Mitochondrial complexome reveals quality-control pathways of protein import.

Mitochondria have crucial roles in cellular energetics, metabolism, signalling and quality control1-4. They contain around 1,000 different proteins that often assemble into complexes and supercomplexes such as respiratory complexes and preprotein translocases1,3-7. The composition of the mitochondrial proteome has been characterized1,3,5,6; however, the organization of mitochondrial proteins into stable and dynamic assemblies is poorly understood for major parts of the proteome1,4,7. Here we report quantitative mapping of mitochondrial protein assemblies using high-resolution complexome profiling of more than 90% of the yeast mitochondrial proteome, termed MitCOM. An analysis of the MitCOM dataset resolves >5,200 protein peaks with an average of six peaks per protein and demonstrates a notable complexity of mitochondrial protein assemblies with distinct appearance for respiration, metabolism, biogenesis, dynamics, regulation and redox processes. We detect interactors of the mitochondrial receptor for cytosolic ribosomes, of prohibitin scaffolds and of respiratory complexes. The identification of quality-control factors operating at the mitochondrial protein entry gate reveals pathways for preprotein ubiquitylation, deubiquitylation and degradation. Interactions between the peptidyl-tRNA hydrolase Pth2 and the entry gate led to the elucidation of a constitutive pathway for the removal of preproteins. The MitCOM dataset-which is accessible through an interactive profile viewer-is a comprehensive resource for the identification, organization and interaction of mitochondrial machineries and pathways.

Development of an advanced electrochemical biosensing platform for E. coli using hybrid metal-organic framework/polyaniline composite.

Because of numerous merits (e.g., the possibility of their synthesis in 1-D, 2-D, and 3-D forms, large surface-to-volume ratio, and flexible framework functionality), metal-organic frameworks (MOFs) are envisaged as excellent media for the development of biosensors for diverse analytes present in environmental media. The present research work, for the first time, reports the development of a Cu-MOF based electrochemical biosensor for highly sensitive detection of E. coli bacteria. In order to realize an MOF-based electrochemically active platform, Cu3(BTC)2 (BTC = 1,3,5-benzenetricarboxylic acid) was mixed with polyaniline (PANI). The spectroscopic/morphological characterizations of the resulting composite were established with the aid of FT-IR, UV-visible spectroscopy, X-ray diffraction, electron microscopy, and surface area analysis. The thin films of Cu3(BTC)2-PANI, on an indium-tin oxide (ITO) substrate, were bio-interfaced with anti-E. coli antibodies for use as a novel biosensing electrode. Based on the electrochemical impedance spectroscopy (EIS) technique of signal measurement, the above sensor exhibited high sensitivity to detect very low concentrations of E. coli (2cfu/mL) in a short response time (~2 min) and was also selective in the presence of other non-specific bacteria. As a novel highlight of the research, this new MOF/PANI based detection platform for E. coli has shown improved performance than many of the previously reported electrochemical biosensors.

A graphene electrode functionalized with aminoterephthalic acid for impedimetric immunosensing of Escherichia coli.

A screen-printed electrode prepared from graphene oxide (GO) has been functionalized with 2-aminoterephthalic acid, followed by the exploitation of this functional material in an electrochemical immunoassay for Escherichia coli (E. coli) by immobilizing the antibody on its surface. The functionalization steps followed a straightforward approach and were proven by various instrumental techniques. The detection of E. coli with antibody immobilized electrodes was performed using electrochemical impedance spectroscopy. The analyses were carried out using the hexacyanoferrate redox couple as the electrochemical probe. The present method has a wide analytical range (from 2.2 × 102 to 2.2 × 108 cfu.mL-1), a low limit of detection (2 cfu.mL-1), fast response (4 min), and good stability (up to 2 months). The analytical performance of the biosensor was comparable to the previously reported electrochemical biosensors for E. coli. As such, the approach of functionalization of graphene with 2-aminoterephthalic acid should be useful to allow the development of other similar sensing systems for other environmentally and clinically important analytes. Graphical abstractSchematic representation of the preparation and the function of an amino-functionalized graphene oxide (NH2-GO) based impedimetric biosensor for the electrochemical detection of E. coli.

CHOROIDAL STRUCTURAL CHANGES AND VASCULARITY INDEX IN STARGARDT DISEASE ON SWEPT SOURCE OPTICAL COHERENCE TOMOGRAPHY.

PURPOSE: To evaluate structural changes in the choroid of patients with Stargardt disease using swept source optical coherence tomography scans. METHODS: A retrospective comparison cohort study was conducted on 39 patients with Stargardt disease, and on 25 age and gender matched-healthy controls. Subfoveal choroidal thickness (SFCT) was computed from the swept source optical coherence tomography machine, and the scans were binarized into luminal area and stromal areas, which were then used to derive choroidal vascularity index (CVI). Choroidal vascularity index and SFCT were analyzed independently using linear mixed effects model. RESULTS: There was no significant difference in SFCT between the 2 groups (347.20 ± 13.61 µm in Stargardt disease vs. 333.09 ± 18.96 µm in the control group, P = 0.548). There was a significant decrease in the CVI among eyes with Stargardt disease as compared with the normal eyes (62.51 ± 0.25% vs. 65.45 ± 0.29%, P < 0.001). There was a negative association between visual acuity and CVI (correlation coefficient = -0.75, P < 0.001) and a positive association between visual acuity and SFCT (correlation coefficient = 0.21, P = 0.035). CONCLUSION: Choroidal vascularity index appears to be a more robust tool compared with SFCT for choroidal changes in Stargardt disease. Choroidal vascularity index can possibly be used as a surrogate marker for disease monitoring. A decrease in CVI was associated with a decrease in visual function in eyes with Stargardt disease.

Use of metabolic monitors in a multidisciplinary Intensive Care Unit: A prospective pilot study of 20 patients.

INTRODUCTION: Caloric intake of critically ill patients are usually calculated using predictive equations. Recent advances in gas exchange measurements have the potential to estimate energy expenditure at the bedside and at different time periods. MATERIALS AND METHODS: Energy needs of critically ill patients were estimated over a period of 3 months using simplistic formula of 25 kcal/kg/day estimated energy expenditure (EEE), Harris-Benedict equation (HBE) (Basal energy expenditure [BEE]) and M-COVX™ metabolic monitor resting energy expenditure (REE) on day 4 of Intensive Care Unit (ICU) admission. Calculations based on HBE were taken as standard, and percentage errors (PE) were calculated for each patient for values derived from simplistic formula and metabolic monitor. Adequacy of nutritional intake in ICU was also assessed. RESULTS: Metabolic monitor could be used in only 20/70 patients. The mean age of patients was 40 years, 65% were males, and average body mass index was 23.69 kg/m(2). Intermittent intolerance to feeds was reported in 50%. Values of REE and EEE were greater than BEE in 70% of patients. A significant difference was reported in values of PE of ≤20% and ≥30%; P = 0.0003 and 0.0001, respectively estimated using REE and EEE. CONCLUSIONS: It is not feasible to use metabolic monitors in all patients. Variability in readings is large and further studies are needed to establish the validity of its measurements. Calculations using simplistic formulas are much closer to values obtained using HBE.

Postoperative monitoring in pregnant patients undergoing surgery for advanced malignancy in last trimester: How long is enough?

Surgery for advanced breast malignancy in the last trimester of pregnancy is uncommon. We present successful management of a 32-year-old woman, 30 weeks pregnant with stage 3 breast malignancy, for surgery followed by normal labor and chemotherapy. Surgery and intraoperative period were uneventful. Patient had uterine contractions 36 h postsurgery, which were managed timely with active interventions and tocolytics. Risk of premature labor following nonobstetric surgery in pregnant patients is estimated to be 8.3%, but majority of the studies have been carried out in second trimester following appendectomy. There is insufficient data in literature regarding the estimation and duration of persistence of risk of premature labor in these patients. No guidelines are available regarding how long they need to be monitored for premature labor. There is some evidence, although little that risk of premature labor persists for 7 days postsurgery. In the absence of convincing studies and guidelines, we recommend postoperative monitoring for at least 7 days in patients undergoing major surgeries for malignancies in last trimester. Multidisciplinary approach is required to manage these patients.

Comparison of Ultrasound Versus Ultrasound With Nerve Stimulation-Guided Obturator Nerve Block to Prevent Adductor Spasm in Patients Undergoing Transurethral Resection of Bladder Tumor: A Randomized Controlled Study.

Background This study aimed to compare ultrasound versus ultrasound with nerve stimulation-guided obturator nerve block (ONB) for the prevention of adductor spasm in patients undergoing transurethral resection of bladder tumor (TURBT). Methodology This randomized controlled study included 240 adult patients in the age group of 30 to 70 years undergoing TURBT for lateral and posterolateral wall bladder tumors who fulfilled the American Society of Anesthesiologists grade I and II criteria. The patients were divided into two groups: group U (n = 120) included patients who underwent ONB using an ultrasound-guided technique and group UN (n = 120) included patients who underwent ONB using ultrasound with the nerve stimulation technique. Block performance time, adductor jerks/spasms, adductor muscle power, and patient and surgeon satisfaction were compared. A P-value <0.05 was considered statistically significant. Results The mean block performance time in group U was significantly less (4.4 ± 0.82 minutes) than in group UN (6.55 ± 0.37 minutes). Compared to group U, group UN had significantly fewer adductor jerks/spasms during the surgery (7.76% vs. 20.35%, p = 0.006), significantly more surgeon satisfaction (92.24% vs. 79.65%, p = 0.006), significantly more patient satisfaction (92.24% vs. 79.65%, p = 0.006), and comparable complications (excessive bleeding and minor bladder injury) and adductor muscle power after the block (p > 0.05). Conclusions ONB using the nerve stimulation technique under ultrasound guidance has a longer mean block performance time, a higher success rate, and higher surgeon satisfaction than ONB under ultrasound guidance only.

Giant Cell Tumor of Mandible : A Case Report.

Giant cell tumor is locally aggressive primary benign neoplasm of bone with tendency of frequent recurrence, metastasis and malignant transformation. Because of the rarity of the disease involving mandible, no definite treatment guideline is established. Surgical treatment is the treatment of choice for giant cell tumor. Due to its proximity to vital structures including skull base, the recurrent disease associated with less invasive procedure could be difficult to manage while more invasive procedure will result in higher morbidity and complex reconstruction. Medical management with denosumab or zoledronic acid has been advocated in surgically unresectable disease. We present a rare case of giant cell tumor of mandible. Patient was a 33 years old lady who presented with gradually progressive painful swelling in chin. After exclusion of distant metastasis by F-18 FDG PET scan, she underwent en-bloc resection of the tumor with free fibula flap reconstruction. During 6 months of follow up visit patient had no recurrence.

Reimagining Carbon Nanomaterial Analysis: Empowering Transfer Learning and Machine Vision in Scanning Electron Microscopy for High-Fidelity Identification.

In this report, we propose a novel technique for identifying and analyzing diverse nanoscale carbon allotropes using scanning electron micrographs. By precisely controlling the quenching rates of undercooled molten carbon through laser irradiation, we achieved the formation of microdiamonds, nanodiamonds, and Q-carbon films. However, standard laser irradiation without proper undercooling control leads to the formation of sparsely located diverse carbon polymorphs, hindering their discovery and classification through manual analyses. To address this challenge, we applied transfer-learning approaches using convolutional neural networks and computer vision techniques to achieve allotrope discovery even with sparse spatial presence. Our method achieved high accuracy rates of 92% for Q-carbon identification and 94% for distinguishing it from nanodiamonds. By leveraging scanning electron micrographs and precise undercooling control, our technique enables the efficient identification and characterization of nanoscale carbon structures. This research significantly contributes to the advancement of the field, providing automated tools for Q-materials and carbon polymorph identification. It opens up new opportunities for the further exploration of these materials in various applications.

Ocular manifestations in renal diseases.

The eyes and kidneys are the targets for end-organ damage in multiple pathologies. Both these organs develop during the same embryonic stage around the fourth to sixth week of gestation, thus sharing a strong correlation between both eye and kidney diseases. Both the eyes and kidneys can be the target of the systemic disease process; however, the eyes can also be affected as a consequence of renal disease or its treatment. Risk factors such as diabetes, hypertension, and smoking are commonly shared between kidney and eye diseases. Ocular manifestations can be predictive of renal disease, and/or patients with renal disease are at higher risk for developing ocular manifestations. Various congenital anomalies of the eyes and kidneys can also present as an oculorenal syndrome. This article summarizes the ocular pathology, which can be seen in renal diseases.

Giant Parathyroid Adenoma: A case Report.

Giant parathyroid adenoma (GPA) is a large tumor weighing more than 3.5 g but can weigh as much as 110 g as compared to parathyroid adenomas which usually weigh in the vicinity of 1 g and measure around 1-2 cm. These mainly present with primary hyperparathyroidism. We describe an interesting case and clinical course of a young woman with giant parathyroid adenoma. A 30-year-old Indian female presented with acute pancreatitis and acute kidney injury and was found to have hypercalcemia and hyperparathyroidism on further evaluation. Her physical examination revealed firm, non-tender, and mobile swelling of size 5 × 2 cm on the left side of her neck. USG neck showed a large heterogenous hypoechoic lesion in the left lower pole region of the left thyroid gland measuring 50 × 24 × 29 mm with a volume of 18 ccs. A parathyroid scan was performed using 99Tc Sesta MIBI tracer which was suggestive of a large adenoma in relation to the left lobe of the thyroid gland. She underwent minimally invasive parathyroidectomy with excision of left parathyroid adenoma under general anesthesia. Intraoperative frozen section confirmed the specimen to be parathyroid tissue. Her parathyroid hormone (PTH) level decreased from 1900 pg/ml in the pre-operative level to 242 pg/ml in the immediate postoperative period.

Aptamers for Viral Detection and Inhibition.

Recent times have experienced more than ever the impact of viral infections in humans. Viral infections are known to cause diseases not only in humans but also in plants and animals. Here, we have compiled the literature review of aptamers selected and used for detection and inhibition of viral infections in all three categories: humans, animals, and plants. This review gives an in-depth introduction to aptamers, different types of aptamer selection (SELEX) methodologies, the benefits of using aptamers over commonly used antibody-based strategies, and the structural and functional mechanism of aptasensors for viral detection and therapy. The review is organized based on the different characterization and read-out tools used to detect virus-aptasensor interactions with a detailed index of existing virus-targeting aptamers. Along with addressing recent developments, we also discuss a way forward with aptamers for DNA nanotechnology-based detection and treatment of viral diseases. Overall, this review will serve as a comprehensive resource for aptamer-based strategies in viral diagnostics and treatment.

Mechanisms and pathways of mitochondrial outer membrane protein biogenesis.

Outer membrane proteins integrate mitochondria into the cellular environment. They warrant exchange of small molecules like metabolites and ions, transport proteins into mitochondria, form contact sites to other cellular organelles for lipid exchange, constitute a signaling platform for apoptosis and inflammation and mediate organelle fusion and fission. The outer membrane contains two types of integral membrane proteins. Proteins with a transmembrane ß-barrel structure and proteins with a single or multiple α-helical membrane spans. All outer membrane proteins are produced on cytosolic ribosomes and imported into the target organelle. Precursors of ß-barrel and α-helical proteins are transported into the outer membrane via distinct import routes. The translocase of the outer membrane (TOM complex) transports ß-barrel precursors across the outer membrane and the sorting and assembly machinery (SAM complex) inserts them into the target membrane. The mitochondrial import (MIM) complex constitutes the major integration site for α-helical embedded proteins. The import of some MIM-substrates involves TOM receptors, while others are imported in a TOM-independent manner. Remarkably, TOM, SAM and MIM complexes dynamically interact to import a large set of different proteins and to coordinate their assembly into protein complexes. Thus, protein import into the mitochondrial outer membrane involves a dynamic platform of protein translocases.

Protein Quality Control at the Mitochondrial Surface.

Mitochondria contain two membranes, the outer and inner membrane. The outer membrane fulfills crucial functions for the communication of mitochondria with the cellular environment like exchange of lipids via organelle contact sites, the transport of metabolites and the formation of a signaling platform in apoptosis and innate immunity. The translocase of the outer membrane (TOM complex) forms the entry gate for the vast majority of precursor proteins that are produced on cytosolic ribosomes. Surveillance of the functionality of outer membrane proteins is critical for mitochondrial functions and biogenesis. Quality control mechanisms remove defective and mistargeted proteins from the outer membrane as well as precursor proteins that clog the TOM complex. Selective degradation of single proteins is also an important mode to regulate mitochondrial dynamics and initiation of mitophagy pathways. Whereas inner mitochondrial compartments are equipped with specific proteases, the ubiquitin-proteasome system is a central player in protein surveillance on the mitochondrial surface. In this review, we summarize our current knowledge about the molecular mechanisms that govern quality control of proteins at the outer mitochondrial membrane.

Randomized controlled trial of trehalose: An efficient autophagic bioprotectant in the management of dry eye disease.

PURPOSE: To compare the therapeutic effect of sodium hyaluronate (SH)-trehalose (Trehalube, Microlabs, Bangalore, India, SH 0.1% and trehalose 3%) or SH (0.1% Hylotears, Raymed, Chandigarh, India) alone in patients with dry eye disease (DED). MATERIALS AND METHODS: Patients were randomized into two groups: SH-trehalose (SH 0.1% and trehalose 3%) or SH (0.1% Hylotears) alone. The Ocular Surface Disease Index (OSDI) questionnaire was used to assess patient's symptoms. Patients were followed up at 4 and 8 weeks, and OSDI score, tear film break-up time (TBUT), tear film height (TFH), Schirmer's test, and conjunctival staining were evaluated at each visit. RESULTS: A total of 384 patients were included in the study, 192 patients in each arm. The mean age of participants was 37.62 + 14.4 years and 225 were women (56%). The improvement in Schirmer's test was significantly better in the SH-trehalose group at 8 weeks (5.26 + 4.3 mm, 95% confidence interval = 4.6-5.9 mm) compared to the SH group (3.71 + 3.9, 95% confidence interval = 3.15-4.28 mm). The TBUT and TFH showed slight improvement at 4 weeks in both groups, but not at 8 weeks. There were no group differences at all-time points in terms of conjunctival staining and OSDI-based grades of DED. CONCLUSION: It was found that treating dry eye with SH-trehalose leads to greater improvement in the Schirmer's values and TBUT after 8 weeks of sustained use in patients with DED, and this was more pronounced in those with severe DED.

Effect of Oral Propranolol on Periocular Infantile Capillary Hemangioma: Outcomes Based on Extent of Involvement.

PURPOSE: To evaluate the efficacy of oral propranolol in the treatment of periocular infantile capillary hemangioma (CHI) based on the involvement of embryological facial placodes and their extent of anatomical involvement. METHODS: Retrospective study of 27 patients. RESULTS: The mean age at the presentation of periocular CHI was 4 months (median, 3 months; range, <1-14 months). There were 11 (41%) males and 16 (59%) females. Based on embryological facial placodes, the involvement was focal in 16 (59%) cases and segmental in 11 (41%) cases. Based on the anatomical distribution, the lesions were preseptal in 4 (15%), postseptal in 13 (48%), and combined in 10 (37%) cases. The duration of use of oral propranolol was 10 months (median, 10 months; range, 4-16 months). Overall, the mean % resolution of periocular CHI was 78% (median, 90%; range, 20%-100%). The mean percentage resolution of focal lesions was 69% (median, 83%; range, 20%-100%), and segmental lesions were 92% (median, 95%; range, 70%-100%). The mean percentage resolution of preseptal component of lesions was 94% (median, 95%; range, 80%-100%) and postseptal component was 74% (median, 85%; range, 20%-100%) over a mean follow-up period of 16 months (median, 15 months; range, 4-37 months). Four (15%) patients exhibited flare-up of lesion after tapering oral propranolol. CONCLUSION: Oral propranolol is effective in the treatment of periocular CHI. Segmental and preseptal lesions respond better to the treatment compared to focal and postseptal lesions.

Advancements in 2D Materials Based Biosensors for Oxidative Stress Biomarkers.

The imbalance in generation of reactive oxygen species and its depletion causes oxidative stress. Because of its importance, there is a need to explore the role of oxidative stress biomarkers. The limitations of the conventional methods cause the researchers look for other alternatives. Biosensors are highly promising candidates for the detection of trace quantities for various analytes with high specificity, selectivity, and quick response time. Nanomaterial based matrices are the most popular choice while fabricating a biosensor. Two-dimensional (2D) materials such as graphene, transitional metal dichalcogenides, and various metal oxides have been used for the biosensing of different oxidative stress analytes. High electron mobility, good optical properties, tunable properties, high yields, easy synthesis, and availability make these materials the first choice. In this review, we have comprehensively discussed various biomarkers associated with oxidative stress. The review will provide information related to different kinds and various synthesis procedures employed for 2D nanomaterials. The major focus of the review is to elaborate the role of 2D nanomaterial based structures for the optical and electrochemical methods for the detection of oxidative stress. This review is an effort to help the researchers better understand various 2D based transducers available for the detection of oxidative stress biomarkers.

Sensitive impedimetric detection of troponin I with metal-organic framework composite electrode.

Metal-organic frameworks (MOFs) are promising materials for biosensing applications due to their large surface to volume ratio, easy assembly as thin films, and better biocompatibility than other nanomaterials. Their application in electrochemical biosensing devices can be realized by integrating them with other conducting materials, like polyaniline (PANI). In the present research, a composite of a copper-MOF (i.e., Cu3(BTC)2) with PANI has been explored to develop an impedimetric sensor for cardiac marker troponin I (cTnI). The solvothermally synthesized Cu3(BTC)2/PANI composite has been coated as a thin layer on the screen-printed carbon electrodes (SPE). This electroconductive thin film was conjugated with anti-cTnI antibodies. The above formed immunosensor has allowed the impedimetric detection of cTnI antigen over a clinically important concentration range of 1-400 ng mL-1. The whole process of antigen analysis could be completed within 5 min. The detection method was specific to cTnI even in the co-presence of other possibly interfering proteins.

County-level demographic, social, economic, and lifestyle correlates of COVID-19 infection and death trajectories during the first wave of the pandemic in the United States.

BACKGROUND: The US COVID-19 epidemic impacted counties differently across space and time, though large-scale transmission dynamics are unclear. The study's objective was to group counties with similar trajectories of COVID-19 cases and deaths and identify county-level correlates of the distinct trajectory groups. METHODS: Daily COVID-19 cases and deaths were obtained from 3141 US counties from January through June 2020. Clusters of epidemic curve trajectories of COVID-19 cases and deaths per 100,000 people were identified with Proc Traj. We utilized polytomous logistic regression to estimate Odds Ratios for trajectory group membership in relation to county-level demographics, socioeconomic factors, school enrollment, employment and lifestyle data. RESULTS: Six COVID-19 case trajectory groups and five death trajectory groups were identified. Younger counties, counties with a greater proportion of females, Black and Hispanic populations, and greater employment in private sectors had higher odds of being in worse case and death trajectories. Percentage of counties enrolled in grades 1-8 was associated with earlier-start case trajectories. Counties with more educated adult populations had lower odds of being in worse case trajectories but were generally not associated with worse death trajectories. Counties with higher poverty rates, higher uninsured, and more living in non-family households had lower odds of being in worse case and death trajectories. Counties with higher smoking rates had higher odds of being in worse death trajectory counties. DISCUSSION: In the absence of clear guidelines and personal protection, smoking, racial and ethnic groups, younger populations, social, and economic factors were correlated with worse COVID-19 epidemics that may reflect population transmission dynamics during January-June 2020. After vaccination of high-risk individuals, communities with higher proportions of youth, communities of color, smokers, and workers in healthcare, service and goods industries can reduce viral spread by targeting vaccination programs to these populations and increasing access and education on non-pharmaceutical interventions.