Generation of myoglobin (MB)-knockout human embryonic stem cell (hESC) line (KAIMRCe002-A-1S) using CRISPR/Cas9 technology.

Myoglobin (MB) is a cytoplasmic hemoprotein that is predominantly expressed in the heart and oxidative myofibers of skeletal muscle. It has been demonstrated that MB binds to oxygen and promotes its diffusion for energy production in the mitochondria. Recently, MB was found to be expressed in different forms of malignant tumors and cancer cell lines. Further studies using gene disruption technology will enhance the understanding of MB's role in human cardiovascular biology and cancers. Here, we describe the generation of a homozygous MB knockout in human embryonic stem cells (hESC-MB-/-) via CRISPR/Cas9 to study MB function in human biology and diseases.

Initial Outcomes of Freehand Transperineal Biopsies Regarding Diagnostic Value and Safety: An Early Experience at King Fahad Specialist Hospital, Buraydah, Saudi Arabia.

BACKGROUND: Prostate cancer is a common type of cancer in Saudi Arabia with a high incidence rate. Trans-rectal ultrasound guided prostatic biopsy (TRUSBx) has been the standard diagnostic study for prostate cancer since a landmark study in 1989 which showed that it is better than digitally directed biopsy sampling of the prostate. As an alternative to TRUSBx, transperineal biopsies (TPBx) have gained popularity as they give a higher accuracy rate and avoid many complications. A new study has been conducted in Riyadh, Saudi Arabia to compare TRUSBx and TPBx showed that TPBx has a significantly higher detection rate of prostate cancer cases compared to TRUSBx (45.1% vs. 29.1%, p=0.003). The aim of this study is to determine the diagnostic value and safety of freehand transperineal prostate biopsy in patients with an elevated prostatic specific antigen (PSA) and/or abnormal digital rectal exam in King Fahad Specialist Hospital KFSH in Buraydah, Qassim region, Saudi Arabia. METHODS: This is an observational retrospective study of all patients (n=39) who underwent transperineal biopsies at KFSH to assess the diagnostic value and safety of the procedure. RESULTS: The mean age of the patients was 70.3 (SD 10.1) years. The most commonly found diagnosis was adenocarcinoma (61.5%), and incidence of complications was detected in (5.1%) of the patients. CONCLUSION: We concluded that the freehand technique TPBx has a high accuracy rate in detecting prostatic cancer. However, the learning curve could be a limiting factor in implementing it. Increasing the number of biopsies could positively affect diagnostic accuracy, especially with our low complication rate in this procedure. A low number of biopsies in the older age group can give an accurate result with a low risk of complications. Although template-guided TPBx and robot-guided TPBx are better options, the freehand technique represents a cost-effective and time-saving alternative. However, more studies are needed to compare the outcome of such a technique.

Utility of Multimodal Treatment Protocols in the Management of Scalp Cutaneous Angiosarcoma.

Cutaneous angiosarcomas are rare, aggressive tumors with high recurrence and poor prognosis. We share our experiences with the challenging surgical management of these lesions, focusing on both ablative and reconstructive outcomes. Methods: Retrospective cross-sectional chart review was conducted of patients diagnosed with scalp cutaneous angiosarcoma between 2005 and 2021. Resectability, defect reconstruction, and survival outcomes were analyzed. Results: Thirty patients were included, 27 (90%) men and three (10%) women, with a mean age at diagnosis of 71.77 ± 7.3 years, and mean follow-up of 429.43 ± 305.6 days. Only 12 patients completed their regular follow-up, while the remaining patients died. There was a median survival of 443.50 days (range, 42-1283) and median time to recurrence of 21 days (range, 30-1690). Multimodal therapy compared with surgery alone had a significantly better overall median survival (468 days versus 71 days; P = 0.021). Defect coverage was obtained in 24 cases (75%) through an anterolateral thigh flap, two patients (6%) with a local transposition flap, and one patient (3%) with a transverse rectus abdominis myocutaneous flap. The remaining three patients received a skin graft. All of the flaps survived, with one requiring a vein graft for venous congestion. Conclusions: Timely multimodal therapy with a histologically safe margin, combined with adjuvant therapy, improves survival and delays recurrence and metastasis, in cutaneous angiosarcoma patients. An anterolateral thigh flap facilitates the coverage of wide defects. Further investigations of advanced treatment modalities such as immunotherapy and/or gene therapy are required to deal with this highly aggressive tumor.

Balancing selection at the human salivary agglutinin gene (DMBT1) driven by host-microbe interactions.

Discovering loci under balancing selection in humans can identify loci with alleles that affect response to the environment and disease. Genome variation data have identified the 5' region of the DMBT1 gene as undergoing balancing selection in humans. DMBT1 encodes the pattern-recognition glycoprotein DMBT1, also known as SALSA, gp340, or salivary agglutinin. DMBT1 binds to a variety of pathogens through a tandemly arranged scavenger receptor cysteine-rich (SRCR) domain, with the number of domains polymorphic in humans. We show that the signal of balancing selection is driven by one haplotype usually carrying a shorter SRCR repeat and another usually carrying a longer SRCR repeat. DMBT1 encoded by a shorter SRCR repeat allele does not bind a cariogenic and invasive Streptococcus mutans strain, in contrast to the long SRCR allele that shows binding. Our results suggest that balancing selection at DMBT1 is due to host-microbe interactions of encoded SRCR tandem repeat alleles.

Ctcf haploinsufficiency mediates intron retention in a tissue-specific manner.

CTCF is a master regulator of gene transcription and chromatin organisation with occupancy at thousands of DNA target sites genome-wide. While CTCF is essential for cell survival, CTCF haploinsufficiency is associated with tumour development and hypermethylation. Increasing evidence demonstrates CTCF as a key player in several mechanisms regulating alternative splicing (AS), however, the genome-wide impact of Ctcf dosage on AS has not been investigated. We examined the effect of Ctcf haploinsufficiency on gene expression and AS in five tissues from Ctcf hemizygous (Ctcf+/-) mice. Reduced Ctcf levels caused distinct tissue-specific differences in gene expression and AS in all tissues. An increase in intron retention (IR) was observed in Ctcf+/- liver and kidney. In liver, this specifically impacted genes associated with cytoskeletal organisation, splicing and metabolism. Strikingly, most differentially retained introns were short, with a high GC content and enriched in Ctcf binding sites in their proximal upstream genomic region. This study provides new insights into the effects of CTCF haploinsufficiency on organ transcriptomes and the role of CTCF in AS regulation.

Diagnosis and management of childhood sleep-disordered breathing. Clinical approach.

Sleep-disordered breathing (SDB) includes disorders of breathing that affect airway patency, which impair children's sleep and lead to negative consequences. Obstructive sleep apnea, hypoventilation and upper airway resistance syndrome are common causes of morbidity and mortality in childhood. These clinical practice guidelines, intended for use by pediatricians and primary care clinicians, provide a clear recommendation for the diagnosis and management of sleep-disordered breathing, focusing on the most serious disorder, obstructive sleep apnea syndrome (OSAS). These clinical guidelines formulate clear recommendations to identify patients with suspected OSAS. Further, the manuscript will highlight the potential consequences of SBD in children, and how to overcome such difficulties, what could be the therapeutic options, a 12 recommendations and what are the future direction for pediatric sleep medicine.

Are Safe Sleep Practice Recommendations For Infants Being Applied Among Caregivers?

Background Sudden infant death syndrome (SIDS) is defined as the sudden unexpected death of an infant, even after investigations and autopsy. SIDS is related to many factors, such as the baby's position and objects in the crib. Adherence to safe sleep recommendations in Saudi Arabia is unclear. This study aims to assess caregivers' implementation of safe sleep practices and if they received any safe sleep education through health care workers. Methods This was a cross-sectional, descriptive study. Inclusion criteria included all infants below the age of one year. Exclusion criteria included infants who were born premature, used ventilation, had a tracheostomy, any anomaly in the upper airway, or underwent spine surgery. A semi-structured questionnaire was used. Data were collected from mothers who had infants visiting the outpatient department of King Abdulaziz University Hospital in Jeddah, Saudi Arabia. An electronic survey was also created and published on a social platform.  Statistical analysis was conducted with the aid of the Statistical Package for Social Sciences (SPSS) software, version 26 (IBM SPSS Statistics, Armonk, NY). Results Among 506 participants, only 22.5% were found to receive education about safe practices from health care providers. Fortunately, most of the infants (63.2%) were found to sleep in a supine position most of the nights. Adherent caregivers to placing the child in a designated baby bed and in a supine position most nights represented 44.86% of the sample. However, when asked about placing any of the following objects in the bed (pillows, blankets, soft toys, hard toys, and electric wires), the percentage of adherence dropped down to only 1.58%. Conclusion There was an obvious non-adherence among caregivers and a possible lack of knowledge of safe sleep recommendations for infants. This highlights the need for optimal education by health care workers and the rule of media and campaigns is obvious and essential to improving their practices and, hopefully, decreasing the risk of SIDS.

Molecular and biochemical investigations of key antioxidant/oxidant molecules in Saudi patients with recurrent miscarriage.

The present study was undertaken to: i) Determine the levels of oxidative stress (OS) markers, malondialdehyde (MDA), superoxide anions (SOA) and hydrogen peroxide (H2O2), in both plasma and placental tissues of recurrent miscarriage (RM) patients in comparison with those of healthy pregnant (HP) and non-pregnant (NP) women; ii) determine the levels of enzymatic antioxidants [glutathione peroxidase (GPx), glutathione reductase (GSR), superoxide dismutase (SOD) and catalase (CAT)], and non-enzymatic antioxidant micronutrients [selenium (Se), zinc (Zn), copper (Cu) and manganese (Mn)] in both plasma and placental tissues of RM patients, in comparison with those of HP and NP women; iii) profile differential expression levels of selected antioxidant and apoptosis-related genes in the placental tissues of RM cases, in relation to those of HP women of matched gestational age, using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). The results revealed highly significant increases of all investigated OS markers in plasma and placental tissues of RM patients compared with those of HP women. Moderate, but significant, increases of OS markers were observed in the plasma of HP patients in relation to those of NP women. The activities of antioxidant enzymes exhibited statistically significant decreases in both plasma and placental tissues of RM patients compared with those of HP women. The significantly reduced level of antioxidant enzymes was also evident in the plasma of HP women as compared with those of NP women. Results of RT-qPCR assays clearly indicated that the expression level of apoptosis-related genes [tumor necrosis factor-related apoptosis-inducing ligand (TRAIL) and S100A8], and pro-inflammatory cytokine genes [tumor necrosis factor-α (TNF-α), interleukin (IL)-6 and IL-8] were significantly upregulated in placental tissue of RM cases in relation to those of HP subjects. By contrast, mRNA transcriptional levels of key antioxidant genes (GPx, SOD, GSR and CAT) were found to be significantly reduced in placental tissue of RM patients in comparison to those of HP women. In conclusion, our data highlight a plausible cause-effect association between the observed increase in placental OS level and depletion of the activity of antioxidant enzymes. This suggests that OS is a contributing factor in the pathogenesis of idiopathic RM.

TAZ induces lung cancer stem cell properties and tumorigenesis by up-regulating ALDH1A1.

Recent studies suggest that lung cancer stem cells (CSCs) may play major roles in lung cancer. Therefore, identification of lung CSC drivers may provide promising targets for lung cancer. TAZ is a transcriptional co-activator and key downstream effector of the Hippo pathway, which plays critical roles in various biological processes. TAZ has been shown to be overexpressed in lung cancer and involved in tumorigenicity of lung epithelial cells. However, whether TAZ is a driver for lung CSCs and tumor formation in vivo is unknown. In addition, the molecular mechanism underlying TAZ-induced lung tumorigenesis remains to be determined. In this study, we provided evidence that constitutively active TAZ (TAZ-S89A) is a driver for lung tumorigenesis in vivo in mice and formation of lung CSC. Further RNA-seq and qRT-PCR analysis identified Aldh1a1, a well-established CSC marker, as critical TAZ downstream target and showed that TAZ induces Aldh1a1 transcription by activating its promoter activity through interaction with the transcription factor TEAD. Most significantly, inhibition of ALDH1A1 with its inhibitor A37 or CRISPR gene knockout in lung cancer cells suppressed lung tumorigenic and CSC phenotypes in vitro, and tumor formation in mice in vivo. In conclusion, this study identified TAZ as a novel inducer of lung CSCs and the first transcriptional activator of the stem cell marker ALDH1A1. Most significantly, we identified ALDH1A1 as a critical meditator of TAZ-induced tumorigenic and CSC phenotypes in lung cancer. Our studies provided preclinical data for targeting of TAZ-TEAD-ALDH1A1 signaling to inhibit CSC-induced lung tumorigenesis in the future.

Recurrent respiratory papillomatosis causing chronic stridor and delayed speech in an 18-month-old boy.

Recurrent respiratory papillomatosis is a relatively uncommon disease that presents clinically with symptoms ranging from hoarseness to severe dyspnea. Human papilloma virus types 6 and 11 are important in the etiology of papillomas and are most probably transmitted from mother to child during birth. Although spontaneous remission is frequent, pulmonary spread and/or malignant transformation resulting in death has been reported. CO2 laser evaporation of papillomas and adjuvant drug therapy using lymphoblastoid interferon-alpha are the most common treatments. However, several other treatments have been tried, with varying success. In the present report, a case of laryngeal papillomatosis presenting with chronic stridor and delayed speech is described.