Sumbawa cattle: a study of growth hormone (GH) gene variants and their association with biometric traits.

The growth hormone (GH) gene plays a vital role in regulating animal metabolism and body size, making it a potential candidate for influencing livestock performance. This study aimed to investigate the polymorphisms within the GH gene and their associations with 10 biometric traits in the Sumbawa cattle population of Indonesia. Biometric trait data and blood samples were collected from 112 Sumbawa cattle individuals, and their GH gene sequences were analyzed using two sets of primers for amplification. Seven single nucleotide polymorphisms (SNPs) were identified in the GH gene: g.442C>T, g.446G>C, g.558C>T, g.649C>A, g.1492C>A, g.1510C>A, and g.1578G>A. All SNPs were located in the intronic region except for SNP g.558C>T, which was found in the coding sequence (CDS) region. The SNP g.558C>T is classified as a synonymous variant. Haplotype analysis revealed a strong linkage disequilibrium between SNPs g.558C>T and g.649C>A. Distributions of genotypes and alleles of all SNPs were in agreement with the Hardy-Weinberg equilibrium (p > 0.05, χ2 < 15.56), except for SNPs g.446G>C and g.1492C>A. The association study showed that the SNP g.442C>T significantly (p < 0.05) affected HL, BL, SH, and PH traits in Sumbawa cattle. Additionally, the g.446G>C and g.558C>T were also found to be associated with PH and CC traits, respectively. The polymorphisms detected in the GH gene could have implications for selection programs to enhance desired biometric traits in Sumbawa cattle. Improving livestock productivity can be done by understanding genetic diversity and its relationship with phenotypic characteristics.

Disclosure of genetic diversity of mackerel fish (Scomberomorus spp.) in Indonesian waters based on the mitochondrial cytochrome oxidase subunit II (COII) gene.

Mackerel fish (Scomberomorus spp.) represents a significant marine fisheries commodity in Indonesia, characterized by its high commercial value and nutritional content. To understand the intraspecific interactions and genetic variability of Scomberomorus spp., a more extensive research of Scomberomorus spp. populations, including both cultivated and wild specimens, is required. This study aimed to explore the genetic diversity of mackerel fish in Indonesian waters, focusing on the mitochondrial DNA (mtDNA) cytochrome oxidase subunit II (COII) gene, which encodes the second subunit of cytochrome c oxidase (complex IV), is essential for aerobic respiration and energy transformation. Muscle tissue samples from 18 individual mackerel fish collected from various regions in Indonesia, including Palembang, Cilacap, Rembang, Banjarmasin, Ambon, and Fak-Fak Regencies, were utilized. The genomic DNA was isolated and amplified using specific primers: CO2TF (5'-ACCGCTCTGTCACTTTCTTC-3') and CO2TR (5'-ATGTCACTAAGGGTGGTTGG-3'). Subsequently, the obtained amplicons were subjected to sequencing. The sequence data were then analyzed using the MEGA11 and DnaSP 6 software. Our findings revealed 120 variable sites within the 691 base pairs of mtDNA COII sequences, resulting in a nucleotide diversity (Pi) of 0.07169. Furthermore, we identified eight haplotypes, demonstrating a haplotype diversity (Hd) of 0.8889. Remarkably, all mackerel samples from Palembang and Cilacap clustered into discrete haplotypes, specifically haplotype 1 and haplotype 2, respectively. Our phylogenetic analysis delineated three distinct clades. Clade I, closely related to Scomberomorus cavalla, encompassed all individuals from Ambon, Palembang, Rembang, and one from Banjarmasin. Clade II, associated with Scomberomorus niphonius, included individuals from Cilacap and two from Banjarmasin. Clade III, linked to Scomberomorus semifasciatus, exclusively consisted of individuals from Fak-Fak (Papua). In conclusion, Indonesian waters harbor diverse genetic variations within Scomberomorus spp., and population relationships based on the mtDNA COII gene exhibit notable complexities. Future research endeavors should focus on further elucidating the diversity and relationships among Scomberomorus spp. in diverse Indonesian populations.

Tracking of thermal microbial shock proteins and genomic variation of Bacillus cereus due to some abiotic stresses.

Bacillus cereus is considered the most potent bacterial strain in terms of the increment in induced proteins during thermal treatment at 52 °C for 90 min. Protein production in food-born microorganism (Bacillus cereus) recovered from contaminated food was investigated in response to heat shock treatment. Bacterial tolerance towards pH, salinity, and temperature at various levels was also investigated. Heat-shock proteins (HSPs) produced when exposed to 52 °C for up to 60 minutes led to significant differences (30%) above the untreated control (37 °C), and the maximum difference was recorded at 52°C at 90 minutes. ISSR detected a higher number of bands/primer than RAPD (13.7 vs. 12.7, respectively), and more polymorphic bands (10.7 vs. 8.4 bands/primer, respectively). The untreated bacterial strain did not grow at pH levels lower than 3, whereas the thermally treated strain grew significantly at pH two. A consistent increase in HSPs was observed, with a gradual increase in salinity of less than 16%. Surprisingly, the gradual increase in temperature did not induce tolerance against higher temperatures. However, a significant growth rate was noticed in response to heat-shocked treatments. The untreated Bacillus cereus demonstrated antibiotic resistance to gentamycin and clindamycin (1.54 and 1.65 cm, respectively), much lower than the corresponding inhibition areas with preheat-treated test pathogen which were recorded (2.37 and 2.49 cm, respectively).

Tracking of thermal microbial shock proteins and genomic variation of Bacillus cereus due to some abiotic stresses / Rastreamento de proteínas de choque microbiano térmico e variação genômica de Bacillus cereus devido a alguns estresses abióticos

Bacillus cereus is considered the most potent bacterial strain in terms of the increment in induced proteins during thermal treatment at 52 °C for 90 min. Protein production in food-born microorganism (Bacillus cereus) recovered from contaminated food was investigated in response to heat shock treatment. Bacterial tolerance towards pH, salinity, and temperature at various levels was also investigated. Heat-shock proteins (HSPs) produced when exposed to 52 °C for up to 60 minutes led to significant differences (30%) above the untreated control (37 °C), and the maximum difference was recorded at 52°C at 90 minutes. ISSR detected a higher number of bands/primer than RAPD (13.7 vs. 12.7, respectively), and more polymorphic bands (10.7 vs. 8.4 bands/primer, respectively). The untreated bacterial strain did not grow at pH levels lower than 3, whereas the thermally treated strain grew significantly at pH two. A consistent increase in HSPs was observed, with a gradual increase in salinity of less than 16%. Surprisingly, the gradual increase in temperature did not induce tolerance against higher temperatures. However, a significant growth rate was noticed in response to heat-shocked treatments. The untreated Bacillus cereus demonstrated antibiotic resistance to gentamycin and clindamycin (1.54 and 1.65 cm, respectively), much lower than the corresponding inhibition areas with preheat-treated test pathogen which were recorded (2.37 and 2.49 cm, respectively).

Bacillus cereus é considerada a cepa bacteriana mais potente em termos de incremento de proteínas induzidas durante o tratamento térmico a 52 °C por 90 min. A produção de proteínas em microorganismos de origem alimentar (Bacillus cereus) recuperados de alimentos contaminados foi investigada em resposta ao tratamento de choque térmico. A tolerância bacteriana ao pH, salinidade e temperatura em vários níveis também foram investigadas. Proteínas de choque térmico (HSPs) produzidas quando expostas a 52 °C por até 60 minutos levaram a diferenças significativas (30%) acima do controle não tratado (37 °C), e a diferença máxima foi registrada a 52 °C em 90 minutos . O ISSR detectou um maior número de bandas/iniciador do que o RAPD (13,7 vs. 12,7, respectivamente) e mais bandas polimórficas (10,7 vs. 8,4 bandas/iniciador, respectivamente). A cepa bacteriana não tratada não cresceu em níveis de pH abaixo de 3, enquanto a cepa tratada termicamente cresceu significativamente em pH dois. Observou-se aumento consistente de HSPs, com aumento gradual da salinidade inferior a 16%. Surpreendentemente, o aumento gradual da temperatura não induziu tolerância a temperaturas mais altas. No entanto, uma taxa de crescimento significativa foi observada em resposta aos tratamentos de choque térmico. O Bacillus cereus não tratado demonstrou resistência antibiótica à gentamicina e clindamicina (1,54 e 1,65 cm, respectivamente), muito menor do que as áreas de inibição correspondentes com patógeno de teste pré-tratado que foram registradas (2,37 e 2,49 cm, respectivamente).

Jatropha curcas L. as a Plant Model for Studies on Vegetative Propagation of Native Forest Plants.

Even though it is a forest native plant, there are already several studies evaluating the small genome of Jatropha curcas L., which belongs to the Euphorbiaceae family, and may be an excellent representative model for the other plants from the same family. Jatropha curcas L. plant has fast growth, precocity, and great adaptability, facilitating silvicultural studies, allowing important information to be obtained quickly, and reducing labor costs. This information justifies the use of the species as a model plant in studies involving the reproduction of native plants. This study aimed to evaluate the possibility of using Jatropha curcas L. as a model plant for studies involving native forest plants and establish possible recommendations for the vegetative propagation of the species using hardwood cuttings. The information collected can be helpful to other native forest plant species, similar to Jatropha curcas L. To this end, the effects of hardwood cutting length (10, 20, and 30 cm) and the part of the hardwood cuttings (basal, middle, and apex) were evaluated. Moreover, the influence of immersing the hardwood cuttings in solutions containing micronutrients (boron or zinc) or plant regulators (2,4-D, GA3) and a biostimulant composed of kinetin (0.09 g L-1), gibberellic acid (0.05 g L-1), and 4-indole-3-butyric acid (0.05 g L-1). The experiments were carried out in duplicates. In one duplicate, sand was used as the substrate, and rooting evaluations were made 77 days after planting. In another duplicate, a substrate composed of 50% soil, 40% poultry litter, and 10% sand was used, and the evaluations of the saplings were performed 120 days after planting. The GA3 solutions inhibited the roots' and sprouts' emissions, while immersion in 2,4-D solution increased the number of primary roots at 77 days after planting. The hardwood cuttings from the basal part of the branch had the best results for producing saplings.

Nucleoside Transport and Nucleobase Uptake Null Mutants in Leishmania mexicana for the Routine Expression and Characterization of Purine and Pyrimidine Transporters.

The study of transporters is highly challenging, as they cannot be isolated or studied in suspension, requiring a cellular or vesicular system, and, when mediated by more than one carrier, difficult to interpret. Nucleoside analogues are important drug candidates, and all protozoan pathogens express multiple equilibrative nucleoside transporter (ENT) genes. We have therefore developed a system for the routine expression of nucleoside transporters, using CRISPR/cas9 to delete both copies of all three nucleoside transporters from Leishmania mexicana (ΔNT1.1/1.2/2 (SUPKO)). SUPKO grew at the same rate as the parental strain and displayed no apparent deficiencies, owing to the cells' ability to synthesize pyrimidines, and the expression of the LmexNT3 purine nucleobase transporter. Nucleoside transport was barely measurable in SUPKO, but reintroduction of L. mexicana NT1.1, NT1.2, and NT2 restored uptake. Thus, SUPKO provides an ideal null background for the expression and characterization of single ENT transporter genes in isolation. Similarly, an LmexNT3-KO strain provides a null background for transport of purine nucleobases and was used for the functional characterization of T. cruzi NB2, which was determined to be adenine-specific. A 5-fluorouracil-resistant strain (Lmex5FURes) displayed null transport for uracil and 5FU, and was used to express the Aspergillus nidulans uracil transporter FurD.

Microbiological investigation study for Apis mellifera yemenitica and Apis mellifera carnica bee venoms on selected bacterial strains.

Bees are one of the ancient and the most social insects worldwide. They are of great economic and medical importance. Bee venom (BV) has many therapeutic effects and has been used since ancient times for the treatment of many diseases. The present study aimed to evaluate and compare the antibacterial effect of BV from two different bee subspecies Apis mellifera yemenitica (A. m. yemenitica) (indigenous strain) and Apis mellifera carnica (A. m. carnica) (carniolan strain) against selected Gram-positive and Gram-negative bacteria. Experimentally, venoms were extracted using an electrical venom collector from honey bee colonies of the subspecies, A. m. yemenitica and A. m. carnica, in Hail, Saudi Arabia. Each venom was tested against selected medically important Gram-negative strains, Salmonella Typhimurium, Pseudomonas aeruginosa, and Escherichia coli, while Staphylococcus aureus was selected as Gram-positive test organism. The minimum inhibitory concentration (MIC) method was used to compare the effect of BV from the two subspecies on the growth of the selected bacterial strains. Results showed that BV from both subspecies could equally inhibit the growth of Salmonella Typhimurium, Pseudomonas aeruginosa, and Escherichia coli at an MIC of 10 mg/ml. However, S. aureus was inhibited by an MIC of 5 and 10 mg/ml of BV from A. m. carnica and A. m. yemenitica, respectively. This suggested that the BV of the carnica subspecie was more inhibitory to this Gram-positive pathogen than its counterpart produced by the yemenitica subspecies. The present study shows that bee venom has a promising antibacterial effect.

Evaluating the responses of some wheat landraces and cultivars cultivated locally in Saudi Arabia to three sources of irrigation water

This study was aimed to investigate the response of some wheat (Triticum aestivum L.) landraces to three irrigation sources, freshwater (FW), well water (WW), and treated municipal wastewater (TMW), regarding the impact on growth, yield, and grains elements contents. The results showed that the various wheat landraces irrigated with treated wastewater were significantly taller (117.3 cm) with multi tillers (22 tiller plant-1), had maximum spikes per plant (2.6), and longer weighty spikes (14.5 cm & 12.1 g). Landraces L1(Burr), L2 (Baldy Burr), C7 (Yecora Rojo), had maximum tillers, L4 (Alssamaa Burr), L5 (Bahaal Burr), L7 (Yecora Rojo) had more spikes per plant and the longer weighty spikes were recorded in L5 Baldy Burr, L1(Burr), heavy spikes were reported in L5 (Bahaal Burr) and L1(Burr). Moreover, these landraces had the highest yield per plant and 1000 grains weight (49.8 g, 12.5 g) respectively. The N, P, K, and Mg contents were increased under TMW, and their levels in landraces and cultivars in order are 3>L2>L4>L5>L1>C6>C7. Even Cu, Fe, Mn, Zn levels were higher in various landraces irrigated with TMW, however, L3, L2, and L4 had maximum contents of all microelements. TMW irrigation enhanced growth, yield, and grain quality in terms of essential elements. The irrigation of landraces L1, L4, L5 with TMW may be a feasible alternative for sustainable wheat production and safe water in arid regions such as Saudi Arabia.

Validation and Performance of FibroScan®-AST (FAST) Score on a Brazilian Population with Nonalcoholic Fatty Liver Disease.

BACKGROUND AND AIM: FAST score has a good performance for diagnosing the composite of NASH + NAS ≥ 4 + F ≥ 2. However, it has not been evaluated in Latin American individuals with nonalcoholic fatty liver disease (NAFLD). We aimed to analyze the performance of the FAST score in a Brazilian NAFLD population. METHODS: Cross-sectional study was held in ≥ 18 years NAFLD patients diagnosed by ultrasonography and submitted to liver biopsy (LB). Liver stiffness (LSM) and CAP measurements were performed with FibroScan®, using M (BMI < 32 kg/m2) or XL probes. Area under receiver operating characteristic (AUROC) curves were calculated as well as sensitivity (S), specificity (Spe), positive predictive value (VPP) and negative predictive value (NPV) for the previously established FAST score cut-offs. RESULTS: Among 287 patients included (75% female; mean age 55 ± 10 years), NASH + NAS ≥ 4 + F ≥ 2 was reported in 30% of LB. For the FAST cut-off of 0.35, the S and NPV to rule out NASH + NAS ≥ 4 + F ≥ 2 were 78.8% and 87.8%, respectively. Regarding the cut-off of 0.67, the Spe and PPV to rule-in NASH + NAS ≥ 4 + F ≥ 2 were 89.1%, 61.8%, respectively. The AUROC of FAST for all included patients was 0.78 (95% CI 0.72-0.84) and for those with ≥ 32 kg/m2 was 0.81 (95% CI 0.74-0.88). CONCLUSION: FAST score has a good performance in a Brazilian NAFLD population, even in patients with higher BMI when the XL probe is adopted. Therefore, FAST can be used as a noninvasive screening tool mainly for excluding the diagnosis of progressive NASH, reducing the number of unnecessary liver biopsies.

Litter Detection with Deep Learning: A Comparative Study.

Pollution in the form of litter in the natural environment is one of the great challenges of our times. Automated litter detection can help assess waste occurrences in the environment. Different machine learning solutions have been explored to develop litter detection tools, thereby supporting research, citizen science, and volunteer clean-up initiatives. However, to the best of our knowledge, no work has investigated the performance of state-of-the-art deep learning object detection approaches in the context of litter detection. In particular, no studies have focused on the assessment of those methods aiming their use in devices with low processing capabilities, e.g., mobile phones, typically employed in citizen science activities. In this paper, we fill this literature gap. We performed a comparative study involving state-of-the-art CNN architectures (e.g., Faster RCNN, Mask-RCNN, EfficientDet, RetinaNet and YOLO-v5), two litter image datasets and a smartphone. We also introduce a new dataset for litter detection, named PlastOPol, composed of 2418 images and 5300 annotations. The experimental results demonstrate that object detectors based on the YOLO family are promising for the construction of litter detection solutions, with superior performance in terms of detection accuracy, processing time, and memory footprint.

Activity of Compounds from Temperate Propolis against Trypanosoma brucei and Leishmania mexicana.

Ethanolic extracts of samples of temperate zone propolis, four from the UK and one from Poland, were tested against three Trypanosoma brucei strains and displayed EC50 values < 20 µg/mL. The extracts were fractionated, from which 12 compounds and one two-component mixture were isolated, and characterized by NMR and high-resolution mass spectrometry, as 3-acetoxypinobanksin, tectochrysin, kaempferol, pinocembrin, 4'-methoxykaempferol, galangin, chrysin, apigenin, pinostrobin, cinnamic acid, coumaric acid, cinnamyl ester/coumaric acid benzyl ester (mixture), 4',7-dimethoxykaempferol, and naringenin 4',7-dimethyl ether. The isolated compounds were tested against drug-sensitive and drug-resistant strains of T. brucei and Leishmania mexicana, with the highest activities ≤ 15 µM. The most active compounds against T. brucei were naringenin 4',7 dimethyl ether and 4'methoxy kaempferol with activity of 15-20 µM against the three T. brucei strains. The most active compounds against L. mexicana were 4',7-dimethoxykaempferol and the coumaric acid ester mixture, with EC50 values of 12.9 ± 3.7 µM and 13.1 ± 1.0 µM. No loss of activity was found with the diamidine- and arsenical-resistant or phenanthridine-resistant T. brucei strains, or the miltefosine-resistant L. mexicana strain; no clear structure activity relationship was observed for the isolated compounds. Temperate propolis yields multiple compounds with anti-kinetoplastid activity.

An approximate analytical formula for estimating the weight of factors affecting the spread of COVID-19: a case study of the first wave.

COVID-19 pandemic has changed the way we live our lives for the foreseen future. To date, there have been over 113 million reported cases and 2.5 million deaths worldwide. Many studies investigated the factors affecting the number of daily cases such as weather conditions, lockdown duration and other factors. In this study, we propose a COVID-19 analytical formula for factors contributing to the number of the new coronavirus daily cases. We have also calculated values of relative weights of those factors. We focus on the first wave data that are publically available. Seven countries were considered including the UK, Italy, Spain, Canada, South Korea, Germany and France. We considered the following factors: temperature, humidity, government expenditure, lockdown hours and the number of daily tests for COVID-19 performed. The weights were calculated based on the hypothesis that a high correlation between recorded data of a given pair of countries implies a high correlation of the pair's COVID-19 proposed analytical formula. The factors are calculated using the brute-force technique. Our results showed that in five out of the seven countries; temperature, humidity, and lockdown duration were the most dominant with values of 26%, 32% and 38%, respectively. In other countries, however, humidity, government expenditure and the daily performed tests for COVID-19 were the most effective factors, with relative values of 35%, 26%, and 28%.

Routine third-trimester ultrasound in low-risk pregnancies and perinatal death: a systematic review and meta-analysis.

OBJECTIVE: This study aimed to determine whether routine third-trimester ultrasounds in low-risk pregnancies decrease the rate of perinatal death compared with regular antenatal care with serial fundal height measurements. DATA SOURCES: This was a systematic review and meta-analysis of randomized control trials to identify relevant studies published from inception to October 2019. The databases used were Ovid, PubMed, Scopus, ClinicalTrials.gov, and the Cochrane Central Register of Controlled Trials using a combination of key words related to "third trimester ultrasound" and "low-risk." STUDY ELIGIBILITY CRITERIA: We included all randomized control trials of singleton, nonanomalous low-risk pregnancies that were randomized to either one or more third-trimester ultrasounds (ultrasound group) or serial fundal height (fundal height group). Exclusion criteria were patients with multiple gestations, maternal medical complications, or fetal abnormalities requiring a third-trimester ultrasound. STUDY APPRAISAL AND SYNTHESIS METHODS: The primary outcome was the rate of perinatal death. The secondary outcomes were rates of fetal growth restriction, suspected large for gestational age, polyhydramnios, oligohydramnios, fetal anomalies, antenatal interventions, stillbirth, neonatal death, cesarean delivery, induction of labor, and other neonatal outcomes. This meta-analysis was performed with the use of the random effects model of DerSimonian and Laird to produce relative risk or mean difference with a corresponding 95% confidence interval. RESULTS: A total of 7 randomized control trials with 23,643 participants (12,343 in the ultrasound group vs 11,300 in the fundal height group) were included. The total rate of perinatal death was similar among the groups (41 of 11,322 [0.4%] vs 34 of 10,285 [0.3%]; relative risk, 1.14; 95% confidence interval, 0.68-1.89). The rate of fetal growth restriction was higher in the ultrasound group (763 of 10,388 [7%] vs 337 of 9021 [4%]; relative risk, 2.11; 95% confidence interval, 1.86-2.39) and the rate of suspected large for gestational age (1060 of 3513 [30%] vs 375 of 3558 [11%]; relative risk, 2.84; 95% confidence interval, 2.6-3.2). Polyhydramnios was also significantly higher in the ultrasound group than the fundal height group (18 of 323 [6%] vs 4 of 322 [1%] relative risk, 3.93; 95% confidence interval, 1.4-11). The rates of the remainder of the secondary outcomes were similar among the groups. CONCLUSION: Routine third-trimester ultrasounds do not decrease the rate of perinatal death compared with serial fundal height in low-risk pregnancies. Ideally, an adequately powered trial is warranted to determine whether perinatal mortality in the fundal height group can be reduced by one-third with third-trimester ultrasound.

Plasma levels of soluble HLA-G and cytokines in papillary thyroid carcinoma before and after thyroidectomy.

BACKGROUND: Increasing evidence shows that chronic inflammation plays an important role in thyroid tumorigenesis. Cytokines as central mediators in inflammatory microenvironment can present both pro-tumour and anti-tumour effects and cytokine release may be influenced by soluble HLA-G (sHLA-G), an immune checkpoint molecule whose expression can also be induced by certain cytokines. AIM: To understand the role of these soluble factors in papillary thyroid cancer (PTC). METHODS: We evaluated plasma levels of sHLA-G and of 13 cytokines using ELISA and flow cytometry, respectively, in PTC patients at two time points: pre- and post-thyroidectomy; and control subjects. RESULTS: Compared with controls, IL-6 levels were increased, while IL-1ß, IFN-α and TGF-ß1 levels were decreased in pre-thyroidectomy PTC patients. IFN-α and TGF-ß1 efficiently discriminated patients from controls and were associated with extrathyroidal extension and lymph node metastasis, respectively. In addition, TNF and IL-13 were associated with male gender, lymph node metastasis and Hashimoto thyroiditis, and sHLA-G with tumour invasion. Compared with pre-thyroidectomy, IL-4, IL-10, TNF, IFN-α and TGF-ß1 levels were increased in post-thyroidectomy. CONCLUSION: There are significant changes in the cytokine profile after surgical removal of the thyroid tumour, and IFN-α e TGF-ß1 showed to be promising cytokines for discriminating PTC patients from controls. We also found that different cytokines are associated with clinicohistopathological characteristics of PTC related to poor prognosis, suggesting that cytokines seem to play an important role in PTC development and management.

Dual-targeted therapeutic strategy combining CSC-DC-based vaccine and cisplatin overcomes chemo-resistance in experimental mice model.

BACKGROUND AND PURPOSE: Emerging evidence suggests that one of the main reasons of chemotherapy treatment failure is the development of multi-drug resistance (MDR) associated with cancer stem cells (CSCs). Our aim is to identify a therapeutic strategy based on MDR-reversing agents. MATERIALS AND METHODS: CSC-enriched Ehrlich carcinoma (EC) cell cultures were prepared by drug-resistant selection method using different concentrations of cisplatin (CIS). Cell cultures following drug exposure were analyzed by flow cytometry for CSC surface markers CD44+/CD24-. We isolated murine bone marrow-derived dendritic cells (DCs) and then used them to prepare CSC-DC vaccine by pulsation with CSC-enriched lysate. DCs were examined by flow cytometry for phenotypic markers. Solid Ehrlich carcinoma bearing mice were injected with the CSC-DC vaccine in conjunction with repeated low doses of CIS. Tumor growth inhibition was evaluated and tumor tissues were excised and analyzed by real-time PCR to determine the relative gene expression levels of MDR and Bcl-2. Histopathological features of tumor tissues excised were examined. RESULTS AND CONCLUSION: Co-treatment with CSC-DC and CIS resulted in a significant tumor growth inhibition. Furthermore, the greatest response of downregulation of MDR and Bcl-2 relative gene expression were achieved in the same group. In parallel, the histopathological observations demonstrated enhanced apoptosis and absence of mitotic figures in tumor tissues of the co-treatment group. Dual targeting of resistant cancer cells using CSC-DC vaccine along with cisplatin represents a promising therapeutic strategy that could suppress tumor growth, circumvent MDR, and increase the efficacy of conventional chemotherapies.

Assessment of Maximum Bite Force in Oral Submucous Fibrosis Patients: A Preliminary Study

Abstract Objective: To determine the maximum bite force (MBF) in oral submucous fibrosis (OSMF) patients and to compare them with that of healthy subjects. Material and Methods: Twenty patients who were clinically confirmed, as OSMF and 20 healthy controls matched for age, gender, and number of intact functional teeth were included in this study. For each subject, age, gender, weight, height and body mass index (BMI) were recorded. The MBF registration was carried out by the two evaluators, who were previously calibrated. Bite force was measured in the first molar region using a force transducer occlusal force meter for each subject seated at the upright position, with Frankfort's plane nearly parallel to the floor, and no head support. The Student's independent t-test was used to determine the statistical significance in relation to mean height, weight, BMI and the presence of number of intact teeth and MBF between the healthy subjects and OSMF individuals. A comparison of grades of OSMF with all variables was carried out by one-way ANOVA test. Results: No significant difference was found in mean age, mean height, weight, BMI and the presence of the number of intact teeth between healthy individuals and OSMF patients. The mean MBF in healthy subjects was 628.23 ± 24.39 N and 635.47 ± 31.22 N in OSMF patients. Even though the healthy subjects reported a higher MBF than OSMF patients did, the difference was statistically non-significant. With regards to sides, no significant difference was observed in mean MBF in healthy subjects and OSMF patients on the right (p=0.7818) and left side (p=0.6154). Conclusion: The healthy subjects reported higher MBF values than OSMF patients did and the difference was statistically non-significant.

The association of HLA-G polymorphisms and the synergistic effect of sMICA and sHLA-G with chronic kidney disease and allograft acceptance.

The HLA-G and MICA genes are stimulated under inflammatory conditions and code for soluble (sMICA and sHLA-G) or membrane-bound molecules that exhibit immunomodulatory properties. It is still unclear whether they would have a synergistic or antagonistic effect on the immunomodulation of the inflammatory response, such as in chronic kidney disease (CKD), contributing to a better prognosis after the kidney transplantation. In this study, we went from genetic to plasma analysis, first evaluating the polymorphism of MICA, NKG2D and HLA-G in a cohort from Southern Brazil, subdivided in a control group of individuals (n = 75), patients with CKD (n = 94), and kidney-transplant (KT) patients (n = 64). MICA, NKG2D and HLA-G genotyping was performed by polymerase chain reaction with specific oligonucleotide probes, Taqman and Sanger sequencing, respectively. Levels of soluble forms of MICA and HLA-G were measured in plasma with ELISA. Case-control analysis showed that the individuals with haplotype HLA-G*01:01/UTR-4 have a lower susceptibility to develop chronic kidney disease (OR = 0.480; p = 0.032). Concerning the group of kidney-transplant patients, the HLA-G genotypes +3010 GC (rs1710) and +3142 GC (rs1063320) were associated with higher risk for allograft rejection (OR = 5.357; p = 0.013 and OR = 5.357, p = 0.013, respectively). Nevertheless, the genotype +3010 GG (OR = 0.136; p = 0.041) was associated with kidney allograft acceptance, suggesting that it is a protection factor for rejection. In addition, the phenotypic analysis revealed higher levels of sHLA-G (p = 0.003) and sMICA (p < 0.001) in plasma were associated with the development of CKD. For patients who were already under chronic pathological stress and underwent a kidney transplant, a high sMICA (p = 0.001) in pre-transplant proved to favor immunomodulation and allograft acceptance. Even so, the association of genetic factors with differential levels of soluble molecules were not evidenced, we displayed a synergistic effect of sMICA and sHLA-G in response to inflammation. This increase was observed in CKD patients, that when undergo transplantation, had this previous amount of immunoregulatory molecules as a positive factor for the allograft acceptance.

A Cone Beam Computed Tomography Study of the Prevalence of Pulp Stones in a Saudi Arabian Adolescent Population

Objective: To assess the prevalence of pulp stones using Cone Beam Computed Tomography (CBCT) in a Saudi Arabian adolescent population. Material and Methods: CBCT scans of 237 individuals compromising of 1018 teeth were examined. All teeth were analyzed in three dimensions and the presence of pulp stones were identified as a round or oval shaped dense structures in the pulp space. The location of involved tooth in terms of arch, side involved and the status of the tooth were documented. Statistical analysis was carried out by applying Chi-square test. The level of significance was set at 5%. Results: Pulp stones were observed in 119 out of the 237 participants, and in 118 teeth out of 1018 teeth examined, with the patient prevalence of 50.2% and tooth prevalence 10.6%. Pulp stones were observed in 76 (51.3%) of males and 43 (48.3%) of females, with no significant difference (p>0.05). A nonsignificant difference was also noted when maxillary and mandibular arches and the left and right sides were compared. Pulp stones were observed more frequently observed in molars and in carious (8.8%) and restored teeth (17.1%). Conclusion: The prevalence of pulp stones was high in molars and in carious and restored teeth. No significant difference was noticed between gender, arch and side of tooth involved with pulp stones.