3D Hydrogel Culture System Recapitulates Key Tuberculosis Phenotypes and Demonstrates Pyrazinamide Efficacy.

The mortality caused by tuberculosis (TB) infections is a global concern, and there is a need to improve understanding of the disease. Current in vitro infection models to study the disease have limitations such as short investigation durations and divergent transcriptional signatures. This study aims to overcome these limitations by developing a 3D collagen culture system that mimics the biomechanical and extracellular matrix (ECM) of lung microenvironment (collagen fibers, stiffness comparable to in vivo conditions) as the infection primarily manifests in the lungs. The system incorporates Mycobacterium tuberculosis (Mtb) infected human THP-1 or primary monocytes/macrophages. Dual RNA sequencing reveals higher mammalian gene expression similarity with patient samples than 2D macrophage infections. Similarly, bacterial gene expression more accurately recapitulates in vivo gene expression patterns compared to bacteria in 2D infection models. Key phenotypes observed in humans, such as foamy macrophages and mycobacterial cords, are reproduced in the model. This biomaterial system overcomes challenges associated with traditional platforms by modulating immune cells and closely mimicking in vivo infection conditions, including showing efficacy with clinically relevant concentrations of anti-TB drug pyrazinamide, not seen in any other in vitro infection model, making it reliable and readily adoptable for tuberculosis studies and drug screening.

Experimental investigation on punch shear strength of poly lactic acid specimens for biomedical applications.

The designed biomedical implants require excellent shear strength primarily for mechanical stability against forces in human body. However, metallic implants undergo stress shielding with release of toxic ions in the body. Thus, Fused Deposition Modeling (FDM) has made significant progress in the biomedical field through the production of customized implants. The mechanical behavior is highly dependent on printing parameters, however, the effect of these parameters on punch shear strength of ASTM D732-02 standard specimens has not been explored. Thus, in the current study, the effect of infill density (IFD), printing speed (PTS), wall thickness (WLT), and layer thickness (LYT) has been investigated on the punch shear strength using Response Surface Methodology. The Analysis of Variance (ANOVA) has been performed for predicting statistical model with 95% confidence interval. During the statistical analysis, the terms with p-value lower than 0.05 were considered significant and the influence of process parameters has been examined using microscopic images. The surface plots have been used for discussing the effect of interactions between printing parameters. The statistical results revealed IFD as the most significant contributing factor, followed by PTS, LYT, and WLT. The study concluded by optimization of printing parameters for obtaining the highest punch shear strength.

Risk-Tailoring Radiotherapy for Endometrial Cancer: A Narrative Review.

Endometrial cancer is the most common gynecologic cancer in the United States and it contributes to the second most gynecologic cancer-related deaths. With upfront surgery, the specific characteristics of both the patient and tumor allow for risk-tailored treatment algorithms including adjuvant radiotherapy and systemic therapy. In this narrative review, we discuss the current radiation treatment paradigm for endometrial cancer with an emphasis on various radiotherapy modalities, techniques, and dosing regimens. We then elaborate on how to tailor radiotherapy treatment courses in combination with other cancer-directed treatments, including chemotherapy and immunotherapy. In conclusion, this review summarizes ongoing research that aims to further individualize radiotherapy regimens for individuals in an attempt to improve patient outcomes.

Experimental investigation of ultrasonic assisted coating on three-point bending behavior of 3D printed polymeric bone plates for biomedical applications.

3D printed Poly Lactic Acid (PLA) bone plates exhibit limited three-point bending strength, restricting their viability in biomedical applications. The application of polydopamine (PDM) enhances the three-point bending strength by undergoing covalent interactions with PLA molecular structure. However, the heavy nature of PDM particles leads to settling at the container base at higher coating solution concentrations. This study investigates the impact of ultrasonic-assisted coating parameters on the three-point bending strength. Utilizing Response Surface Methodology (RSM) for statistical modeling, the study examines the influence of ultrasonic vibration power (UP), coating solution concentration (CC), and submersion time (TIME). RSM optimization recommended 100 % UP, 6 mg/ml CC, and 150 min TIME, resulting in maximum three-point bending strength of 83.295 MPa. Microscopic images from the comparative analysis revealed non-uniform coating deposition with mean thickness of 6.153 µm under normal coating. In contrast, ultrasonic-assisted coating promoted uniform deposition with mean thickness of 18.05 µm. The results demonstrate that ultrasonic-assisted coating induces PDM particle collision, preventing settling at the container base, and enhances three-point bending strength by 7.27 % to 23.24 % compared to the normal coating condition. This study emphasizes on the potential of ultrasonic-assisted coating to overcome the limitations of direct immersion coating technique.

Clinical and immunological predictors of post-rituximab paradoxical pemphigus flare: A prospective cohort study.

Background Paradoxical flare of pemphigus following rituximab infusion has been reported previously, however, its incidence or risk factors have not been studied in detail. Objectives To evaluate the clinical and immunological predictors associated with post-rituximab paradoxical pemphigus flare. Materials and Methods This was a prospective cohort study including adult patients with pemphigus vulgaris or foliaceus who were treated with rituximab. Patients were administered 1000 mg of intravenous rituximab on days 0 and 14 (Rheumatoid arthritis (RA) protocol), with or without oral prednisolone and/or conventional immunosuppressive agents. Baseline clinical and immunological predictors of post-rituximab pemphigus flares were assessed. Results Fifty patients (mean age 40.44 ± 12.36 years) with a mean pemphigus disease area index (PDAI) score of 27.8 ± 15.48 were administered rituximab. Post-rituximab flare occurred in 10 (20%) patients after a mean of 14.1 ± 4.33 days after the first rituximab infusion. The mean baseline PDAI score (36.4 ± 11.7 vs. 25.6 ± 15.7, P = 0.02) and serum anti-Dsg1 levels (1216.8 ± 850.1 vs. 592 ± 562.12 RU/mL, P = 0.03) were statistically significantly higher in patients experiencing a flare. Using ROC-curve analysis, a PDAI score of 328 (OR 8.3, 95% CI 1.5-44.7) was 80% sensitive and 67.5% specific in predicting post-rituximab flare, while serum anti-Dsg1 level of 31137.78 RU/ml had a sensitivity of 60% and specificity of 85%. There was no significant difference in terms of affected body surface area, type of pemphigus, starting prednisolone dose, oral immunosuppressive adjuvant, serum anti-Dsg3, serum anti-AchRM3, and peripheral CD19+ B cell population. Limitations Our study is limited by a relatively small sample size. Immunological factors were not evaluated at the time of pemphigus flare. Though these unexpected pemphigus flares are likely to be associated with rituximab infusion, the possibility of spontaneous disease exacerbation cannot be entirely excluded. Conclusions Patients with more severe pemphigus or high serum anti-Dsg1 are at risk of post-rituximab paradoxical flare, and may benefit from rituximab administration under close monitoring.

Chronic sexually acquired reactive arthritis secondary to Chlamydiatrachomatis.

Reactive arthritis is included in the spectrum of seronegative spondyloarthritides, occurring secondary to triggers of genitourinary and gastrointestinal tract infections. We describe two cases of sexually acquired reactive arthritis secondary to genital infection by Chlamydia trachomatis, diagnosed by in-house polymerase chain reaction performed on the first void urine. Both patients were managed with a combined approach of short course antibiotics, immunosuppressive agents, biologicals and surgical intervention.

Assessment of Trachyspermum ammi essential oil against Aspergillus flavus, aflatoxin B1 contamination, and post-harvest quality of Sorghum bicolor.

The present investigation explored the antifungal effectiveness of Trachyspermum ammi essential oil (TAEO) against Aspergillus flavus, aflatoxin B1 (AFB1) contamination, and its mechanism of action using biochemical and computational approaches. The GC-MS result revealed the chemical diversity of TAEO with the highest percentage of γ-terpinene (39 %). The TAEO exhibited minimum inhibitory concentration against A. flavus growth (0.5 µL/mL) and AFB1 (0.4 µL/mL) with radical scavenging activity (IC50 = 2.13 µL/mL). The mechanism of action of TAEO was associated with the alteration in plasma membrane functioning, antioxidative defense, and carbon source catabolism. The molecular dynamic result shows the multi-regime binding of γ-terpinene with the target proteins (Nor1, Omt1, and Vbs) of AFB1 biosynthesis. Furthermore, TAEO exhibited remarkable in-situ protection of Sorghum bicolor seed samples against A. flavus and AFB1 contamination and protected the nutritional deterioration. Hence, the study recommends TAEO as a natural antifungal agent for food protection against A. flavus mediated biodeterioration.

Diversity in morpho-pomological attributes and biochemical profiling of bael (Aegle marmelos (L.) Correa) genotypes of North-Western India.

Bael is a medicinal cum fruit tree with multipurpose utility and propagated mostly through seeds. The present study aimed to assess and analyse the morpho-pomological and biochemical traits of eighty seedlings grown bael genotypes comparison with two commercial cultivars (NB-5 and NB-9) of bael. The significant differences were detected among the genotypes based on the measured morpho-pomological and biochemical traits. The morpho-pomological and biochemical traits of bael exhibited variation ranging from 6.17% to 133.65%. Trunk girth ranged from 29.50 to 63.40 cm and tree spread (N-S) varied 1.00-6.30 m. Fruit length ranged from 4.60 to 12.05 cm and fruit width ranged from 4.64 to 11.72 cm. Moreover, fruit weight ranged from 56.33 to 917.65 g and pulp percentage varied from 58.64 to 81.38%. Soluble Solid Content ranged from 25.90 to 36.77 0brix and ascorbic acid varied from 14.38 to 25.45 mg/100 g. Fruit length was positively correlated with fruit width, fruit weight, pulp percentage, seed length, seed diameter and number of seeds per fruit, while it was negatively correlated with fruit surface and total number of fruit per plant. Principal component analysis showed that 76.66% of the variability observed was explained by the 13 components. Ward cluster analysis using Euclidean distance classified the genotypes into two main clusters. These findings contribute to a better understanding of the diversity and relationships among the studied genotypes, aiding future breeding and selection programs for improved bael cultivation.

Head and Neck Cancer Patient Population, Management, and Oncologic Outcomes from the COVID-19 Pandemic.

The COVID-19 pandemic precipitated drastic changes in cancer care. Its impact on the U.S. head and neck cancer population has yet to be fully understood. This study aims to understand the impact of pandemic-related changes on the head and neck cancer population. An observational study of head and neck cancer patients at a single institution during the spring of 2020 and 2019 was performed. Clinical characteristics and survival outcomes were analyzed. In 2020, 54 head and neck cancer patients were evaluated in the department of radiation oncology vs. 74 patients seen in 2019; 42% of the patients were female in 2019 versus 24% in 2020 (p = 0.036). The median follow-up time was 19.4 and 31 months for 2020 and 2019, respectively. After adjusting for stage, the relapse-free survival probability at 6 and 12 months was 79% and 69% in 2020 vs. 96% and 89% in 2019, respectively (p = 0.036). There was no significant difference in the overall survival, with 94% and 89% in 2020 and 2019, respectively (p = 0.61). Twenty-one percent of patients received induction chemotherapy in 2020 versus 5% in 2019 (p = 0.011); significantly more treatment incompletions occurred in 2020, 9% vs. 0% in 2019 (p = 0.012). Moreover, the stage-adjusted RFS differed between cohorts, suggesting head and neck cancer patients seen during the initial wave of COVID-19 may experience worse oncologic outcomes.

Dose and fractionation regimen for brachytherapy boost in cervical cancer in the US.

PURPOSE: Curative-intent radiotherapy for locally advanced and select early stage cervical cancer in the US includes external beam radiotherapy (EBRT) with brachytherapy. Although there are guidelines for brachytherapy dose and fractionation regimens, there are limited data on practice patterns. This study aims to evaluate the contemporary utilization of cervical cancer brachytherapy in the US and its association with patient demographics and facility characteristics. METHODS: We retrospectively analyzed clinical covariates of cervical cancer patients diagnosed and treated in 2018-2020 with curative-intent radiotherapy from the 2020 National Cancer Database. Associations between patient and institutional factors with the number of brachytherapy fractions were identified with logistic regression. Factors with association (p < 0.10) were then included in a multivariable logistic regression model. All tests were two-sided with significance <0.05 unless specified otherwise. RESULTS: Among the eligible 2517 patients, 97.3% received HDR or LDR and is further analyzed. More patients received HDR than LDR brachytherapy (98.9% vs 1.1%) and intracavitary than interstitial brachytherapy (86.4% vs 13.6%). The most common number of HDR fractions prescribed were 5 (51.0%), 4 (32.9%), and 3 (8.6%). After adjusting for the other variables in the model, ethnicity, private insurance status, overall insurance status, and facility type were the only factors that were significantly associated with the number of brachytherapy factions (p < 0.0001, p = 0.028, p = 0.001, and p < 0.0001, respectively, n = 2184). CONCLUSIONS: In the US, various HDR brachytherapy regimens are utilized depending on patient and institutional factors. Future research may optimize cervical cancer brachytherapy by correlating specific dose and fractionation regimens with patient outcomes.

Essential oils as green promising alternatives to chemical preservatives for agri-food products: New insight into molecular mechanism, toxicity assessment, and safety profile.

Microbial food spoilage caused by food-borne bacteria, molds, and associated toxic chemicals significantly alters the nutritional quality of food products and makes them unpalatable to the consumer. In view of potential adverse effects (resistance development, residual toxicity, and negative effects on consumer health) of some of the currently used preservative agents and consumer preferences towards safe, minimally processed, and chemical-free products, food industries are looking for natural alternatives to the chemical preservatives. In this context, essential oils (EOs) showed broad-range antimicrobial effectiveness, low toxicity, and diverse mechanisms of action, and could be considered promising natural plant-based antimicrobials. The existing technical barriers related to the screening of plants, extraction methods, characterization, dose optimization, and unpredicted mechanism of toxicity in the food system, could be overcome using recent scientific and technological advancements, especially bioinformatics, nanotechnology, and mathematical approaches. The review focused on the potential antimicrobial efficacy of EOs against food-borne microbes and the role of recent scientific technology and social networking platform in addressing the major obstacles with EOs-based antimicrobial agents. In addition, a detailed mechanistic understanding of the antimicrobial efficacy of EOs, safety profile, and risk assessment using bioinformatics approaches are summarized to explore their potential application as food preservatives.

A Novel Method for Prediction of Raised Intracranial Pressure Through Automated ONSD and ETD Ratio Measurement From Ocular Ultrasound.

The paper presents a novel framework for the prediction of the raised Intracranial Pressure (ICP) from ocular ultrasound images of traumatic patients through automated measurement of Optic Nerve Sheath Diameter (ONSD) and Eyeball Transverse Diameter (ETD). The measurement of ONSD using an ocular ultrasound scan is non-invasive and correlates with the raised ICP. However, the existing studies suggested that the ONSD value alone is insufficient to indicate the ICP condition. Since the ONSD and ETD values may vary among patients belonging to different ethnicity/origins, there is a need for developing an independent global biomarker for predicting raised ICP condition. The proposed work develops an automated framework for the prediction of raised ICP by developing algorithms for the automated measurement of ONSD and ETD values. It is established that the ONSD and ETD ratio (OER) is a potential biomarker for ICP prediction independent of ethnicity and origin. The OER threshold value is determined by performing statistical analysis on the data of 57 trauma patients obtained from the AIIMS, New Delhi. The automated OER is computed and compared with the conventionally measured ICP by determining suitable correlation coefficients. It is found that there is a significant correlation of OER with ICP (r = .81, p ≤ .01), whereas the correlation of ONSD alone with ICP is relatively less (r = .69, p = .004). These correlation values indicate that OER is a better parameter for the prediction of ICP. Further, the threshold value of OER is found to be 0.21 for predicting raised ICP conditions in this study. Scatter plot and Heat map analysis of OER and corresponding ICP reveal that patients with OER ≥ 0.21, have ICP in the range of 17 to 35 mm Hg. In the data available for this research work, OER ranges from 0.17 to 0.35.

WATCHMEN: a case report highlighting the closure of a bilobed left atrial appendage using two WATCHMAN-FLX devices.

Background: Left atrial appendage occlusion (LAAO) performed percutaneously has emerged as a widely accepted method for stroke prevention, offering a viable alternative to anticoagulation. Numerous studies have demonstrated the effectiveness and safety of this procedure. However, in certain cases, the use of a single LAAO device may not adequately achieve optimal closure due to variations in the anatomy of the left atrial appendage (LAA). Case summary: In this manuscript, we highlight the successful closure of a bilobed LAA with a large ostium utilizing two WATCHMAN™ FLX devices and using the double sheath technique. The aim was to achieve optimal closure and address the unique anatomical characteristics of the patient's LAA. Discussion: The utilization of two LAAO devices in bilobed appendage anatomy, where a single device may not be sufficient, is possible, although it poses a challenge because of the lack of technical expertise and limited published evidence. Transoesophageal imaging can serve as a valuable tool for assessing the precise anatomy of the LAA and guide the selection and placement of the occlusion devices.

Enrichment of Native and Recombinant Extracellular Vesicles of Mycobacteria.

Most bacteria, including mycobacteria, generate extracellular vesicles (EVs). Since bacterial EVs (bEVs) contain a subset of cellular components, including metabolites, lipids, proteins, and nucleic acids, several groups have evaluated either the native or recombinant versions of bEVs for their protective potency as subunit vaccine candidates. Unlike native EVs, recombinant EVs are molecularly engineered to contain one or more immunogens of interest. Over the last decade, different groups have explored diverse approaches for generating recombinant bEVs. However, here, we report the design, construction, and enrichment of recombinant mycobacterial EVs (mEVs) in mycobacteria. Towards that, we use Mycobacterium smegmatis (Msm), an avirulent soil mycobacterium as the model system. We first describe the generation and enrichment of native EVs of Msm. Then, we describe the design and construction of recombinant mEVs that contain either mCherry, a red fluorescent reporter protein, or EsxA (Esat-6), a prominent immunogen of Mycobacterium tuberculosis. We achieve this by separately fusing mCherry and EsxA N-termini with the C-terminus of a small Msm protein Cfp-29. Cfp-29 is one of the few abundantly present proteins of MsmEVs. The protocol to generate and enrich recombinant mEVs from Msm remains identical to the generation and enrichment of native EVs of Msm.

3D printing for spine pathologies: a state-of-the-art review.

Three-Dimensional Printing has advanced throughout the years in the field of biomedical science with applications, especially in spine surgeries. 3D printing has the ability of fabricating highly complex structures with ease and high dimensional accuracy. The complexity of the spine's architecture and the inherent dangers of spinal surgery bring the evaluation of 3D printed models into consideration. This article summarizes the benefits of 3D printing based models for application in spine pathology. 3D printing technique is extensively used for fabrication of anatomical models, surgical guides and patient specific implants (PSI). The 3D printing based anatomical models assist in preoperative planning and training of students. Furthermore, 3D printed models can be used for improved communication and understanding of patients about the spinal disorders. The use of 3D printed surgical guides help in the stabilization of the spine during surgery, improving post procedural outcomes. Improved surgical results can be achieved by using PSIs that are tailored for patient specific needs. Finally, this review discusses the limitations and potential future scope of 3D printing in spine pathologies. 3D printing is still in its infancy, and further research would provide better understanding of the technology's true potential in spinal procedures.