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The Food and Drug Administration (FDA) has an expectation that products filed for marketing authorization have to include data that are representative of the US patient population. Any foreign clinical data that is submitted has to represent an ethnically diverse population that is generated utilizing qualified Principal Investigators (PIs) and conducted according to Good Clinical Practices (GCP) requirements outlined in 21 CFR 312.120, Foreign clinical studies not conducted under an IND. With the recent passing of the Omnibus Legislation, FDA now also has the authority to require Diversity Plans for all Phase 3 clinical trials of new drugs or “as appropriate, another pivotal study of a new drug.” The FDA has released a guidance document, “Diversity Plans to Improve Enrollment of Participants from Underrepresented Racial and Ethnic Populations in Clinical Trials” (April 2022), for the industry with expectations to update the document in 2023 now that legislation is passed. This whitepaper discusses the FDA guidance and expectations of sponsors with regards to foreign clinical data and the intersection with enrolling ethnically diverse populations in clinical studies.

Abstract Background Thymic epithelial cells (TECs) are responsible for shaping the repertoires of T cells, where their postnatal regeneration depends on a subset of clonogenic TECs. Despite the implications for regenerative medicine, their cultivation and expansion remain challenging. Primary explant cell culture is a technique that allows the seeding and expansion of difficult-to-culture cells. Here, we report a reliable and simple culture system to obtain functional TECs and thymic interstitial cells (TICs). Methods To establish primary thymic explants, we harvested 1 mm cleaned fragments of thymus from 5-week-old C57/BL6 mice. Tissue fragments of a complete thymic lobe were placed in the center of a Petri dish with 1 mL of DMEM/F-12 medium supplemented with 20% fetal bovine serum (FBS) and 1% penicillin‒streptomycin. To compare, thymic explants were also cultivated by using serum-free DMEM/F-12 medium supplemented with 10% KnockOut™. Results We obtained high numbers of functional clonogenic TECs and TICs from primary thymic explants cultivated with DMEM/F-12 with 20% FBS. These cells exhibited a highly proliferative and migration profile and were able to constitute thymospheres. Furthermore, all the subtypes of medullary TECs were identified in this system. They express functional markers to shape T-cell and type 2 innate lymphoid cells repertoires, such as Aire, IL25, CCL21 and CD80. Finally, we also found that ≥ 70% of lineage negative TICs expressed high amounts of Aire and IL25. Conclusion Thymic explants are an efficient method to obtain functional clonogenic TECs, all mTEC subsets and different TICs Aire+IL25+ with high regenerative capacity.

Denosumab is a fully human MAB that acts as a potent anti-resorptive by inhibiting activation of osteoclasts by inhibiting the receptor activator of nuclear factor-kappa B (RANK) ligand. Hypocalcaemia has been reported as one of the serious adverse sequelae of use of denosumab. We present a case of refractory hypocalcaemia following administration of a single dose of denosumab in a patient with metastatic castrate-resistant prostate cancer. The patient’s serum calcium and vitamin D concentrations and renal function were normal prior to denosumab administration. Serum alkaline phosphatase (ALP) level was however elevated pre-morbidly consistent with known bone metastases. The patient was treated with high-dose oral and IV calcium without any appreciable response in serum calcium. During his 30-day hospital admission, he demonstrated disease progression with development of new liver metastases and bone marrow involvement. Normocalcaemia was not achieved despite 1 month of aggressive therapy. Given the patient was asymptomatic and prognosis guarded, he was eventually discharged for ongoing supportive care under the palliative care team.

Rubus fruits are rich in nutrition with a wide range of phytochemical and consumed by humans, but many of its wild varieties are not commercially useful. Phytochemical exploration of these wild fruits could improve the commercial usage of these Rubus species.This study focuses to analyze the phytochemical, antioxidant and cytotoxicity of three wild Rubus ellipticus, R. niveus and R. fairholmianus fruits comparatively, which has not revealed. The in vitro antioxidant studies indicated the ethyl acetate and methanol extracts of studied Rubus species competent in scavenging different stable and generated radical. The correlation and regression analysis also statistically supported the contents in the extracts could act against various radicals. Correlation between phytochemical constituent and radical scavenging activity found to be highly positive. The results acquired from this investigation revealed that they are equally competent and could be recommended as dietary supplements or as nutraceuticals. Keywords: Anticancer, Antioxidant, Flavonoids, Nutraceuticals, Phenolics

Abstract In this work, we provide specific clinical examples to demonstrate basic practical techniques involved in image segmentation, computer-aided design, and 3D printing. A step-by-step approach using United States Food and Drug Administration cleared software is provided to enhance surgical intervention in a patient with a complex superior sulcus tumor. Furthermore, patient-specific device creation is demonstrated using dedicated computer-aided design software. Relevant anatomy for these tasks is obtained from CT Digital Imaging and Communications in Medicine images, leading to the generation of 3D printable files and delivery of these files to a 3D printer.

INTRODUCTION: The diverse clinical phenotype of hemoglobin E (HbE)/β- thalassemia has not only confounded clinicians in matters of patient management but has also led scientists to investigate the complex mechanisms involved in maintaining the delicate red cell environment where, even with apparent similarities of α- and β-globin genotypes, the phenotype tells a different story. The BTB and CNC homology 1 (BACH1) protein is known to regulate α- and β-globin gene transcriptions during the terminal differentiation of erythroid cells. With the mutations involved in HbE/β-thalassemia disorder, we studied the role of BACH1 in compensating for the globin chain imbalance, albeit for fine-tuning purposes. METHODS: A total of 47 HbE/β-thalassemia samples were analyzed using real-time quantitative polymerase chain reaction and correlated with age, sex, red blood cell parameters, globin gene expressions, and some clinical data. RESULTS: The BACH1 expression among the β-thalassemia intermedia patients varied by up to 2-log differences and was positively correlated to age; α -, β-, and γ-globin gene expression level; and heme oxygenase 1 protein. BACH1 was also negatively correlated to reticulocyte level and had a significant correlation with splenectomy. DISCUSSION AND CONCLUSION: This study indicates that the expression of BACH1 could be elevated as a compensatory mechanism to decrease the globin chain imbalance as well as to reduce the oxidative stress found in HbE/β-thalassemia.

A single-photon CMOS image sensor (CIS) design based on pinned photodiode (PPD) with multiple charge transfers and sampling is described. In the proposed pixel architecture, the photogenerated signal is sampled non-destructively multiple times and the results are averaged. Each signal measurement is statistically independent and by averaging, the electronic readout noise is reduced to a level where single photons can be distinguished reliably. A pixel design using this method was simulated in TCAD and several layouts were generated for a 180-nm CMOS image sensor process. Using simulations, the noise performance of the pixel was determined as a function of the number of samples, sense node capacitance, sampling rate and transistor characteristics. The strengths and limitations of the proposed design are discussed in detail, including the trade-off between noise performance and readout rate and the impact of charge transfer inefficiency (CTI). The projected performance of our first prototype device indicates that single-photon imaging is within reach and could enable ground-breaking performances in many scientific and industrial imaging applications.

In the present study, the anti-oxidant and cytotoxic effects of the different solvent extracts of Pogostemon mollis were analyzed. The phenolic, tannin and flavonoid contents were highest in the ethyl acetate extract and analogous to the antioxidant activity results. The extracts showed activities similar to the standard anti-oxidants. The extent to which the extracts protect free radical damage on DNA was evaluated and showed good genoprotective effects. Bacterial cells and fungal spores and hyphae showed visible damages due to the treatment of ethyl acetate extract. Finally, in the cytotoxic analysis, IC50 value was calculated based on the absorbance value of different concentrations. It concluded that P. mollis is a prospective candidate for the various therapeutic applications especially its ethyl acetate extract.

Right Ventricular to Left Ventricular (RV/LV) diameter ratio has been shown to be a prognostic biomarker for patients suffering from acute Pulmonary Embolism (PE). While Computed Tomography Pulmonary Angiography (CTPA) images used to confirm a clinical suspicion of PE do include information of the heart, a numerical RV/LV diameter ratio is not universally reported, likely because of lack in training, inter-reader variability in the measurements, and additional effort by the radiologist. This study designs and validates a completely automated Computer Aided Detection (CAD) system to compute the axial RV/LV diameter ratio from CTPA images so that the RV/LV diameter ratio can be a more objective metric that is consistently reported in patients for whom CTPA diagnoses PE.The CAD system was designed specifically for RV/LV measurements. The system was tested in 198 consecutive CTPA patients with acute PE. Its accuracy was evaluated using reference standard RV/LV radiologist measurements and its prognostic value was established for 30-day PE-specific mortality and a composite outcome of 30-day PE-specific mortality or the need for intensive therapies. The study was Institutional Review Board (IRB) approved and HIPAA compliant.The CAD system analyzed correctly 92.4% (183/198) of CTPA studies. The mean difference between automated and manually computed axial RV/LV ratios was 0.03±0.22. The correlation between the RV/LV diameter ratio obtained by the CAD system and that obtained by the radiologist was high (r=0.81). Compared to the radiologist, the CAD system equally achieved high accuracy for the composite outcome, with areas under the receiver operating characteristic curves of 0.75 vs. 0.78. Similar results were found for 30-days PE-specific mortality, with areas under the curve of 0.72 vs. 0.75.An automated CAD system for determining the CT derived RV/LV diameter ratio in patients with acute PE has high accuracy when compared to manual measurements and similar prognostic significance for two clinical outcomes.

Englerin A is a structurally unique natural product reported to selectively inhibit growth of renal cell carcinoma cell lines. A large scale phenotypic cell profiling experiment (CLiP) of englerin A on ¬over 500 well characterized cancer cell lines showed that englerin A inhibits growth of a subset of tumor cell lines from many lineages, not just renal cell carcinomas. Expression of the TRPC4 cation channel was the cell line feature that best correlated with sensitivity to englerin A, suggesting the hypothesis that TRPC4 is the efficacy target for englerin A. Genetic experiments demonstrate that TRPC4 expression is both necessary and sufficient for englerin A induced growth inhibition. Englerin A induces calcium influx and membrane depolarization in cells expressing high levels of TRPC4 or its close ortholog TRPC5. Electrophysiology experiments confirmed that englerin A is a TRPC4 agonist. Both the englerin A induced current and the englerin A induced growth inhibition can be blocked by the TRPC4/C5 inhibitor ML204. These experiments confirm that activation of TRPC4/C5 channels inhibits tumor cell line proliferation and confirms the TRPC4 target hypothesis generated by the cell line profiling. In selectivity assays englerin A weakly inhibits TRPA1, TRPV3/V4, and TRPM8 which suggests that englerin A may bind a common feature of TRP ion channels. In vivo experiments show that englerin A is lethal in rodents near doses needed to activate the TRPC4 channel. This toxicity suggests that englerin A itself is probably unsuitable for further drug development. However, since englerin A can be synthesized in the laboratory, it may be a useful chemical starting point to identify novel modulators of other TRP family channels.

To test the hypothesis that wide area detector face transplant surgical planning CT angiograms with simulated lower radiation dose and iterative reconstruction (AIDR3D) are comparable in image quality to those with standard tube current and filtered back projection (FBP) reconstruction.The sinograms from 320-detector row CT angiography of four clinical candidates for face transplantation were processed utilizing standard FBP, FBP with simulated 75, 62, and 50% tube current, and AIDR3D with corresponding dose reduction. Signal-to-noise ratio (SNR) and contrast-to-noise ratio (CNR) were measured at muscle, fat, artery, and vein. Image quality for each reconstruction strategy was assessed by two independent readers using a 4-point scale.Compared to FBP, the median SNR and CNR for AIDR3D images were higher at all sites for all 4 different tube currents. The AIDR3D with simulated 50% tube current achieved comparable SNR and CNR to FBP with standard dose (median muscle SNR: 5.77 vs. 6.23; fat SNR: 6.40 vs. 5.75; artery SNR: 43.8 vs. 45.0; vein SNR: 54.9 vs. 55.7; artery CNR: 38.1 vs. 38.6; vein CNR: 49.0 vs. 48.7; all p-values >0.19). The interobserver agreement in the image quality score was good (weighted κ = 0.7). The overall score and the scores for smaller arteries were significantly lower when FBP with 50% dose reduction was used. The AIDR3D reconstruction images with 4 different simulated doses achieved a mean score ranging from 3.68 to 3.82 that were comparable to the scores from images reconstructed using FBP with original dose (3.68-3.77).Simulated radiation dose reduction applied to clinical CT angiography for face transplant planning suggests that AIDR3D allows for a 50% reduction in radiation dose, as compared to FBP, while preserving image quality.

The fluid-filled lung exists in relative hypoxia in utero (∼25 mm Hg), but at birth fills with ambient air where the partial pressure of oxygen is ∼150 mm Hg. The impact of this change was studied in mouse lung with microarrays to analyze gene expression one day before, and 2, 6, 12 and 24 hours after birth into room air or 10% O(2). The expression levels of >150 genes, representing transcriptional regulation, structure, apoptosis and antioxidants were altered 2 hrs after birth in room air but blunted or absent with birth in 10% O(2). Kruppel-like factor 4 (Klf4), a regulator of cell growth arrest and differentiation, was the most significantly altered lung gene at birth. Its protein product was expressed in fibroblasts and airway epithelial cells. Klf4 mRNA was induced in lung fibroblasts exposed to hyperoxia and constitutive expression of Klf4 mRNA in Klf4-null fibroblasts induced mRNAs for p21(cip1/Waf1), smooth muscle actin, type 1 collagen, fibronectin and tenascin C. In Klf4 perinatal null lung, p21(cip1/Waf1)mRNA expression was deficient prior to birth and associated with ongoing cell proliferation after birth; connective tissue gene expression was deficient around birth and smooth muscle actin protein expression was absent from myofibroblasts at tips of developing alveoli; p53, p21(cip1/Waf1) and caspase-3 protein expression were widespread at birth suggesting excess apoptosis compared to normal lung. We propose that the changing oxygen environment at birth acts as a physiologic signal to induce lung Klf4 mRNA expression, which then regulates proliferation and apoptosis in fibroblasts and airway epithelial cells, and connective tissue gene expression and myofibroblast differentiation at the tips of developing alveoli.

Production of silver nanoparticles from the leaf extracts of Pimenta dioica is reported for the first time in this paper. Three different sets of leaves were utilized for the synthesis of nanoparticles—fresh, hot-air oven dried, and sun-dried. These nanoparticles were characterized using UV-Vis spectroscopy and AFM. The results were diverse in that different sizes were seen for different leaf conditions. Nanoparticles synthesized using sun-dried leaves (produced using a particular ratio (1 : 0.5) of the leaf extract sample and silver nitrate (1 mM), resp.) possessed the smallest sizes. We believe that further optimization of the current green-synthesis method would help in the production of monodispersed silver nanoparticles having great potential in treating several diseases.

Objectives To describe the initial dilemmas, mental stress, adaptive measures implemented and how the healthcare team collectively coped while providing healthcare services in a large slum in India, during the COVID-19 pandemic.Setting Community Health Division, Bangalore Baptist Hospital, Bangalore.Study design We used mixed methods research with a quantitative (QUAN) paradigm nested in the primary qualitative (QUAL) design. QUAL methods included ethnography research methods, in-depth interviews and focus group discussions.Participants A healthcare team of doctors, nurses, paramedical and support staff. Out of 87 staff, 42 participated in the QUAL methods and 64 participated in the QUAN survey.Results Being cognizant of the extreme vulnerability of the slums, the health team struggled with conflicting thoughts of self-preservation and their moral obligation to the marginalised section of society. Majority (75%) of the staff experienced fear at some point in time. Distracting themselves with hobbies (20.3%) and spending more time with family (39.1%) were cited as a means of emotional regulation by the participants in the QUAN survey. In the QUAL interviews, fear of death, the guilt of disease transmission to their loved ones, anxiety about probable violence and stigma in the slums and exhaustion emerged as the major themes causing stress among healthcare professionals. With positive cognitive reappraisal, the health team collectively designed and implemented adaptive interventions to ensure continuity of care. They dealt with the new demands by positive reframing, peer support, distancing, information seeking, response efficacy, self-efficacy, existential goal pursuit, value adherence and religious coping.Conclusion The novel threat of the COVID-19 pandemic threw insurmountable challenges potentiating disastrous consequences; slums becoming a threat to themselves, threat to the health providers and a threat for all. Perhaps, a lesson we could learn from this pandemic is to incorporate ‘slum health’ within universal healthcare.