The mammalian sperm factor phospholipase C zeta is critical for early embryo division and pregnancy in humans and mice.

STUDY QUESTION: Are sperm phospholipase C zeta (PLCζ) profiles linked to the quality of embryogenesis and pregnancy? SUMMARY ANSWER: Sperm PLCζ levels in both mouse and humans correlate with measures of ideal embryogenesis whereby minimal levels seem to be required to result in successful pregnancy. WHAT IS KNOWN ALREADY: While causative factors underlying male infertility are multivariable, cases are increasingly associated with the efficacy of oocyte activation, which in mammals occurs in response to specific profiles of calcium (Ca2+) oscillations driven by sperm-specific PLCζ. Although sperm PLCζ abrogation is extensively linked with human male infertility where oocyte activation is deficient, less is clear as to whether sperm PLCζ levels or localization underlies cases of defective embryogenesis and failed pregnancy following fertility treatment. STUDY DESIGN, SIZE, DURATION: A cohort of 54 couples undergoing fertility treatment were recruited at the assisted reproductive technology laboratory at the King Faisal Hospital and Research Centre, Riyadh, Kingdom of Saudi Arabia. The recruitment criteria for males was a minimum sperm concentration of 5×106 sperm/ml, while all female patients had to have at least five oocytes. Sperm PLCζ analysis was performed in research laboratories, while semen assessments were performed, and time-lapse morphokinetic data were obtained, in the fertility clinic as part of routine treatment. The CRISPR/Cas9 system was concurrently used to induce indels and single-nucleotide mutations within the Plcζ gene to generate strains of Plcζ mutant mice. Sperm PLCζ was evaluated using immunofluorescence and immunoblotting with an antibody of confirmed consistent specificity against PLCζ. PARTICIPANTS/MATERIALS, SETTING, METHODS: We evaluated PLCζ profiles in sperm samples from 54 human couples undergoing fertility treatment in the context of time-lapse morphokinetic analysis of resultant embryos, correlating such profiles to pregnancy status. Concurrently, we generated two strains of mutant Plcζ mice using CRISPR/Cas9, and performed IVF with wild type (WT) oocytes and using WT or mutant Plcζ sperm to generate embryos. We also assessed PLCζ status in WT and mutant mice sperm in the context of time-lapse morphokinetic analysis and breeding outcomes. MAIN RESULTS AND THE ROLE OF CHANCE: A significant (P ≤ 0.05) positive relationship was observed between both PLCζ relative fluorescence and relative density with the times taken for both the second cell division (CC2) (r = 0.26 and r = 0.43, respectively) and the third cell division (S2) (r = 0.26). Examination of localization patterns also indicated significant correlations between the presence or absence of sperm PLCζ and CC2 (r = 0.27 and r = -0.27, respectively; P ≤ 0.025). Human sperm PLCζ levels were at their highest in the ideal times of CC2 (8-12 h) compared to time ranges outside the ideal timeframe (<8 and >12 h) where levels of human sperm PLCζ were lower. Following assignment of PLCζ level thresholds, quantification revealed a significantly higher (P ≤ 0.05) rate of successful pregnancy in values larger than the assigned cut-off for both relative fluorescence (19% vs 40%, respectively) and relative density (8% vs 54%, respectively). Immunoblotting indicated a single band for PLCζ at 74 kDa in sperm from WT mice, while a single band was also observed in sperm from heterozygous of Plcζ mutant mouse sperm, but at a diminished intensity. Immunofluorescent analysis indicated the previously reported (Kashir et al., 2021) fluorescence patterns in WT sperm, while sperm from Plcζ mutant mice exhibited a significantly diminished and dispersed pattern at the acrosomal region of the sperm head. Breeding experiments indicated a significantly reduced litter size of mutant Plcζ male mice compared to WT mice, while IVF-generated embryos using sperm from mutant Plcζ mice exhibited high rates of polyspermy, and resulted in significantly reduced numbers of these embryos reaching developmental milestones. LIMITATIONS, REASONS FOR CAUTION: The human population examined was relatively small, and should be expanded to examine a larger multi-centre cohort. Infertility conditions are often multivariable, and it was not possible to evaluate all these in human patients. However, our mutant Plcζ mouse experiments do suggest that PLCζ plays a significant role in early embryo development. WIDER IMPLICATIONS OF THE FINDINGS: We found that minimal levels of PLCζ within a specific range were required for optimal early embryogenesis, correlating with increased pregnancy. Levels of sperm PLCζ below specific thresholds were associated with ineffective embryogenesis and lower pregnancy rates, despite eliciting successful fertilization in both mice and humans. To our knowledge, this represents the first time that PLCζ levels in sperm have been correlated to prognostic measures of embryogenic efficacy and pregnancy rates in humans. Our data suggest for the first time that the clinical utilization of PLCζ may stand to benefit not just a specific population of male infertility where oocyte activation is completely deficient (wherein PLCζ is completely defective/abrogated), but also perhaps the larger population of couples seeking fertility treatment. STUDY FUNDING/COMPETING INTEREST(S): J.K. is supported by a faculty start up grant awarded by Khalifa University (FSU-2023-015). This study was also supported by a Healthcare Research Fellowship Award (HF-14-16) from Health and Care Research Wales (HCRW) to J.K., alongside a National Science, Technology, and Innovation plan (NSTIP) project grant (15-MED4186-20) awarded by the King Abdulaziz City for Science and Technology (KACST) for J.K. and A.M.A. The authors declare no conflicts of interest. TRIAL REGISTRATION NUMBER: N/A.

Bioengineered Organoids Offer New Possibilities for Liver Cancer Studies: A Review of Key Milestones and Challenges.

Hepatic cancer is widely regarded as the leading cause of cancer-related mortality worldwide. Despite recent advances in treatment options, the prognosis of liver cancer remains poor. Therefore, there is an urgent need to develop more representative in vitro models of liver cancer for pathophysiology and drug screening studies. Fortunately, an exciting new development for generating liver models in recent years has been the advent of organoid technology. Organoid models hold huge potential as an in vitro research tool because they can recapitulate the spatial architecture of primary liver cancers and maintain the molecular and functional variations of the native tissue counterparts during long-term culture in vitro. This review provides a comprehensive overview and discussion of the establishment and application of liver organoid models in vitro. Bioengineering strategies used to construct organoid models are also discussed. In addition, the clinical potential and other relevant applications of liver organoid models in different functional states are explored. In the end, this review discusses current limitations and future prospects to encourage further development.

Epidemiology, clinical presentation, and outcome of mpox: A study of 381 cases in Saudi Arabia.

Objectives: There are limited data on the clinical and epidemiological aspects of mpox in Saudi Arabia. This study investigates the characteristics of Mpox cases from May to September 2023. Methods: A total of 381 cases of Mpox were included in this study, diagnosed based on a combination of clinical symptoms and laboratory testing. Results: The majority of mpox cases were males (91.1%), with a mean age of (±SD) of 32.4 (±8.3) years; 356 (93.4%) did not report travel, 277 (72.7%) denied engaging in extra-marital sex, and 379 (99.5%) were not linked to secondary cases. Fever was reported in 371 (97.4%), whereas headache was present in 314 (82.4%). Cough (1.3%) and conjunctivitis (0.5%) were rare. The most commonly affected areas in terms of lesions were the palms and soles (297 cases, 78%), followed by the genitals (206 cases, 54%), face (198 cases, 52%), and mouth (160 cases, 42%). Of the 1325 identified contacts, 1134 (85.5%) were hospital contacts, and 191 (14.5%) were community contacts, and 393 (29.6%) were high-risk contact. Of the high-risk contacts, 284 (72.3%) accepted post-exposure vaccination. The genotyped samples were all subclade IIb (formerly the West Africa clade). Conclusions: This study provides valuable insights into mpox characteristics in Saudi Arabia. The genome of monkeypox virus belonged to subclade IIb of the West Africa clade. Further analysis of the global tree sublineage is needed.

Conjugated Linoleic Acid-Carboxymethyl Chitosan Polymeric Micelles to Improve the Solubility and Oral Bioavailability of Paclitaxel.

Oral delivery, the most common method of therapeutic administration, has two significant obstacles: drug solubility and permeability. The challenges of current oral medicine delivery are being tackled through an emerging method that uses structures called polymeric micelles. In the present study, polymeric micelles were developed using conjugates of linoleic acid-carboxymethyl chitosan (LA-CMCS) for the oral delivery of paclitaxel (PCL). The developed micelles were evaluated by particle size, zeta potential, Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC), and thermogravimetric analysis (TGA). When PCL was contained within micelles, its solubility increased by almost 13.65 times (around 60 µg/mL). The micelles' zeta potentials were -29 mV, their polydispersity indices were 0.023, and their particle diameters were 93 nm. Micelles showed PCL loading and entrapment efficiencies of 67% and 61%, respectively. The sustained release qualities of the PCL release data from micelles were good. In comparison to the pure PCL suspension, the permeability of the PCL from micelles was 2.2 times higher. The pharmacokinetic data revealed that PCL with LA-CMCS micelles had a relative bioavailability of 239.17%, which was much greater than the PCL in the suspension. The oral bioavailability of PCL was effectively increased by LA-CMCS micelles according to an in vivo study on animals. The polymer choice, maybe through improved permeability, plays an essential role when assessing oral bioavailability enhancement and solubility improvement (13.65 times). The outcomes demonstrated that PCL's solubility and pharmacokinetics were improved in the micelles of the LA-CMCS conjugate.

Unveiling the Clinical Spectrum of Post-COVID-19 Conditions: Assessment and Recommended Strategies.

SARS-CoV-2 caused the pandemic of the rapidly evolving COVID-19. As of December 6, 2023, there were 765,152,854 COVID-19-recovering cases. Long-term consequences known as "long COVID" and "post-COVID-19 conditions" (PCCs) or "post-acute COVID-19 syndrome" are being reported more frequently in a subset of recovering patients. Systemic, neuropsychiatric, cardio-respiratory, and gastrointestinal symptoms are the most prevalent. The management of PCCs poses unique challenges due to the lack of official guidelines and the complex nature of the illness. This abstract highlights key principles derived from recent reviews and expert recommendations to provide healthcare professionals with a comprehensive approach to manage post-COVID-19 patients. Preventive medicine plays a crucial role in managing PCCs. While no specific medications are available for treatment, preventive measures such as COVID-19 vaccination, adherence to precautionary measures, regular consultations with medical professionals, monitoring symptoms and progress, and seeking information on symptom management are essential to assist patients in their recovery and improve their quality of life. Medical management requires transparent goal-setting and collaborative decision-making based on the patient's symptoms, comorbidities, and treatment objectives. Treatment plans for post-COVID-19 patients should focus on patient education, using registries and calendars to track symptoms and triggers, providing support and reassurance, and offering holistic support through peer networks and supportive psychotherapy techniques. Symptomatic and rehabilitative care, including well-established symptom management techniques, physical rehabilitation programs, and addressing mental health and well-being, are vital components of post-COVID-19 management. Lifestyle factors such as stress reduction, nutrition, and sleep should be incorporated into managing underlying medical conditions in post-COVID-19 patients. Regular follow-up visits and referrals to specialists are recommended to monitor the patient's progress and address specific organ system involvement or additional care needs. In summary, for the effective management of PCCs, a holistic approach should include preventive measures, patient education, supportive psychotherapy, symptomatic and rehabilitative care, medical management, counseling on lifestyle elements, and appropriate follow-up plans. However, it is crucial to stay updated with evolving guidelines and recommendations from healthcare authorities to provide the most effective and evidence-based care to post-COVID-19 patients.

Using Liver Organoids as Models to Study the Pathobiology of Rare Liver Diseases.

Liver organoids take advantage of several important features of pluripotent stem cells that self-assemble in a three-dimensional culture matrix and reproduce many aspects of the complex organization found within their native tissue or organ counterparts. Compared to other 2D or 3D in vitro models, organoids are widely believed to be genetically stable or docile structures that can be programmed to virtually recapitulate certain biological, physiological, or pathophysiological features of original tissues or organs in vitro. Therefore, organoids can be exploited as effective substitutes or miniaturized models for the study of the developmental mechanisms of rare liver diseases, drug discovery, the accurate evaluation of personalized drug responses, and regenerative medicine applications. However, the bioengineering of organoids currently faces many groundbreaking challenges, including a need for a reasonable tissue size, structured organization, vascularization, functional maturity, and reproducibility. In this review, we outlined basic methodologies and supplements to establish organoids and summarized recent technological advances for experimental liver biology. Finally, we discussed the therapeutic applications and current limitations.

The economic burden of viral severe acute respiratory infections in the Kingdom of Saudi Arabia: A nationwide cost-of-illness study.

Objectives: The authors conducted a study to estimate the rising disease and economic burden of viral severe acute respiratory infections and their management, including COVID-19, respiratory syncytial virus, and influenza types A and B, in early and delayed diagnosis scenarios in the Kingdom of Saudi Arabia from a national perspective. Methods: This cross-sectional study was conducted in May 2022 using a de novo Excel-based universal cost-of-illness calculator model. The study used primary data, such as expert interviews, as well as secondary data from a thorough literature search. Results: The total economic burden of viral severe acute respiratory infections in patients with an early diagnosis was lower than those with a delayed diagnosis among patients with complications, both from the payer's (United States dollar [USD] 3846 million vs USD 4726 million) and societal (USD 4048 million vs USD 5020 million, respectively) perspectives. The major cost driver of the total economic burden for both early and delayed diagnosis was disease management costs: 49% (USD 1880 million) and 58% (USD 2730 million), respectively. Conclusions: In the Kingdom of Saudi Arabia, the total economic burden for COVID-19, influenza (epidemic phase), and respiratory syncytial virus was higher with a delayed diagnosis vs an early diagnosis, emphasizing the importance of using a broader diagnostic method.

Epidemiology of tuberculosis in Saudi Arabia following the implementation of end tuberculosis strategy: Analysis of the surveillance data 2015-2019.

OBJECTIVES: To analyze the evolution of tuberculosis (TB) epidemiology in Saudi Arabia in the 5 years following the implementation of the end-TB Strategy. METHODS: A retrospective analysis of surveillance data, reported by the national tuberculosis control program from 2015-2019, was carried out. The annual incidence and the percentage of yearly changes were calculated and compared to the World Health Organization (WHO) milestones, which anticipate a 4-5% annual decline. Additionally, various other epidemiological indicators of TB were examined. RESULTS: The national TB incidence declined from 10.55% per 100,000 in 2015 to 8.76% per 100,000 in 2019, aligning with the WHO's 2019 milestone estimated between 8.59-8.96% per 100,000. While Makkah Region (40.3%) and Riyadh (24.6%) accounted for the majority of cases, Jazan region consistently exhibited the highest incidence throughout the study period. Demographic features shifted towards a younger age category, male, and native dominance. There was a consistent decrease in resistance and intermediate sensitivity to all first-line anti-TB drugs, associated with a substantial decrease in both polydrug resistance (from 4.7-1.9%; p<0.001) and multidrug resistance (from 4.4-2.4%; p=0.008). CONCLUSION: The figures of TB incidence TB in Saudi Arabia between 2015-2019 has met the WHO end-TB milestones, predicting successful progress toward the 2035 goal.

Comparison of Impairments, Activity Limitations, Balance, and Quality of Life between Patients with and without Meniscus Repair or Partial Meniscectomy Post-ACL Reconstruction.

(1) Background: The anterior cruciate ligament (ACL) is a crucial ligament in the knee joint. This study compares the differences in knee range of motion (ROM), knee proprioception error, balance, function, and quality of life (QOL) among participants with and without meniscus repair or partial meniscectomy nine months post ACL reconstruction. (2) Methods: In this cross-sectional study, 57 male participants were selected through convenience sampling from a tertiary care hospital. Knee flexion and extension ROM were assessed using a digital goniometer; a digital inclinometer was used to assess knee proprioception error; the Y balance test was used to evaluate balance; the lower extremity functional scale (LEFS) was used to assess activity; and QOL was assessed using the ACLQOL questionnaire. (3) Results: There were no significant differences in outcomes except balance. The YB composite score had a moderate negative correlation with knee proprioception error with an R-value of -0.372 **. (4) Conclusions: Nine to 12 months post ACL reconstruction, the isolated ACL reconstruction participants had better lower-quarter single-leg balance than those who underwent ACL reconstruction and meniscal repair or partial meniscectomy. The remaining parameters, like knee ROM, knee proprioception error, LEFS score, and ACLQOL scores, were similar between these two groups.

Whole Liver Derived Acellular Extracellular Matrix for Bioengineering of Liver Constructs: An Updated Review.

Biomaterial templates play a critical role in establishing and bioinstructing three-dimensional cellular growth, proliferation and spatial morphogenetic processes that culminate in the development of physiologically relevant in vitro liver models. Various natural and synthetic polymeric biomaterials are currently available to construct biomimetic cell culture environments to investigate hepatic cell-matrix interactions, drug response assessment, toxicity, and disease mechanisms. One specific class of natural biomaterials consists of the decellularized liver extracellular matrix (dECM) derived from xenogeneic or allogeneic sources, which is rich in bioconstituents essential for the ultrastructural stability, function, repair, and regeneration of tissues/organs. Considering the significance of the key design blueprints of organ-specific acellular substrates for physiologically active graft reconstruction, herein we showcased the latest updates in the field of liver decellularization-recellularization technologies. Overall, this review highlights the potential of acellular matrix as a promising biomaterial in light of recent advances in the preparation of liver-specific whole organ scaffolds. The review concludes with a discussion of the challenges and future prospects of liver-specific decellularized materials in the direction of translational research.

Viral aetiology of severe acute respiratory illness among patients admitted during the 2022 peri-Hajj period.

Introduction: Severe acute respiratory illness (SARI) among pilgrims continues to be an important healthcare issue. The aim of this study was to describe the viral aetiology of patients admitted to hospitals in the holy cities of Makkah and Madinah during the 2022 peri-Hajj period. Methods: This is a retrospective analysis of patients admitted to hospitals with SARI. Patients were tested with multiplex polymerase chain reaction for the most common viral aetiologies. Results: In total, 179 cases of SARI were identified during the study period. Of these, 101 (56.4%) were males, 78 (43.6%) were females, and 78 (43.6%) were Saudi. The mean age was 58.60 years (standard deviation 20.5) years. The most common age group was ≥65 years (n=68, 36%), followed by 55-59 years (n=37, 19%). The most common comorbidities were diabetes mellitus (n=67, 36%), hypertension (n=65, 35%) and chronic lung disease (n=34, 18%). Eighty-five (47.5%) patients tested negative and 94 (42.5%) tested positive for various viral aetiologies. The most frequently detected viruses were severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) (n=28, 15%) and influenza (n=22, 12%); of the influenza cases, 16 were influenza A (6 (43%) were H3N2), and six were influenza B. The only case of Middle East respiratory syndrome coronavirus (MERS-CoV) was in a citizen, and none of the visitors or residents had MERS-CoV. Of the total cases, 27 (14%) died during the follow-up period. In a binary regression analysis, only age was associated with mortality (P=0.002). Conclusion: The most commonly detected viruses among patients admitted to hospital with SARI were SARS-CoV-2 and influenza. It is important to continue surveillance of admitted and non-admitted patients in different Hajj periods to identify any shift in the aetiologic agents.

Distribution of Primary Healthcare Centers Between 2017 and 2021 Across Saudi Arabia.

BACKGROUND AND AIM: Access to adequate healthcare systems is seen as a fundamental human right. Therefore, healthcare services must be distributed and accessible in the most efficient way possible to those who need them the most. Primary healthcare centers are the backbone of any healthcare system as they provide essential healthcare services to the general population. Inequalities in the distribution of primary healthcare centers can lead to disparities in healthcare access and outcomes. This study aimed to assess the equity of primary healthcare centers distribution, we analyzed and evaluated the allocation of primary healthcare centers in Saudi Arabia from 2017 to 2021. METHOD: This cross-sectional study utilized data from the Health Statistical Yearbook published by the Ministry of Health, Saudi Arabia, during the period of 2017-2021. The number of primary healthcare centers per 10,000 population was calculated for the 20 health regions. We used the Gini index to measure inequality in the distribution of primary healthcare centers. The Pearson coefficient was calculated to assess the correlation between the number of primary healthcare centers and the population in each health region. RESULT:  The overall ratio of primary healthcare centers to population decreased from 0.72 to 0.62 between 2017 and 2021. The Gini index showed relative equality in the distribution of primary healthcare centers from 2017 to 2021 with values between 0.2 and 0.3. There was a positive correlation between the population and the primary healthcare centers in Jeddah, Tabouk, and the Northern Region. However, in Riyadh, Makkah, Taif, Madinah, Qaseem, Eastern Region, Al-Ahsa, Aseer, Hail, Jazan, Najran, Al-Baha, and Al-Jouf, the correlation was found to be negative. CONCLUSION:  From 2017 to 2021, primary healthcare facilities are distributed fairly throughout 20 health regions of Saudi Arabia. However, there are still some disparities between provinces, and efforts must be made to ensure that primary healthcare centers are distributed equitably across the country to improve healthcare access and outcomes for all.

Trauma and Injuries Pattern During Hajj, 1443 (2022): A Cross-Sectional Study.

Introduction Trauma and injuries are common among pilgrims during Hajj, the biggest mass gathering event. Trauma and injury causes vary from falling and pressing in crowds to being burned by boiled water and road traffic accidents (RTA). Time to reach the hospital during highly condensed areas in Hajj are challenges for the public health authorities and the healthcare system to achieve optimum control, management, and outcome. This study aims to explore the pattern of trauma and injuries during Hajj as it is crucial to improve future preventive measures and care quality. Methods A cross-sectional questionnaire-based study was conducted in one hospital in each of the Mena and Arafat (Al-Mashaar's areas) in Makkah City, Saudi Arabia, from July 8 to 10, 2022. Data was collected through interviews with patients who visit the hospitals or enter the emergency department and are diagnosed with trauma or injury during the Hajj season of 1443 Hijri date (2022). Results A total of 264 people volunteered to participate in the survey. The mean age by years was 43.5 ± 10.7, and the majority (56%) were between 41 and 64. There were multiple nationalities - the most common nationality was Egyptian (25%), followed by Saudi (10%). The commonest type of trauma was cutting wounds (50%), and the commonest cause was falling (39%), followed by foot twisting (31%). There were 142 cases in Arafat and 122 cases in Mena in the study duration. Tissue contusions are higher in Arafat. Fractures (5%) were in both areas but higher in Mena with burns and sprains. Friction blister injuries were only in Mena and were statistically associated with walking barefoot (p<0.01), which was associated with Egyptians (p<0.05). Also, thigh chafing is only in Mena, while eye traumas and abrasion are only in Arafat. There were four causes of injury that are statistically significantly associated with the area (p<0.05): foot twisting in Arafat, pressing in overcrowding, stoning, and burning in Mena. Moreover, all the RTA cases (n=4) were in Arafat, and all the stoning and burning by boiling water were in Mena. Admission was only for burning (n=2) and falling (n=2) cases and only in Mena emergency hospital; otherwise, all trauma cases were discharged after receiving management - no deaths among the study sample. Injuries in Mena are likely to happen in the evening and night (n=91), while in Arafat, it is more likely in two periods (n=113), in the early morning and afternoon. This difference is statistically significant between the two areas (p<0.05). Most pilgrims (n=129/253) reach the hospital in 16 to 30 minutes. A statistically significant association exists between the duration and the area (p<0.05). Most patients in Arafat (88%) reach the hospital in less than 30 minutes, while only 50% take the same duration in Mena. Conclusion The Hajj season of 1443 H (2022) has a similar trauma pattern and improved outcomes compared to previous seasons. Discovering and digging into the causes of traumas and injuries should be optimized in future research for better control and customized prevention measures. Establishing new and remodeling current prevention measures is recommended for more control.

Partial Obstruction of the Endotracheal Tube by a Part of the Metallic Stylet; Case Report and Review of the Literature.

Endotracheal intubation is common in the emergency department, intensive care units, and operating rooms. It involves the insertion of an endotracheal tube (ETT) through the mouth or nose into the trachea to maintain a patent airway and facilitate mechanical ventilation. Using a stylet during intubation can guide the ETT through the vocal cords. However, complications can arise when the stylet is not removed after successful intubation. Herewith, we reported a patient who was two years old and suffered from respiratory failure. However, in the first 12 hours, we observed a foreign body in the trachea, a small end of a metal stylet immediately removed by bronchoscope. This case demonstrates that multiple uses of a stylet, especially by a single user, can result in impaction of the stylet in the ETT during intubation, requiring force when the stylet is withdrawn after intubation, which can result in breakage, shearing, and retention of the stylet or plastic sheath in the lumen of the ETT.

ß-catenin attenuation leads to up-regulation of activating NKG2D ligands and tumor regression in BrafV600E-driven thyroid cancer cells.

Introduction: BRAFV600E mutations frequently occur in papillary thyroid cancer (PTC). ß-catenin, encoded by CTNNB1, is a key downstream component of the canonical Wnt signaling pathway and is often overexpressed in PTC. BRAFV600E-driven PTC tumors rely on Wnt/ß-catenin signaling to sustain growth and progression. Methods: In the present study, we investigated the tumorigenicity of thyroid cancer cells derived from BRAFV600E PTC mice following Ctnnb1 ablation (BVE-Ctnnb1null). Results: Remarkably, the tumorigenic potential of BVE-Ctnnb1null tumor cells was lost in nude mice. Global gene expression analysis of BVE-Ctnnb1null tumor cells showed up-regulation of NKG2D receptor activating ligands (H60a, H60b, H60c, Raet1a, Raet1b, Raet1c, Raet1d, Raet1e, and Ulbp1) and down-regulation of inhibitory MHC class I molecules H-2L and H-2K2 in BVE-Ctnnb1null tumor cells. In vitro cytotoxicity assay demonstrated that BVE-Ctnnb1wt tumor cells were resistant to NK cell-mediated cytotoxicity, whereas BVE-Ctnnb1null tumor cells were sensitive to NK cell-mediated killing. Furthermore, the overexpression of any one of these NKG2D ligands in the BVE-Ctnnb1wt cell line resulted in a significant reduction of tumor growth in nude mice. Conclusions: Our results indicate that active ß-catenin signaling inhibits NK cell-mediated immune responses against thyroid cancer cells. Targeting the ß-catenin signaling pathway may have significant therapeutic benefits for BRAF-mutant thyroid cancer by not only inhibiting tumor growth but also enhancing host immune surveillance.

Antigen Unmasking Is Required to Clinically Assess Levels and Localisation Patterns of Phospholipase C Zeta in Human Sperm.

Mammalian oocyte activation is initiated by intracellular calcium (Ca2+) oscillations, driven by the testis-specific phospholipase C zeta (PLCζ). Sperm PLCζ analysis represents a diagnostic measure of sperm fertilisation capacity. The application of antigen unmasking/retrieval (AUM) generally enhanced the visualisation efficacy of PLCζ in mammalian sperm, but differentially affected the PLCζ profiles in sperm from different human males. It is unclear whether AUM affects the diagnosis of PLCζ in human sperm. Herein, we examined whether the application of AUM affected the correlation of PLCζ profiles with sperm parameters and fertilisation capacity. PLCζ fluorescence levels and localisation patterns were examined within the sperm of males undergoing fertility treatment (55 patients aged 29-53) using immunofluorescence in the absence/presence of AUM. The changes in PLCζ profiles following AUM were examined in relation to sperm health and fertilisation outcome. AUM enhanced the observable levels and specific localisation patterns of PLCζ in relation to both optimal sperm parameters and fertilisation outcome, without which significant differences were not observed. The extent of the change in levels and localisation ratios of PLCζ was also affected to a larger degree in terms of the optimal parameters of sperm fertility and fertilisation capacity by AUM. Collectively, AUM was essential to accurately assesses PLCζ in human sperm in both scientific and clinical contexts.

Mammalian-specific decellularized matrices derived bioink for bioengineering of liver tissue analogues: A review.

The absolute shortage of compatible liver donors and the growing number of potential recipients have led scientists to explore alternative approaches to providing tissue/ organ substitutes from bioengineered sources. Bioartificial regeneration of a fully functional tissue/organ replacement is highly dependent on the right combination of engineering tools, biological principles, and materiobiology horizons. Over the past two decades, remarkable achievements have been made in hepatic tissue engineering by converging various advanced interdisciplinary research approaches. Three-dimensional (3D) bioprinting has arisen as a promising state-of-the-art tool with strong potential to fabricate volumetric liver tissue/organ equivalents using viscosity- and degradation-controlled printable bioinks composed of hydrous microenvironments, and formulations containing living cells and associated supplements. Source of origin, biophysiochemical, or thermomechanical properties and crosslinking reaction kinetics are prerequisites for ideal bioink formulation and realizing the bioprinting process. In this review, we delve into the forecast of the potential future utility of bioprinting technology and the promise of tissue/organ- specific decellularized biomaterials as bioink substrates. Afterward, we outline various methods of decellularization, and the most relevant studies applying decellularized bioinks toward the bioengineering of in vitro liver models. Finally, the challenges and future prospects of decellularized material-based bioprinting in the direction of clinical regenerative medicine are presented to motivate further developments.

Magnetic Resonance Imaging Features of Rhino-Orbito-Cerebral Mucormycosis in Post-COVID-19 Patients: Radio-Pathological Correlation.

There has been a notable increase in rhino-orbito-cerebral mucormycosis (ROCM) post-coronavirus disease 2019 (COVID-19), which is an invasive fungal infection with a fatal outcome. Magnetic resonance imaging (MRI) is a valuable tool for early diagnosis of ROCM and assists in the proper management of these cases. This study aimed to describe the characteristic MRI findings of ROCM in post-COVID-19 patients to help in the early diagnosis and management of these patients. This retrospective descriptive study was conducted at a single hospital and included 52 patients with COVID-19 and a histopathologically proven ROCM infection who were referred for an MRI of the paranasal sinuses (PNS) due to sino-orbital manifestations. Two radiologists reviewed all the MR images in consensus. The diagnosis was confirmed by histopathological examination. The maxillary sinus was the most commonly affected PNS (96.2%). In most patients (57.7%), multiple sinuses were involved with the black turbinate sign on postcontrast images. Extrasinus was evident in 43 patients with orbital involvement. The pterygopalatine fossa was involved in four patients. Three patients had cavernous sinus extension, two had pachymeningeal enhancement, and one had epidural collection. The alveolar margin was affected in two patients, and five patients had an extension to the cheek. The awareness of radiologists by the characteristic MRI features of ROCM in post-COVID-19 patients helps in early detection, early proper management, and prevention of morbid complications.

The Epidemiology of Work-Related Injuries in Saudi Arabia Between 2016 and 2021.

Background Workplace safety has increased in many developed nations, but work-related injuries (WRIs) are on the rise. Globally, approximately 2.9 million fatal work-related injuries and diseases occurred in 2021, up from 1.1 million in 1999. Method This work employs a cross-sectional study using the open data published by the General Organization for Social Insurance (GOSI) on an annual basis for the years 2016 to 2021. The data from one year in each category were compared with a reference year by using the index value method and simple linear regression. Additionally, the researchers calculated the injury-to-worker ratio for each year. Aim The aim of this study is to assess the trending epidemiology of work-related injuries in Saudi Arabia from 2016 to 2021 among insured workers and compare this trend to previous studies. Result A total of 1,009 work-related deaths wererecorded over the study period, averaging 168 per year. In comparison with non-Saudi workers, the Saudi workers' shares of injuries rose from 5.3% in 2016 to 10% in 2021. Injuries in the construction sector comprised 42%-48% of all injuries, followed by the commerce sector. In 2021, the highest percentage of injuries involved service occupations (47.5%), followed by the engineering sector. Between 2016 and 2018, the most common injury cause was falls (28.5% in 2018), and from 2020 to 2021, exposure to inanimate mechanical forces caused the most injuries (46%). Conclusion The study revealed that the trend of work-related injuries had decreased during the study period, consistent with the results of similar previous studies.

The First Genomic Characterization of the Chikungunya Virus in Saudi Arabia.

BACKGROUND: Chikungunya is an arboviral infection caused by the Chikungunya virus (CHIKV) transmitted to humans by mosquitoes of Aedes spp. CHIKV has been confined to African countries and South-East Asia up to 2004, but since then, the pathogen has become more global, and its high morbidity rate has become more visible. Saudi Arabia is not an endemic region of CHIKV, and the virus's origin is not yet fully understood. This study aimed to characterize the genome of CHIKV from samples detected in Jeddah in 2018. METHOD: Twenty-two sets of primers were designed to amplify near-full length genome of CHIKV. RT-PCR was conducted from clinical samples. Two samples were used for studying near complete genome sequence while the remaining samples were used to study the E1 gene. Different bioinformatics tools were utilized. RESULTS: Phylogenetic analysis showed that the CHIKV strains clustered with strains isolated from Kenya during 2017-2018 and belonged to ECSA genotype. E1: L136F, K211E and I317V mutations were identified in our strains. Also, E2: M74I, A76T, and V264A mutations were documented. Additionally, the capsid N79S substitution was also detected. CONCLUSION: The genome of CHIKV was analyzed for the first time in Saudi Arabia to better understand the origin of the CHIKV and its genetic diversity, which showed high similarity with IE-a subclade of CHIKV strains detected in Mombasa (Kenya) indicating its possible origin.