respiratory chain-mapped metabolites (MRCMs), whose perturbations theoretically provide an overall reflection on mitochondrial complex activity. By introducing N-dimethyl-p-phenylenediamine and N-methyl-p-phenylenediamine as a pair of mass spectrometry probes, an ultraperformance liquid chromatography-tandem mass spectrometry method with high sensitivity (LLOQ as low as 0.2 fmol) was developed to obtain accurate quantitative data of MRCMs. Machine learning was then combined to capture the relationship between MRCMs and mitochondrial complex activity. Using Complex I as a proof-of-concept, we identified NADH, alanine, and phosphoenolpyruvate as metabolites associated with Complex I activity based on the whole-cell level. The effectiveness of using their concentrations to reflect Complex I activity was further validated in external data sets. Hence, by capturing the relationship between metabolites and mitochondrial complex activity at the whole-cell level, this study explores a novel analytical paradigm for the interrogation of mitochondrial complex activity, offering a favorable complement to existing methods particularly when sample quantities, type, and treatment timeliness pose challenges. More importantly, it shifts the focus from individual metabolic reactions within mitochondria to a more comprehensive view of an interactive metabolic network, which should serve as a promising direction for future research into the functional architecture between mitochondrial complexes and metabolites.

Surgical site infection and antimicrobial use following caesarean section at QECH in Blantyre, Malawi: a prospective cohort study

BackgroundSurgical site infections (SSIs) are one of the most common healthcare-associated infections and preventable complication of surgical procedure; continue to threaten public health with significant effects on the patients and health care human and financial resources. Therefore, this study aimed to determine the incidence of SSIs, risk factors and common microorganisms associated with SSI and assess the practice of antimicrobial use in women following Caesarean Section (CS) at Queen Elizabeth Central Hospital (QECH).MethodsThis was a hospital-based quantitative prospective study design involving pregnant women who underwent a CS between February, 2023 and July, 2023 at QECH with 30 day-follow-ups. Wound specimens (wound swabs) were collected from all infected CS wounds and processed at QECH main laboratory, and susceptibility testing was conducted using the Kirby-Bauer disk diffusion method with results reported only as susceptible, intermediate, or resistant and the collected data was analyzed using Stata.ResultsThe overall cumulative incidence of SSI recorded at QECH during the study period was 9.61% (20 cases out of 208). Of these, 19 (95%) of them reported superficial SSI following CS. The mean age was 26.1 years with a standard deviation of 6.2. All pregnant women who underwent for CS received antibiotic prophylaxis. This study revealed that 138 (66.35%) patients received both preoperative antibiotics (ceftriaxone) and post-CS antibiotics without knowing the specific bacterial organism isolated. This study revealed that ruptured membrane had twice the incidence of SSIs compared to intact membrane (χ2 = 2.0922), though not statistically significant. The majority of patients with SSIs (n = 12, 60%) were readmitted and 5 (25%) out of 20 with SSIs had antimicrobial resistance following susceptibility testing. Staphylococcus aureus was the most common organism (3, 60%) and other bacterial isolates included were Enterobacteriaceae and Acinetobacter baumanni.ConclusionThe incidence of SSIs and inappropriate antimicrobial use following CS remains a challenge at QECH. Therefore, due to increased number of SSIs following CS with relative emergence of AMR ensure intensive infection prevention and control practices, establishing AMS program and routine surveillance of SSIs at QECH.

Permeability-Controlled Probe for Ratiometric Detection of Plasma Membrane Integrity and Late Apoptosis.

The destruction of plasma membrane integrity is closely related to immune response, neuronal injury, cell apoptosis, and other pathological events. However, the construction of ratiometric fluorescent probes capable of detecting plasma membrane integrity remains a significant challenge, hindering in-depth studies on related biomedical areas. Herein, a polarity-responsive fluorescent probe was constructed for the ratiometric detection of cell membrane integrity for the first time. The probe targeted intact plasma membranes in healthy cells and relocated into the cytoplasm to give significantly red-shifted fluorescence after plasma membrane damage. Molecular simulations revealed that the high transmembrane barrier and amphipathic nature of the probe were responsible for its targeting ability. With the probe, the ratiometric detection of late apoptosis stage was realized for the first time, and the membrane damage of tumor cells induced by UV irradiation, toxins, and antitumor drugs was visualized. The effect of formaldehyde on membrane integrity was evaluated using a probe, and cysteine was proved to be a potential detoxifier to counteract the toxicity of formaldehyde.

Primary and revision myringoplasty in children: Long-term outcome and analysis of the factors influencing the results

ObjectiveAssess the long-term outcome of pediatric myringoplasty. MethodsTympanoplasty type I, myringoplasty, was performed on 85 children (91 consecutive operations, 74 primary and 17 revisions) under 16 years of age. The perforations were sequela either to acute or chronic inflammatory middle ear disease. Medial grafting technique was employed with temporalis fascia. Adenoidectomy was performed earlier on all but five children. The preoperative observation period exceeded one year for all patients. The mean follow-up was 5.4 (SD 3.6) years. ResultsThe long-term graft take rate was 84% for primary myringoplasty, and 53% for revision operations. The re-perforations were associated with postoperative discharge, subtotal or total perforation and revision surgery. In the cohort, sixty ears (66%) were completely healthy (intact tympanic membrane in normal position without adhesions and with good mobility) after follow-up. Spontaneous healing took place in 37% after re-perforation. The preoperative ventilation tube treatment did not affect the outcome and there were no differences between age groups. Preoperative sonotubometry or Valsalva test results did not correlate with outcome. Four ears needed a ventilation tube during the follow-up due to poor ventilation. The mean pure tone thresholds improved significantly after operation. ConclusionsMyringoplasty in children is a reliable procedure without age restrictions. One year of preoperative observation excludes most unstable ears, and high tendency of spontaneous healing after re-perforation suggests the need for a lengthy follow-up before revision surgery.

A limbal stem cell deficiency murine model with residual limbal stem cells.

Bilateral limbal stem cell deficiency (LSCD) is a significant cause of corneal blindness and is more difficult to treat, as compared with unilateral LSCD because no source of autologous limbal stem cells (LSCs) remains in these patients. Thus, bilateral patients could be candidates for treatment with allogeneic LSC transplants that require long-term systemic immunosuppression therapy. Thus, if possible, for the correct candidates, using autologous LSCs could be a preferred treatment. Recent in vivo laser confocal microscopic examination of the ocular surface in situ , combined with impression cytology, has indicated that some patients diagnosed with a complete bilateral LSCD possess residual LSCs. However, it remains unknown whether these residual LSCs still have stem cell potential due to the lack of animal models that mimic this pathology. The goal of the current study is to make a complete LSCD model that possesses evidence of residual LSCs. We induced complete LSCD in mice using two methods: (1) removed the corneal epithelium and the epithelial basement membrane using a rotating burr, and (2) removed the corneal epithelium using 20% ethanol but retained an intact epithelial basement membrane. A complete LSCD was defined by a lack of CK12-positive corneal epithelial cells and the presence of infiltrating CK19-positive conjunctival epithelial cells. Corneas were examined for wound closure, corneal opacity, LSC exhaustion, and inflammation. We observed that complete LSCD mice without an intact epithelial basement membrane resulted in few residual LSCs. By contrast, complete LSCD mice that retained the epithelial basement membrane were accompanied by a reduced inflammatory response plus a significant number of residual LSCs. This model will allow future studies to determine the function of residual LSCs in complete LSCD.

Intracellularly Gelated Macrophages Loaded with Probiotics for Therapy of Colitis.

Probiotics therapy has garnered significant attention in the treatment of inflammatory bowel disease (IBD). However, a large number of oral administrated probiotics are inactivated after passing through the gastric acid environment, and their ability to colonize in the intestine is also weak. Herein, this study develops a novel probiotics formulation (GM-EcN) by incorporating Escherichia coli Nissle 1917 (EcN) into intracellularly gelated macrophages (GM). Intracellular hydrogel is designed to load and prevent EcN from digestion in gastric juice, and GM acts as a macrophage-like carrier to carry the attached probiotics to colonize in the inflammatory intestine. In addition, hydrogel serves as an ideal cytoskeletal structure to maintain the intact cell morphology and membrane structure of GM, comparable to source macrophages. Due to the receptor-ligand interaction, inflammation-related membrane proteins enable GM as a cell sponge to sequestrate and neutralize multiple inflammatory cytokines. In vivo treatment demonstrates that GM-EcN efficiently alleviates IBD symptoms and enhances gut microbiota recovery.

Histopathologic and ultrastructural findings in oak (Quercus spp.) toxicity on 2 beef cattle farms in Colorado.

Natural oak toxicity, a phenomenon sporadically reported in the United States, is due to consumption of any part of most oak trees (Quercus spp.). Ruminants, mainly cattle, are disproportionately susceptible to oak toxicity. Toxicity is attributed to degradation of the oak plant hydrolysable tannins by rumen microbes and enzymes into absorbable low-molecular-weight metabolites, which are postulated to bind and damage endothelial cells by unknown mechanisms. The clinical manifestations of acute toxicosis are nonspecific or broadly suggestive of renal disease due to acute tubular injury. Here we document the clinical, gross, histopathologic, and novel ultrastructural features of natural acute oak nephrotoxicity in 3 beef calves on 2 farms in Colorado, USA. Gross postmortem findings included perirenal edema with renomegaly and hemorrhagic gastroenteritis. Histologically, renal tubular epithelial necrosis was severe, with hemorrhage and intratubular hyaline casts. Transmission electron microscopy revealed extensive involvement of proximal and distal convoluted tubules, with predominantly intact basement membranes, and glomerular and interstitial endothelial injury and necrosis. The ultrastructural details of toxic nephropathy and vasculopathy induced by oak metabolites in natural cases of bovine oak toxicosis have not been described previously, to our knowledge.

The disease burden of untreated chronic otitis media in Indigenous children from remote communities does not improve over time. A longitudinal study.

In Australia, middle ear disease disproportionately affects Indigenous children, leading to poor hearing outcomes. This study aimed to determine the natural history of untreated chronic otitis media in Indigenous children in remote South Australia. Baseline and 3 year follow-up data was collected from Indigenous children aged 5-18 years living on the Anangu Pitjantjatjara Yankunytjatjara Lands. 4-frequency pure-tone audiometry was used to determine hearing levels. Middle ear pathology was determined by video-otoscopy with tympanometry and classified at baseline as group 1 (normal), group 2 (abnormal with intact tympanic membrane), or group 3 (perforated tympanic membrane). A total of 253 children were included in this study. Children in group 1 (20.6 ± 1.5 dBHL; Mean ± SD) had significantly better hearing outcomes at 3 year follow-up than children with abnormal ears (groups 2 and 3) (23.8 ± 7.0 dBHL), P < 0.001. The difference was greatest for group 1 versus 3 (27.6 ± 8.4 dBHL), P < 0.001, followed by 1 versus 2 (22.4 ± 5.8 dBHL), P = 0.009, and between 2 (22.4 ± 5.8 dBHL) and 3 (27.6 ± 8.4 dBHL), P < 0.001. Hearing in Indigenous children with untreated middle ear pathology remains poor at follow-up compared to those without pathology. Intervention is therefore critical to prevent persisting poor hearing outcomes.

High cooling rate of 60°C/min around ice nucleation during cryopreservation compromises chicken sperm viability.

The present study compares two protocols for the cryopreservation of chicken semen. Both protocols had an initial low cooling rate in the first step, followed by higher cooling rates around ice nucleation (Protocol 1) or following the dissipation of the latent heat of fusion (Protocol 2) in the second step. Semen ejaculates obtained from 12 roosters were diluted with Rootex with 6% dimethylformamide and frozen following either Protocol 1 (from +5°C to -10°C at 5°C/min and from -10°C to -130°C at 60°C/min) or Protocol 2 (from +5°C to -35°C at 7°C/min and from -35°C to -140°C at 60°C/min). Compared with fresh semen, following both protocols, cryopreservation resulted in reduced post-thaw sperm quality (p < .001). Post-thaw percentage of sperm with an intact plasma membrane was greater using Protocol 2 than Protocol 1 (p < .05). The results suggest that high cooling rates around the time of ice nucleation are not recommendable.

Markers of epithelial-mesenchymal transformation in placentas from very early preterm births

Premature birth complicates one in ten births in the world, two-thirds of which are caused by spontaneous labor and premature rupture of membranes (PROM). The opinion about the inflammatory origin of preterm birth is generally accepted, more and more attention is devoted to the phenomena of epithelial-mesenchymal transformation (EMT).The objective: to study markers of cell proliferation and EMT in samples of placentas from very early preterm, early preterm and term births.Materials and methods. Placenta samples were examined from 101 women who had labour at 22–27 weeks of gestation on the background of PROM (I group), 102 women who had labour at 22–27 weeks on the background of intact fetal membranes (II group), 100 women who had labour at 28-34 weeks on the background of PROM (III group) and 102 women in labour – at 28-34 weeks on the background of intact membranes (IV group). Samples of 60 placentas from timely normal births were included in the control group.The presence of proliferation factor Ki-67 was determined in placenta samples by immunohistochemical method, cytokeratin-18 content was determined in 10 placentas of each group, and vimentin – in 10 amniotic membrane samples. Regarding the proliferation antigen Ki-67, the placentas were divided according to the detection of the sign in the distal, proximal and basal villi. The expression of cytokeratin and vimentin in syncytium and amnion cells was measured in conventional units and compared between groups.The statistical evaluation of the obtained differences was carried out using the Student’s test.Results. Proliferation antigen Ki-67 was detected in 61.9% of all placentas, including term births. In the groups of very early preterm birth (PB), they were detected mainly in villi of distal localization – in 44.6% in the group of PROM and in 51.0% – in the group of labour on the background of intact membranes. In the case of deliveries on the background of intact membranes in extremely premature terms, only 22.5% of the samples had a positive antigen in the basal villi, on the background of PROM in these terms – in 36.7%, in the groups of early PB – in 79.4% and 74.0% respectively.Distal Ki-67 expression in such placentas was a rare finding – 10.0% in the III group, 11.8% – in the IV group, and 8.3% – in placentas from term deliveries.In all gestational periods, the expression of cytokeratin in amniotic membranes in placentas from births on the background of PROM is statistically lower than in placentas from births on the background of intact membranes.In the case of very early PB, the greater expression of vimentin in the distal villi (0.324±0.001 units in the I group and 0.356±0.007 units in the II group) than in the intermediate villi (0.234±0.004 units and 0.248±0.002 units, respectively) and basal ones (0.178±0.002 units and 0.189±0.006 units, respectively). This once again confirms that the inflammatory reaction and the associated EMT at birth before 28 weeks are mainly of fetal origin.In placentas from early PB, the pattern is the opposite – from 0.345±0.007 units and 0.369±0.009 units in III and IV groups, respectively, in basal villi to 0.257±0.004 units and 0.239±0.005 – in intermediate villi and 0.178±0.009 units and 0.165±0.005 units – in the distal ones.Conclusions. 1. The expression of the inflammatory proliferation marker Ki-67 was detected in two thirds of the studied placentas regardless of the term of delivery, but very early PBs are characterized by the predominant localization of the marker in the distal villi, while in early PBs – in the basal and proximal villi. 2. The expression of EMT markers indicates the fetal origin of the inflammatory response in very early PB – less cytokeratin in syncytial cells and more vimentin in distal chorionic villi. In placentas from early PB and term deliveries, a more pronounced expression of vimentin in the basal vessels of the chorion dominates.

The Association Between Maternal ABO Blood Group and the Occurrence of Spontaneous Preterm Birth: A Retrospective Population-Based Cohort Study.

There is limited and inconsistent evidence that imply a relationship between ABO blood types and rate of preterm birth (PTB). We aim to examine the association between maternal ABO blood group and PTB rate. A retrospective-study conducted at a university teaching institution on data collected between 2013 and 2019. Women who delivered a viable neonate at ≥ 24 weeks without major malformations were included. Indicated PTBs were excluded. PTB and early PTB were defined as deliveries that occurred < 37 and < 34 weeks respectively. PTB was further divided into 3 subgroups according to etiology: membranes rupture, intact membranes, and placental abruption regardless of membranes' status. The primary outcome was spontaneous PTB rate. Of 19,301 women included, PTB and early PTB rates were 7.3% (1,418/19,301) and 2.3% (440/19,301) respectively. Rates of PTB in blood groups A, B, O, and AB, were 7.3%, 6.9%, 7.5%, and 7.5% respectively (p = 0.68). There was no significant difference according to etiology. Rates of early PTB were also comparable (p = 0.63). After adjustment for demographic and obstetric variables, blood type was associated with increased placental abruption rate among women who had early PTB (p = 0.038). Placental abruption rate was significantly higher in group A (22.5%) compared to group B (14.1%), (adjusted p = 0.04) and group O (14.0%), (adjusted p = 0.01). The rate in group AB was 17.1%, (adjusted p = 0.85). In conclusion, no association was found between a particular blood group and PTB rate. Women with group A, admitted in early PTB, had an increased risk that the underlying etiology was placental abruption.

Comprehensive management of chronic ear disease: Consecutive patient analysis at a tertiary children's hospital

ObjectiveTo characterize medical complexity and advanced interventions necessary to treat pediatric patients with chronic ear disease. MethodsA retrospective case review of children who received surgery for chronic otitis media, cholesteatoma, or tympanic membrane perforation at a tertiary children's hospital from 2020 to 2024 was performed. The frequency and type of medical complexities as well as distribution and success of advanced otologic interventions were recorded. ResultsOperations were performed on 80 ears. Forty-five percent were in medically complex patients. Twenty-five percent had a syndrome, 13 % had American Society for Anesthesiologists Severity Class 3 or more, and 38 % attended multi-disciplinary programs. Advanced audiology testing protocols were utilized in 53 % of patients, and multiple children received complex sleep and airway operations. Advanced surgical techniques were required in 58 %. Post-operatively, an intact tympanic membrane was achieved in 95 % of operated ears (p < 0.01). Residual and recurrent cholesteatoma were each present in 5 % of cases, with mean follow-up of 11.6 months. Otorrhea reduced from 64 % of patients pre-operatively to 2 % post-operatively (p < 0.01), including cessation in all individuals who reported social/bullying issues related to ear drainage/odor. Unrestricted water exposure was achieved in 99 % of individuals (p < 0.01). Patients showed a mean improvement after surgery of 7.6 dB, as measured by the change in air-bone gap (p < 0.05). Conclusions: Comprehensive treatment of otologic conditions included care that was holistic of all medical needs, integrated with other services, developmentally appropriate, encompassing of non-otologic procedures, technically advanced, effective, and functionally-focused. Surgical teams should endeavor together to provide this combination of services.

Topological Heterogeneity of Protein Kinase C Modulators in Human T-Cells Resolved with In-Cell Dynamic Nuclear Polarization NMR Spectroscopy.

Phorbol ester analogs are a promising class of anticancer therapeutics and HIV latency reversing agents that interact with cellular membranes to recruit and activate protein kinase C (PKC) isoforms. However, it is unclear how these esters interact with membranes and how this might correlate with the biological activity of different phorbol ester analogs. Here, we have employed dynamic nuclear polarization (DNP) NMR to characterize phorbol esters in a native cellular context. The enhanced NMR sensitivity afforded by DNP and cryogenic operation reveals topological heterogeneity of 13C-21,22-phorbol-myristate-acetate (PMA) within T cells utilizing 13C-13C correlation and double quantum filtered NMR spectroscopy. We demonstrate the detection of therapeutically relevant amounts of PMA in T cells down to an upper limit of ∼60.0 pmol per million cells and identify PMA to be primarily localized in cellular membranes. Furthermore, we observe distinct 13C-21,22-PMA chemical shifts under DNP conditions in cells compared to model membrane samples and homogenized cell membranes, that cannot be accounted for by differences in conformation. We provide evidence for distinct membrane topologies of 13C-21,22-PMA in cell membranes that are consistent with shallow binding modes. This is the first of its kind in-cell DNP characterization of small molecules dissolved in the membranes of living cells, establishing in-cell DNP-NMR as an important method for the characterization of drug-membrane interactions within the context of the complex heterogeneous environment of intact cellular membranes. This work sets the stage for the identification of the in-cell structural interactions that govern the biological activity of phorbol esters.

Regulatory effects of vitamin E nano-emulsion on blood metabolites, immunological variables, testicular architecture, and sperm ultrastructure of heat-stressed V-line rabbit bucks

The study investigated the beneficial effects of vitamin E-loaded nano-emulsion (NEVE) on hemato-biochemical changes, oxidative stress, inflammatory responses, immunological variables, and semen quality in heat-stressed rabbit bucks. Forty adult V-line rabbit bucks were randomly assigned to four groups and fed diets containing NEVE at 0 (NEVE0), 50 (NEVE50), 100 (NEVE100), and 200 (NEVE200) mg/kg for 12 weeks. The NEVE had a mean particle diameter of 32.6 nm, a polydispersity index of 0.6, and a zeta potential of −31.1 mV. The dietary NEVE significantly improved the hematological, blood protein, lipid profile, and liver functions of rabbit bucks (p < 0.05). The NEVE200 treated group showed higher levels of nitric oxide, TAC, and SOD in blood serum and seminal plasma compared to the control group (p < 0.05). Meanwhile, levels of MDA, IL-4, and TNF-α were significantly shrunken in response to the dietary treatment (p < 0.0001), while the IgA and IgG were improved in the NEVE100 and NEVE200 groups. The NEVE200 exhibited higher progressive motility, ejaculate volume, viability, and better libido (lower reaction time) than the untreated group (p < 0.05). TEM images illustrated that NEVE200 exhibited higher sperm cells with a normal structure, intact plasma membranes, and acrosomes than the other groups. Moreover, testicular architecture was improved with NEVE supplementation. Overall, the dietary addition of NEVE at 100 or 200 mg/kg could be an effective strategy to improve the health and semen quality of rabbit bucks during hot climates.

Enhancing patient care: a multimodality strategy for dilated cardiomyopathy in Chagas disease

Chagas disease is a systemic illness characterized by acute and chronic phases. If untreated, it can lead to dysfunction of vital organs, notably the heart, ultimately resulting in heart failure. Transmission primarily occurs through the feces of triatomine insects carrying the protozoan parasite Trypanosoma cruzi, either via a bite wound or intact mucous membranes. Diagnosis of Chagas disease involves serological tests, electrocardiographic findings, and imaging studies. A 58-year-old male patient from Peru with chronic dilated cardiomyopathy underwent evaluation at a tertiary care hospital. Given the uncertain etiology, a comprehensive diagnostic approach was adopted, emphasizing the pivotal role of cardiovascular magnetic resonance imaging and computed tomography angiography in managing chronic cardiomyopathy of Chagas disease. Leveraging these imaging modalities together could augment our ability to evaluate myocardial inflammation and tailor therapeutic strategies accordingly.