Group IIA Research Articles

PLA2G2A Phospholipase Promotes Fatty Acid Synthesis and Energy Metabolism in Pancreatic Cancer Cells with K-ras Mutation.

Oncogenic K-ras is often activated in pancreatic ductal adenocarcinoma (PDAC) due to frequent mutation (>90%), which drives multiple cellular processes, including alterations in lipid metabolism associated with a malignant phenotype. However, the role and mechanism of the altered lipid metabolism in K-ras-driven cancer remains poorly understood. In this study, using human pancreatic epithelial cells harboring inducible K-rasG12D (HPNE/K-rasG12D) and pancreatic cancer cell lines, we found that the expression of phospholipase A2 group IIA (PLA2G2A) was upregulated by oncogenic K-ras. The elevated expression of PLA2G2A was also observed in pancreatic cancer tissues and was correlated with poor survival of PDAC patients. Abrogation of PLA2G2A by siRNA or by pharmacological inhibition using tanshinone I significantly increased lipid peroxidation, reduced fatty acid synthase (FASN) expression, and impaired mitochondrial function manifested by a decrease in mitochondrial transmembrane potential and a reduction in ATP production, leading to the inhibition of cancer cell proliferation. Our study suggests that high expression of PLA2G2A induced by oncogenic K-ras promotes cancer cell survival, likely by reducing lipid peroxidation through its ability to facilitate the removal of polyunsaturated fatty acids from lipid membranes by enhancing the de novo fatty acid synthesis and energy metabolism to support cancer cell proliferation. As such, PLA2G2A might function as a downstream mediator of K-ras and could be a potential therapeutic target.

Analysis of changes in matrix metalloproteinases-9 and tissue inhibitors-1 levels in newly diagnosed pulmonary tuberculous with different profile of drug-resistance

Objective — to analyze changes in MMP-9 and TIMP-1 parameters in patients with newly diagnosed destructive pulmonary TB with different pathogen resistance profile depending on the course of the tuberculosis process.
 Materials and methods. The study included 124 patients with newly diagnosed pulmonary TB with destruction and bacterial excretion, who were treated according to the order of the Ministry of Health of Ukraine N 620 in 2014—2017. The patients were divided into 2 groups: Group I (n = 84) — multidrug-resistant TB (MDR-TB); Group II (n = 40) — drug-susceptible TB. Groups Ia (n = 56) and IIa (n = 28) included patients with effective treatment; groups Ib (n = 28) and IIb (n = 12) included patients with ineffective treatment. The level of MMP-9 and TIMP-1 was studied in blood serum by ELISA. Blood was taken in the morning on an empty stomach at the beginning of treatment, after 2 and 3 months.
 Results and discussion. The level of MMP-9 increased in both groups for 3 months. However, the dynamics of the increase was more intensive in the group of drug-susceptible TB. The level of TIMP-1 also increased within 3 months in both groups. This dynamics was more intensive in Group II. In the course of treatment, more intensive dynamics of the ratio decrease MMP-9/TMP-1 was observed in Group II with a decrease of 25 % compared with 16.6 % in Group I.In the group of patients with MDR-TB, the ratio of MMP-9/TIMP-1 at the beginning of treatment was lower in Group Ia than in Group Ib (p < 0.001). Its decrease within 3 months was more pronounced in Group Ib (by 25 %) compared with Group Ia (7.5 %). In the group of patients with drug-susceptible tuberculosis at the beginning of treatment, higher levels of the MMP-9/TIMP-1 ratio were observed in Group IIb. Against the background of anti-TB treatment for 3 months. There was also a decrease in the ratio in both groups (IIa and IIb). However, the dynamics of the decrease was more pronounced in Group IIb (by 41 %), compared with group IIa (by 10 %).
 Conclusions. In dynamics, against the background of antimycobacterial therapy, a more rapid onset of lung healing processes was observed in patients with drug-susceptible pulmonary TB compared with patients with MDR-TB. This is evidenced by the obtained indicators of MMP-9, TIMP-1 and their ratio. Thus, the level of TIMP-1, which is an inhibitor of MMP-9, was higher after 3 months of treatment and showed more intensive growth dynamics in Group II, that is, in patients with drug-susceptible TB, which indicates a favorable course of the disease in this group, and an earlier onset of healing processes of lung tissue destruction.When patients were divided into groups of effective and ineffective treatment, a more intensive decrease in the ratio of MMP-9 to TIMP-1 was observed in patients with an unfavorable course of the tuberculosis process, which indicates more significant changes in the connective tissue of the lungs in patients from these groups.

Hydrolysis of polyhydroxy polyunsaturated fatty acid-glycerophosphocholines by Group IIA, V, and X secretory phospholipases A2.

It is widely accepted that unesterified polyunsaturated ω-6 and ω-3 fatty acids (PUFA) are converted through various lipoxygenases, cyclooxygenases, and cytochrome P450 enzymes to a range of oxygenated derivatives (oxylipins), among which the polyhydroxides of unesterified PUFA have recently been recognized as cell signaling molecules with anti-inflammatory and pro-resolving properties, known as specialized pro-resolving mediators (SPMs). This study investigates the mono-, di-, and trihydroxy 16:0/PUFA-GPCs, and the corresponding 16:0/SPM-GPC, in plasma lipoproteins. We describe the isolation and identification of mono-, di-, and trihydroxy AA, EPA, and DHA-GPC in plasma LDL, HDL, HDL3, and acute phase HDL using normal phase LC/ESI-MS, as previously reported. The lipoproteins contained variable amounts of the polyhydroxy-PUFA-GPC (0-10nmol/mg protein), likely the product of lipid peroxidation and the action of various lipoxygenases and cytochrome P450 enzymes on both free fatty acids and the parent GPCs. Polyhydroxy-PUFA-GPC was hydrolyzed to variable extent (20%-80%) by the different secretory phospholipases A2 (sPLA2 s), with Group IIA sPLA2 showing the lowest and Group X sPLA2 the highest activity. Surprisingly, the trihydroxy-16:0/PUFA-GPC of APHDL was largely absent, while large amounts of unidentified material had migrated in the free fatty acid elution area. The free fatty acid mass spectra were consistent with that anticipated for branched chain polyhydroxy fatty acids. There was general agreement between the masses determined by LC/ESI-MS for the polyhydroxy PUFA-GPC and the masses calculated for the GPC equivalents of resolvins, protectins, and maresins using the fatty acid structures reported in the literature.

Assessing sexual dysfunction in patients with external genital endometriosis

Introduction. External genital endometriosis (EGE) is a chronic, relapsing disease that affects various aspects of sexual function, the quality of sexual and interpersonal relationships, physical and psycho-emotional health and leads to social maladjustment and anxiety-depressive disorders, resulting in progressively decreased quality of life (QoL).Aim: to analyze an impact of sexual disorders on QoL of women with painful and painless EGEs.Materials and Methods. A prospective comparative randomized interventional study involved 160 patients (110 and 50 women with painful and painless EGE, respectively) of reproductive age, with laparoscopically and morphologically verified diagnosis according to the revised American Fertility Society scoring system (R-AFS, 1996). Pain syndrome and sexual dysfunction were assessed using specialized questionnaires: visual analogue scale (VAS), PainDETECT, and Female Sexual Function Index (FSFI). After surgical treatment, 8 patients refused to participate in the study (6 with painful EGE and 2 with painless EGE). Thus, only 152 women passed all stages of the study. The patients were divided into 4 groups depending on the presence and/or absence of pain syndrome and the tactics of the rehabilitation period ("active" or "passive"): group IA consisted of 49 (47.1 %) women with pain and "active" rehabilitation; group IБ – 55 (52.9 %) women with pain syndrome and "passive" rehabilitation tactics; group IIA – 23 (47.9 %) patients without pain syndrome with "active" rehabilitation; group IIБ – 25 (52.1 %) patients without pain syndrome with "passive" rehabilitation tactics.Results. Women with painful vs. painless EGE were found to have a more severe course of the disease and a markedly decreased quality of sexual life. It was observed that patients with "active" rehabilitation tactics (groups IA and IIA) had intensity of the pain syndrome and the neuropathic pain component significantly decreased, improved parameters of the quality of sexual life, sexual desire and sexual excitability compared to patients with "passive" rehabilitation tactics applied with the measures within the framework of the National Clinical Guidelines (groups IБ and IIБ).Conclusion. A significantly decreased quality of sexual life of EGE patients indicates a need for assessing and correcting sexual disorders as well as introducing early onset of rehabilitation measures by using a multidisciplinary approach.

Post-transplantation cyclophosphamide as graft versus host disease prophylaxis in patients with acute leukemia received fully matched allogeneic HSCT or haplo-identical HSCT

Post-transplant Cyclophosphamide (PT-Cy) has proved efficacy as GVHD prophylaxis regimen after HSCT. However, experiences are limited with controversial results. We herein, assess the efficacy of PT-Cy compared to Methotrexate regimen. Eighty patients with acute leukemia received a fully matched allogeneic HSCT or a Haplo-identical HSCT were analyzed. Group I (Historical group) included 40 patients received Methotrexate, Cyclosporine and MMF. They were transplanted with fully matched allogeneic transplantation. Group II included 40 patients received PT-Cy, Cyclosporine and MMF. They were subdivided to 2 subgroups. Subgroup IIA included 22 patients received fully matched allo- HSCT and subgroup IIB included 18 patients received Haplo- HSCT. Group IIA showed significantly lower incidence of cGVHD when compared to group I with an incidence of 22.7% and 67% respectively (P = 0.002). Group IIA was associated with reduced risk of extensive cGVHD compared to MTX group (P =0.003). No significant differences were found in the incidence of aGVHD, relapse rates, relapse or non-relapse related mortalities, OS and DFS data among the different groups (P>0.05). In conclusion, PT-Cy with addition of IS drugs has statistically significant difference in reducing the incidence of cGvHD in both fully matched and Haplo SCT with less hepatic and renal toxicity.

Multimodal regulation of the osteoclastogenesis process by secreted group IIA phospholipase A2.

Increasing evidence points to the involvement of group IIA secreted phospholipase A2 (sPLA2-IIA) in pathologies characterized by abnormal osteoclast bone-resorption activity. Here, the role of this moonlighting protein has been deepened in the osteoclastogenesis process driven by the RANKL cytokine in RAW264.7 macrophages and bone-marrow derived precursor cells from BALB/cJ mice. Inhibitors with distinct selectivity toward sPLA2-IIA activities and recombinant sPLA2-IIA (wild-type or catalytically inactive forms, full-length or partial protein sequences) were instrumental to dissect out sPLA2-IIA function, in conjunction with reduction of sPLA2-IIA expression using small-interfering-RNAs and precursor cells from Pla2g2a knock-out mice. The reported data indicate sPLA2-IIA participation in murine osteoclast maturation, control of syncytium formation and resorbing activity, by mechanisms that may be both catalytically dependent and independent. Of note, these studies provide a more complete understanding of the still enigmatic osteoclast multinucleation process, a crucial step for bone-resorbing activity, uncovering the role of sPLA2-IIA interaction with a still unidentified receptor to regulate osteoclast fusion through p38 SAPK activation. This could pave the way for the design of specific inhibitors of sPLA2-IIA binding to interacting partners implicated in osteoclast syncytium formation.

Association between body mass index and fecal calprotectin levels in children and adolescents with irritable bowel syndrome.

Irritable bowel syndrome (IBS) is a common pediatric functional gastrointestinal disorder. It is characterized by recurrent abdominal pain and changes in bowel habits and is more prevalent in obese patients. We investigated the association between obesity and IBS in pediatric patients through fecal calprotectin testing.Patients under 18 years of age with IBS who underwent fecal calprotectin testing from January 2015 through April 2020 were retrospectively investigated. The patients were divided into groups based on body mass index (BMI): group I (BMI < 85th percentile) and group II (BMI ≥ 85th percentile). Group II was divided into group IIa, overweight (85th percentile ≤ BMI < 95th percentile), and group IIb, obese (BMI ≥ 95th percentile).Among 277 included patients, 202 (72.9%) were in group I, and 75 (27.1%) were in group II (mean calprotectin levels, 75.60 ± 103.48 vs 45.89 ± 66.57 µg/g, respectively; P = .006). There were significant differences in mean calprotectin levels between groups I and IIa (75.60 ± 103.48 vs 45.45 ± 63.38 µg/g, respectively; P = .028) and groups I and IIb (75.60 ± 103.48 vs 46.22 ± 69.59 µg/g, respectively; P = .025). There was a significant difference in mean calprotectin levels between groups I and II (85.69 ± 142.13 vs 32.04 ± 28.17 µg/g, respectively; P = .029) among patients between 6 and 12 years of age but not among adolescents aged between 12 and 18 years (P = .139). Fecal calprotectin was lower when moderate-to-severe fatty livers were observed by ultrasound compared with normal livers (68.52 ± 97.22 vs 18.53 ± 18.56 µg/g, respectively; P = .017).Fecal calprotectin levels were higher in normal-weight pediatric IBS patients than in their obese counterparts, and this difference was more prominent in younger patients. In young children, IBS symptoms are thought to be influenced more by factors other than intestinal inflammation.

Identification of a Dual Inhibitor of Secreted Phospholipase A2 (GIIA sPLA2) and SARS-CoV-2 Main Protease.

The development of novel agents to combat COVID-19 is of high importance. SARS-CoV-2 main protease (Mpro) is a highly attractive target for the development of novel antivirals and a variety of inhibitors have already been developed. Accumulating evidence on the pathobiology of COVID-19 has shown that lipids and lipid metabolizing enzymes are critically involved in the severity of the infection. The purpose of the present study was to identify an inhibitor able to simultaneously inhibit both SARS-CoV-2 Mpro and phospholipase A2 (PLA2), an enzyme which plays a significant role in inflammatory diseases. Evaluating several PLA2 inhibitors, we demonstrate that the previously known potent inhibitor of Group IIA secretory PLA2, GK241, may also weakly inhibit SARS-CoV-2 Mpro. Molecular mechanics docking and molecular dynamics calculations shed light on the interactions between GK241 and SARS-CoV-2 Mpro. 2-Oxoamide GK241 may represent a lead molecular structure for the development of dual PLA2 and SARS-CoV-2 Mpro inhibitors.

Epiphysiodesis of the first ray as a method of treat-ment of adolescent hallux valgus

Presentation of short-term results from the application of epiphysiodesis of the first ray as a method of treatment of adolescent hallux valgus.&#x0D; Material and method: The operative technique was introduced in USHATO in January 2018 like a operative procedure for treatment of mild and moderate deformity of adolescent hallux valgus (AHV). The technique includes percutaneous access to the growth zones in the area of ​​the first metatarsal bone and proximal phalanx and partial epiphysiodesis of the growth complex. The study presents 12 cases in 9 children with a mean age of 10.1 years (min. 5 years, max. 13 years). Most of the patients were female - 77.78%. According to the three-dimensional Stantrock classification and the HV severity classification, the twelve cases fell into group II-A and moderate severity. Three angular parameters hallux valgus angle(HVA), intra-metatarsal angle (IMA), distal-metatarsal articulation angle (DMAA), were used to monitor the results, and the measurements were performed preoperatively, intraoperatively and postoperatively.&#x0D; The American Orthopaedic Foot and Ankle Society (AOFAS), was used to report clinical outcomes.&#x0D; Results: Patients were followed up for a mean period of 17.58 months. For this period we do not established postoperative recurrences, and all cases remain in the same group according Stantrock classification and the HV severity classification.&#x0D; In the three monitored radiographic parameters we report clinical, but not statistical improvement: HVA by an average of 1.19°, IMA by an average of 0.55° and loss of correction in DMAA by 0.9°. For the follow-up period, we report a statistical improvement in the clinical parameters reported on the AOFAS scale (p &lt;0.4).&#x0D;

Phenotypic and Genotypic Characterization of Phytophthora infestans Isolates Associated with Tomato and Potato Crops in Colombia.

Late blight disease, caused by the plant pathogen Phytophthora infestans, is one of the major threats for tomato and potato crops. Monitoring the populations of P. infestans is important to determine if there are changes in the sensitivity to fungicides and host preference. In this study, microsatellite markers and mitochondrial haplotypes were used to assess the genotype of isolates of P. infestans collected from tomato and potato plants in Colombia. Furthermore, sensitivity to the three fungicides cymoxanil (penetrant fungicide), mefenoxam, and fluopicolide (systemic fungicides), and tomato-potato host preference, were evaluated. Mitochondrial haplotyping showed that isolates collected on tomato were from the genetic groups Ia and Ib, while isolates collected on potatoes belonged to group IIa. Microsatellite analyses showed that isolates from tomato form two groups, including the Ib mitochondrial haplotype (which is genetically close to the US-1 clonal lineage) and the Ia haplotype (related to the EC-3 lineage), whereas Colombian isolates from potato formed a separate group. Furthermore, differences in sensitivity to fungicides were observed. Eighty-one percent of the isolates tested were resistant to mefenoxam with an EC50 >10 μg ml-1. Forty-two percent of the isolates showed an intermediate resistance to cymoxanil. The EC50 values ranged between 1 and 10 μg ml-1. For fluopicolide, 90% of the isolates were sensitive, with EC50 <1 μg ml-1. Host preference assays showed that potato isolates infected both host species. Thus, isolates that infect potatoes may pose a risk for tomato crops nearby.

Sinapicacid Inhibits Group IIA Secretory Phospholipase A2 and Its Inflammatory Response in Mice.

Human Group IIA secreted phospholipase A2 (sPLA2-IIA) enzyme plays a crucial role in several chronic inflammatory diseases such asasthma, atherosclerosis, gout, bronchitis, etc. Several studies showed that the antioxidants exert an anti-inflammatory function by inhibiting the sPLA2-IIA enzyme. Hence, the present study evaluated an antioxidant molecule, sinapic acid, for sPLA2-IIA inhibition as an anti-inflammatory function. Initially, the antioxidant efficacy of sinapic acid was evaluated, and it showed greater antioxidant potency. Further, sinapic acid inhibited 94.4 ± 4.83% of sPLA2-IIA activity with an IC50 value of 4.16 ± 0.13 µM. The mode of sPLA2-IIA inhibition was examined by increasing the substrate concentration from 30 to 120nM and the calcium concentration from 2.5 to 15 mM, which did not change the level of inhibition. Further, sinapic acid altered the intrinsic fluorescence and distorted the far UltraViolet Circular Dichroism (UV-CD) spectra of the sPLA2-IIA, indicating the direct enzyme-inhibitor interaction. Sinapic acid reduced the sPLA2-IIA mediated hemolytic activity from 94 ± 2.19% to 12.35 ± 2.57% and mouse paw edema from 171.75 ± 2.2% to 114.8 ± 1.98%, demonstrating the anti-inflammatory efficiency of sinapic acid by in situ and in vivo methods, respectively. Finally, sinapic acid reduced the hemorrhagic effect of Vipera russelli venom hemorrhagic complex-I (VR-HC-I) as an anti-hemorrhagic function. Thus, the above experimental results revealed the sinapic acid potency to be an antioxidant, anti-inflammatory and anti-hemorrhagic molecule, and therefore, it appears to be a promising therapeutic agent.

MeWRKY IIas, Subfamily Genes of WRKY Transcription Factors From Cassava, Play an Important Role in Disease Resistance.

Cassava (Manihot esculenta Crantz) is an important tropical crop for food, fodder, and energy. Cassava bacterial blight (CBB) caused by Xanthomonas axonopodis pv. manihotis (Xam) occurs in all cassava growing regions and threatens global cassava production. WRKY transcription factor family plays the essential roles during plant growth, development, and abiotic or biotic stress. Particularly, previous studies have revealed the important role of the group IIa WRKY genes in plant disease resistance. However, a comprehensive analysis of group IIa subfamily in cassava is still missing. Here, we identified 102 WRKY members, which were classified into three groups, I, II, and III. Transient expression showed that six MeWRKY IIas were localized in the nucleus. MeWRKY IIas transcripts accumulated significantly in response to SA, JA, and Xam. Overexpression of MeWRKY27 and MeWRKY33 in Arabidopsis enhanced its resistance to Pst DC3000. In contrast, silencing of MeWRKY27 and MeWRKY33 in cassava enhanced its susceptibility to Xam. Co-expression network analysis showed that different downstream genes are regulated by different MeWRKY IIa members. The functional analysis of downstream genes will provide clues for clarifying molecular mechanism of cassava disease resistance. Collectively, our results suggest that MeWRKY IIas are regulated by SA, JA signaling, and coordinate response to Xam infection.

Quality of life and psychoemotional status in patients with external genital endometriosis

Introduction. Chronic pain syndrome with external genital endometriosis (EGE) causes profound psychoemotional changes, anxiety disorders, decreased physical and social activity, general well-being and mood, sexual dysfunction, and as a consequence, deteriorated quality of patients' life.Aim: to study an influence of psychoemotional disorders on the quality of life of women with painful and painless forms of EGE.Material and Methods. There were enrolled 160 patients (110 women with painful and 50 with painless forms of EGE) of reproductive age with laparoscopically and morphologically verified diagnosis, according to the Revised American Fertility Society score (R-AFS) classification. Pain syndrome and psychoemotional status were assessed using various quality of life assessment questionnaires: Visual Analog Scale (VAS), Endometriosis Health Profile-30 (EHP-30), Hospital Anxiety and Depression Scale (HADS). After surgical treatment, 8 patients refused to participate in the work (6 with painful form of EGE and 2 with painless form). Thus, 152 women passed all stages of the study. The patients were divided into 4 groups depending on the presence and/or absence of pain syndrome and tactics of the rehabilitation period ("active" or "passive"): group IA consisted of 49 (47.1 %) women with pain syndrome and "active" rehabilitation; group IБ – 55 (52.9 %) women with pain syndrome and "passive" rehabilitation tactics; group IIA – 23 (47.9 %) patients without pain syndrome and with "active" rehabilitation; group IIБ – 25 (52.1 %) patients without pain syndrome and with "passive" rehabilitation tactics.Results. It was found that painful vs. painless form of EGE differs by a more severe course of the disease and markedly decreased quality of life. In a comparative analysis of the four groups it was noted that women with "active" tactics of rehabilitation measures (IA and IIA) had a decrease in pain syndrome, improved emotional, social and sexual activity, decreased depression and anxiety scores as compared to the patients (IБ and IIБ) who underwent a set of rehabilitation measures within the National clinical guidelines.Conclusion. Timely diagnostics and correction of psychoemotional disorders will allow to develop a differentiated approach to provide specialized medical and psychological care and improve the quality of life of patients with EGE.

Genome-Wide Analysis and Exploration of WRKY Transcription Factor Family Involved in the Regulation of Shoot Branching in Petunia.

The WRKY transcription factors (TFs) participate in various physiological, growth and developmental processes of plants. In our study, a total of 79 WRKY family members were identified and classified into three groups (Group I, Group IIa–e, and Group III) based on phylogenetic and conservative domain analyses. Conserved motif analysis showed that seven WRKYGQK domains changed. The promoter sequence analysis suggested that there were multiple stress- and hormone-related cis-regulatory elements in the promoter regions of PhWRKY genes. Expression patterns of PhWRKYs based on RNA-seq data revealed their diverse expression profiles in five tissues and under different treatments. Subcellular localization analysis showed that PhWRKY71 was located in the nucleus. In addition, overexpression of PhWRKY71 caused a significant increase in branch number. This indicated that PhWRKY71 played a critical role in regulating the shoot branching of Petunia hybrida. The above results lay the foundation for further revealing the functions of PhWRKY genes.

Group IIA secreted phospholipase A2 inhibition by elemolic acid as a function of anti-inflammatory activity

Human group IIA secreted phospholipase A2 (GIIA) is a key enzyme in inflammatory reactions, worsening the condition of several chronic inflammatory diseases. The natural inhibitors of GIIA potentially block the production of inflammatory mediators. In the present study, elemolic acid, a triterpenoid from Boswellia serrata inhibited the GIIA enzyme in a concentration-dependent manner with IC50 value of 5.70 ± 0.02 µM. The mode of GIIA inhibition was studied by increasing the concentration of the substrate from 30 to 120 nM, and calcium from 2.5 to 15 mM, the level of inhibition was not changed. The inhibitor-enzyme interaction was examined by fluorimetry and Circular Dichroism (CD) studies; elemolic acid altered intrinsic fluorescence intensity and shifted far UV- CD spectra of GIIA enzyme, suggesting the direct interaction with GIIA. Elemolic acid neutralized the GIIA mediated indirect hemolytic activity from 94.5 to 9.8% and reduced GIIA induced mouse paw edema from 171.75 to 113.68%. Elemolic acid also reduced the hemorrhagic effect of GIIA along with Vipera russelii neurotoxic non-enzymatic peptide -VNTx-II (VR-HC-I). Thus, the elemolic acid has been proven as a potent inhibitor of GIIA enzyme and modulated the GIIA induced inflammatory response by in situ and in vivo methods.

Beneficial effects of ascorbic acid and calcium in fluoride induced changesin some biochemical parameters in testes of Swiss Albino Mice

The present investigation was undertaken to elucidate the effects of sodium fluoride on mice understanding the mechanism of fluoride action on metabolism of testis through sequential biochemical changes in level of different parameters and its reversibility by ascorbic acid or calcium and combined effects of both ascorbic and calcium. The healthy, adult Swiss albino mice (Mus musculus) were divided into two equal groups. Group I (fed on standard diet), Group IIA (NaF 5ppm and 50ppm), Group IIB (withdrawal of treatment), Group II C Recovery was also observed in with simultaneously administration of A.A +Ca respectively. Animals from each group were autopsied by cervical dislocation at each post treatment intervals of 10, 20, and 30 days. The changes in the biochemical parameters were dose and duration dependent. Total protein, acid phosphatase, ascorbic acid DNA and RNA decreased while, alkaline phosphatase, showed an increase in all experimental groups. Whereas after the cessation of the treatment recovery was observed and it was more pronounced with ascorbic acid+ calcium. The experimental findings indicate that both testicular protein and DNA synthesis decreased suggesting that fluoride in some way interfere with the RNA metabolism and consequently interfere with the synthesis of specific testicular enzymes etc.

Old but New: Group IIA Phospholipase A2 as a Modulator of Gut Microbiota.

Among the phospholipase A2 (PLA2) superfamily, the secreted PLA2 (sPLA2) family contains 11 mammalian isoforms that exhibit unique tissue or cellular distributions and enzymatic properties. Current studies using sPLA2-deficient or -overexpressed mouse strains, along with mass spectrometric lipidomics to determine sPLA2-driven lipid pathways, have revealed the diverse pathophysiological roles of sPLA2s in various biological events. In general, individual sPLA2s exert their specific functions within tissue microenvironments, where they are intrinsically expressed through hydrolysis of extracellular phospholipids. Recent studies have uncovered a new aspect of group IIA sPLA2 (sPLA2-IIA), a prototypic sPLA2 with the oldest research history among the mammalian PLA2s, as a modulator of the gut microbiota. In the intestine, Paneth cell-derived sPLA2-IIA acts as an antimicrobial protein to shape the gut microbiota, thereby secondarily affecting inflammation, allergy, and cancer in proximal and distal tissues. Knockout of intestinal sPLA2-IIA in BALB/c mice leads to alterations in skin cancer, psoriasis, and anaphylaxis, while overexpression of sPLA2-IIA in Pla2g2a-null C57BL/6 mice induces systemic inflammation and exacerbates arthritis. These phenotypes are associated with notable changes in gut microbiota and fecal metabolites, are variable in different animal facilities, and are abrogated after antibiotic treatment, co-housing, or fecal transfer. These studies open a new mechanistic action of this old sPLA2 and add the sPLA2 family to the growing list of endogenous factors capable of affecting the microbe–host interaction and thereby systemic homeostasis and diseases.

Comparative evaluation of micro-leakage and shear bond strength between a self adhesive flowable composite resin and self etch based flowable composite restorative material

Background: An in vitro evaluation and comparing the microleakage and shear bond strength between a self- adhesive flowable composite resin (Dyad Flow) and self-etch based flowable composite restorative material ( Nano Filtek Z 350XT) Materials and methods: A total of sixty specimens were prepared from extracted teeth, which were divided into two groups. For microleakage analysis, Group I consisting of thirty teeth which were further divided into two subgroups of fifteen teeth each that is Group IA and Group IB, on which standard class V cavity was prepared and restored with Dyad Flow and Nano Filtek Z 350XT respectively. The prepared specimens were immersed in 0.6% aqueous Rhodamine B dye for 24 hours. The sectioned samples were examined under a Confocal Microscope at 10 X magnification and microleakage scores were analyzed statistically using Mann-Whitney U test. For shear bond strength evaluation, Group II consisting of thirty teeth, which were subdivided into two subgroups of fifteen teeth each that is Group IIA and Group IIB.

Transforming growth factors TGF- β1, TGF- β2 and TGF- β3 in the tissue of nasal polyps in different phenotypes of chronic rhinosinusitis with nasal polyps

Chronic rhinosinusitis with nasal polyps (CRSP) is a heterogenous disease. We have earlier detected differences in severity of clinical manifestations, cellular infiltration degree shown in nasal polyps, of eosinophil-to-neutrophil ratio, efficacy of intranasal glucocorticosteroid baseline therapy, and various inflammatory patterns for several cytokines on the mRNA expression level in different phenotypes with isolated CRSP cases, CRSP associated with respiratory allergy (RA), or non-allergic bronchial asthma.The purpose of this work was to study the cytokines of TGF-â family in the tissues of nasal polyps in patients with different CRSP phenotypes. The research involved 292 patients suffering from CRSP divided into 3 phenotypic groups. Group I consisted of patients with isolated CRSP free of associated BA and/or sensitization to atopic allergens. Group II included patients with CRSP combined with respiratory allergy and was further divided into two subgroups. I.e., Group 2a comprised patients with CRSP, allergic BA (aBA), and allergic rhinitis (AR), while the patients with CRSP, AR, and non-allergic BA were placed to the group 2b. The patients suffering from CRSP complicated with non-allergic BA were allocated to the group III. The patients with hypertrophic rhinitis served as control. The levels of TGF-â1, TGF-â2, and TGF-â3 proteins (pg/mg) were measured by means of multiplex immunoassay approach in supernates of tissue homogenates from nasal polyps removed by surgery, and in posterior parts of inferior nasal conchae. The total protein level was determined in tissue supernatant, with cytokine contents recalculated for the mg/ml protein concentration for standardization of measurements.In the control group, trace concentrations of all three growth factors were detected. Significant difference in protein contents was found for the studied cytokines, depending on CRSP phenotype. The levels of TGF-â1 and TGF-â2 were statistically lower in isolated CRSP than in other groups of comorbid CRSP patients. TGF-â1 and TGF-â2 concentrations were significantly lower in CRSP + allergic BA group IIa than in CRSP + nonallergic BA and CRSP + RA groups. The amount of TGF-â3 cytokine was maximal in CRSP + non-allergic BA group III compared to the patients with isolated CRSP of group I and CRSP + non-allergic BA group 2a.Conclusions.The high level of all three TGF-â isoforms in patients with CRSP compared to the control group suggested a high potential of mucous membranes of paranasal sinuses for active tissue remodeling followed by nasal polype formation.Different mechanisms were presumed for development of local pathological process in different clinical phenotypes of CRSP, depending on the comorbid pathology, especially, BA or respiratory disorders.Minimal TGF-â1 and TGF-â2 levels were detected in isolated CRSP.The highest concentrations of TGF-â1, TGF-â2, and TGF-â3 were discovered in the patients with CRSP accompanied by non-allergic BA as compared to the groups with isolated CRSP and CRSP+allergic BA.5. Determination of TGF-â1, TGF-â2, and TGF-â3 levels can serve as an additional criterion for differentiating between the mechanisms of mucous membrane damage in local pathological process in tissues of comorbid patients with different CRSP phenotypes.

Creatine kinase-(MB) and hepcidin as candidate biomarkers for early diagnosis of pulmonary tuberculosis: a proof-of-concept study in Lambaréné, Gabon

BackgroundThe present study aimed to evaluate the diagnostic utility of creatine kinase-MB (CK-MB), hepcidin (HEPC), phospholipase A2 group IIA (PLa2G2A), and myosin-binding protein C (MYBPC1) for tuberculosis (TB). These four biomarkers are differentially regulated between quiescent Mycobacterium tuberculosis (Mtb) infected individuals (non-progressors to TB disease) and Mtb-infected TB disease progressors 6 months before the onset of symptoms.MethodsWe enrolled samples from patients experiencing moderate-to-severe pulmonary infections diseases including 23 TB cases confirmed by smear microscopy and culture, and 34 TB-negative cases. For each participant, the serum levels of the four biomarkers were measured using ELISA.ResultsThe levels of CK-MB and HEPC were significantly reduced in patients with active TB disease. CK-MB median level was 2045 pg/ml (1455–4000 pg/ml) in active TB cases and 3245 pg/ml (1645–4000 pg/ml) in non-TB pulmonary diseases. Using the receiver operating characteristic curve (ROC) analysis, HEPC and CK-MB had the Area Under the Curve (AUC) of 79% (95% CI 67–91%) and 81% (95% CI 69–93%), respectively. Both markers correlated with TB diagnosis as a single marker. PLa2G2A and MYBPC1 with AUCs of 48% (95% CI 36–65%) and 62% (95% CI 48–76%) did not performed well as single biomarkers. The three markers’model (CK-MB-HEPC-PLa2G2A) had the highest diagnostic accuracy at 82% (95% CI 56–82%) after cross-validation.ConclusionCK-MB and HEPC levels were statistically different between confirmed TB cases and non-TB cases. This study yields promising results for the rapid diagnosis of TB disease using a single marker or three biomarkers model.