Inhibition of SARS-CoV-2 infection by Porphyromonas gingivalis and the oral microbiome.

The COVID-19 pandemic persists despite the availability of vaccines, and it is therefore crucial to develop new therapeutic and preventive approaches. In this study, we investigated the potential role of the oral microbiome in SARS-CoV-2 infection. Using an in vitro SARS-CoV-2 pseudovirus infection assay, we found a potent inhibitory effect exerted by Porphyromonas gingivalis on SARS-CoV-2 infection mediated by known P. gingivalis compounds such as phosphoglycerol dihydroceramide (PGDHC) and gingipains as well as by unknown bacterial factors. We found that the gingipain-mediated inhibition of infection is likely due to cytotoxicity, while PGDHC inhibited virus infection by an unknown mechanism. Unidentified factors present in P. gingivalis supernatant inhibited SARS-CoV-2 likely via the fusion step of the virus life cycle. We addressed the role of other oral bacteria and found certain periodontal pathogens capable of inhibiting SARS-CoV-2 pseudovirus infection by inducing cytotoxicity on target cells. In the human oral cavity, we observed the modulatory activity of oral microbial communities varied among individuals in that some saliva-based cultures were capable of inhibiting while others were enhancing infection. These findings contribute to our understanding of the complex relationship between the oral microbiome and viral infections, offering potential avenues for innovative therapeutic strategies in combating COVID-19.

Strategies for improving the financing of family medicine program: Evidence from Iran.

Background and Aims: The impact of health financing on the performance of the entire health system, including access, quality, and efficiency of healthcare, has been emphasized in the Astana Declaration, and the need to strengthen primary healthcare (PHC) and policy integration has been emphasized. After about two decades, the family medicine (FM) program in Iran is still facing great challenges. The aim of this study is to explore strategies for strengthening financing of the FM program in Iran, a vital component of PHC. Methods: A qualitative study was conducted in 2021. Purposeful sampling was used to select 34 policymakers, managers, and experts from various levels of the Ministry of Health, Iran universities of medical sciences, plan and budget organization of Iran, and health insurance organization in Iran. Thirty-four semistructured interviews were conducted to collect data, which were analyzed by content analysis. Results: Through the analysis of interviews, our study has identified five strategies (identification and management of sustainable resources, pooling of sustainable resources, modeling of service provision, payment system model and its implementation process, and FM management structure), and 13 actions for strengthening financing of the FM program in Iran. Conclusion: Our study has identified five strategies and 13 actions for strengthening the financing of the FM program in Iran. These strategies and actions should be considered by policymakers during the review of the FM program in Iran. Without implementation of the suggested strategies and action, allocated resources may be wasted.

Extracellular Release of Citrullinated Vimentin Directly Acts on Osteoclasts to Promote Bone Resorption in a Mouse Model of Periodontitis.

Elevated osteoclast (OC)-mediated bone resorption, a common pathological feature between periodontitis and rheumatoid arthritis (RA), implicates a possible mutually shared pathogenesis. The autoantibody to citrullinated vimentin (CV), a representative biomarker of RA, is reported to promote osteoclastogenesis (OC-genesis). However, its effect on OC-genesis in the context of periodontitis remains to be elucidated. In an in vitro experiment, the addition of exogenous CV upregulated the development of Tartrate-resistant acid phosphatase (TRAP)-positive multinuclear OCs from mouse bone marrow cells and increased the formation of resorption pits. However, Cl-amidine, an irreversible pan-peptidyl arginine deiminase (PAD) inhibitor, suppressed the production and secretion of CV from RANKL-stimulated OC precursors, suggesting that the citrullination of vimentin occurs in OC precursors. On the other hand, the anti-vimentin neutralizing antibody suppressed in vitro Receptor activator of nuclear factor kappa-Β ligand (RANKL)-induced OC-genesis. The CV-induced upregulation of OC-genesis was abrogated by the Protein kinase C (PKC)-δ inhibitor Rottlerin, accompanied by the downmodulation of OC-genesis-related genes, including Osteoclast stimulatory transmembrane protein (OC-STAMP), TRAP and Matrix Metallopeptidase 9 (MMP9) as well as extracellular signal-regulated kinase (ERK) mitogen-activated protein (MAP)-kinase phosphorylation. Elevated levels of soluble CV and vimentin-bearing mononuclear cells were found in the bone resorption lesions of periodontitis induced in mice in the absence of an anti-CV antibody. Finally, local injection of anti-vimentin neutralizing antibody suppressed the periodontal bone loss induced in mice. Collectively, these results indicated that the extracellular release of CV promoted OC-genesis and bone resorption in periodontitis.

Soluble Sema4D cleaved from osteoclast precursors by TACE suppresses osteoblastogenesis.

Bone remodelling is mediated by orchestrated communication between osteoclasts and osteoblasts which, in part, is regulated by coupling and anti-coupling factors. Amongst formally known anti-coupling factors, Semaphorin 4D (Sema4D), produced by osteoclasts, plays a key role in downmodulating osteoblastogenesis. Sema4D is produced in both membrane-bound and soluble forms; however, the mechanism responsible for producing sSema4D from osteoclasts is unknown. Sema4D, TACE and MT1-MMP are all expressed on the surface of RANKL-primed osteoclast precursors. However, only Sema4D and TACE were colocalized, not Sema4D and MT1-MMP. When TACE and MT1-MMP were either chemically inhibited or suppressed by siRNA, TACE was found to be more engaged in shedding Sema4D. Anti-TACE-mAb inhibited sSema4D release from osteoclast precursors by ~90%. Supernatant collected from osteoclast precursors (OC-sup) suppressed osteoblastogenesis from MC3T3-E1 cells, as measured by alkaline phosphatase activity, but OC-sup harvested from the osteoclast precursors treated with anti-TACE-mAb restored osteoblastogenesis activity in a manner that compensates for diminished sSema4D. Finally, systemic administration of anti-TACE-mAb downregulated the generation of sSema4D in the mouse model of critical-sized bone defect, whereas local injection of recombinant sSema4D to anti-TACE-mAb-treated defect upregulated local osteoblastogenesis. Therefore, a novel pathway is proposed whereby TACE-mediated shedding of Sema4D expressed on the osteoclast precursors generates functionally active sSema4D to suppress osteoblastogenesis.

Simultaneous effects of MWCNT and SiO2 on the rheological behavior of cooling oil and sensitivity analysis.

Many scholars are attracted to nano-lubricants due to their unique properties. In the present study, the rheological behavior of a new generation of lubricants has been investigated. SiO2 nanoparticles with an average diameter of 20-30 nm along with MWCNT with an internal diameter of 3-5 nm and the external diameter of 5-15 nm has been dispersed in 10W40 engine oil as base-lubricant and MWCNTs-SiO2 (20%-80%)/10W40 hybrid nano-lubricant has been produced. Nano-lubricant behavior agrees with the Herschel-Bulkley model and is of Bingham pseudo-plastic type below 55 ° C . Also, in the temperature of 55 ° C , nano-lubricant behavior has changed to Bingham dilatant. The viscosity is increased by 32% in the proposed nano-lubricant compared to base lubricant (Dynamics Viscosity Enhancement). Finally, a new correlation with a precision index of R-squared >0.9800, Adj. R-squared >0.9800, and the maximum margin of deviation of 2.72% has been presented, increasing the applicability of this nano-lubricant. Eventually, the sensitivity analysis of nano-lubricant has been conducted, studying the comparative effect of volume fraction and temperature on viscosity.

Assessment of SARS-CoV-2 entry in gingival epithelial cells expressing CD147.

SARS-CoV-2, the causative agent of the debilitating COVID-19, is mainly transmitted by first infecting nose and lung epithelial cells. The mouth is also believed to be a viral portal site since certain types of oral epithelial cells were shown to express ACE2 receptor. However, it is unclear whether oral epithelial cells are directly infected by SARS-CoV-2. In this study, we addressed whether epithelial cells of the oral gingiva were susceptible to infection. Interestingly, we found that KRT5+ and KRT18+ gingival epithelial cells do not express ACE2 but highly express TMPRSS2 and Furin as well as CD147, which was proposed to be an alternative receptor for SARS-CoV-2. However, using SARS-CoV-2 pseudoviruses containing the spike protein, we observed that gingival epithelial cells were not susceptible to infection due to the lack of ACE2 expression and the inability of CD147 to mediate viral entry. These results strongly suggest that epithelial cells from the gingiva are not susceptible to SARS-CoV-2 and CD147 is not a receptor for the SARS-CoV-2 virus. The susceptibility of oral cells from other oral structures under healthy and pathological conditions still needs to be confirmed to better understand the role of the oral cavity in COVID-19 infection and transmission.

Challenges in the implementation of an electronic referral system: A qualitative study in the Iranian context.

Referral chain reduces patient costs and results in more efficient use of workforce and equipment. Despite the implementation of the Family Physician Programme in Iran, the referral system does not function as intended. Therefore, this study was conducted to explain the challenges in the implementation of an electronic referral system in Iran. This qualitative study was performed in Golestan province in northern Iran. The participants were 46 family physicians, specialist physicians, healthcare managers, experts, and patients who were selected through purposive sampling. Data were collected using in-depth individual interviews and were analysed using content analysis. Implementation of the electronic referral system faces management challenges (planning, organisation, motivation, control), administrative challenges (scheduling, patient load and waiting time, visit and diagnosis, feedback), and educational, policy, sociocultural, technological, and economic challenges. Policymakers can adopt appropriate strategies to improve the quantity and quality of their electronic referral system according to the challenges identified herein.

Assessing related factors to fasting blood sugar and glycosylated hemoglobin in patients with type 2 diabetes simultaneously by a multivariate longitudinal marginal model.

The multivariate marginal model can be used to simultaneously examine the factors affecting both FBS and HbA1c using longitudinal data. The model fitted to multivariate longitudinal data should prevent redundant parameter estimation in order to have greater efficiency. In this study, a multivariate marginal model is used to simultaneously investigate the factors affecting both FBS and HbA1c with longitudinal data for patients with type 2 diabetes in Northern Iran. The present research is a retrospective cohort study. Overall, 500 medical records with complete information were reviewed. The multivariate marginal model is used to determine the factors associated with FBS and HbA1c using longitudinal data. Data have been analyzed in R-3.4.0 using 'mmm2' package. Given that the coefficients for the interactions of rtype with the intercept, time, family history of diabetes, history of hypertension, history of smoking, insulin therapy, systolic/diastolic blood pressure and duration of disease at first visit are significantly different from zero (P < 0.05), the effect of the independent variables on the two response variables is different and different coefficients should be used for each. Therefore, the interactions of these variables with rtype are kept in the final model. The coefficients for the interactions of rtype with sex, age at first visit, history of high cholesterol, and weight are not significantly different from zero (P > 0.05), indicating that their effect on the two response variables is similar and only one coefficient should be used for each. We examined the similarity of coefficients when fitting the longitudinal multivariate model for the relationship between FBS/HbA1c and sex, age, history of high blood cholesterol, and body weight. If an independent variable has similar effects on both responses, only one coefficient should be estimated, which will increase the efficiency of the model and the reliability of the results.

Designing and Launching Health Observatory Dashboard of Islamic Republic of Iran.

Background: Health observatory dashboard can assist in promoting the quality of academic and governmental services by generator high-quality information. The aim of this research is to describe the stages of designing and launching the national public health dashboard. Methods: This study was conducted with a qualitative approach and designing a web application using C#, ASP.NET and JQuery languages. The required data were gathered via 2 sources: (1) reviewing existing documents, and (2) gathering expert opinions. Results: The dashboard is developed in 3 sections, including a conceptual model of the indicators, a page for selecting the indicators, and metadata of each indicator. The indicators are demonstrated in 3 classes based on data sources (surveys and routine data collection), health effects (mortality, morbidity, risk factors, service coverage, social factors affecting health, health system functions, financial protection, population indicators, and macro indicators), as well as a plan, including the health reform plan. The page for selecting the indicators includes 190 major indicators encompassing the 3 mentioned areas. The metadata of each indicator includes the indicator name, its definition, its last figure, its source, the section for descriptive and comparative diagrams (the indicator's trend, provincial distribution, and international comparison of the indicator), and policy options. Conclusion: The Health Observatory System of Iran has been launched. The credibility of this system and user satisfaction depends on implementation of the health observatory calendar, qualitative control of the path of the recorded data, and national determination of policymakers.

Relationship of Blood Group with Level of Cooperation of Pediatric Dental Patients.

Acquaintance with the behavior of children in dental office setting is highly important in treatment success. People with different blood groups often have different behaviors. Thus, the blood group may aid in prediction of behavior of pediatric dental patients. This study is aimed at assessing the relationship of the blood group with level of cooperation of pediatric dental patients. This cross-sectional study was conducted on 130 children between 4 and 6 years of age. The blood group of children was recorded according to their identification card or by collecting an intraoral blood sample during pulpotomy and using the respective kit. To assess the level of cooperation of children, their behavior was videotaped during the procedure, and the videos were assessed by two pedodontists. The behavior of children was scored according to the Venham scale. Data were analyzed by SPSS 26 and Chi-square, Fisher's exact test, and Mann-Whitney U test. The blood group was A in 35.7%, B in 5.8%, AB in 3.2%, and O in 53.5%. Children with blood group O had maximum cooperation (52.6) while those with blood group B had minimum cooperation. Pairwise comparisons of the groups regarding the Venham scale revealed significant differences between blood groups A and B (P = 0.0001) and also B and O (P = 0.005). Pairwise comparisons of the groups regarding the level of cooperation also revealed significant differences between blood groups A and B (P = 0.0001) and B and O (P = 0.019). Blood group B may be correlated with certain behavioral traits such as dental fear and anxiety and the resultant poor cooperation.

Locally Secreted Semaphorin 4D Is Engaged in Both Pathogenic Bone Resorption and Retarded Bone Regeneration in a Ligature-Induced Mouse Model of Periodontitis.

It is well known that Semaphorin 4D (Sema4D) inhibits IGF-1-mediated osteogenesis by binding with PlexinB1 expressed on osteoblasts. However, its elevated level in the gingival crevice fluid of periodontitis patients and the broader scope of its activities in the context of potential upregulation of osteoclast-mediated periodontal bone-resorption suggest the need for further investigation of this multifaceted molecule. In short, the pathophysiological role of Sema4D in periodontitis requires further study. Accordingly, attachment of the ligature to the maxillary molar of mice for 7 days induced alveolar bone-resorption accompanied by locally elevated, soluble Sema4D (sSema4D), TNF-α and RANKL. Removal of the ligature induced spontaneous bone regeneration during the following 14 days, which was significantly promoted by anti-Sema4D-mAb administration. Anti-Sema4D-mAb was also suppressed in vitro osteoclastogenesis and pit formation by RANKL-stimulated BMMCs. While anti-Sema4D-mAb downmodulated the bone-resorption induced in mouse periodontitis, it neither affected local production of TNF-α and RANKL nor systemic skeletal bone remodeling. RANKL-induced osteoclastogenesis and resorptive activity were also suppressed by blocking of CD72, but not Plexin B2, suggesting that sSema4D released by osteoclasts promotes osteoclastogenesis via ligation to CD72 receptor. Overall, our data indicated that ssSema4D released by osteoclasts may play a dual function by decreasing bone formation, while upregulating bone-resorption.

Investigating the Factors Affecting the Need for Unilateral Space Maintainer for First Primary Molars in Late Mixed Dentition.

Background: Premature loss of deciduous teeth can lead to loss of space and have a negative effect on occlusion. The use of space maintainers can reduce the severity of problems such as crowding. However, the literature is controversial regarding the effects of early loss of primary first molars The aim of this study was to determine the factors affecting the need for unilateral space maintainer for the first deciduous molars in late mixed dentition. In this cross-sectional study, fifty children between 6 to 8 years who had lost a primary first molar unilaterally later than 6 months ago were randomly selected. Midline deviation, molar and canine relationships at both sides, facial growth pattern, and the amount of space loss were all assessed. Data were analyzed using SPSS version 25 via one-sample t-test, paired t-test, and linear regression (alpha = 0.05). Methods: In this cross-sectional study, 47 children aged 8 to 10 years with early unilaterally loss of first primary molar in the past 6 months were randomly selected. First, the type of occlusion based on the angle molar relationship and the growth pattern of face based on the Hall and Farkas and midline were assessed. Anterior crowding was measured. SPSS 25 program and Chi-square, t-test, ANOVA, and linear regression were used to analyze the data. A significance level of 0.05 was considered. Results: The results showed that the amount of space loss is 0.56 mm (maxilla = 0.54 and mandible = 0.58), which is not clinically significant, and there is no need for space maintainer. Increasing age (p = 0.021) and increasing the percentage ratio of facial pattern (p = 0.009) significantly reduced the space loss and increased the duration of tooth loss (p = 0.002), and molar relationship in the control side (p = 0.05) and increasing the canine to lateral distance (p = 0.016) significantly increased the space loss. Other factors such as crowding, midline deviation, and canine relationship on the control side did not have significant effects on space loss. Conclusion: Space loss due to extraction of the first primary molars in late mixed dentition was neither statistically nor clinically significant. However, in cases of severe crowding, the vertical growth pattern of the face, and molar relationship, further studies are needed, and follow-up of patients is recommended.

Patient-specific finite element analysis of heart failure and the impact of surgical intervention in pulmonary hypertension secondary to mitral valve disease.

Pulmonary hypertension (PH), a chronic and complex medical condition affecting 1% of the global population, requires clinical evaluation of right ventricular maladaptation patterns under various conditions. A particular challenge for clinicians is a proper quantitative assessment of the right ventricle (RV) owing to its intimate coupling to the left ventricle (LV). We, thus, proposed a patient-specific computational approach to simulate PH caused by left heart disease and its main adverse functional and structural effects on the whole heart. Information obtained from both prospective and retrospective studies of two patients with severe PH, a 72-year-old female and a 61-year-old male, is used to present patient-specific versions of the Living Heart Human Model (LHHM) for the pre-operative and post-operative cardiac surgery. Our findings suggest that before mitral and tricuspid valve repair, the patients were at risk of right ventricular dilatation which may progress to right ventricular failure secondary to their mitral valve disease and left ventricular dysfunction. Our analysis provides detailed evidence that mitral valve replacement and subsequent chamber pressure unloading are associated with a significant decrease in failure risk post-operatively in the context of pulmonary hypertension. In particular, right-sided strain markers, such as tricuspid annular plane systolic excursion (TAPSE) and circumferential and longitudinal strains, indicate a transition from a range representative of disease to within typical values after surgery. Furthermore, the wall stresses across the RV and the interventricular septum showed a notable decrease during the systolic phase after surgery, lessening the drive for further RV maladaptation and significantly reducing the risk of RV failure.

Dual-Function Semaphorin 4D Released by Platelets: Suppression of Osteoblastogenesis and Promotion of Osteoclastogenesis.

Effects of the antiosteoblastogenesis factor Semaphorin 4D (Sema4D), expressed by thrombin-activated platelets (TPs), on osteoblastogenesis, as well as osteoclastogenesis, were investigated in vitro. Intact platelets released both Sema4D and IGF-1. However, in response to stimulation with thrombin, platelets upregulated the release of Sema4D, but not IGF-1. Anti-Sema4D-neutralizing monoclonal antibody (mAb) upregulated TP-mediated osteoblastogenesis in MC3T3-E1 osteoblast precursors. MC3T3-E1 cells exposed to TPs induced phosphorylation of Akt and ERK further upregulated by the addition of anti-sema4D-mAb, suggesting the suppressive effects of TP-expressing Sema4D on osteoblastogenesis. On the other hand, TPs promoted RANKL-mediated osteoclastogenesis in the primary culture of bone-marrow-derived mononuclear cells (BMMCs). Among the known three receptors of Sema4D, including Plexin B1, Plexin B2 and CD72, little Plexin B2 was detected, and no Plexin B1 was detected, but a high level of CD72 mRNA was detected in RANKL-stimulated BMMCs by qPCR. Both anti-Sema4D-mAb and anti-CD72-mAb suppressed RANKL-induced osteoclast formation and bone resorptive activity, suggesting that Sema4D released by TPs promotes osteoclastogenesis via ligation to a CD72 receptor. This study demonstrated that Sema4D released by TPs suppresses osteogenic activity and promotes osteoclastogenesis, suggesting the novel property of platelets in bone-remodeling processes.

Knowledge Translation Improvement Strategies in Universities of Medical Sciences in Iran: A Qualitative Study.

BACKGROUND: Recently, one of the challenges in the health system of the country is the need for research contributing to policy-making. Therefore, it is crucial to develop activities in the field of knowledge Translation (KT). This study aimed to propose KT improvement strategies in universities of medical sciences in Iran. METHODS: In this qualitative study, 18 semi-structured interviews were conducted with key informants from the medical universities in Iran during January-July 2018. The transcribed documents were analyzed using the Gale framework analysis approach. Data organization was carried out using MAXQDA version 10 software. RESULTS: According to framework analysis, six KT improvement strategies were identified including improving the abilities and skills of researchers, improving the processes and quality of knowledge production, revising policies and laws, improving the prerequisites, culture-building, and promoting the use of evidence. CONCLUSION: Given the challenges and strategies outlined in this study, it seems that the mechanism of KT and its effects on improving health plans for policymakers and researchers has not been elucidated yet. Therefore, considerable changes in prerequisites, knowledge production processes, academic procedures, policies and laws are necessary for implementing KT in universities of medical sciences in Iran.

A policy analysis of the national phenylketonuria screening program in Iran.

BACKGROUND: Phenylketonuria (PKU) screening is a public health measure taken to diagnose and treat the patients with PKU to prevent severe neurological disorders in them. The present study was aimed at analyzing the policies of the national PKU screening (NaPS) program in Iran. METHODS: PKU screening program policies were analyzed in compliance with the policy triangle model. Document review and 38 semi-structured interviews were used for data collection. Document review data were analyzed using content analysis, and interview data were analyzed using framework analysis. RESULTS: The national PKU screening (NaPS) program was a decision made at the genetics department of Ministry of Health and Medical Education (MOHME) in Iran. Many internal and external stakeholders were involved in it and valid evidence was used to formulate the policies. Despite some opposition and insufficient support, the program was implemented due to the continuous persistence of parents, interested executives, formulated valid content and a top-down approach. The main barriers included rapid substitution of managers, shortage of Phe-free milk, little awareness of patients' families, social stigma, and inadequate co-operation of some hospital administrators. CONCLUSIONS: The policy triangle framework contributed to explaining the different components of the PKU screening program. A successful PKU screening program requires more stability of senior managers in MOHME, enough human resources and Phe-free milk, educating patients' families, and commitment of hospitals administrators. Meanwhile, all the stakeholders need to be involved in the program effectively.

Lifetime Cost Effectiveness of Phenylketonuria Screening National Program in Iran.

BACKGROUND: Phenylketonuria (PKU) is an autosomal recessive disorder that screening and timely control of this disorder can prevent the adverse effects. Regarding the high prevalence of PKU in Iran, the PKU screening program was started in Iran in 2006. This study was conducted to determine the cost-effectiveness of PKU screening in Iran. METHODS: An economic evaluation was performed among screened and treated individuals compared to no screening in 2013. The study population included 1356132 newborns that screened for PKU diagnosis. Lifetime costs, quality-adjusted life year (QALY) gains and incremental cost-effectiveness ratio (ICER) were calculated from the perspective of government. A discount rate of 3% was considered for both QALYs and costs. A one-way sensitivity analysis was used for assessing the robustness of the results. RESULTS: The discounted lifetime cost of intervention and non-intervention were $59528953.8 and $85295501.6 respectively. Therefore, the total estimated cost saving was $25766547.84. PKU screening produces an ICER of $1844420 per QALY gained. CONCLUSION: Screening and early treatment for PKU is highly cost-effective. Therefore, the screening can improve quality of life of the patients and increase financial saving in health system.

Predifferentiated Gingival Stem Cell-Induced Bone Regeneration in Rat Alveolar Bone Defect Model.

Cleft alveolus, a common birth defect of the maxillary bone, affects one in 700 live births every year. This defect is traditionally restored by autogenous bone grafts or allografts, which may possibly cause complications. Cell-based therapies using the mesenchymal stem cells (MSCs) derived from human gingiva (gingiva-derived mesenchymal stem cells [GMSCs]) is attracting the research interest due to their highly proliferative and multilineage differentiation capacity. Undifferentiated GMSCs expressed high level of MSC-distinctive surface antigens, including CD73, CD105, CD90, and CD166. Importantly, GMSCs induced with osteogenic medium for a week increased the surface markers of osteogenic phenotypes, such as CD10, CD92, and CD140b, indicating their osteogenic potential. The objective of this study was to assess the bone regenerative efficacy of predifferentiated GMSCs (dGMSCs) toward an osteogenic lineage in combination with a self-assembling hydrogel scaffold PuraMatrix™ (PM) and/or bone morphogenetic protein 2 (BMP2), on a rodent model of maxillary alveolar bone defect. A critical size maxillary alveolar defect of 7 mm × 1 mm × 1 mm was surgically created in athymic nude rats. The defect was filled with either PM/BMP2 or PM/dGMSCs or the combination of three (PM/dGMSCs/BMP2) and the bone regeneration was evaluated at 4 and 8 weeks postsurgery. New bone formation was evaluated by microcomputed tomography and histology using Hematoxylin and Eosin staining. The results demonstrated the absence of spontaneous bone healing, either at 4 or 8 weeks postsurgery in the defect group. However, the PM/dGMSCs/BMP2 group showed significant enhancement in bone regeneration at 4 and 8 weeks postsurgery, compared with the transplantation of individual material/cells alone. Apart from developing the smallest critical size defect, results showed that PM/dGMSCs/BMP2 could serve as a promising option for the regeneration of bone in the cranio/maxillofacial region in humans.

5-[Aryloxypyridyl (or nitrophenyl)]-4H-1,2,4-triazoles as novel flexible benzodiazepine analogues: Synthesis, receptor binding affinity and lipophilicity-dependent anti-seizure onset of action.

A new series of 5-(2-aryloxy-4-nitrophenyl)-4H-1,2,4-triazoles and 5-(2-aryloxy-3-pyridyl)-4H-1,2,4-triazoles, possessing C-3 thio or alkylthio substituents, was synthesized and evaluated for their benzodiazepine receptor affinity and anti-seizure activity. These analogues revealed similar to significantly superior affinity to GABAA/benzodiazepine receptor complex (IC50 values of 0.04-4.1 nM), relative to diazepam as the reference drug (IC50 value of 2.4 nM). To determine the onset of anti-seizure activity, the time-dependent effectiveness of i.p. administration of compounds on pentylenetetrazole induced seizure threshold was studied and a very good relationship was observed between the lipophilicity (cLogP) and onset of action of studied analogues (r2 = 0.964). The minimum effective dose of the compounds, determined at the time the analogues showed their highest activity, was demonstrated to be 0.025-0.1 mg/kg, relative to diazepam (0.025 mg/kg).

Differential gene expression analysis and cytological evidence reveal a sexual stage of an amoeba with multiparental cellular and nuclear fusion.

Sex is a hallmark of eukaryotes but its evolution in microbial eukaryotes is poorly elucidated. Recent genomic studies revealed microbial eukaryotes possess a genetic toolkit necessary for sexual reproduction. However, the mechanism of sexual development in a majority of microbial eukaryotes including amoebozoans is poorly characterized. The major hurdle in studying sex in microbial eukaryotes is a lack of observational evidence, primarily due to its cryptic nature. In this study, we used a tractable fusing amoeba, Cochliopodium, to investigate sexual development using stage-specific Differential Gene Expression (DGE) and cytological analyses. Both DGE and cytological results showed that most of the meiosis and sex-related genes are upregulated in Cochliopodium undergoing fusion in laboratory culture. Relative gene ontology (GO) category representations in unfused and fused cells revealed a functional skew of the fused transcriptome toward DNA metabolism, nucleus and ligases that are suggestive of a commitment to sexual development. However, the GO categories of unfused cells were dominated by metabolic pathways and other processes indicative of a vegetative phase. Our study provides strong evidence that the fused cells represent a sexual stage in Cochliopodium. Our findings have further implications in understanding the evolution and mechanism of inheritance involving multiparents in other eukaryotes with a similar reproductive strategy.