Circulating Placental Alkaline Phosphatase Expressing Exosomes in Maternal Blood Showed Temporal Regulation of Placental Genes

Background: Analysis of placental genes could unravel maternal-fetal complications. However, inaccessibility to placental tissue during early pregnancy has limited this effort. We tested if exosomes (Exo) released by human placenta in the maternal circulation harbor crucial placental genes.Methods: Placental alkaline phosphate positive exosomes (ExoPLAP) were enriched from maternal blood collected at the following gestational weeks; 6–8th (T1), 12–14th (T2), 20–24th (T3), and 28th−32nd (T4). Nanotracking analysis, electron microscopy, dynamic light scattering, and immunoblotting were used for characterization. We used microarray for transcriptome and quantitative PCR (qPCR) for gene analysis in ExoPLAP.Results: Physical characterization and presence of CD63 and CD9 proteins confirmed the successful ExoPLAP enrichment. Four of the selected 36 placental genes did not amplify in ExoPLAP, while 32 showed regulations (n = 3–8/time point). Most genes in ExoPLAP showed significantly lower expression at T2–T4, relative to T1 (p < 0.05), such as NOS3, TNFSF10, OR5H6, APOL3, and NEDD4L. In contrast, genes, such as ATF6, NEDD1, and IGF2, had significantly higher expression at T2–T4 relative to T1. Unbiased gene profiling by microarray also confirmed expression of above genes in ExoPLAP-transcriptome. In addition, repeated measure ANOVA showed a significant change in the ExoPLAP transcriptome from T2 to T4 (n = 5/time point).Conclusion: Placental alkaline phosphate positive exosomes transcriptome changed with gestational age advancement in healthy women. The transcriptome expressed crucial placental genes involved in early embryonic development, such as actin cytoskeleton organization, appropriate cell positioning, DNA replication, and B-cell regulation for protecting mammalian fetuses from rejection. Thus, ExoPLAP in maternal blood could be a promising source to study the placental genes regulation for non-invasive monitoring of placental health.

Response of Broiler Chickens to Oral Administration of Ocimum Gratissimum Leaf Extracts

The study was carried out on one hundred and ninety-five day-old broiler chicks of cobb500 to study the effect of Ocimum gratissimum leaf extracts on growth performance, blood profile, microbial population in the faecal and gut samples. The chicks were randomly selected into five groups with thirty-nine chicks per group and each group replicated thrice. The groups were: control (synthetic antibiotics), extracts from 200 g fresh leaf, 400 g fresh leaf, 40 g air-dried leaf and 80 g aid-dried leaf per litre of water respectively. Data collected were subjected to one-way Analysis of Variance. The growth performance except mortality was similar (P>0.05) across the groups. Albumin, urea, cholesterol, alkaline phosphate and sodium were influenced (P<0.05) at the starter phase while only alkaline phosphate was significantly (P<0.05) highest in birds on 400 g of fresh leaf extract at the finisher phase. Neutrophil was highest (P<0.05) in birds on 400 g of fresh leaf extract while those on 200 g had higher values of lymphocytes and eosinophil at starter phase. At finisher phase, birds on antibiotics and 80 g of air-dried leaf extract had higher (P<0.05) white blood cell. Faecal total microbial population was least (<0.05) in birds on antibiotics and 200 g fresh Ocimum gratissimum leaf extract at starter phase. The study concluded that the adoption of Ocimum gratissimum leaf extract as prophylactic treatment against bacteria should be encouraged among poultry farmers.

Fatigue Behavior of an AM50 Die-Casting Alloy Anodized by Plasma Electrolytic Oxidation

While an anodizing process is essential for magnesium alloys to be used under corrosive environments, it sometimes stimulates a fatigue fracture that initiates at the interface between the coating layer and the substrate. In this study, a plasma electrolyte oxidation (PEO) technique was employed to provide excellent adhesion between the anodizing layer and the AM50 die-cast by applying an extremely high dielectric discharge in an alkaline phosphate electrolyte, and its effect on corrosion and fatigue behaviors was investigated. The stress intensity factor at the fatigue limit was estimated to be 0.28 MPam0.5. The specimen anodized using the PEO technique exhibits enhanced strength and corrosion resistance compared to the unanodized counterpart. Furthermore, it shows a relative fatigue life in spite of the thick anodizing layer because the crack initiates from the interface, not from the pore near the interface.

Effects Of Maytenus senegalensis (L) and Cassia alata (L) Extracts on the Liver Enzymes of Albino Rats, Infected with Schistosomes Cercariae

This reaearch aim was to evaluate some liver enzymes of albino rats subjected to administration of methanol and water extracts of Maytenus senegalensis and Cassia alata after infection with schistosome cercariae. As curative agents, the extracts were well tolerated by rats. The total protein (69.0-77.9g/l and 71.8- 79.1g/l) for methanolic extracts of and (64.3-74.3g/l and 70.3- 77.5g/l) for water extracts for M. senegalensis and C. alata respectively. The Alkaline phosphate values range from (145-226iu/l and 189-207iu/l) for methanol and (143-234iu/l and195-207iu/l) for water as for M. senegalensis and C. alata extracts. The Aspartate aminotransferase values were (7.7-8.7iu/l) for M. senegalensis and (8.0- 8.6iu/l) for C. alata with methanol extraxts and (7.1-8.8iu/l) for M. senegalensis and (8.4-8.9iu/) for C. alata water extracts. The Alkaline aminotransferase values range from (5.1-6.3iu/l) for M. senegalensis and (3.5-6.4iu/l) for C. alata in methanol and (4.2-6.3iu/l) for M. senegalensis and (4.1-6.4iu/l for C. alata with water extracts. The Total Bilirubin values of both methanol and water extracts for M. senegalensis (6.3-18.9μm/l) and (9.0-12.7μm/l) for C. alata and (4.2-6.3μm/l) for M. senegalensis and (4.1-6.4μm/l) for C. alata respectively were all within the normal values. This indicates that all these extracts were well tolerated. However the 9g/kg body weight of the water leaf extracts, although having normal values were toxic to other organs such as the lungs.

Safety warning for ChAdOx1 nCov-19 vaccine in patients with sickle cell disease

Vaccines against acute respiratory syndrome Coronavirus 2(SARS-CoV2) are critical weapons to control the spread of the deadly Coronavirus 2019(COVId-19) virus worldwide. Although these vaccines are generally safe, their widespread use has produced reports of rare complications, including vaccine-induced immune thrombotic thrombocytopenia (VIITT), particularly in connection with ChAdOx1 nCov-19. We have identified three cases of sickle cell disease (SCD) experiencing a severe vaso-occlusive crisis (VOC) shortly after the vaccine. Despite being stable for a long time, they had fever with tachycardia, along with a significant rise in WBC, liver enzymes, particularly alkaline phosphate, with a remarkable drop in hemoglobin, and platelets and one of them probably had fatal TTP like syndrome. Given these findings, physicians and patients should exercise caution when taking this type of vaccine and be aware of these safety concerns.

Effect of Allium fistulosum Extracts on the Stimulation of Longitudinal Bone Growth in Animal Modeling Diet-Induced Calcium and Vitamin D Deficiencies

Allium fistulosum is a perennial plant species grown worldwide belonging to the family Liliaceae. In Korean medicine, it is referred to as Chongbaek (CB), and it is prescribed for symptoms associated with the common cold due to its antipyretic properties. This study examined the effects of aqueous (CBW) and 30% ethanol (CBE) extracts on bone growth using a calcium- and vitamin D-deficient animal model. In an in vitro experiment, the alkaline phosphate activities of the extracts were examined using MC3T3-E1 and MG63 cells, and both the aqueous and ethanolic extracts had significant alkaline phosphate activities. In vivo, a serum analysis indicated that the CB extracts promoted bone growth based on the osteogenic markers ALP, calcium, osteocalcin, and collagen type 1 and increased the bone mineral content (BMC), bone mineral density (BMD), and growth plate length. Overall, our results indicate that both CBW and CBE of A. fistulosum can be utilized to facilitate bone growth and increase BMD in children and adolescents by lengthening the growth plate without adverse side effects, such as metabolic disorders or the release of obesity-inducing hormones.

Effect of a Simvastatin-Impregnated Chitosan Scaffold on Cell Growth and Osteoblastic Differentiation

This study aims to evaluate the effect of chitosan (CS) scaffold, alone, and the potential synergistic effect when impregnated with simvastatin (SIM), on immortalized human bone-marrow mesenchymal stem cells (hbMMSCs) compared to CollaCote (CL). CS scaffolds were fabricated and seeded with immortalized hBMMSCs. Samples were divided into control groups (negative with no added material and positive with CL added) and four experimental groups: CS alone, CS/SIM 0.01, 0.03, and 0.05 mg, respectively. Cell viability, osteoblastic differentiation and calcium deposition were investigated via AlamarBlue, alkaline phosphate activity assays and alizarin red S staining at 1 and 14 days, respectively. At day one, no significant difference was noted between the groups regarding cell viability. However, all CS/SIM groups showed significant cutback at day 14 in cell proliferation compared to CS alone and CL groups (p < 0.001). All groups supported osteoblastic differentiation with no significant difference. Alkaline phosphate activity increased in both time periods in the CS/SIM 0.05 mg group compared to the other SIM groups, with no significant difference among the experimental groups. Chitosan scaffold is a bioactive compatible material capable of regenerative potential of hBMMSCs and a promising material to be used for perforation repair.

Estimation of Alanine Transaminase (ALT), Alkaline Phosphate (ALP) and Aspartate (AST) Irrespective of Dietary Supplementation, Body Mass Index and Nature of Exercise

Purpose: This research study was conducted for the assessment of liver enzymes i.e. Alanine Transaminase (ALT), Alkaline Phosphate (ALP) and Aspartate (AST) irrespective of dietary supplementation, body mass index and nature of the exercise. Methods: A randomized control trails were taken and thus Sixty (60) subjects ageing 20 to 30 years, (Twenty (20) from low-intensity exercise (EXG-II), Twenty (20) from high-intensity exercise (EXG-III) and Twenty (20) subjects as a control group (CG-III) were included in the study by using International Physical Activity Scale (IPAQ).5ml blood samples were collected from the subjects for the determination of liver functions and blood redox status. Each blood sample was marked with a separate identification mark. After the collection of blood samples, three particular liver enzymes tests i.e. ALT, ALP and AST were performed. The results of the samples test proceeded through the Statistical Package for Social Science (SPSS) Version 23. Results: The P-Value of the low-intensity exercise group (EXG-II) in term of ALT was less than the high-intensity exercise group ( EXG-III) ( p >.05), The P-Value of the low-intensity exercise group (EXG-II) in term of ALP was less than the high-intensity exercise group ( EXG-III) ( p >.05). The P-Value of low-intensity exercise group (EXG-II) in term of AST was less than high-intensity exercise group (EXG-III) (p >.05). Conclusion: Based on findings, it was concluded that low-intensity exercise with dietary supplementation positively affects the functions of the liver i.e. Alanine Transaminase (ALT), Alkaline Phosphate (ALP) and Aspartate (AST) as compared to high-intensity exercise. Similarly high-intensity exercise with dietary supplementation also positively influence as compared to performers of high-intensity exercise without dietary supplementation.

An Uncommon Case of Hypophophatasia Presenting With Galactorrhea, Diagnosed at a Later Age Than Expected

Hypophosphatasia is a rare genetic disorder which causes accumulation of inorganic pyrophosphate that in turn inhibits mineralization. It is caused by a mutation in the non-specific alkaline phosphatase gene (TNSALP). We will review the diagnosis during adulthood when the presentation is characterized by poor healing, recurrent metatarsal stress fractures and bone pain. We are reporting the case of a 38-year-old white female with history of Melanoma referred to the Pituitary Clinic due to galactorrhea. During the encounter it was noted that she had history of disseminated joint pain, myalgias, periodontal complications as well as biochemical evidence of low alkaline phosphatase. At age three she had premature teeth loss, and clumsy gait, which later resolved. She was evaluated for this and diagnosed with odontohypophophatasia at which time no treatment was initiated. As an adult patient had two non-traumatic ankle fractures with associated frail teeth, chipping, and cracking without significant trauma. At around 36 years of age, patient started developing worsening joint pain, stiffness, myalgias and 35 pounds weight gain. Family History is remarkable for her father, having history of low alkaline phosphate and odontogenic infections with no joint or muscle pain. Paternal grandmother had history of dental infections and frail teeth. The patient has four sons; her oldest son has low alkaline phosphate with no clinical symptoms. Patient’s biochemical analysis was unremarkable except for low alkaline phosphate range between 25–38 U/L (40–125 U/L) and elevated Vitamin B6 69.5 ng/mL (2.1–21.7 ng/mL). The physical exam was unremarkable. Brain MRI showed a 3 mm microadenoma with normal pituitary hormonal workup, including diluted prolactin. Mammogram did not show any abnormalities. It was determined that the pituitary microadenoma was non-functioning, and galactorrhea was not hormonal mediated. The patient was referred to a genetic specialist for evaluation of Hypophosphatasia. Testing for TNSALP mutation was done which showed an ALP Mutation variant c.1172G&gt;A (p. Arg391His). She was started on asfotase alfa (Strensiq) an enzyme replacement for Hypophosphatasia, in November 2019. Hypophosphatasia is a rare condition often diagnosed during early life. Due to wide number of possible mutations in the TNSALP gene, the clinical presentation can vary between patients and diagnosis can be elusive. In our patient, despite being diagnosed with odontohypophophatasia at an early age, and having many periodontal complications, hypophosphatasia was not confirmed until more than 30 years after the initiation of symptoms. In the management of Hypophosphatasia, having an accurate timely diagnosis can provide patients with early intervention of advanced treatment modalities such as Strensiq that can improve symptoms.