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Shazia Bano,1–3 Samina Nazir,2 Alia Nazir,1 Saeeda Munir,3 Tariq Mahmood,2 Muhammad Afzal,1 Farzana Latif Ansari,4 Kehkashan Mazhar3 1Department of Physics, The Islamia University of Bahawalpur, Bahawalpur, 2Nanosciences and Technology Department, National Centre for Physics, 3Institute of Biomedical and Genetic Engineering (IBGE), 4Pakistan Council for Science and Technology, Islamabad, Pakistan Abstract: Superparamagnetic iron oxide nanoparticles (SPIONs) have the potential to be used as multimodal imaging and cancer therapy agents due to their excellent magnetism and ability to generate reactive oxygen species when exposed to light. We report the synthesis of highly biocompatible SPIONs through a facile green approach using fruit peel extracts as the biogenic reductant. This green synthesis protocol involves the stabilization of SPIONs through coordination of different phytochemicals. The SPIONs were functionalized with polyethylene glycol (PEG)-6000 and succinic acid and were extensively characterized by X-ray diffraction analysis, field emission scanning electron microscopy, energy-dispersive X-ray spectroscopy, atomic force microscopy, Rutherford backscattering spectrometry, diffused reflectance spectroscopy, fluorescence emission, Fourier-transform infrared spectroscopy, ultraviolet-visible spectroscopy, and magnetization analysis. The developed SPIONs were found to be stable, almost spherical with a size range of 17–25 nm. They exhibited excellent water dispersibility, colloidal stability, and relatively high R2 relaxivity (225 mM-1 s-1). Cell viability assay data revealed that PEGylation or carboxylation appears to significantly shield the surface of the particles but does not lead to improved cytocompatibility. A highly significant increase of reactive oxygen species in light-exposed samples was found to play an important role in the photokilling of human cervical epithelial malignant carcinoma (HeLa) cells. The bio-SPIONs developed are highly favorable for various biomedical applications without risking interference from potentially toxic reagents. Keywords: green approach, biocompatible, multifunctional, bio synthesis, surface modification, microwave incubation, MRI contrast agent, photosensitive SPIONs more...

Shazia Bano,1–3,* Samina Nazir,2,* Saeeda Munir,3 Mohamed Fahad AlAjmi,4 Muhammad Afzal,1 Kehkashan Mazhar3 1Department of Physics, The Islamia University of Bahawalpur, 2Nanosciences and Technology Department, National Centre for Physics, Islamabad, 3Institute of Biomedical and Genetic Engineering, Islamabad, Pakistan; 4College of Pharmacy, King Saud University, Riyadh, Kingdom of Saudi Arabia *These authors contributed equally to this work Abstract: We report “smart” nickel oxide nanoparticles (NOPs) as multimodal cancer therapy agent. Water-dispersible and light-sensitive NiO core was synthesized with folic acid (FA) connected bovine serum albumin (BSA) shell on entrapped doxorubicin (DOX). The entrapped drug from NOP-DOX@BSA-FA was released in a sustained way (64 hours, pH=5.5, dark conditions) while a robust release was found under red light exposure (in 1/2 hour under λmax=655 nm, 50 mW/cm2, at pH=5.5). The cell viability, thiobarbituric acid reactive substances and diphenylisobenzofuran assays conducted under light and dark conditions revealed a high photodynamic therapy potential of our construct. Furthermore, we found that the combined effect of DOX and NOPs from NOP-DOX@BSA-FA resulted in cell death approximately eightfold high compared to free DOX. We propose that NOP-DOX@BSA-FA is a potential photodynamic therapy agent and a collective drug delivery system for the systemic administration of cancer chemotherapeutics resulting in combination therapy. Keywords: light-triggered drug release, cancer, bovine serum albumin, multi-model therapy more...

Abstract Background Histotripsy is a novel, non-invasive, non-ionizing, and non-thermal ablation technique that disrupts tumors using acoustic cavitation. We report the first use of Histotripsy as bridging therapy prior to liver transplant for hepatocellular carcinoma (HCC) treated with histotripsy. Case presentation A 59-year-old woman presented with Metabolic-Associated Steatotic Liver Disease (MASLD) cirrhosis (labMELD = 14), hepatic encephalopathy, and a single 2 cm OPTN V lesion in the left lateral segment consistent with HCC. The patient underwent histotripsy treatment of the lesion as bridging therapy before receiving liver transplantation. Histopathology analysis of the explanted liver showed total necrosis of the treated area, with no residual viable tissue tumor. Conclusion This case demonstrates the potential utility of histotripsy as an effective bridging therapy for patients with combined cirrhosis and hepatocellular carcinoma (HCC) awaiting liver transplantation, with complete tumor necrosis on explant pathology demonstrating its therapeutic efficacy. more...

Photocatalytic reduction of carbon dioxide (CO2) presents a pivotal solution to address meteorological and ecological challenges. Currently, metal-organic frameworks (MOFs) with their crystalline porosity, adjustable structures, and diverse chemical functionalities have garnered significant attention in the realm of photocatalytic CO2 reduction. This review provides a brief introduction to CO2 reduction and MOF material and their applications in CO2 reduction. Then, it undertakes a comprehensive examination of MOFs, summarizing their key attributes, including porosity, large surface area, structural multifunctionalities, and responsiveness to visible light, along with an analysis of heterojunctions and their methods of preparation. Additionally, it elucidates the fundamental principle of photocatalysis and CO2 reduction, encompassing both half and overall reactions. Furthermore, the classification of MOF-based materials is explored, along with the proposed mechanism for CO2 reduction and an update on recent developments in this field. Finally, this review outlines the challenges and potential opportunities for utilizing MOFs in CO2 reduction, offering valuable insights to scholars seeking innovative approaches not only to enhance CO2 reduction but also to advance other photocatalytic processes. more...

OBJECTIVES Cutaneous metastases are a very rare presentation of ovarian tumors (OV). They occur in about 3.5 percent of cases. The prognosis of cutaneous metastases is not good despite recent treatment modalities. Researchers are looking for much more personalized treatments. The patient had a history of ovarian tumor, for which she had a hysterectomy and bilateral salpingo-oophorectomy. She presented with multiple cutaneous eruptions. The histopathology shows high-grade serous carcinoma of the ovaries. Immunochemistry is positive for cytokeratin 7, WT1, and CAX. She had developed left leg swelling, generalized edema, and complaints of dysuria and difficulty defecating. Many targeted therapies have been developed, such as inhibiting growth factor receptors and immunological treatments that induce cell death, opening the door to precision medicine. more...

Native pulmonary valve endocarditis is a rare phenomenon as native valve endocarditis tends to typically affect the left sided heart valves. However, the right-sided heart valves can be affected in patients with a history of intravenous drug use, whereby the tricuspid valve is most commonly affected. We present two cases who were diagnosed with native pulmonary valve endocarditis in the absence of congenital heart disease. In the first case, the native pulmonary valve endocarditis was probably a derivative of compounding factors of an enlarged underlying pulmonary artery and staphylococcal bacteraemia. In the second case, a common causal organism of native valve endocarditis following dental treatment and the resultant echocardiography findings was of significant interest. In summary, native pulmonary valve endocarditis is relatively rare complication in the adult population, especially in the absence of congenital heart disease. [ABSTRACT FROM AUTHOR] more...

Asthenoteratozoospermia is one of the most severe types of qualitative sperm defects. Most cases are due to mutations in genes encoding the components of sperm flagella, which have an ultrastructure similar to that of motile cilia. Coiled-coil domain containing 103 (CCDC103) is an outer dynein arm assembly factor, and pathogenic variants of CCDC103 cause primary ciliary dyskinesia (PCD). However, whether CCDC103 pathogenic variants cause severe asthenoteratozoospermia has yet to be determined. Whole-exome sequencing (WES) was performed for two individuals with nonsyndromic asthenoteratozoospermia in a consanguineous family. A homozygous CCDC103 variant segregating recessively with an infertility phenotype was identified (ENST00000035776.2, c.461A>C, p.His154Pro). CCDC103 p.His154Pro was previously reported as a high prevalence mutation causing PCD, though the reproductive phenotype of these PCD individuals is unknown. Transmission electron microscopy (TEM) of affected individuals' spermatozoa showed that the mid-piece was severely damaged with disorganized dynein arms, similar to the abnormal ultrastructure of respiratory ciliary of PCD individuals with the same mutation. Thus, our findings expand the phenotype spectrum of CCDC103 p.His154Pro as a novel pathogenic gene for nonsyndromic asthenospermia. more...

Background: Increased afterload affects many of the flow dependent metrics assessed during transthoracic echocardiography (TTE) especially in the evaluation valvular disease. A single timepoint blood pressure (BP) may not accurately reflect the afterload present at the time of flow-dependent imaging and quantification. We assessed the magnitude of change in BP at discrete timepoints during routine TTE. Method: We conducted a prospective study where participants underwent automated BP measurement while undergoing a clinically indicated TTE. The first reading was obtained right after the patient lay supine and subsequent readings were taken at 10-min intervals during image acquisition. Result: We included 50 participants (66% were male, with a mean age of 64 years). After 10 min, 40 (80%) participants had a drop in systolic BP of >10 mmHg. Compared to the baseline, there was a significant drop in systolic BP (mean decrease 20.0 ± 12.8 mmHg; P more...

Non-obstructive azoospermia (NOA) is a severe factor of male infertility; it affects approximately 1% of the global male population and accounts for 40% of male infertility cases. However, the majority of NOA cases remain idiopathic. This is the first study using whole-exome sequencing (WES) to identify a novel missense mutation in the DND1 gene (c.212A>C, p. E71A) from a Pakistani family, that includes three males with NOA. This mutation is predicted to cause DND1 protein misfolding and weaken the DND1 interaction with NANOS2, a significant regulator in primordial germ cell development. Our study identified a DND1 pathogenic mutation in NOA patients and highlighted its critical role in male fertility in humans. more...

Introduction: Pediatric trauma centers (PTCs) were created to address the unique needs of injured children with the expectation that outcomes would be improved. However, prior studies to evaluate the impact of PTCs have had conflicting results. Our study was conducted to further clarify this question. We hypothesize that severely injured children ≤ 14 years of age have better outcomes at PTCs and that better survival may be due to higher emergency department (ED) survival rates than at adult trauma centers (ATCs). Methods: A retrospective analysis of severely injured children (ISS>15) ≤18 years of age entered into the National Trauma Data Bank (NTDB) between 2011 and 2012 was performed. Subjects were stratified into 2 age cohorts; young children (0-14 years) and adolescents (15-18 years). Primary outcomes were emergency department (ED) and in-patient (IP) mortality. Secondary outcomes included in-hospital complications, hospital and ICU length of stay, and ventilator days. Outcome differences were assessed using multilevel logistic and negative binomial regression analyses. Results: A total of 10,028 children were included. Median ISS was 22 (Interquartile range 17-29). Adjusting for confounders on multivariate analysis, children ≤ 14 had lower odds of ED (0.42[CI 0.25-0.71], p=0.001) and IP mortality (0.73[CI 0.5-0.9], p=0.02) at PTCs. There were no differences in odds of ED mortality (0.81 [CI 0.5-1.3], p=0.4) or IP mortality (1.01 [CI 0.8-1.2], p=0.88) for adolescents between centers. There were no differences in complication rates between PTCs and ATCs (OR 0.86 [CI 0.69-1.06], p=1.7) but children were more likely to be discharged to home and have more ICU and ventilator free days if treated at a PTC. Conclusion: Young children but not adolescents have better ED survival at PTCs compared to ATCs. more...

Estimation of an optimal controller is a fundamental problem in control engineering and is widely known as Optimization. Numerous computation and numerical techniques have been evolved during the past years for the estimation of the optimal solution. Optimal control of a discrete-time system is concerned with optimizing a given objective function using "Homogenous Polynomial Lyapunov Function (HPLF)". This research focuses upon the design of optimal Guaranteed Cost Controller (GCC) for discretetime uncertain system using HPLF. The uncertainties are assumed to be norm-bounded uncertainties. The effect of actuator saturation is also incorporated in the system. Sufficient conditions for the existence of HPLF are derived in terms of Linear Matrix Inequalities (LMI). The LMI approach has the advantage that it can be solved efficiently using Convex Optimization. LMI's combined with HPLF helps to design the guaranteed cost controller which minimizes the cost by minimizing cost function. Furthermore, the state trajectories and the invariant set are also shown for the observation of the overall performance. [ABSTRACT FROM AUTHOR] more...

Male infertility is a prevalent disorder distressing an estimated 70 million people worldwide. Despite continued progress in understanding the causes of male infertility, idiopathic sperm abnormalities such as multiple morphological abnormalities of sperm flagella (MMAF) still account for about 30% of male infertility. Recurrent mutations in DNAH1 have been reported to cause MMAF in various populations, but the underlying mechanism is still poorly explored. This study investigated the MMAF phenotype of two extended consanguineous Pakistani families without manifesting primary ciliary dyskinesia symptoms. The transmission electron microscopy analysis of cross-sections of microtubule doublets revealed a missing central singlet of microtubules and a disorganized fibrous sheath. SPAG6 staining, a marker generally used to check the integration of microtubules of central pair, further confirmed the disruption of central pair in the spermatozoa of patients. Thus, whole-exome sequencing (WES) was performed, and WES analysis identified two novel mutations in the DNAH1 gene that were recessively co-segregating with MMAF phenotype in both families. To mechanistically study the impact of identified mutation, we generated Dnah1 mice models to confirm the in vivo effects of identified mutations. Though Dnah1△iso1/△iso1 mutant mice represented MMAF phenotype, no significant defects were observed in the ultrastructure of mutant mice spermatozoa. Interestingly, we found DNAH1 isoform2 in Dnah1△iso1/△iso1 mutant mice that may be mediating the formation of normal ultrastructure in the absence of full-length protein. Altogether we are first reporting the possible explanation of inconsistency between mouse and human DNAH1 mutant phenotypes, which will pave the way for further understanding of the underlying pathophysiological mechanism of MMAF. more...

Introduction The annual mortality and national expense of the opioid crisis continue to rise in the USA (130 deaths/day, $50 billion/year). Opioid use disorder usually starts with the prescription of opioids for a medical condition. Its risk is associated with greater pain intensity and coping strategies characterised by pain catastrophising. Non-pharmacological analgesics in the hospital setting are critical to abate the opioid epidemic. One promising intervention is virtual reality (VR) therapy. It has performed well as a distraction tool and pain modifier during medical procedures; however, little is known about VR in the acute pain setting following traumatic injury. Furthermore, no studies have investigated VR in the setting of traumatic brain injury (TBI). This study aims to establish the safety and effect of VR therapy in the inpatient setting for acute traumatic injuries, including TBI.Methods and analysis In this randomised within-subjects clinical study, immersive VR therapy will be compared with two controls in patients with traumatic injury, including TBI. Affective measures including pain catastrophising, trait anxiety and depression will be captured prior to beginning sessions. Before and after each session, we will capture pain intensity and unpleasantness, additional affective measures and physiological measures associated with pain response, such as heart rate and variability, pupillometry and respiratory rate. The primary outcome is the change in pain intensity of the VR session compared with controls.Ethics and dissemination Dissemination of this protocol will allow researchers and funding bodies to stay abreast in their fields through exposure to research not otherwise widely publicised. Study protocols are compliant with federal regulation and University of Maryland Baltimore’s Human Research Protections and Institutional Review Board (protocol number HP-00090603). Study results will be published on completion of enrolment and analysis, and deidentified data can be shared by request to the corresponding author.Trial registration number NCT04356963; Pre-results. more...

The emergence of recent SARS-CoV-2 has become a global health issue. This single-stranded positive-sense RNA virus is continuously spreading with increasing morbidities and mortalities. The proteome of this virus contains four structural and sixteen nonstructural proteins that ensure the replication of the virus in the host cell. However, the role of phosphoprotein (N) in RNA recognition, replicating, transcribing the viral genome, and modulating the host immune response is indispensable. Recently, the NMR structure of the N-terminal domain of the Nucleocapsid Phosphoprotein has been reported, but its precise structural mechanism of how the ssRNA interacts with it is not reported yet. Therefore, here, we have used an integrated computational pipeline to identify the key residues, which play an essential role in RNA recognition. We generated multiple variants by using an alanine scanning strategy and performed an extensive simulation for each system to signify the role of each interfacial residue. Our analyses suggest that residues T57A, H59A, S105A, R107A, F171A, and Y172A significantly affected the dynamics and binding of RNA. Furthermore, per-residue energy decomposition analysis suggests that residues T57, H59, S105 and R107 are the key hotspots for drug discovery. Thus, these residues may be useful as potential pharmacophores in drug designing. more...

Atrial fibrillation is the most common arrhythmia worldwide with increasing frequency noted with age. Hyperthyroidism is a well-known cause of atrial fibrillation with a 16%–60% prevalence of atrial fibrillation in patients with known hyperthyroidism Ross et al. (2016). While hyperthyroidism as a causative factor of atrial fibrillation is well established, this literature review aims to answer several questions on this topic including: 1. The relationship of atrial fibrillation to hyperthyroidism 2. Atrial fibrillation as a predictor of hyperthyroidism 3. The pathophysiology of thyrotoxic atrial fibrillation 4. Subclinical hyperthyroidism and the relationship with atrial fibrillation 5. Cardioversion and Catheter ablation of hyperthyroid patients with atrial fibrillation 6. Thrombotic risk of hyperthyroid patients with atrial fibrillation 7. Management of Thyrotoxic Atrial fibrillation 8. Pharmacological rhythm control in patients with hyperthyroidism and atrial fibrillation 9. Treatment of Hyperthyroidism to prevent atrial fibrillation 10. Clinical Implications of Hyperthyroidism and Atrial Fibrillation more...

ABSTRACT: OBJECTIVES: The purpose of this study was to know about the risk factors of malnutrition and the incidence of risk factors among known cases of malnutrition admitted in tertiary care hospitals of Peshawar. METHODOLOGY: A descriptive cross-sectional study was done. Study was conducted in tertiary care hospitals of Peshawar from March 2019 to July 2019. Study included 100 patients of protein calorie malnutrition less than five years of age. Internationally recognized Gomez classification was used to label patient as malnourished. Risk factors of primary malnutrition including socioeconomic status, ignorance of weaning, poverty, lack of immunization and primary care, maternal illiteracy and risk factors of secondary malnutrition including infections, congenital diseases, malabsorptive disorders and metabolic disorders were assessed. Data was collected through a questionnaire. Tables and graphs were used to determine the frequency of risk factors for protein calorie malnutrition. RESULTS: Among 100 patients of PCM 59 were male and 41 were females. After assessment of both genders, risk factors that show close association with protein calorie malnutrition were low socioeconomic status, maternal illiteracy, lack of family planning, poor weaning and repeated infections. Among these risk factors the most frequently observed risk factors for protein calorie malnutrition were low socioeconomic status and repeated infections. CONCLUSION: Low socioeconomic condition and repeated infections are the leading cause of protein calorie malnutrition. KEYWORDS: Malnutrition, Gomez Classification, Frequency, Risk Factors more...

This study investigated the effects of four different strainer types on flow characteristics (orifice coefficient (k), exponent coefficient (n), individual flow rate deviation (φ) and discharge coefficient (Cd)) of different nozzle types. The volumetric flow rates of anti-drift (AD) and multi-range (LU) flat-fan nozzles of three different orifice sizes were determined at different operational pressures (1.5, 3.0, 4.0, 6.0 and 8.0 bars). In each treatment, the nozzles were used together with cup screen strainer of 50-mesh, slotted strainer, cylindrical strainers of 40-, 50-, and 80-meshes, and ball-check strainers of 50- and 80-meshes. The flow rate measurements were also obtained without strainers. The relation between the flow rate (Q) and spray pressure (P) for each nozzle combination (nozzle type, strainer type, and orifice size) was presented using the power regression model . The “k” coefficient, which is the slope of the line referring to the relation between the nozzle flow rate and spray pressure, was evaluated as a comparison parameter between the nozzle combinations. The “k” mean values of the nozzle types using with ball-check strainers were lower than those of the without strainer, cup screen, slotted and cylindrical strainers. This result showed that the volumetric flow rate decreased with respect to the other nozzle combinations operated at the same operational pressure. Thus, the deviation rate from the nominal flow rate of the nozzles used with the ball-check strainers exceeded the acceptable deviation limit of ±10%. As the “n” coefficients of the LU and AD nozzles used with cup screen, slotted, cylindrical strainers and without strainers were close to 0.50, the ball-check strainers resulted in increasing the “n” coefficient of the nozzles. The “n” coefficient of the nozzles used with the ball-check strainers of 50- and 80- meshes was determined as 0.586 and 0.608 for the AD nozzle, respectively and, 0.576 and 0.584 for the LU nozzle, respectively. The ball-check strainers dramatically decreased the discharge coefficient (Cd) of the nozzles compared to the other strainers and the usage without strainer. For the cup screen, slotted, cylindrical strainers and the usage without strainer, the Cd means ranged from 0.67 to 0.77 for the AD nozzle, and 0.91 to 0.94 for the LU nozzle. The Cd means of the nozzles used with the ball-check strainers of 50- and 80- meshes were determined as 0.39 and 0.34 for the AD nozzle, respectively, and 0.56 and 0.53 for the LU nozzle, respectively. more...

ABSTRACT. Almeida D.O., Tortely R., Nascimento E.R., Khan M., Pereira V.L.A & Babapoor S. [Use of RT-PCR and elisa techniques for the diagnostic of infectious bronchitis virus in broilers at slaughter.] Uso das técnicas de RT-PCR e elisa no diagnóstico da bronquite infecciosa em frangos de corte ao abate. Revista Brasileira de Medicina Veterinária, 37(1):55-59, 2015. Departamento de Saúde Coletiva Veterinária e Saúde Pública, Universidade Federal Fluminense, Rua Vital Brazil Filho, 64, Santa Rosa, Niterói, RJ 24230-340, Brasil. E-mail: elmiro@vm.uff.br Avian infectious bronchitis (IB) is a viral, acute and highly contagious disease caused by infectious bronchitis virus (IBV). The disease affects broilers improvement and performance causing major economic losses in the world poultry industry. Generally, IBV infections can be diagnosed by detection of IBV itself or the specific antibody response. This study aimed to detect IBV in broilers under Sanitary Inspection by RT-PCR and ELISA, comparing the results and relating them with the average weight of the flock. Samples were collected from 40 vaccinated broiler flocks under Sanitary Inspection at slaughter. Ten birds from every flock were randomly selected and blood samples were collected for ELISA as well three birds had their trachea and caecal tonsils collected for RT- -PCR test. From 40 flocks, 30 were IBV positive by ELISA and 26 by RT-PCR, in which 15 were detected simultaneously in trachea and caecal tonsil and 5 in each sample. There was no agreement between ELISA and RT-PCR results regarding IBV diagnosis as well positivity in both tests was not statistically significant with the average weight of the flock. There was an improvement on IBV diagnosis when caecal tonsils and tracheas were used instead of just one of them. Considering the only vaccine serotype allowed by Brazilian government is the Mass serotype and its persistence in broilers would only be detected up to 28 days, possibly the studied broilers were challenge with a field strain. more...

ABSTRACT OBJECTIVE: to study the efficacy and safety of using appendix as cath­eterizable conduit based on Mitrofanoff procedure in urinary diversion. METHODOLOGY: This prospective observational study was conducted at department of surgery, Hayatabad medical complex and Khyber teaching hospital from 1st January 2005 till 31st December 2011. Total number of 56 patients (38 males and 18 females) from 5-60 years of age were included in the study. Mitrofanoff principle was used to achieve clean intermittent catheterization and urinary continence. Patients were followed for two years with, initially 3 months and then 6 months intervals. All the data was collected on structured proforma and was analyzed on SPSS. RESULTS: Out of 56 total patient, primary diagnosis was transitional cell carcinoma in 23 (41%) case, exstrophy-epispadias in 10(17.9%), urethral injury in 7(12.5%) cases and miscellaneous in 16 (28.5%) cases. Augmentation cystoplasty was performed in 9 (16%) and 33(58.9%) patients had neobladder formation in conjunction with Mitrofanoff pro­cedure. Urinary continence was achieved in 51(91%) patients. Mortality rate was 3.5% (n=2/56) and 3(5.3%) patients were lost to follow up. Complications were observed in 18 (35.2%) cases. Stomal stenosis was most common complication in 7(12.5%) patients, followed by incisional hernia, wound infection, suprapubic fistula and intestinal obstruction in 2 (3.9%) patients each. Only 7 (13.7%) needed surgical management, rest of 11(21.5%) were treated conservatively. CONCLUSION: Mitrofanoff procedure provides immense advantage to patients needing urinary diversion, as it is associated with far less complications, higher continence rate and greatly improved level of independence in bladder management. KEY WORDS: Radical cystectomy, Urinary diversion, Mitrofanoff prin­ciple. more...

In this study, the size and shape properties of twelve different bean cultivars (eleven varieties and a genotype) were determined using image processing method on digital photography and the cultivars were grouped depending on their properties. The highest mean values of geometric diameter, projected area, length, width and shape factor in the study were determined in the ‘Çayırlı’ genotype. ‘Terzibaba’ cultivar, more spherical shape at width orientation compared to other cultivars, had the lowest value of thickness (4.6 mm) and more circular at the width orientation among the cultivars. The highest mean of thickness (6.0 mm) were measured in ‘Elkoca-05’ and ‘Noyanbey-98’ cultivars. The sphericity mean of ‘Karacaşehir-90’ (70%) was found higher than those of the other cultivars, while the geometric mean diameter, projected area, length and width values had the lowest mean. ’Yunus-90’ and ‘Göynük-98’ cultivars had the lowest sphericity mean values of 58.9% and 58.7%, respectively. According to the results of Principal Component Analysis (PCA), the most important variables of the first component were mass, geometric mean diameter, surface area, volume, projected area, equivalent diameter, perimeter and length. According to the physical properties of bean cultivars ‘Aras-98’, ‘Elkoca-05’, ‘Göynük-98’, ‘Noyanbey-98’, ‘Yakutiye-98’ and ‘Yunus-90’ were oblongus; ‘Karacaşehir-90’ and ‘Önceler-98’ were ellipticus; ‘Akman-98’, ‘Kantar-05’ and ‘Terzibaba’ were subcompressus and ‘Çayırlı’ genotype was compressus. The size and shape features of the bean cultivars are considerably crucial parameters in terms of designing the seed metering mechanism of seed drills, and transportation, separating and sizing systems. So, the size and shape data of bean cultivars are of mostly importance to engineers, machine manufacturers and machine designers. more...

Vanes throwing the particles of fertilizer or granular material onto a field are one of the most important machine parts influencing the fertilizer distribution pattern in spinning disc broadcasters. Manufacturers prefer mostly straight-shaped vanes in spinning disc broadcasters, probably because their production is easy. In previous studies, therefore, many researchers studied the spinning disc broadcaster with the straight-shaped vanes only. In this study, the effect of the combinations of the straight-shaped vane with different shaped vanes on the fertilizer distribution uniformity in single-spinning disc broadcaster was investigated with various flow rates using triple superphosphate. The combinations used in the study were the straight-shaped vane in combination with forward-curved-5, forward-curved-10, back-curved-5, and back-curved-10 vane, and ‘no combination’ consisted of the straight-shaped vane only. The circular orifices of 30, 40, and 50 mm diameters were used at the bottom of hopper to obtain the different flow rates of the fertilizer. The results of the study showed that the combination of the different shaped vanes on the spinning disc had a significant effect on fertilizer distribution uniformity. ‘No combination’ used commonly in practice did not give the best fertilizer distribution uniformity. The best fertilizer distribution uniformity was obtained from the combination of the straight-shaped vane with forward-curved-5 vane shape for all the flow rates. The CV values obtained from this combination were 10.31%, 15.49%, and 19.92% for the orifices with diameters of 30, 40, and 50 mm, respectively. more...

Some physical such as length, width, thickness, mass, geometric mean diameter, sphericity and surface area and mechanical properties such as deformation at cracking, cracking force, energy absorbed and hardness of nuts and kernels were determined at three orientations first time for two walnut cultivars, namely Mara?-18 and Yalova-1 widely grown in Turkey. For nuts, the average length (45.48 mm), mass (12.70 g), geometric mean diameter (36.83 mm) and surface area (42.66 cm2) were the highest in cv. Yalova-1 while width (34.18 mm), thickness (33.74 mm), shell thickness (1.48 mm) and sphericity (87.41%) were the highest in cv. Mara?-18. For all the loading orientations, the values of deformation at cracking, cracking force, energy absorbed and hardness of cv. Mara?-18 were higher than those of cv. Yalova-1. The effect of loading orientation to kernel extraction quality was found insignificant for both walnut cultivars. more...

The vagus nerves are important carriers of sensory information from the viscera to the central nervous system. Emerging evidence suggests that sensory signaling through the right, but not the left, vagus nerve evokes striatal dopamine release and reinforces appetitive behaviors. However, the extent to which differential gene expression within vagal sensory neurons contributes to this asymmetric reward-related signaling remains unknown. Here, we use single-cell RNA sequencing to identify genes that are differentially expressed between the left and right nodose ganglia (NG) to identify candidate genes likely to contribute to vagus-mediated reward signaling. We find that a group of neurons expressing Chrna3 (nicotinic acetylcholine receptor subunit 3) and Cckar (cholecystokinin A receptor) is preferentially expressed in the right NG of both rats and mice. This result suggests that differential expression of gut-innervating nutrient sensors in NG neurons may contribute to asymmetric encoding of interoceptive rewards by the vagus nerves. more...

Autoimmune diseases such as rheumatoid arthritis (RA) can promote states of chronic inflammation with accompanying tissue destruction and pain. RA can cause inflammatory synovitis in peripheral joints, particularly within the hands and feet, but can also sometimes trigger temporomandibular joint (TMJ) arthralgia. To better understand the effects of ongoing inflammation-induced pain signaling, dorsal root ganglia (DRGs) were acquired from individuals with RA for transcriptomic study. We conducted RNA sequencing from the L5 DRGs because it contains the soma of the sensory neurons that innervate the affected joints in the foot. DRGs from 5 RA patients were compared with 9 non-arthritic controls. RNA-seq of L5 DRGs identified 128 differentially expressed genes (DEGs) that were dysregulated in the RA subjects as compared to the non-arthritic controls. The DRG resides outside the blood brain barrier and, as such, our initial transcriptome analysis detected signs of an autoimmune disorder including the upregulated expression of immunoglobulins and other immunologically related genes within the DRGs of the RA donors. Additionally, we saw the upregulation in genes implicated in neurogenesis that could promote pain hypersensitivity. Overall, our DRG analysis suggests that there are upregulated inflammatory and pain signaling pathways that can contribute to chronic pain in RA., (© 2024. This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply.) more...

The nasal, oropharyngeal, and bronchial mucosa are primary contact points for airborne pathogens like Mycobacterium tuberculosis (Mtb), SARS-CoV-2, and influenza virus. While mucosal surfaces can function as both entry points and barriers to infection, mucosa-associated lymphoid tissues (MALT) facilitate early immune responses to mucosal antigens. MALT contains a variety of specialized epithelial cells, including a rare cell type called a microfold cell (M cell) that functions to transport apical antigens to basolateral antigen-presenting cells, a crucial step in the initiation of mucosal immunity. M cells have been extensively characterized in the gastrointestinal (GI) tract in murine and human models. However, the precise development and functions of human airway M cells is unknown. Here, using single-nucleus RNA sequencing (snRNA-seq), we generated an atlas of cells from the human adenoid and identified 16 unique cell types representing basal, club, hillock, and hematopoietic lineages, defined their developmental trajectories, and determined cell-cell relationships. Using trajectory analysis, we found that human airway M cells develop from progenitor club cells and express a gene signature distinct from intestinal M cells. Surprisingly, we also identified a heretofore unknown epithelial cell type demonstrating a robust interferon-stimulated gene signature. Our analysis of human adenoid cells enhances our understanding of mucosal immune responses and the role of M cells in airway immunity. This work also provides a resource for understanding early interactions of pathogens with airway mucosa and a platform for development of mucosal vaccines. more...

Diabetic neuropathy is frequently accompanied by pain and loss of sensation attributed to axonal dieback. We recovered dorsal root ganglia (DRGs) from 90 organ donors, 19 of whom had medical indices for diabetic painful neuropathy (DPN). Nageotte nodules, dead sensory neurons engulfed by non-neuronal cells, were abundant in DPN DRGs and accounted for 25% of all neurons. Peripherin-and Nav1.7-positive dystrophic axons invaded Nageotte nodules, forming small neuroma-like structures. Using histology and spatial sequencing, we demonstrate that Nageotte nodules are mainly composed of satellite glia and non-myelinating Schwann cells that express SPP1 and are intertwined with sprouting sensory axons originating from neighboring neurons. Our findings solve a 100-year mystery of the nature of Nageotte nodules linking these pathological structures to pain and sensory loss in DPN., Competing Interests: Competing interests: TJP is a co-founder of 4E Therapeutics. The authors declare no other competing interests related to this work. more...

Gene expression is influenced by chromatin architecture via controlled access of regulatory factors to DNA. To better understand gene regulation in the human dorsal root ganglion (hDRG) we used bulk and spatial transposase-accessible chromatin technology followed by sequencing (ATAC-seq). Using bulk ATAC-seq, we detected that in females diverse differentially accessible chromatin regions (DARs) mapped to the X chromosome and in males to autosomal genes. EGR1/3 and SP1/4 transcription factor binding motifs were abundant within DARs in females, and JUN, FOS and other AP-1 factors in males. To dissect the open chromatin profile in hDRG neurons, we used spatial ATAC-seq. The neuron cluster showed higher chromatin accessibility in GABAergic, glutamatergic, and interferon-related genes in females, and in Ca 2+ - signaling-related genes in males. Sex differences in transcription factor binding sites in neuron-proximal barcodes were consistent with the trends observed in bulk ATAC-seq data. We validated that EGR1 expression is biased to female hDRG compared to male. Strikingly, XIST , the long-noncoding RNA responsible for X inactivation, hybridization signal was found to be highly dispersed in the female neuronal but not non-neuronal nuclei suggesting weak X inactivation in female hDRG neurons. Our findings point to baseline epigenomic sex differences in the hDRG that likely underlie divergent transcriptional responses that determine mechanistic sex differences in pain., Competing Interests: Conflict of Interest Statement The authors declare no financial conflicts of interest related to this work. more...

Sensitization of spinal nociceptive circuits plays a crucial role in neuropathic pain. This sensitization depends on new gene expression that is primarily regulated via transcriptional and translational control mechanisms. The relative roles of these mechanisms in regulating gene expression in the clinically relevant chronic phase of neuropathic pain are not well understood. Here, we show that changes in gene expression in the spinal cord during the chronic phase of neuropathic pain are substantially regulated at the translational level. Downregulating spinal translation at the chronic phase alleviated pain hypersensitivity. Cell-type-specific profiling revealed that spinal inhibitory neurons exhibited greater changes in translation after peripheral nerve injury compared to excitatory neurons. Notably, increasing translation selectively in all inhibitory neurons or parvalbumin-positive (PV + ) interneurons, but not excitatory neurons, promoted mechanical pain hypersensitivity. Furthermore, increasing translation in PV + neurons decreased their intrinsic excitability and spiking activity, whereas reducing translation in spinal PV + neurons prevented the nerve injury-induced decrease in excitability. Thus, translational control mechanisms in the spinal cord, particularly in inhibitory neurons, play a role in mediating neuropathic pain hypersensitivity., Competing Interests: Declaration of Interests: Hien T. Zhao and Bethany Fitzsimmons are full-time employees and shareholders of Ionis Pharmaceutical, Inc. Other authors declare no competing interests. more...

Bone pain is a presenting feature of bone cancers such as osteosarcoma (OS), relayed by skeletal-innervating peripheral afferent neurons. Potential functions of tumor-associated sensory neurons in bone cancers beyond pain sensation are unknown. To uncover neural regulatory functions, a chemical-genetic approach in mice with a knock-in allele for TrkA was used to functionally perturb sensory nerve innervation during OS growth and disease progression. TrkA inhibition in transgenic mice led to significant reductions in sarcoma-associated sensory innervation and vascularization, tumor growth and metastasis, and prolonged overall survival. Single-cell transcriptomics revealed that sarcoma denervation was associated with phenotypic alterations in both OS tumor cells and cells within the tumor microenvironment, and with reduced calcitonin gene-related peptide (CGRP) and vascular endothelial growth factor (VEGF) signaling. Multimodal and multi-omics analyses of human OS bone samples and human dorsal root ganglia neurons further implicated peripheral innervation and neurotrophin signaling in OS tumor biology. In order to curb tumor-associated axonal ingrowth, we next leveraged FDA-approved bupivacaine liposomes leading to significant reductions in sarcoma growth, vascularity, as well as alleviation of pain. In sum, TrkA-expressing peripheral neurons positively regulate key aspects of OS progression and sensory neural inhibition appears to disrupt calcitonin receptor signaling (CALCR) and VEGF signaling within the sarcoma microenvironment leading to significantly reduced tumor growth and improved survival. These data suggest that interventions to prevent pathological innervation of osteosarcoma represent a novel adjunctive therapy to improve clinical outcomes and survival. more...

Diabetic peripheral neuropathy (DPN) is a prevalent complication of diabetes mellitus that is caused by metabolic toxicity to peripheral axons. We aimed to gain deep mechanistic insight into the disease process using bulk and spatial RNA sequencing on tibial and sural nerves recovered from lower leg amputations in a mostly diabetic population. First, our approach comparing mixed sensory and motor tibial and purely sensory sural nerves shows key pathway differences in affected nerves, with distinct immunological features observed in sural nerves. Second, spatial transcriptomics analysis of sural nerves reveals substantial shifts in endothelial and immune cell types associated with severe axonal loss. We also find clear evidence of neuronal gene transcript changes, like PRPH, in nerves with axonal loss suggesting perturbed RNA transport into distal sensory axons. This motivated further investigation into neuronal mRNA localization in peripheral nerve axons generating clear evidence of robust localization of mRNAs such as SCN9A and TRPV1 in human sensory axons. Our work gives new insight into the altered cellular and transcriptomic profiles in human nerves in DPN and highlights the importance of sensory axon mRNA transport as an unappreciated potential contributor to peripheral nerve degeneration., Competing Interests: Conflict-of-interest statement: The authors declare no conflicts of interest. more...

Irritable bowel syndrome and related disorders of gut-brain interaction (DGBI) are common and exhibit a complex, poorly understood etiology that manifests as abnormal gut motility and pain. Risk factors such as biological sex, stressors during critical periods, and inflammation are thought to influence DGBI vulnerability by reprogramming gut-brain circuits, but the specific cells affected are unclear. Here, we used a model of early life stress to understand cellular mechanisms in the gut that produce DGBIs. Our findings identify enteric glia as a key cellular substrate in which stress and biological sex converge to dictate DGBI susceptibility. Enteric glia exhibit sexual dimorphism in genes and functions related to cellular communication, inflammation, and disease susceptibility. Experiencing early life stress has sex-specific effects on enteric glia that cause a phenotypic switch in male glia toward a phenotype normally observed in females. This phenotypic transformation is followed by physiological changes in the gut, mirroring those observed in DGBI in humans. These effects are mediated, in part, by alterations to glial prostaglandin and endocannabinoid signaling. Together, these data identify enteric glia as a cellular integration site through which DGBI risk factors produce changes in gut physiology and suggest that manipulating glial signaling may represent an attractive target for sex-specific therapeutic strategies in DGBIs. more...