Phosphatidylinositol (4,5)-bisphosphate drives the formation of EGFR and EphA2 complexes.

Receptor tyrosine kinases (RTKs) regulate many cellular functions and are important targets in pharmaceutical development, particularly in cancer treatment. EGFR and EphA2 are two key RTKs that are associated with oncogenic phenotypes. Several studies have reported functional interplay between these receptors, but the mechanism of interaction is still unresolved. Here we utilize a time-resolved fluorescence spectroscopy called PIE-FCCS to resolve EGFR and EphA2 interactions in live cells. We tested the role of ligands and found that EGF, but not ephrin A1 (EA1), stimulated hetero-multimerization between the receptors. To determine the effect of anionic lipids, we targeted phospholipase C (PLC) activity to alter the abundance of phosphatidylinositol (4,5)-bisphosphate (PIP 2 ). We found that higher PIP 2 levels increased homo-multimerization of both EGFR and EphA2, as well as hetero-multimerization. This study provides a direct characterization of EGFR and EphA2 interactions in live cells and shows that PIP 2 can have a substantial effect on the spatial organization of RTKs.

Establishment of the mid-sagittal reference plane for three-dimensional assessment of facial asymmetry: a systematic review : Establishment of the mid-sagittal reference plane: a systematic review.

OBJECTIVE: To systematically review the literature for mid-sagittal plane establishment approaches to identify the most effective method for constructing the mid-sagittal plane for the evaluation of facial asymmetry. MATERIALS AND METHODS: Six electronic databases (PubMed, Medline (via Ovid), EMBASE (via Ovid), Cochrane Library, Web of Science, and Scopus) and grey literature were searched for the studies that computed the mid-sagittal reference plane three-dimensionally, using a combination of MeSH terms and keywords. The methodological quality and the level of evidence for the included studies were analyzed using QUADAS-2 and GRADE, respectively. RESULTS: The preliminary search yielded 6746 records, of which 42 articles that met the predefined inclusion criteria were included in the final analysis. All the included articles reported the construction of the mid-sagittal reference plane (MSP) using varied methods. The risk of bias and concerns regarding the applicability of the included studies were judged to be 'low'. The level of evidence was determined to be 'low' for the effectiveness of the technique and 'moderate' for the ease of clinical applicability. CONCLUSION: Despite methodological heterogeneity, this review substantiates the comparable efficacy of cephalometric and morphometric MSP construction methods. A fully automated morphometric MSP holds promise as a viable option for routine clinical use. Nevertheless, future prospective studies with an emphasis on the impact, accuracy, and clinical applicability of MSP construction techniques in cases of facial asymmetry are required. CLINICAL RELEVANCE: The present review will assist clinicians in selecting the most suitable method for MSP construction, leading to improved treatment planning and ultimately more favorable treatment outcomes.

Echoes of images: multi-loss network for image retrieval in vision transformers.

This paper introduces a novel approach to enhance content-based image retrieval, validated on two benchmark datasets: ISIC-2017 and ISIC-2018. These datasets comprise skin lesion images that are crucial for innovations in skin cancer diagnosis and treatment. We advocate the use of pre-trained Vision Transformer (ViT), a relatively uncharted concept in the realm of image retrieval, particularly in medical scenarios. In contrast to the traditionally employed Convolutional Neural Networks (CNNs), our findings suggest that ViT offers a more comprehensive understanding of the image context, essential in medical imaging. We further incorporate a weighted multi-loss function, delving into various losses such as triplet loss, distillation loss, contrastive loss, and cross-entropy loss. Our exploration investigates the most resilient combination of these losses to create a robust multi-loss function, thus enhancing the robustness of the learned feature space and ameliorating the precision and recall in the retrieval process. Instead of using all the loss functions, the proposed multi-loss function utilizes the combination of only cross-entropy loss, triplet loss, and distillation loss and gains improvement of 6.52% and 3.45% for mean average precision over ISIC-2017 and ISIC-2018. Another innovation in our methodology is a two-branch network strategy, which concurrently boosts image retrieval and classification. Through our experiments, we underscore the effectiveness and the pitfalls of diverse loss configurations in image retrieval. Furthermore, our approach underlines the advantages of retrieval-based classification through majority voting rather than relying solely on the classification head, leading to enhanced prediction for melanoma - the most lethal type of skin cancer. Our results surpass existing state-of-the-art techniques on the ISIC-2017 and ISIC-2018 datasets by improving mean average precision by 1.01% and 4.36% respectively, emphasizing the efficacy and promise of Vision Transformers paired with our tailor-made weighted loss function, especially in medical contexts. The proposed approach's effectiveness is substantiated through thorough ablation studies and an array of quantitative and qualitative outcomes. To promote reproducibility and support forthcoming research, our source code will be accessible on GitHub.

The ß2-adrenergic receptor associates with CXCR4 multimers in human cancer cells.

While the existence and functional role of class C G-protein-coupled receptors (GPCR) dimers is well established, there is still a lack of consensus regarding class A and B GPCR multimerization. This lack of consensus is largely due to the inherent challenges of demonstrating the presence of multimeric receptor complexes in a physiologically relevant cellular context. The C-X-C motif chemokine receptor 4 (CXCR4) is a class A GPCR that is a promising target of anticancer therapy. Here, we investigated the potential of CXCR4 to form multimeric complexes with other GPCRs and characterized the relative size of the complexes in a live-cell environment. Using a bimolecular fluorescence complementation (BiFC) assay, we identified the ß2 adrenergic receptor (ß2AR) as an interaction partner. To investigate the molecular scale details of CXCR4-ß2AR interactions, we used a time-resolved fluorescence spectroscopy method called pulsed-interleaved excitation fluorescence cross-correlation spectroscopy (PIE-FCCS). PIE-FCCS can resolve membrane protein density, diffusion, and multimerization state in live cells at physiological expression levels. We probed CXCR4 and ß2AR homo- and heteromultimerization in model cell lines and found that CXCR4 assembles into multimeric complexes larger than dimers in MDA-MB-231 human breast cancer cells and in HCC4006 human lung cancer cells. We also found that ß2AR associates with CXCR4 multimers in MDA-MB-231 and HCC4006 cells to a higher degree than in COS-7 and CHO cells and in a ligand-dependent manner. These results suggest that CXCR4-ß2AR heteromers are present in human cancer cells and that GPCR multimerization is significantly affected by the plasma membrane environment.

Cone Beam Computed Tomography-Based Evaluation of the Accuracy of Implant Surgery with Conventional (Free Hand) Implant Placement vs Computer Fabricated 3D Guide Implant Placement.

Introduction: Dental implants have been used in a variety of conventional technique-based forms for many years which had its own drawbacks. With the advent of cone beam CT, proper surgical and prosthetic planning is possible now a days. To achieve ideal implant placement, good prosthetic fabrication and overall successful prognosis computer fabricated guide aided surgery have been developed. Aim and Objective: The aim of this study was to compare and evaluate the accuracy of implant placement in partially edentulous patients with conventional free hand technique and computer fabricated guide of implant placement by comparing pre- and post-CBCT data. Methods: The present split mouth study design was conducted with forty sample size on twenty randomly selected patients who were treated with bilateral partially edentulous sites requiring dental implants. Patients were treated with both conventional (free hand) technique and computer fabricated 3D guide aided technique of implant placement. Comparison of accuracy of implant placement was done by comparing the pre- and postoperative CBCT data in terms of mean coronal deviation, mean apical deviation and mean angular deviation. Results and Conclusion: The results showed that there is no statistically significant difference between mean coronal deviation, mean apical deviation and mean angular deviation of planned and placed implants in both conventional technique (free hand technique) and computer fabricated 3D guide aided implant placement technique. Hence, this study concluded that conventional technique of implant placement is equally efficient in comparison with computer fabricated guide aided surgery in terms of accuracy of implant placement.

Comparison of surgical site infection (SSI) between negative pressure wound therapy (NPWT) assisted delayed primary closure and conventional delayed primary closure in grossly contaminated emergency abdominal surgeries: a randomized controlled trial.

PURPOSE: NPWT has been tried in many surgical fields, including colorectal, thoracic, vascular, and non-healing wounds, for the prevention of SSI. However, its efficacy in the prevention of SSI-grade IV closed abdominal wounds is yet to be explored. METHODS: All patients with grade IV abdominal wounds were included in the study. They were randomized into the conventional arm and the VAC arm after confirming the diagnosis intra-operatively. The sheath was closed, and the skin was laid open in the postoperative period. In the VAC arm, the NPWT dressing was applied on postoperative day (POD)-1 and removed on POD-5. In the conventional arm, only regular dressing was done postoperatively. The skin was closed with a delayed primary intention on POD-5 in both arms. The sutures were removed after 7 to 10 days of skin closure. RESULTS: The rate of SSI (10% in the VAC arm vs. 37.5% in the conventional arm, p-value = 0.004) was significantly lower in the VAC arm, as were the rates of seroma formation (2.4% in the VAC arm vs. 20% in the conventional arm, p = 0.014) and wound dehiscence (7.3% vs. 30%, p = 0.011). The conventional arm had a significant delay in skin closure beyond POD5 due to an increased rate of SSI, which also led to a prolonged hospital stay (5 days in the VAC arm vs. 6.5 days in the conventional arm, p-value = 0.005). CONCLUSION: The VAC dressing can be used routinely in grade IV closed abdominal wounds to reduce the risk of SSI and wound dehiscence.

Can smartphones be used for routine dental clinical application? A validation study for using smartphone-generated 3D facial images.

OBJECTIVES: To compare the accuracy of smartphone-generated three-dimensional (3D) facial images to that of direct anthropometry (DA) and 3dMD with the aim of assessing the validity and reliability of smartphone-generated 3D facial images for routine clinical applications. MATERIALS AND METHODS: Twenty-five anthropometric soft-tissue facial landmarks were labelled manually on 22 orthognathic surgery patients (11 males and 11 females; mean age 26.2 ± 5.3 years). For each labelled face, two imaging operations were performed using two different surface imaging systems: 3dMDface and Bellus3D FaceApp. Next, 42 inter-landmark facial measurements amongst the identified facial landmarks were measured directly on each labelled face and also digitally on 3D facial images. The measurements obtained from smartphone-generated 3D facial images (SGI) were statistically compared with those from DA and 3dMD. RESULTS: SGI had slightly higher measurement values than DA and 3dMD, but there was no statistically significant difference between the mean values of inter-landmark measures across the three methods. Clinically acceptable differences (≤3 mm or ≤5°) were observed for 67 % and 74 % of measurements with good agreement between DA and SGI, and 3dMD and SGI, respectively. An overall small systematic bias of ± 0.2 mm was observed between the three methods. Furthermore, the mean absolute difference between DA and SGI methods was highest for linear (1.41 ± 0.33 mm) as well as angular measurements (3.07 ± 0.73°). CONCLUSIONS: SGI demonstrated fair trueness compared to DA and 3dMD. The central region and flat areas of the face in SGI are more accurate. Despite this, SGI have limited clinical application, and the panfacial accuracy of the SGI would be more desirable from a clinical application standpoint. CLINICAL SIGNIFICANCE: The usage of SGI in clinical practice for region-specific macro-proportional facial assessment involving central and flat regions of the face or for patient education purposes, which does not require accuracy within 3 mm and 5° can be considered.

131I dose coefficients for a reference population using age-specific models.

Age-specific dose coefficients are required to assess internal exposure to the general public. This study utilizes reference age-specific biokinetic models of iodine to estimate the total number of nuclear disintegrations ã(rS,τ) occurring in source regions (rS) during the commitment time (τ). Age-specific S values are estimated for 35 target regions due to131I present in 22rSusing data from 10 paediatric reference computational phantoms (representing five ages for both sexes) published recently by the International Commission of Radiation Protection (ICRP). Monte Carlo transport simulations are performed in FLUKA code. The estimated ã(rS,τ) and S values are then used to compute the committed tissue equivalent dose HT(τ) for 27 radiosensitive tissues and dose coefficients e(τ) for all five ages due to inhalation and ingestion of131I. The derived ã(rS,τ) values in the thyroid source are observed to increase with age due to the increased retention of iodine in the thyroid. S values are found to decrease with age, mainly due to an increase in target masses. Generally, HT(τ) values are observed to decrease with age, indicating the predominant behaviour of S values over ã(rS,τ). On average, ingestion dose coefficients are 63% higher than for inhalation in all ages. The maximum contribution to dose coefficients is from the thyroid, accounting for 96% in the case of newborns and 98%-99% for all other ages. Furthermore, the estimated e(τ) values for the reference population are observed to be lower than previously published reference values from the ICRP. The estimated S, HT(τ) and e(τ) values can be used to improve estimations of internal doses to organs/whole body for members of the public in cases of131I exposure. The estimated dose coefficients can also be interpolated for other ages to accurately evaluate the doses received by the general public during131I therapy or during a radiological emergency.

Genome size, genetic diversity, and phenotypic variability imply the effect of genetic variation instead of ploidy on trait plasticity in the cross-pollinated tree species of mulberry.

Elucidation of genome size (GS), genetic and phenotypic variation is the fundamental aspect of crop improvement programs. Mulberry is a cross-pollinated, highly heterozygous tree eudicot, and comprised of huge ploidy variation with great adaptability across the world. However, because of inadequate information on GS, ploidy-associated traits, as well as the correlation between genetic and phenotypic variation hinder the further improvement of mulberry. In this present research, a core set of 157 germplasm accessions belonging to eight accepted species of Morus including promising functional varieties were chosen to represent the genetic spectrum from the whole germplasm collection. To estimate the GS, accessions were subjected to flow cytometry (FCM) analysis and the result suggested that four different ploidies (2n = 2x, 3x, 4x, and 6x) with GS ranging from 0.72±0.005pg (S-30) to 2.89±0.015pg (M. serrata), accounting~4.01 fold difference. The predicted polyploidy was further confirmed with metaphase chromosome count. In addition, the genetic variation was estimated by selecting a representative morphologically, diverse population of 82 accessions comprised of all ploidy variations using simple sequence repeats (SSR). The estimated average Polymorphism Information Content (PIC) and expected heterozygosity showed high levels of genetic diversity. Additionally, three populations were identified by the model-based population structure (k = 3) with a moderate level of correlation between the populations and different species of mulberry, which imply the effect of genetic variation instead of ploidy on trait plasticity that could be a consequence of the high level of heterozygosity imposed by natural cross-pollination. Further, the correlation between ploidies, especially diploid and triploid with selected phenotypic traits was identified, however, consistency could not be defined with higher ploidy levels (>3x). Moreover, incite gained here can serve as a platform for future omics approaches to the improvement of mulberry traits.

On-demand (SOS) analgesia through Transversus Abdominis Plane (TAP) catheter route for post-operative pain relief in Emergency Laparotomies-a non-randomised interventional study (STAPLE trial).

INTRODUCTION: This study aimed to compare the efficacy and safety of on-demand bupivacaine infusion via transversus abdominis plane (TAP) catheter in emergency laparotomy patients. METHODS: A non-randomised interventional study was conducted on patients undergoing emergency midline laparotomy. The intervention group received an on-demand infusion of 10 ml 0.5% bupivacaine through TAP catheters, whilst the control group received standard analgesic care. The primary outcome was the amount of rescue analgesic consumption. Secondary outcomes included the post-operative, measured by visual analogue scores (VAS), side effects, time to first flatus, post-operative nausea and vomiting, and pulmonary complications. RESULTS: One-hundred-twenty patients (58 in the TAP-SOS group, 62 in the control group) were included in the final analysis. The TAP-SOS group showed significantly reduced rescue analgesic requirement by 91% (p < 0.001) and lower VAS scores at 3, 6, 12, and 24 h (adjusted p < 0.00). Time to out-of-bed mobilisation was significantly shorter in the TAP-SOS group by 12.47 h (p < 0.001), and post-operative pulmonary complications were lower by 75% (p < 0.05). There were no significant differences in bowel recovery, catheter-related complications, or post-operative morbidity. No incidences of catheter-site infection were reported on follow-up; however, the catheter tip-culture was positive in 3 (5.17%) patients. CONCLUSION: On-demand bupivacaine infusion through a TAP catheter effectively reduced post-operative pain and opioid requirements in emergency laparotomy patients without complications. If an epidural is not an option, the TAP-SOS approach can be a helpful adjunct in implementing the ERAS protocol in an emergency since it allows for early ambulation and better pain management.

Comparison of laparoscopic port site skin closure techniques (CLOSA): transcutaneous suturing versus subcuticular sutures versus adhesive strips: a prospective single-blinded randomized control trial.

BACKGROUND: Cosmesis is an essential aspect of laparoscopic surgery. Various methods of skin closure techniques have been described. We conducted a study to evaluate the cosmesis and patient satisfaction with the scars three months after laparoscopic surgery using transcutaneous suture (TS) vs. adhesive strips (AS) and subcuticular suturing (SS). METHODS: A randomized, controlled, prospective study was conducted at AIIMS, Bhubaneswar. The included patients were randomly assigned among the three arms. The time for skin closure was measured. Wounds were assessed till discharge, at 14 days, one month, and three months. Cosmesis was measured by the Hollander Wound evaluation scale (HWES) for each incision separately, and patient satisfaction by a 10- point Visual analog scale (VAS). RESULTS: One hundred six patients were assessed for eligibility, and 90 patients were randomized. Three-month follow-up data was obtained from 83 patients (92.22%). Baseline characteristics were similar among the groups. Cosmetic outcome was assessed in 312 incisions across 83 patients, and 206 (66.03%) incisions had an HWE Score of 0, but there was no significant difference (p = 0.86). Patient satisfaction was highest in the TS group (TS = 1.29, SS = 1.79, AS = 2.04, p = 0.03). Time for skin closure was the least in the AS arm (41.4 secs, p = 0.00). Skin dehiscence was significantly more in the AS arm. Four (4.44%) patients had port site infections. CONCLUSION: This study demonstrates that skin closure by transcutaneous, subcuticular, or adhesive strip methods had comparable cosmetic outcomes at three months. However, the transcutaneous closure method showed better patient satisfaction and minimal post-operative complications.

Harnessing the potential of probiotics in the treatment of alcoholic liver disorders.

In the current scenario, prolonged consumption of alcohol across the globe is upsurging an appreciable number of patients with the risk of alcohol-associated liver diseases. According to the recent report, the gut-liver axis is crucial in the progression of alcohol-induced liver diseases, including steatosis, steatohepatitis, fibrosis, cirrhosis, and hepatocellular carcinoma. Despite several factors associated with alcoholic liver diseases, the complexity of the gut microflora and its great interaction with the liver have become a fascinating area for researchers due to the high exposure of the liver to free radicals, bacterial endotoxins, lipopolysaccharides, inflammatory markers, etc. Undoubtedly, alcohol-induced gut microbiota imbalance stimulates dysbiosis, disrupts the intestinal barrier function, and trigger immune as well as inflammatory responses which further aggravate hepatic injury. Since currently available drugs to mitigate liver disorders have significant side effects, hence, probiotics have been widely researched to alleviate alcohol-associated liver diseases and to improve liver health. A broad range of probiotic bacteria like Lactobacillus, Bifidobacteria, Escherichia coli, Sacchromyces, and Lactococcus are used to reduce or halt the progression of alcohol-associated liver diseases. Several underlying mechanisms, including alteration of the gut microbiome, modulation of intestinal barrier function and immune response, reduction in the level of endotoxins, and bacterial translocation, have been implicated through which probiotics can effectively suppress the occurrence of alcohol-induced liver disorders. This review addresses the therapeutic applications of probiotics in the treatment of alcohol-associated liver diseases. Novel insights into the mechanisms by which probiotics prevent alcohol-associated liver diseases have also been elaborated.

Cystine rather than cysteine is the preferred substrate for ß-elimination by cystathionine γ-lyase: implications for dietary methionine restriction.

Dietary methionine restriction (MR) increases longevity by improving health. In experimental models, MR is accompanied by decreased cystathionine ß-synthase activity and increased cystathionine γ-lyase activity. These enzymes are parts of the transsulfuration pathway which produces cysteine and 2-oxobutanoate. Thus, the decrease in cystathionine ß-synthase activity is likely to account for the loss of tissue cysteine observed in MR animals. Despite this decrease in cysteine levels, these tissues exhibit increased H2S production which is thought to be generated by ß-elimination of the thiol moiety of cysteine, as catalyzed by cystathionine ß-synthase or cystathionine γ-lyase. Another possibility for this H2S production is the cystathionine γ-lyase-catalyzed ß-elimination of cysteine persulfide from cystine, which upon reduction yields H2S and cysteine. Here, we demonstrate that MR increases cystathionine γ-lyase production and activities in the liver and kidneys, and that cystine is a superior substrate for cystathionine γ-lyase catalyzed ß-elimination as compared to cysteine. Moreover, cystine and cystathionine exhibit comparable Kcat/Km values (6000 M-1 s-1) as substrates for cystathionine γ-lyase-catalyzed ß-elimination. By contrast, cysteine inhibits cystathionine γ-lyase in a non-competitive manner (Ki ~ 0.5 mM), which limits its ability to function as a substrate for ß-elimination by this enzyme. Cysteine inhibits the enzyme by reacting with its pyridoxal 5'-phosphate cofactor to form a thiazolidine and in so doing prevents further catalysis. These enzymological observations are consistent with the notion that during MR cystathionine γ-lyase is repurposed to catabolize cystine and thereby form cysteine persulfide, which upon reduction produces cysteine.

Pharmacologic improvement of CFTR function rapidly decreases sputum pathogen density, but lung infections generally persist.

BackgroundLung infections are among the most consequential manifestations of cystic fibrosis (CF) and are associated with reduced lung function and shortened survival. Drugs called CF transmembrane conductance regulator (CFTR) modulators improve activity of dysfunctional CFTR channels, which is the physiological defect causing CF. However, it is unclear how improved CFTR activity affects CF lung infections.MethodsWe performed a prospective, multicenter, observational study to measure the effect of the newest and most effective CFTR modulator, elexacaftor/tezacaftor/ivacaftor (ETI), on CF lung infections. We studied sputum from 236 people with CF during their first 6 months of ETI using bacterial cultures, PCR, and sequencing.ResultsMean sputum densities of Staphylococcus aureus, Pseudomonas aeruginosa, Stenotrophomonas maltophilia, Achromobacter spp., and Burkholderia spp. decreased by 2-3 log10 CFU/mL after 1 month of ETI. However, most participants remained culture positive for the pathogens cultured from their sputum before starting ETI. In those becoming culture negative after ETI, the pathogens present before treatment were often still detectable by PCR months after sputum converted to culture negative. Sequence-based analyses confirmed large reductions in CF pathogen genera, but other bacteria detected in sputum were largely unchanged. ETI treatment increased average sputum bacterial diversity and produced consistent shifts in sputum bacterial composition. However, these changes were caused by ETI-mediated decreases in CF pathogen abundance rather than changes in other bacteria.ConclusionsTreatment with the most effective CFTR modulator currently available produced large and rapid reductions in traditional CF pathogens in sputum, but most participants remain infected with the pathogens present before modulator treatment.Trial RegistrationClinicalTrials.gov NCT04038047.FundingThe Cystic Fibrosis Foundation and the NIH.

Transfer Learning Based Lightweight Ensemble Model for Imbalanced Breast Cancer Classification.

Automated classification of breast cancer can often save lives, as manual detection is usually time-consuming & expensive. Since the last decade, deep learning techniques have been most widely used for the automatic classification of breast cancer using histopathology images. This paper has performed the binary and multi-class classification of breast cancer using a transfer learning-based ensemble model. To analyze the correctness and reliability of the proposed model, we have used an imbalance IDC dataset, an imbalance BreakHis dataset in the binary class scenario, and a balanced BACH dataset for the multi-class classification. A lightweight shallow CNN model with batch normalization technology to accelerate convergence is aggregated with lightweight MobileNetV2 to improve learning and adaptability. The aggregation output is fed into a multilayer perceptron to complete the final classification task. The experimental study on all three datasets was performed and compared with the recent works. We have fine-tuned three different pre-trained models (ResNet50, InceptionV4, and MobilNetV2) and compared it with the proposed lightweight ensemble model in terms of execution time, number of parameters, model size, etc. In both the evaluation phases, it is seen that our model outperforms in all three datasets.

Ligation of Intersphincteric Fistulous Tract vs Endorectal Advancement Flap for High-Type Fistula in Ano: A Randomized Controlled Trial.

BACKGROUND: This study aimed to compare the postoperative outcomes and success rate of the endorectal advancement flap and ligation of intersphincteric fistulous tract (LIFT) in high-type fistula in ano. STUDY DESIGN: This randomized control trial included patients with high-type fistula in ano of cryptoglandular origin. The primary endpoint was complete fistula healing at the end of 6 months. However, the patients were followed up for 2 years. Other parameters studied were perioperative complications, duration of surgery, postoperative pain, hospital stay in hours, continence, and quality of life at 6 months. RESULTS: A total of 84 patients were recruited (42 in each group). The healing rate in the LIFT arm was better than that in the endorectal advancement flap arm (76.2.% vs 54.7%, p = 0.039). Four patients in the endorectal advancement flap group and two in the LIFT group had flatus incontinence at the end of 6 months, but all were continent at 2 years. At the end of the first week, the Visual Analog Scale score and quality of life at 6 months were better in the LIFT arm (3.7 ± 1.16 vs 4.7 ± 0.81 and 0.7 vs 0.6, p < 0.05). The mean duration of surgery was significantly less in the LIFT group (46.43 ± 9.32 vs 89.29 ± 10.90 minutes). None had any postoperative complications, and >80% were discharged within 24 hours. CONCLUSIONS: The shorter operative duration, better quality of life at 6 months, and higher healing rate make LIFT a superior treatment option for high fistula in ano. However, studies with a large sample size will be needed to verify these results.

Genome Capture Sequencing Selectively Enriches Bacterial DNA and Enables Genome-Wide Measurement of Intrastrain Genetic Diversity in Human Infections.

Within-host evolution produces genetic diversity in bacterial strains that cause chronic human infections. However, the lack of facile methods to measure bacterial allelic variation in clinical samples has limited understanding of intrastrain diversity's effects on disease. Here, we report a new method termed genome capture sequencing (GenCap-Seq) in which users inexpensively make hybridization probes from genomic DNA or PCR amplicons to selectively enrich and sequence targeted bacterial DNA from clinical samples containing abundant human or nontarget bacterial DNA. GenCap-Seq enables accurate measurement of allele frequencies over targeted regions and is scalable from specific genes to entire genomes, including the strain-specific accessory genome. The method is effective with samples in which target DNA is rare and inhibitory and DNA-degrading substances are abundant, including human sputum and feces. In proof-of-principle experiments, we used GenCap-Seq to investigate the responses of diversified Pseudomonas aeruginosa populations chronically infecting the lungs of people with cystic fibrosis to in vivo antibiotic exposure, and we found that treatment consistently reduced intrastrain genomic diversity. In addition, analysis of gene-level allele frequency changes suggested that some genes without conventional resistance functions may be important for bacterial fitness during in vivo antibiotic exposure. GenCap-Seq's ability to scalably enrich targeted bacterial DNA from complex samples will enable studies on the effects of intrastrain and intraspecies diversity in human infectious disease. IMPORTANCE Genetic diversity evolves in bacterial strains during human infections and could affect disease manifestations and treatment resistance. However, the extent of diversity present in vivo and its changes over time are difficult to measure by conventional methods. We developed a novel approach, GenCap-Seq, to enrich microbial DNA from complex human samples like sputum and feces for genome-wide measurements of bacterial allelic diversity. The approach is inexpensive, scalable to encompass entire targeted genomes, and works in the presence of abundant untargeted nucleic acids and inhibiting substances. We used GenCap-Seq to investigate in vivo responses of diversified bacterial strains to antibiotic treatment. This method will enable new ideas about the effects of intrastrain diversity on human infections to be tested.

Biopolymeric nanoparticles based effective delivery of bioactive compounds toward the sustainable development of anticancerous therapeutics.

Nowadays, effective cancer therapy is a global concern, and recent advances in nanomedicine are crucial. Cancer is one of the major fatal diseases and a leading cause of death globally. Nanotechnology provides rapidly evolving delivery systems in science for treating diseases in a site-specific manner using natural bioactive compounds, which are gaining widespread attention. Nanotechnology combined with bioactives is a very appealing and relatively new area in cancer treatment. Natural bioactive compounds have the potential to be employed as a chemotherapeutic agent in the treatment of cancer, in addition to their nutritional benefits. Alginate, pullulan, cellulose, polylactic acid, chitosan, and other biopolymers have been effectively used in the delivery of therapeutics to a specific site. Because of their biodegradability, biopolymeric nanoparticles (BNPs) have received a lot of attention in the development of new anticancer drug delivery systems. Biopolymer-based nanoparticle systems can be made in a variety of ways. These systems have developed as a cost-effective and environmentally friendly solution to boost treatment efficacy. Effective drug delivery systems with improved availability, increased selectivity, and lower toxicity are needed. Recent research findings and current knowledge on the use of BNPs in the administration of bioactive chemicals in cancer therapy are summarized in this review.

Acute Pulmonary Exacerbation Phenotypes in Patients with Cystic Fibrosis.

Rationale: The etiology of cystic fibrosis (CF) pulmonary exacerbations (PEx) is likely multifactorial with viral, bacterial, and non-infectious pathways contributing. Objectives: To determine whether viral infection status and CRP (C-reactive protein) can classify subphenotypes of PEx that differ in outcomes and biomarker profiles. Methods: Patients were recruited at time of admission for a PEx. Nasal swabs and sputum samples were collected and processed using the respiratory panel of the FilmArray multiplex polymerase chain reaction (PCR). Serum and plasma biomarkers were measured. PEx were classified using serum CRP and viral PCR: "pauci-inflammatory" if CRP < 5 mg/L, "non-viral with systemic inflammation" if CRP ⩾ 5 mg/L and no viral infection detected by PCR and "viral with systemic inflammation" if CRP ⩾ 5 mg/L and viral infection detected by PCR. Results: Discovery cohort (n = 59) subphenotype frequencies were 1) pauci-inflammatory (37%); 2) non-viral with systemic inflammation (41%); and 3) viral with systemic inflammation (22%). Immunoglobulin G, immunoglobulin M, interleukin-10, interleukin-13, serum calprotectin, and CRP levels differed across phenotypes. Reduction from baseline in forced expiratory volume in 1 second as percent predicted (FEV1pp) at onset of exacerbation differed between non-viral with systemic inflammation and viral with systemic inflammation (-6.73 ± 1.78 vs. -13.5 ± 2.32%; P = 0.025). Non-viral with systemic inflammation PEx had a trend toward longer duration of intravenous antibiotics versus pauci-inflammation (18.1 ± 1.17 vs. 14.8 ± 1.19 days, P = 0.057). There were no differences in percent with lung function recovery to <10% of baseline FEV1pp. Similar results were seen in local and external validation cohorts comparing a pauci-inflammatory to viral/non-viral inflammatory exacerbation phenotypes. Conclusions: Subphenotypes of CF PEx exist with differences in biomarker profile, clinical presentation, and outcomes.