Organoid models of human and mouse ductal pancreatic cancer.

Pancreatic cancer is one of the most lethal malignancies due to its late diagnosis and limited response to treatment. Tractable methods to identify and interrogate pathways involved in pancreatic tumorigenesis are urgently needed. We established organoid models from normal and neoplastic murine and human pancreas tissues. Pancreatic organoids can be rapidly generated from resected tumors and biopsies, survive cryopreservation, and exhibit ductal- and disease-stage-specific characteristics. Orthotopically transplanted neoplastic organoids recapitulate the full spectrum of tumor development by forming early-grade neoplasms that progress to locally invasive and metastatic carcinomas. Due to their ability to be genetically manipulated, organoids are a platform to probe genetic cooperation. Comprehensive transcriptional and proteomic analyses of murine pancreatic organoids revealed genes and pathways altered during disease progression. The confirmation of many of these protein changes in human tissues demonstrates that organoids are a facile model system to discover characteristics of this deadly malignancy.

The Rho guanine dissociation inhibitor α inhibits skeletal muscle Rac1 activity and insulin action.

The molecular events governing skeletal muscle glucose uptake have pharmacological potential for managing insulin resistance in conditions such as obesity, diabetes, and cancer. With no current pharmacological treatments to target skeletal muscle insulin sensitivity, there is an unmet need to identify the molecular mechanisms that control insulin sensitivity in skeletal muscle. Here, the Rho guanine dissociation inhibitor α (RhoGDIα) is identified as a point of control in the regulation of insulin sensitivity. In skeletal muscle cells, RhoGDIα interacted with, and thereby inhibited, the Rho GTPase Rac1. In response to insulin, RhoGDIα was phosphorylated at S101 and Rac1 dissociated from RhoGDIα to facilitate skeletal muscle GLUT4 translocation. Accordingly, siRNA-mediated RhoGDIα depletion increased Rac1 activity and elevated GLUT4 translocation. Consistent with RhoGDIα's inhibitory effect, rAAV-mediated RhoGDIα overexpression in mouse muscle decreased insulin-stimulated glucose uptake and was detrimental to whole-body glucose tolerance. Aligning with RhoGDIα's negative role in insulin sensitivity, RhoGDIα protein content was elevated in skeletal muscle from insulin-resistant patients with type 2 diabetes. These data identify RhoGDIα as a clinically relevant controller of skeletal muscle insulin sensitivity and whole-body glucose homeostasis, mechanistically by modulating Rac1 activity.

Streptomyces avermitilis MICNEMA2022: a new biorational strain for producing abamectin as an integrated nematode management agent.

BACKGROUND: Abamectin (ABA) is considered a powerful insecticidal and anthelmintic agent. It is an intracellular product of Streptomyces avermitilis; is synthesized through complicated pathways and can then be extracted from mycelial by methanol extraction. ABA serves as a biological control substance against the root-knot nematode Meloidogyne incognita. This investigation is intended to reach a new strain of S. avermitilis capable of producing ABA effectively. RESULTS: Among the sixty actinobacterial isolates, Streptomyces St.53 isolate was chosen for its superior nematicidal effectiveness. The mycelial-methanol extract of isolate St.53 exhibited a maximum in vitro mortality of 100% in one day. In the greenhouse experiment, the mycelial-methanol extract demonstrated, for the second-stage juveniles (J2s), 75.69% nematode reduction and 0.84 reproduction rate (Rr) while for the second-stage juveniles (J2s), the culture suspension demonstrated 75.38% nematode reduction and 0.80 reproduction rate (Rr). Molecular identification for St.53 was performed using 16 S rRNA gene analysis and recorded in NCBI Genbank as S. avermitilis MICNEMA2022 with accession number (OP108264.1). LC-MS was utilized to detect and identify abamectin in extracts while HPLC analysis was carried out for quantitative determination. Both abamectin B1a and abamectin B1b were produced and detected at retention times of 4.572 and 3.890 min respectively. CONCLUSION: Streptomyces avermitilis MICNEMA2022 proved to be an effective source for producing abamectin as a biorational agent for integrated nematode management.

Characterization of Candida species isolated from clinical specimens: insights into virulence traits, antifungal resistance and molecular profiles.

BACKGROUND: Candida species have emerged as a significant cause of opportunistic infections. Alongside the expression of various virulence factors, the rise of antifungal resistance among Candida species presents a considerable clinical challenge. AIM: This study aimed to identify different Candida species isolated from clinical specimens, evaluate their antifungal sensitivity patterns, identify key genes regulating virulence mechanisms using multiplex PCR and to assess any correlation between their virulence profiles and antifungal resistance patterns. METHOD: A total of 100 Candida spp. was isolated from 630 different clinical specimens and identified to the species level. Their antifungal susceptibility was phenotypically evaluated in accordance with CLSI guidelines using the Vitek-2 Compact System. Virulence markers, including biofilm formation capacity, protease production, melanin production, coagulase production and hemolysin production, were also phenotypically detected. The genetic determinants for biofilm formation and extracellular hydrolytic enzymes were assessed using a multiplex PCR assay. RESULTS: The prevalence of Candida spp. was 15.9%, with C. albicans (48%) and C. glabrata (16%) being the most common. C. albicans showed the highest virulence, with strong biofilm formation, and high proteinase and melanin production. Multiplex PCR revealed Hlp in 22.0%, Hwp in 80.0%, Als in 56.0%, and Sap genes in 56.0% of isolates. Virulence genes were more common in C. albicans than in non-albicans Candida (NAC). Resistance patterns significantly correlated with virulence profiles, with notable associations between flucytosine resistance and the presence of Hlp and Hwp genes. CONCLUSION: The significant correlation between virulent markers such as germination, coagulase, hemolysin production and resistance patterns among different Candida isolates is crucial for predicting the severity and outcomes of Candida infections. This understanding aids in guiding tailored treatment strategies.

A recurrent KCNK4 variant in a dominant pedigree with hypertrichosis and gingival fibromatosis syndrome: Variable phenotypic expressivity and insights on patients' dental management.

Abnormal hyperpolarization of the KCNK4 gene, expressed in the nervous system, brain, and periodontal ligament fibroblasts, leads to impaired neurotransmitter sensitivity, cardiac arrhythmias, and endocrine dysfunction, as well as, progressive cell proliferation. De novo gain of function variants in the KCNK4 gene were reported to cause a recognizable syndrome characterized by facial dysmorphism, hypertrichosis, epilepsy, intellectual/developmental delay, and gingival overgrowth (FHEIG, OMIM# 618381). FHEIG is extremely rare with only three reported cases in the literature. Herein, we describe the first inherited KCNK4 variant (c.730G>C, p.Ala244Pro) in an Egyptian boy and his mother. Variable phenotypic expressivity was noted as the patient presented with the full-blown picture of the syndrome while the mother presented only with hypertrichosis and gingival overgrowth without any neurological manifestations. The c.730G>C (p.Ala244Pro) variant was described before in a single patient and when comparing the phenotype with our patient, a phenotype-genotype correlation seems likely. Atrial fibrillation and joint laxity are new associated findings noted in our patient extending the clinical phenotype of the syndrome. Dental management was offered to the affected boy and a dramatic improvement was noted as the patient regained his smile, restored the mastication function, and resumed his psychological stability.

Association of ACE1 I/D polymorphism and susceptibility to COVID-19 in Egyptian children and adolescents.

BACKGROUND: Given the sparse data on the renin-angiotensin system (RAS) and its biological effector molecules ACE1 and ACE2 in pediatric COVID-19 cases, we investigated whether the ACE1 insertion/deletion (I/D) polymorphism could be a genetic marker for susceptibility to COVID-19 in Egyptian children and adolescents. METHODS: This was a case-control study included four hundred sixty patients diagnosed with COVID-19, and 460 well-matched healthy control children and adolescents. The I/D polymorphism (rs1799752) in the ACE1 gene was genotyped by polymerase chain reaction (PCR), meanwhile the ACE serum concentrations were assessed by ELISA. RESULTS: The ACE1 D/D genotype and Deletion allele were significantly more represented in patients with COVID-19 compared to the control group (55% vs. 28%; OR = 2.4; [95% CI: 1.46-3.95]; for the DD genotype; P = 0.002) and (68% vs. 52.5%; OR: 1.93; [95% CI: 1.49-2.5] for the D allele; P = 0.032). The presence of ACE1 D/D genotype was an independent risk factor for severe COVID-19 among studied patients (adjusted OR: 2.6; [95% CI: 1.6-9.7]; P < 0.001. CONCLUSIONS: The ACE1 insertion/deletion polymorphism may confer susceptibility to SARS-CoV-2 infection in Egyptian children and adolescents. IMPACT: Recent studies suggested a crucial role of renin-angiotensin system and its biological effector molecules ACE1 and ACE2 in the pathogenesis and progression of COVID-19. To our knowledge, ours is the first study to investigate the association of ACE1 I/D polymorphism and susceptibility to COVID-19 in Caucasian children and adolescents. The presence of the ACE1 D/D genotype or ACE1 Deletion allele may confer susceptibility to SARS-CoV-2 infection and being associated with higher ACE serum levels; may constitute independent risk factors for severe COVID-19. The ACE1 I/D genotyping help design further clinical trials reconsidering RAS-pathway antagonists to achieve more efficient targeted therapies.

Comparative Assessment of Water Models in Protein-Glycan Interaction: Insights from Alchemical Free Energy Calculations and Molecular Dynamics Simulations.

Accurate computational simulations of protein-glycan dynamics are crucial for a comprehensive understanding of critical biological mechanisms, including host-pathogen interactions, immune system defenses, and intercellular communication. The accuracy of these simulations, including molecular dynamics (MD) simulation and alchemical free energy calculations, critically relies on the appropriate parameters, including the water model, because of the extensive hydrogen bonding with glycan hydroxyl groups. However, a systematic evaluation of water models' accuracy in simulating protein-glycan interaction at the molecular level is still lacking. In this study, we used full atomistic MD simulations and alchemical absolute binding free energy (ABFE) calculations to investigate the performance of five distinct water models in six protein-glycan complex systems. We evaluated water models' impact on structural dynamics and binding affinity through over 5.8 µs of simulation time per system. Our results reveal that most protein-glycan complexes are stable in the overall structural dynamics regardless of the water model used, while some show obvious fluctuations with specific water models. More importantly, we discover that the stability of the binding motif's conformation is dependent on the water model chosen when its residues form weak hydrogen bonds with the glycan. The water model also influences the conformational stability of the glycan in its bound state according to density functional theory (DFT) calculations. Using alchemical ABFE calculations, we find that the OPC water model exhibits exceptional consistency with experimental binding affinity data, whereas commonly used models such as TIP3P are less accurate. The findings demonstrate how different water models affect protein-glycan interactions and the accuracy of binding affinity calculations, which is crucial in developing therapeutic strategies targeting these interactions.

High Mobility Group Box 1 Gene Polymorphism and Serum High Mobility Group Box 1, Interleukin 1 Beta, and Alpha-Klotho Crosstalk in Severe COVID-19 Patients.

AIM: To evaluate the serum levels of HMGB1, IL1ß, and α-klotho in COVID-19 patients with different disease severity, investigate their association with clinicopathological parameters, and to assess HMGB1 rs1045411 polymorphism and its relation with clinical severity. METHODS: 120 COVID-19 patients (89 critically ill, 15 severe, and 16 moderately severe) along with 80 healthy control were enrolled.The serum levels of HMGB1,IL1ß, and α-klotho were determined by ELISA. The HMGB1 rs1045411 polymorphism was detected by RT- PCR. RESULTS: The serum levels of HMGB1, IL1ß, and α-klotho were significantly higher in critically ill COVID-19 patients compared to other groups. The HMGB1rs1045411 polymorphism revealed a significant decrease in the percentage of T/T genotypes in COVID-19 patients compared to controls. The (ROC) analysis showed moderate diagnostic potential for serum HMGB1, IL1ß, and α-klotho. CONCLUSION: The serum HMGB1, IL1ß, and α-klotho may be severity markers and promising therapeutic targets for COVID-19 patients.

Caffeine intake and its association with nutrition, sleep, and physical activity among schoolchildren in the United Arab Emirates: a national cross-sectional study.

PURPOSE: Recent surveys indicate a significant increase in total caffeine intake among schoolchildren. Limited research has been published concerning the total intake of caffeine among schoolchildren in the Middle East and Gulf Cooperation Council (GCC) countries, including the UAE. METHODS: This cross-sectional survey estimated the total caffeine intake from foods and beverages among 10,275 schoolchildren in the UAE. Caffeine intakes were related to the European Food Safety Authority's level of no safety concern (3.0 mg/kg BW) and level of effects on sleep (1.4 mg/kg BW). RESULTS: More than half (56.2%) of the students consumed more than 100 mg (the upper limit allowed) of caffeine from dietary sources. High intake of caffeine (> 100 mg/day) was significantly associated (p = 0.001) with reduced sleep duration, difficulty falling asleep, infrequent exercising, using smart devices for more than 2 h a day, getting a lower GPA, skipping breakfast, eating fewer servings of vegetables than recommended, frequent consumption of fast food and more frequent snack consumption. CONCLUSION: Excessive intake of caffeine from different dietary sources coexists with unhealthy dietary and lifestyle behaviors and sleep problems. Tailoring educational programs and intervention strategies is warranted to correct the unhealthy intake of caffeine and the associated unhealthy dietary and lifestyle behaviors among schoolchildren in the UAE.

The epigenetic determinants for systemic juvenile idiopathic arthritis phenotyping and treatment response.

BACKGROUND: Determining the role of epigenetics in systemic juvenile idiopathic arthritis (SJIA) provides an opportunity to explore previously unrecognized disease pathways and new therapeutic targets. AIM: We aimed to identify the clinical significance of microRNAs (miRNA-26a, miRNA-223) in SJIA. MATERIALS AND METHODS: This cross-sectional study was conducted on a group of children with SJIA attending to pediatric rheumatology clinic, at Mansoura University Children's Hospital (MUCH) from December 2021 to November 2022. Patient demographics, and clinical, and laboratory data were collected with the measurement of microRNAs by quantitative real-time PCR. The Mann-Whitney, Kruskal-Wallis, and Spearman correlation tests were used for variable comparison and correlations, besides the receiver operating characteristic (ROC) curve for microRNAs disease activity and treatment non-response discrimination. RESULTS: Forty patients were included in the study. On comparison of miRNA-26a, and miRNA-223 levels to the clinical, assessment measures, and laboratory features, miRNA-26a was statistically higher in cases with systemic manifestations versus those without. Similarly, it was higher in children who did not fulfill the Wallace criteria for inactive disease and the American College of Rheumatology (ACR) 70 criteria for treatment response. Meanwhile, miRNA-223 was not statistically different between cases regarding the studied parameters. The best cut-off value for systemic juvenile arthritis disease activity score-10 (sJADAS-10) and the ability of miRNA-26a, and miRNA-223 to discriminate disease activity and treatment non-response were determined by the (ROC) curve. CONCLUSION: The significant association of miRNA-26a with SJIA features points out that this molecule may be preferentially assessed in SJIA disease activity and treatment non-response discrimination.

Novel Bis(2-cyanoacrylamide) Linked to Sulphamethoxazole: Synthesis, DNA Interaction, Anticancer, ADMET, Molecular Docking, and DFT Studies.

In the light of advancement and potential extensive use of medication design and therapy, new bis(cyanoacrylamides) incorporating sulphamethoxazole derivatives (7 a-7 f) were synthesized and confirmed by different spectral tools. In vitro anticancer activity towards different human cancer cells (HCT116, MDA-MB-231 and A549) was assessed using MTT assay. Among all derivatives, 4C- and 6C-spacer derivatives (7 e and 7 f) had the most potent growth inhibitory activities against HCT116 cells with IC50 values of 39.7 and 28.5 µM, respectively. 7 e and 7 f induced apoptosis and suppressed migration of HCT116 cells. These compounds also induced a significant increase in caspase-3 and CDH1 activities, and a downregulation of Bcl2 using ELISA. pBR322 DNA cleavage activities of cyanoacrylamides were determined using agarose gel electrophoresis. Furthermore, 7 e and 7 f showed good DNA and BSA binding affinities using different spectroscopic techniques. Furthermore, molecular docking for 7 e and 7 f was performed to anticipate their binding capabilities toward various proteins (Bcl2, CDH1 and BSA). The docking results were well correlated with those of experimental results. Additionally, density functional theory and ADMET study were performed to evaluate the molecular and pharmacokinetic features of 7 e and 7 f, respectively. Thus, this work reveals promising antitumor lead compounds that merit future research and activity enhancement.

Exploring the synergistic effect of chromium (Cr) tolerant Pseudomonas aeruginosa and nano zero valent iron (nZVI) for suppressing Cr uptake in Aloe Vera.

Chromium (Cr) contamination of soil has substantially deteriorated soil health and has interfered with sustainable agricultural production worldwide and therefore, its remediation is inevitable. Inoculation of plant growth promoting rhizobacteria (PGPR) in association with nanotechnology has exerted broad based impacts in agriculture, and there is an urgent need to exploit their synergism in contaminated soils. Here, we investigated the effect of co-application of Cr-tolerant "Pseudomonas aeruginosa CKQ9" strain and nano zerovalent iron (nZVI) in improving the phytoremediation potential of aloe vera (Aloe barbadensis L.) under Cr contamination. Soil was contaminated by using potassium dichromate (K2Cr2O7) salt and 15 mg kg-1 contamination level in soil was maintained via spiking and exposure to Cr lasted throughout the duration of the experiment (120 days). We observed that the co-application alleviated the adverse impacts of Cr on aloe vera, and improved various plant attributes such as plant height, root area, number of leaves and gel contents by 51, 137, 67 and 49% respectively as compared to control treatment under Cr contamination. Similarly, significant boost in the activities of various antioxidants including catalase (124%), superoxide dismutase (87%), ascorbate peroxidase (36%), peroxidase (89%) and proline (34%) was pragmatic under contaminated soil conditions. In terms of soil Cr concentration and its plant uptake, co-application of P. aeruginosa and nZVI also reduced available Cr concentration in soil (50%), roots (77%) and leaves (84%), while simultaneously increasing the relative production index by 225% than un-inoculated control. Hence, integrating PGPR with nZVI can be an effective strategy for enhancing the phytoremediation potential of aloe vera.

Combined effect of PGPR and nanotechnology in the bioremediation of toxic contaminants is well reported in literature. Most of these reports comprise the use of hyperaccumulator plants for phytoextraction of heavy metals. However, phytostabilization potential of hyperaccumulators is still un-explored. Current study investigated the role of PGPR and Fe-NPs in suppressing the uptake of Cr in aloe vera, a hyperaccumulator plant.

Evaluation of health services and treatment alliance among extensively hospitalized patients due to severe self-harm - results from the Extreme Challenges project.

BACKGROUND: Extensive psychiatric hospitalization due to repeated severe self-harm (SH), is a poorly researched area, but a challenge within health services (HS). Recent studies have demonstrated high levels of involuntary treatment among patients with severe personality disorder (PD) and complex comorbidity. Keeping focus on extensively hospitalized SH patients, this study aimed to investigate patients' and clinicians' evaluation of HS and treatment alliance. METHOD: A cross-sectional study with an inpatient sample (age >18 years) with frequent (>5) or long (>4 weeks) psychiatric hospital admissions last year due to SH or SA recruited from 12 hospitals across health regions (N = 42). Evaluation included patient and clinician report. RESULTS: A minority of the patients (14%) were satisfied with HS before the current admission, 45% (patients) and 20% (clinicians) found the current admission helpful, and 46% (patients) and 14% (clinicians) worried about discharge. Treatment complaints were received in 38% of the cases. Outpatient mental HS were available after discharge for 68% and a majority of clinicians indicated satisfactory contact across HS. More intensive or specialized formats were unusual (structured outpatient treatment 35%, day treatment 21%, ambulatory services 32%, planned inpatient services 31%). Mutual problem understanding, aims, and confidence in therapists during the hospital stay were limited (patient-rated satisfactory mutual problem understanding: 39%, aims of stay: 50%, confidence: 50%). Patient and therapist alliance-ratings were in concordance for the majority. CONCLUSION: The study highlights poor HS satisfaction, poor patient-therapist coherence, limited treatment alliance and limited follow-up in structured treatments addressing SH or intermediary supportive ambulatory/day/inpatient services.

Petroleum wastewater: Environmental protection, treatment, and safe reuse: An overview.

Oil petroleum production consumes about 1.0-7.2 bbl. The needed water for such production ranges between 0.47 and 7.2 L water to 1.0 L crude. Between 80 and 90% of the consumed water is disposed of as wasted effluents. Consequently, there is an important connection between petroleum production and the contamination of the environment and surface water in addition to their ecotoxicological effects. The objective of the present review is to through light on the hazardous impact of petroleum wastewater on the environment and water ways. The present study presents several wastewater treatment technologies in handling the petroleum produced water (PPW) and reducing the hazardous impact to the environment. Safe reuse is also presented including simple, advanced, and environmentally friendly techniques. The reported treatment technologies are divided into five main categories: membrane technologies, biological treatment processes, electro-chemical coagulation, physical/chemical treatment processes (dissolved air flotation (DAF)/air flotation (IAF), adsorption, and chemical flocculation), and catalytic oxidation including chemicals such as advanced and Fenton oxidation processes (AOPs). The analysis and observation of each treatment process are also presented. Implementing of these processes in sequential and/or in combined to avoid the drawbacks of any poor treatment are discussed. The present review discusses; also, in detail each of these treatment technologies and their efficiency including the observation and conclusions of each one. The study shows; also; how the final treated effluent can be reused for non-potable purposes as an additional water resource according to the degree of decontamination. An additional advantage of treatment is protection of both the environment and the water ways by avoiding any discharge of such hazardous wastewater.

Metformin Loaded Zein Polymeric Nanoparticles to Augment Antitumor Activity against Ehrlich Carcinoma via Activation of AMPK Pathway: D-Optimal Design Optimization, In Vitro Characterization, and In Vivo Study.

Metformin (MET), an antidiabetic drug, is emerging as a promising anticancer agent. This study was initiated to investigate the antitumor effects and potential molecular targets of MET in mice bearing solid Ehrlich carcinoma (SEC) as a model of breast cancer (BC) and to explore the potential of zein nanoparticles (ZNs) as a carrier for improving the anticancer effect of MET. ZNs were fabricated through ethanol injection followed by probe sonication method. The optimum ZN formulation (ZN8) was spherical and contained 5 mg zein and 30 mg sodium deoxycholate with a small particle size and high entrapment efficiency percentage and zeta potential. A stability study showed that ZN8 was stable for up to three months. In vitro release profiles proved the sustained effect of ZN8 compared to the MET solution. Treatment of SEC-bearing mice with ZN8 produced a more pronounced anticancer effect which was mediated by upregulation of P53 and miRNA-543 as well as downregulation of NF-κB and miRNA-191-5p gene expression. Furthermore, ZN8 produced a marked elevation in pAMPK and caspase-3 levels as well as a significant decrease in cyclin D1, COX-2, and PGE2 levels. The acquired findings verified the potency of MET-loaded ZNs as a treatment approach for BC.

A pedicled buccal periosteal flap for the closure of oro-antral fistula.

BACKGROUND: An oroantral fistula is a communication between the maxillary antrum and oral cavity. This pathological communication is formed mainly due to dental extraction of maxillary premolars and molars. Adequate management should include closing the oroantral fistula and eliminating sinus infections to prevent recurrence and sinusitis. PURPOSE: This study aimed to evaluate the effectiveness of using the pedicled buccal periosteal flap for closing an oroantral fistula without changing the native intraoral structure. PATIENTS & METHODS: Patients with oroantral fistulas were included in this study. The patients were examined clinically by Valsalva test and cheek-blowing test, the hole was probed, and the extent of the underlying bone defect was determined radiographically using computed tomography preoperatively. All patients underwent surgical closure of oroantral fistula using a pedicled buccal periosteal flap. RESULTS: All 10 patients obtained satisfactory results with marked improvement in the function of the maxillary sinus and complete healing of oroantral fistula with no recurrence except in Case No. 5, who had a recurrence of the oroantral fistula, also there was no statistically significant difference between the vestibular depth preoperatively and postoperatively. CONCLUSION: A pedicled buccal periosteal flap is a novel technique for oroantral fistula closure as it preserves vestibular depth with a tension-free closure flap and harbors the advantages of the regenerative potential of the periosteum. REGISTRATION DATE: 14/8/2023 REGISTRATION NUMBER: NCT05987943.

Bone lid technique versus standard technique for treatment of mandibular lesions.

BACKGROUND: Jaw lesions are frequent in the oral and maxillofacial areas. Different methods for enucleating jaw lesions in the oral and maxillofacial sites have been proposed, including the bone lid technique. PURPOSE: The aim of this study was to compare the clinical and radiographic results of the bone lid technique employing a piezoelectric surgery to the traditional technique in individuals with mandibular lesions. MATERIALS AND METHODS: A randomized controlled trial was conducted on 24 patients with mandibular lesions. They were randomly allocated into two groups (n = 12 for each group). Group I: the mandibular lesion was excised with bone lid technique using a piezoelectric device, followed by the fixation of the bony window after its repositioning. Group II: the lesion was excised with the traditional method using rotatory burs. Pain, soft tissue healing, bone exposure, bone lid integration, and the volume of the residual bone defect were all assessed clinically and radiographically after one week, one month, and six months. RESULTS: All patients in both groups showed adequate soft tissue healing except for one case in group I experienced wound dehiscence and bone lid exposure. The bone lid group reported significantly less pain than the usual approach at the 3rd and 7th days. After six months, the volume of bone defect filling was considerably higher in the bone lid group compared to the conventional group. CONCLUSION: The bone lid technique was an effective procedure in the management of mandibular lesions compared to the standard method. Besides, this technique provides better bone healing and reduces bone loss. TRIAL REGISTRATION: This clinical trial was registered at clinicaltrials.gov on 14/8/2023 and had registration number NCT05987930.

Promising biotherapeutic prospects of different probiotics and their derived postbiotic metabolites: in-vitro and histopathological investigation.

BACKGROUND: Probiotics and their derived postbiotics, as cell-free supernatants (CFS), are gaining a solid reputation owing to their prodigious health-promoting effects. Probiotics play a valuable role in the alleviation of various diseases among which are infectious diseases and inflammatory disorders. In this study, three probiotic strains, Lactiplantibacillus plantarum, Lacticaseibacillus rhamnosus, and Pediococcus acidilactici, were isolated from marketed dietary supplements. The antimicrobial activity of the isolated probiotic strains as well as their CFS was investigated. The neutralized CFS of the isolated probiotics were tested for their antibiofilm potential. The anti-inflammatory activity of the isolated Lactobacillus spp., together with their CFS, was studied in the carrageenan-induced rat paw edema model in male Wistar rats. To the best of our knowledge, such a model was not previously experimented to evaluate the anti-inflammatory activity of the CFS of probiotics. The histopathological investigation was implemented to assess the anti-inflammatory prospect of the isolated L. plantarum and L. rhamnosus strains as well as their CFS. RESULTS: The whole viable probiotics and their CFS showed variable growth inhibition of the tested indicator strains using the agar overlay method and the microtiter plate assay, respectively. When tested for virulence factors, the probiotic strains were non-hemolytic lacking both deoxyribonuclease and gelatinase enzymes. However, five antibiotic resistance genes, blaZ, ermB, aac(6')- aph(2"), aph(3'')-III, and vanX, were detected in all isolates. The neutralized CFS of the isolated probiotics exhibited an antibiofilm effect as assessed by the crystal violet assay. This effect was manifested by hindering the biofilm formation of the tested Staphylococcus aureus and Pseudomonas aeruginosa clinical isolates in addition to P. aeruginosa PAO1 strain. Generally, the cell cultures of the two tested probiotics moderately suppressed the acute inflammation induced by carrageenan compared to indomethacin. Additionally, the studied CFS relatively reduced the inflammatory changes compared to the inflammation control group but less than that observed in the case of the probiotic cultures treated groups. CONCLUSIONS: The tested probiotics, along with their CFS, showed promising antimicrobial and anti-inflammatory activities. Thus, their safety and their potential use as biotherapeutics for bacterial infections and inflammatory conditions are worthy of further investigation.

Therapeutic properties of Helicobacter pylori-derived vacuolating cytotoxin A in an animal model of chronic allergic airway disease.

BACKGROUND: It has previously been shown that the Helicobacter pylori (H. pylori)-derived molecule vacuolating cytotoxin A (VacA) could be suitable for the treatment of allergic airway disease. The therapeutic activity of the protein, which acts through modulation of dendritic cells (DC) and regulatory T cells (Tregs), was demonstrated in murine short-term acute models. The aim of this study is to further evaluate the therapeutic potential of VacA by determining the effectiveness of different application routes and the suitability of the protein for treating the chronic phase of allergic airway disease. METHODS: VacA was administered by the intraperitoneal (i.p.), oral (p.o.) or intratracheal (i.t.) routes, and long-term therapeutic effectiveness, allergic airway disease hallmarks, and immune phenotype were analyzed in murine models of acute and chronic allergic airway disease. RESULTS: Administration of VacA via the i.p., p.o or i.t. routes was associated with a reduction in airway inflammation. The i.p. route showed the most consistent effect in reducing airway inflammation and i.p. treatment with VacA was the only treatment that significantly reduced mucus cell hyperplasia. In a murine model of chronic allergic airway disease, both short- and long-term treatment with VacA showed a therapeutic effect, with a reduction in a variety of asthma hallmarks, including bronchoalveolar lavage eosinophilia, lung inflammation and goblet cell metaplasia. Short-term treatment was associated with induction of Tregs, while repetitive long-term administration of VacA influenced immunological memory in the lung. CONCLUSIONS: In addition to showing therapeutic efficacy in short-term models, treatment with VacA also appeared to be effective in suppressing inflammation in a chronic airway disease model. The observation that treatment was effective after administration via several different routes highlights the potential of VacA as a therapeutic agent with different routes of administration in humans.

CHST3-related skeletal dysplasia in 14 patients: Identification of 8 novel variants and further expansion of the phenotypic spectrum.

Biallelic variants in CHST3 gene result in congenital dislocation of large joints, club feet, short stature, rhizomelia, kypho-scoliosis, platyspondyly, epiphyseal dysplasia, flared metaphysis, in addition to minor cardiac lesions and hearing loss. Herein, we describe 14 new patients from 11 unrelated Egyptian families with CHST3-related skeletal dysplasia. All patients had spondyloepiphyseal changes that were progressive with age in addition to bifid distal ends of humeri which can be considered a diagnostic key in patients with CHST3 variants. They also shared peculiar facies with broad forehead, broad nasal tip, long philtrum and short neck. Rare unusual associated findings included microdontia, teeth spacing, delayed eruption, prominent angulation of the lumbar-sacral junction and atrial septal defect. Mutational analysis revealed 10 different homozygous CHST3 (NM_004273.5) variants including 7 missense, two frameshift and one nonsense variant. Of them, the c.384_391dup (p.Pro131Argfs*88) was recurrent in two families. Eight of these variants were not described before. Our study presents the largest series of patients with CHST3-related skeletal dysplasia from the same ethnic group. Furthermore, it reinforces that lethal cardiac involvement is a critical clinical finding of the disorder. Therefore, we believe that our study expands the phenotypic and mutational spectrum, and also highlights the importance of performing echocardiography in patients harboring CHST3 variants.