Immunomodulatory Expression of Cathelicidins Peptides in Pulp Inflammation and Regeneration: An Update.

The role of inflammatory mediators in dental pulp is unique. The local environment of pulp responds to any changes in the physiology that are highly fundamental, like odontoblast cell differentiation and other secretory activity. The aim of this review is to assess the role of cathelicidins based on their capacity to heal wounds, their immunomodulatory potential, and their ability to stimulate cytokine production and stimulate immune-inflammatory response in pulp and periapex. Accessible electronic databases were searched to find studies reporting the role of cathelicidins in pulpal inflammation and regeneration published between September 2010 and September 2020. The search was performed using the following databases: Medline, Scopus, Web of Science, SciELO and PubMed. The electronic search was performed using the combination of keywords "cathelicidins" and "dental pulp inflammation". On the basis of previous studies, it can be inferred that LL-37 plays an important role in odontoblastic cell differentiation and stimulation of antimicrobial peptides. Furthermore, based on these outcomes, it can be concluded that LL-37 plays an important role in reparative dentin formation and provides signaling for defense by activating the innate immune system.

Clinical features and haematological parameters among malaria patients in Mangaluru city area in the southwestern coastal region of India.

The aim of this study was to assess the clinical profile, severity and complications of patients suffering from malaria in Mangaluru, a southwestern coastal city in India. A total of 579 patients, who were treated at the District Wenlock Hospital, Mangaluru, and 168 healthy controls were recruited in this study. The clinical profile, haematological and biochemical parameters, and disease complications were assessed. The majority of patients were treated as outpatients and patients who had severe clinical conditions were admitted to the hospital for treatment and supportive care. Among the total 579 patients recruited in this study, the distribution of P. vivax, P. falciparum and mixed infections were 364 (62.9%), 150 (25.9%) and 65 (11.2%), respectively. Among these, 506 (87.4%) had mild malaria, whereas 73 (12.6%) had severe malaria. Overall, the clinical features and severity of malaria in P. vivax and mixed infection patients were comparable to P. falciparum patients, albeit with some significant differences. The clinical complications in severe malaria cases included thrombocytopenia (50.7%), metabolic acidosis (30.1%), severe anaemia (26.0%), jaundice (21.9%), hepatic dysfunction (15.1%), acute renal failure (6.8%), haematuria (8.2%), hypotension (9.6%), cerebral malaria (1.4%) and acute respiratory distress syndrome (1.4%). All the patients with severe malaria recruited in our study were successfully treated and discharged. Majority of patients had mild malaria, likely due to seeking treatment soon after experiencing symptoms and/or having preexisting immune protection. However, a significant number of patients had severe malaria and required hospital admission indicating that there is a substantial need for creating awareness among vulnerable immigrant population. Implementing effective surveillance and vector control measures in malaria hotspot locations in the city and educating people about preventive measures are likely to reduce the malaria burden in this endemic region.

Malaria prevalence in Mangaluru city area in the southwestern coastal region of India.

BACKGROUND: Malaria is highly prevalent in many parts of India and the Indian subcontinent. Mangaluru, a city in the southwest coastal region of Karnataka state in India, and surrounding areas are malaria endemic with 10-12 annual parasite index. Despite high endemicity, to-date, very little has been reported on the epidemiology and burden of malaria in this area. METHODS: A cross-sectional surveillance of malaria cases was performed among 900 febrile symptomatic native people (long-time residents) and immigrant labourers (temporary residents) living in Mangaluru city area. During each of dry, rainy, and end of rainy season, blood samples from a group of 300 randomly selected symptomatic people were screened for malaria infection. Data on socio-demographic, literacy, knowledge of malaria, and treatment-seeking behaviour were collected to understand the socio-demographic contributions to malaria menace in this region. RESULTS: Malaria is prevalent in Mangaluru region throughout the year and Plasmodium vivax is predominant species compared to Plasmodium falciparum. The infection frequency was found to be high during rainy season. Infections were markedly higher in males than females, and in adults aged 16-45 years than both younger and older age groups. Also, malaria incidence was high among immigrants compared to native population. In both groups, infection rate was directly correlated with their literacy level, knowledge on malaria, dwelling environment, and protective measures used. There was also a significant difference in treatment-seeking behaviour between these two groups. CONCLUSIONS: Malaria incidences in Mangaluru region are predominantly localized to certain hotspot areas within the city, where socioeconomically underprivileged and immigrant labourers are densely populated. These areas have inadequate sanitation and constant water stagnation, harbouring high vector density and contributing to high infection incidences. Additionally, people in these areas seldom practice preventive measures such as using bed nets. The high incidences of malaria in adults are due to minimal cloth wearing, and long working hours stretching to late evenings in places with high vector density. Instituting heightened preventive public measures by governments and creating awareness on using preventive protective and environmental hygienic measures through educational programmes may substantially reduce the risk of contracting infections in these areas and spreading to other areas.

Comparison of salivary calcium, phosphate and alkaline phosphatase levels in children with early childhood caries after administration of milk, cheese and GC tooth mousse: an in vivo study.

BACKGROUND AND OBJECTIVES: This study compares the Salivary Calcium, Phosphate and Alkaline Phosphataselevels in children with Early Childhood Caries after administration of Milk, Cheese and GC Tooth Mousse to a control group of caries resistant children. STUDY DESIGN: 90 kindergarten children both males and females aged 5 years, from the South Canara region were included in the study. Based on the dmfs score, children were divided into 3 groups: Control group, ECC group and S-ECC group. The Salivary Calcium, Phosphate and Alkaline Phosphatase levels in the sample were assessed before and after administration of Milk, Cheese and GC Tooth Mousse at three different intervals, i.e within 5 minutes, 30 minutes and 60 minutes by using Spectrophotometry. RESULTS: The mean Salivary Calcium levels were higher in caries free group whereas Phosphate and Alkaline Phosphatase were lower in the caries free group which was statistically highly significant (p < 0.001). The Tooth Mousse group showed higher bioavailability of calcium and phosphate which was statistically highly significant (p < 0.001). Salivary Calcium and Phosphate levels within 5 minutes after application of Milk, Cheese and Tooth Mousse were higher than at 30 and 60 minutes. Salivary Alkaline Phosphatase levels were lower than the baseline values at all the 3 intervals after administration of Milk, Cheese and Tooth Mousse and was statistically not significant (p > 0.05). CONCLUSION: Saliva should be saturated with Calcium and Phosphate to affect their bioavailability in amounts adequate for remineralizaton. Milk, Cheese and GC Tooth Mousse applicaton were equally beneficial in saturating the saliva with adequate amount of Calcium and Phosphate.

Delineating the role of single-nucleotide polymorphism of CYP19 gene on aromatase activity in South Indian women with polycystic ovary syndrome.

BACKGROUND: Polycystic ovary syndrome is a common multifactorial endocrinopathy disorder affecting 5-15% of reproductive women worldwide. The CYP19 gene encodes key enzyme aromatase involved in androgen-to-estrogen conversion which plays a crucial role in the pathophysiology of the syndrome. Very few studies have been done in the Indian population; hence, we investigated whether CYP19 gene rs2414096 SNP is associated with PCOS and hyperandrogenism susceptibility in Karnataka women. METHODS: Three-hundred subjects including 150 PCOS and 150 age-matched controls were involved in the current case-control study. Sex hormones and biochemical estimation were performed by ELISA. Sanger sequencing and PCR-RFLP were used to genotype the SNP rs2414096. Genotypic-phenotypic association was studied. Statistical analysis was performed. RESULTS: The GG genotype was more common in patients, while the GA genotype was more common in control women. LH/FSH was significantly increased in GG genotype in PCOS when compared with AA and GA genotypes. Variations of CYP19 rs2414096 were not statistically significant with PCOS. CONCLUSION: CYP19 rs2414096 polymorphism was not associated with PCOS; however, the homozygous wild GG genotype may exhibit reduced aromatase activity with subsequent hyperandrogenism implicating endocrine abnormalities.

FOK l Vitamin D Receptor Gene Polymorphism and Risk of Dental Caries: A Case-Control Study.

The prevalence of dental caries in individuals who practice good oral hygiene increasingly indicates that other etiological factors, such as genetic factors, may be responsible for occurrence of caries, and its prevalence in younger individuals, such as adolescents, is an early manifestation of their genetic makeup. Therefore, there is a need to investigate the correlation of various genetic factors with the occurrence of dental caries in populations. Thus, this study assessed the relationship between single nucleotide polymorphism (rs2228570) in the vitamin D receptor gene and dental caries susceptibility. After obtaining ethical approval (NU/CEC/2020/0339), 377 adults, aged 18-40 years, were included in this study. Among the participants consenting to participate, salivary samples were collected, and an oral examination was conducted using the World Health Care Oral Health Survey Format 2013. The DMFT and PUFA index scores were recorded along with basic demographic details. The subjects were categorized as caries-free (controls, DMFT = 0) and caries-active (cases). The case group was further divided into the high-risk group (DMFT ≤ 10), moderate-risk group (DMFT = 4-9), and low-risk group (DMFT = 1-3). Saliva samples were used for vitamin D level analysis and DNA isolation. Polymerase chain reaction-based restriction fragment length polymorphism analysis using Fok1 digestion was performed on the isolated DNA. Salivary vitamin D levels were markedly higher in the caries-free group than in the caries-active group (p < 0.001). The T allele of rs2228570 was significantly associated with having active caries, while the C allele was associated with being caries-free. Individuals with the rs2228570 TC genotype had 2.814-fold increased likelihood, and individuals with the TT genotype had 3.116- fold increased likelihood of being caries-active. This finding is important in terms of patient counselling, as well as possibly in terms of prevention and treatment of caries.

A Comparative Evaluation of Antimicrobial Efficacy of Novel Surfactant-Based Endodontic Irrigant Regimen's on Enterococcus faecalis.

Introduction: Irrigants play an important role in the eradication of microorganisms in the complex root canal system. Sodium hypochlorite (NaOCl), chlorhexidine (CHX), and iodine potassium iodide (IKI) have certain limitations despite their routine clinical use. Surfactant irrigants with antimicrobial properties can be a milestone in endodontics to overcome the drawbacks of conventional irrigants. The aim of this study was to evaluate the antimicrobial efficacy of surfactants in comparison with routine endodontic irrigants on Enterococcus faecalis. Materials and Methods: Primary irrigants NaOCl, CHX, IKI were prepared at concentrations of 5%, 2.5%, 2%, and 1%; while mixture of tetracycline acid and detergent (MTAD) (100%) served as control group. Surfactants such as cetrimide (CTR) and sodium dodecyl sulfate (SDS) were prepared at concentrations of 0.5%, 1%, and 2%. The direct contact assay was used to evaluate the antimicrobial efficacy after 5 min. Statistical analysis was performed using one-way ANOVA and Mann-Whitney U-test. Results: In the primary group, only 5% CHX was significant over MTAD (P < 0.05). In the surfactant group, all concentrations of CTR and SDS (except 0.5%) were significant (P < 0.05) in killing E. faecalis over MTAD. Two percent CHX in combination with 0.5% CTR and 1% SDS had an effective kill percentage over 2% CHX and MTAD alone. Conclusion: Surfactant irrigant regimens can be used as adjuncts with CHX to overcome its clinical limitations and potentiate its substantivity, thereby enhancing clinical success in endodontics.

Impact of oxidative stress in response to malarial infection during pregnancy: Complications, histological changes, and pregnancy outcomes.

Background and Objectives: Pregnancy malaria is a major underestimated global public health problem. To understand the involvement of oxidative stress (OS) in the pathophysiology of placental malaria, OS biomarkers, malondialdehyde (MDA), uric acid (UA), and superoxide dismutase (SOD) levels were analyzed and correlated to placental histopathological changes and pregnancy outcomes. Methods: A hospital-based study was conducted in Mangaluru, Karnataka, India, to analyze the changes in hematological parameters and the serum OS biomarker levels. Histological analysis of placenta, associated complications, and pregnancy outcomes were compared using Kruskal-Wallis test, and pairwise comparison between two groups was made by Mann-Whitney U-test. Correlations were calculated by Pearson's and Spearman's rank correlations. Results: Among 105 pregnant women, 34 were healthy controls and the infected group comprised of Plasmodium Vivax (Pv) (n = 48), Plasmodium falciparum (Pf) (n = 13), and mixed (n = 10) malaria infections. Of 71 infected cases, 67.6% had mild malaria, whereas 32.4% had severe malaria. The white blood cell and C-reactive protein levels were found to increase, whereas hemoglobin, red blood cell, and platelet levels decreased during both types of malarial infections. The MDA and UA values increased and SOD levels decreased particularly during severe Pf infections. Histological changes such as syncytial knots, syncytial ruptures, and fibrinoid necrosis were observed particularly during Pf infections and leukocyte infiltration was observed in Pv malaria. Conclusion: Evaluation of MDA, UA, and SOD levels can serve as an indicator of OS during pregnancy malaria. The OS during pregnancy may lead to complications such as severe anemia, pulmonary edema, intra uterine growth retardation, premature delivery, and low birth weight, not only during Pf but also in Pv malaria. It is important to create awareness among rural and immigrant population residing in Mangaluru and its surroundings about required preventive measures and free government-supported antenatal care services.

Fatty acids and their role in type-2 diabetes (Review).

Age, lifestyle and diet are major risk factors for the onset of type 2 diabetes mellitus (T2DM). Insulin resistance (IR) and ß-cell dysfunction underlie the pathophysiology of T2DM. Diabetic populations are also prone to lipid and lipoprotein abnormalities as an indirect effect of IR on key metabolic enzymes. However, recent studies suggested that lipid changes may not only be a consequence of impaired glucose metabolism but also a causative factor. Fatty acids (FAs) influence translocation of glucose transporters and insulin receptor binding and signalling, in addition to cell membrane fluidity and permeability. It is thus suggested that FAs may have an essential role in the development of IR and T2DM. Specific combinations of FAs within phospholipids and triglycerides were indicated to exhibit the strongest associations with the risk of T2DM. The aim of the present review was to investigate the role of FAs in the pathogenesis of T2DM, as it has yet to be fully elucidated.

Estimation of Serum MicroRNA 21 as a Tumour Marker in Carcinoma Oesophagus.

Worldwide, carcinoma of the oesophagus is the sixth leading reason for deaths due to malignancy. MicroRNAs (MiRs) are a cluster of small non-coding RNAs that have potent stability in stored RNA samples. This strengthens their potential as biomarkers. This study aimed to isolate MiR 21 from serum samples of carcinoma oesophagus patients and healthy subjects for determining its strength of association in various stages of the carcinoma oesophagus. This is a case-control study with 1:1 matched cases and controls. We included 40 patients of carcinoma oesophagus and 40 healthy individuals. Serum separated from venous blood samples of cases and controls was stored at - 80 °C. Total RNA was extracted, and the real-time polymerase chain reaction was used to measure the serum MiR 21. On statistical comparison, we found a marked increase in MiR 21 in cases of carcinoma oesophagus when compared with the controls (p < 0.0001). This study also showed a steady rise in the MiR 21 levels with the disease severity (p < 0.0001). Based on the result of this study, we conclude that MiR 21 levels can be used as a novel marker for assessing disease severity in carcinoma oesophagus.

Y chromosome microdeletion and cytogenetic findings in male infertility: A cross-sectional descriptive study.

BACKGROUND: Infertility affects about 15% of couples worldwide, and the male factor alone is responsible for approximately 50% of the cases. Genetic factors have been found to play important roles in the etiology of azoospermia and severe oligospermia conditions that affect 30% of individuals seeking treatment at infertility clinics. OBJECTIVE: To determine the frequency of chromosomal abnormalities and Y chromosome microdeletion in infertile men. MATERIALS AND METHODS: A total of 100 infertile men with abnormal semen parameters were included in this study from 2014 to 2018. Chromosomal analysis was carried out using standard G-banding using Trypsin Giemsa protocol. Multiplex polymerase chain reaction was used to determine the Y microdeletion frequency. RESULTS: All participants were aged between 22 and 48 yr with a mean and standard deviation of 35.5 ± 5.1. Of the 100 subjects included in the study, three had Klinefelter syndrome-47,XXY, one had balanced carrier translocation-46,XY,t(2;7)(q21;p12), one with the balanced carrier translocation with inversion of Y chromosome 45,XY,der(13;14)(q10;q10),inv(Y), one had polymorphic variant of chromosome 15, one had Yqh-, and another had an inversion of chromosome 9. Y chromosome microdeletion of Azoospermia factor c region was observed in 2% of the cases. To the best of our knowledge, the current study is the first reported case with unique, balanced carrier translocation of chromosome 2q21 and 7p21. CONCLUSION: The present study emphasizes the importance of routine cytogenetic screening and Y microdeletion assessment for infertile men, which can provide specific and better treatment options before undergoing assisted reproductive technology during genetic counseling.

Exploring the differentially expressed genes in human lymphocytes upon response to ionizing radiation: a network biology approach.

PURPOSE: The integration of large-scale gene data and their functional analysis needs the effective application of various computational tools. Here we attempted to unravel the biological processes and cellular pathways in response to ionizing radiation using a systems biology approach. MATERIALS AND METHODS: Analysis of gene ontology shows that 80, 42, 25, and 35 genes have roles in the biological process, molecular function, the cellular process, and immune system pathways, respectively. Therefore, our study emphasizes gene/protein network analysis on various differentially expressed genes (DEGs) to reveal the interactions between those proteins and their functional contribution upon radiation exposure. RESULTS: A gene/protein interaction network was constructed, which comprises 79 interactors with 718 interactions and TP53, MAPK8, MAPK1, CASP3, MAPK14, ATM, NOTCH1, VEGFA, SIRT1, and PRKDC are the top 10 proteins in the network with high betweenness centrality values. Further, molecular complex detection was used to cluster these associated partners in the network, which produced three effective clusters based on the Molecular Complex Detection (MCODE) score. Interestingly, we found a high functional similarity from the associated genes/proteins in the network with known radiation response genes. CONCLUSION: This network-based approach on DEGs of human lymphocytes upon response to ionizing radiation provides clues for an opportunity to improve therapeutic efficacy.

Acquired antibody responses against merozoite surface protein-119 antigen during Plasmodium falciparum and P.vivax infections in South Indian city of Mangaluru.

Merozoite surface protein-1 (MSP-1) of malaria parasites has been extensively studied as a malaria vaccine candidate and the antibody response to this protein is an important indicator of protective immunity to malaria. Mangaluru city and its surrounding areas in southwestern India are endemic to malaria with Plasmodium vivax being the most widespread and prevalent species although P. falciparum also frequently infects. However, no information is available on the level of protective immunity in this population. In this regard, a prospective hospital-based study was performed in malarial patients to assess antibody responses against the 19-kDa C-terminal portion of P. vivax and P. falciparum MSP-1 (MSP-119). Serum samples from 51 healthy endemic controls and 267 infected individuals were collected and anti-MSP-119 antibody levels were analyzed by ELISA. The possible association between the antibody responses and morbidity parameters such as malarial anemia and thrombocytopenia was investigated. Among the 267 infected cases, 144 had P. vivax and 123 had P. falciparum infections. Significant levels of anti-MSP-119 antibody were observed both in P. vivax (123/144; 85.4%) and P. falciparum (108/123; 87.9%) infected individuals. In both type of infections, the major antibody isotypes were IgG1 and IgG3. The IgG levels were found to be increased in patients with severe anemia and thrombocytopenia. The antibody levels were also higher in infected individuals who had several previous infections, although antibodies produced during previous infections were short lived. The predominance of cytophilic anti-MSP-119 IgG1 and IgG3 antibodies suggests the possibility of a dual role of Pv MSP-119 and Pf MSP-119 during malarial immunity and pathogenesis.

Plasma fatty acids composition and estimated delta desaturases activity in women with breast cancer.

INTRODUCTION: Fatty acids (FAs) are the vital constituents of membrane structures. De novo synthesis of FAs includes an enzymatic complex of FA synthase and delta desaturases. These enzymes are overexpressed in tumors, and inhibition of these enzymes is gaining interest. Our aim was to determine if delta desaturase activities are altered in breast cancer (BC) cases and if altered whether delta desaturase activities differ among BC genotypes. MATERIALS AND METHODS: In this observational comparative study, 50 women with BC and 30 control women were recruited for the study. Gas chromatography-flame ionization detector was used to measure the plasma FA levels. Desaturase activities were assessed as product-to-precursor FA ratios. The Wilcoxon signed-rank test was used to compare between two groups, and P ≤ 0.05 was considered as statistically significant. RESULTS: The FA analysis revealed higher levels of monounsaturated FAs (MUFAs) and linolenic acid metabolites (C18:3n-6, C20:4n-6) in BC patients, whereas C20:5n-3 was higher in controls. The Delta 9 desaturase (D9D) and D6D were higher in BC cases suggesting greater conversion saturated FA to MUFA and linoleic acid to its metabolites. D9D-16 activity was statistically significant (P = 0.03) in BC women, particularly in estrogen-receptor-positive patients. CONCLUSION: There is limited evidence to substantiate the link between diet and cancer. The current study showed there is an altered lipid desaturase activity. Nutritional intervention and drugs that target the FA pathway may provide a new approach to prevent and treat BC.

A comparative evaluation of the DNA damage in the serum of chronic periodontitis patients with and without diabetes mellitus type II.

CONTEXT: Periodontal disease is an immunoinflammatory disease that is initiated by the interaction between microbial plaque and the periodontal tissues. There is very limited data available on the assessment of DNA damage with relation to periodontal diseases. Therefore, a need for a study in this area was felt. AIMS: To evaluate the DNA damage in the serum of chronic periodontitis patients and chronic periodontitis with diabetes mellitus (DM) type II patients and to compare it with healthy controls, to assess whether periodontitis can have systemic effects beyond the periodontium. SETTINGS AND DESIGN/SUBJECTS AND METHODS: This cross-sectional study was conducted involving 150 subjects in the age group of 30-60 years, from October 2010 to May 2015. A blood sample of 5 ml of venous blood was collected from each of the study subjects, from the antecubital vein. Fresh blood was used to assess the DNA damage. The DNA damage was estimated using the alkaline single-cell gel (comet) assay. RESULTS: The DNA damage to the cells was calculated by assessing the percentage of "DNA in tail." The results showed that the values were higher in the periodontitis with diabetes group, as compared to the periodontitis and control group. When the Olive moment was calculated, the values were higher in the periodontitis with diabetes group as compared with the other two groups. Although the values were seen to be higher, it was not statistically significant. CONCLUSIONS: The results obtained from this study although statistically insignificant suggest that the DNA damage was higher in chronic periodontitis as compared with healthy control. There was a potentiated difference of the values in patients with DM type II when compared to chronic periodontitis alone.

Estimation of Sialic Acid and IL10 Levels in Stage 1 and 2 Periodontitis Patients.

OBJECTIVE: The role of biomarkers in staging and grading periodontal disease has become detrimental in relation to the overall treatment plan. This study aimed at evaluating and comparing the role of sialic acid and IL10 in the early and moderate stages of periodontitis. MATERIALS AND METHODS: Patients were selected according to the assessment of pocket depth and radiographic bone loss. Bone loss was calculated as <15% for stage 1 and 15-33% for stage 2. Salivary samples were collected using spit technique 2 hrs post consumption of food. The unstimulated saliva was collected in a sterile graduated container every minute for 5-8 minutes. IL10 estimation was done using ELISA, and sialic acid estimation was done using the diphenylamine method. The variables for the three groups were assessed using ANOVA, and intragroup comparisons for quantitative data were evaluated using the post hoc Bonferroni test (P < 0.05). RESULTS: On comparing sialic acid levels among the three groups, stage 2 showed the highest mean (8.61) compared with the other two groups and was highly significant (P < 0.001). On the contrary, IL10 when compared to stage 1 and 2 periodontitis revealed insignificant change. CONCLUSION: The value of IL10 was higher as patients progressed from health to periodontitis.

Association between Inflammatory Cytokine Levels and Thrombocytopenia during Plasmodium falciparum and P. vivax Infections in South-Western Coastal Region of India.

BACKGROUND: Thrombocytopenia is a most commonly observed complication during malaria infections. Inflammatory cytokines such as IL-1, IL-6, and IL-10 have been documented in malaria induced thrombocytopaenia. This study was aimed to understand the possible relationship between inflammatory cytokines across varying degrees of thrombocytopenia during P. vivax, P. falciparum, and mixed infections. METHODS: A hospital-based cross sectional study was conducted at District Wenlock Hospital in Mangaluru, a city situated along the south-western coastal region of Arabian Sea in India. In this study, blood samples from 627 malaria patients were analyzed for infected parasite species, clinical conditions, platelet levels, and key cytokines that are produced in response to infection; samples from 176 uninfected healthy individuals were used as controls. RESULTS: The results of our study showed a high prevalence of malarial thrombocytopenia (platelets <150 ×103/µl) in this endemic settings. About 62.7% patients had mild-to-moderate levels of thrombocytopenia and 16% patients had severe thrombocytopenia (platelets <50 × 103/µl). Upon comparison of cytokines across varying degrees of thrombocytopenia, irrespective of infecting species, the levels of TNF-α and IL-10 were significantly higher during thrombocytopenia, whereas IL-6 levels were considerably lower in severe thrombocytopenia patients suffering from P. vivax or P. falciparum infections. The severe clinical complications observed in patients with malarial thrombocytopenia included severe anemia (17.5%), acute renal failure (12.7%), jaundice (27.0%), metabolic acidosis (36.5%), spontaneous bleeding (3.2%), hypoglycemia (25.4%), hyperparasitemia (4.8%), acute respiratory distress syndrome (1.6%), pulmonary edema (19.0%), and cerebral malaria (1.6%) in various combinations. CONCLUSION: Overall, the results of our study suggest that inflammatory cytokines influence the transformation of mild forms of thrombocytopenia into severe forms during malarial infections. Further studies are needed to understand the association of inflammatory cytokine responses with severe malaria complications and thrombocytopenia.

Association between inflammatory cytokine levels and anemia during Plasmodium falciparum and Plasmodium vivax infections in Mangaluru: A Southwestern Coastal Region of India.

BACKGROUND AND OBJECTIVES: Dysregulated production of inflammatory cytokines might play important role in anemia during malaria infection. The objective of this study was to assess the extent of anemia due to malaria, associated complications, and inflammatory cytokines (tumor necrosis factor alpha [TNF-α], interleukin [IL]-6, and IL-10) across varying anemic intensity during malaria infections. MATERIALS AND METHODS: A hospital-based cross-sectional study was conducted at District Wenlock hospital in Mangaluru city. Samples from 627 patients and 168 healthy controls (HC) were analyzed for level of hemoglobin (Hb), red blood cells (RBCs), and inflammatory cytokines. The blood cell parameters and inflammatory cytokines levels across varying intensity of anemia were analyzed using Kruskal-Wallis test and pair-wise comparison between two groups were by Mann-Whitney U-test. Correlations were calculated by Pearson's and Spearman rank correlations. RESULTS: Compared to HC, Hb, and RBC levels were significantly lower in infected patients. On comparison with mild anemia patients (Hb 8-10.9 g/dL), the levels of TNF-α and IL-6 were significantly elevated, whereas IL-10 levels were lower during severe anemia (SA) (Hb <5 g/dL). In this endemic setting, we found a strong negative association between Hb levels and parasitemia, Hb and TNF-α, and positive relationship with IL-10; anemic patients also had significantly high TNF-α/IL-10 ratios. SA was associated with complications such as acute renal failure (16.0%), jaundice (16.0%), metabolic acidosis (24.0%), hypoglycemia (12.0%), hyperparasitemia (4.0%), and hepatic dysfunction (16.0%). CONCLUSIONS: Contrary to its benign reputation, Plasmodium vivax (Pv) infections can also result in severe malarial anemia (SMA) and its associated severe complications similar to Plasmodium falciparum infections. Dysregulated inflammatory cytokine responses play an important role in the pathogenesis of SMA, especially during Pv infections.

Malaria Severity in Mangaluru City in the Southwestern Coastal Region of India.

Dakshina Kannada district in the Southwestern region of Karnataka state, India, including Mangaluru city is endemic to malaria. About 80% of malaria infections in Mangaluru and its surrounding areas are caused by Plasmodium vivax and the remainder is due to Plasmodium falciparum. Malaria-associated clinical complications significantly occur in this region. Here, we report the pathological conditions of 41 cases of fatal severe malaria, admitted to the district government hospital in Mangaluru city during January 2013 through December 2016. The results of clinical, hematological, and biochemical analyses showed that most of these severe malaria cases were associated with thrombocytopenia, anemia, metabolic acidosis, acute respiratory distress, and single or multi-organ dysfunction involving liver, kidney, and brain. Of the 41 fatal malaria cases, 24, 10, and seven patients had P. vivax, P. falciparum, and P. vivax and P. falciparum mixed infections, respectively. These data suggest that besides P. falciparum that is known to extensively cause severe and fatal malaria illnesses, P. vivax causes fatal illnesses substantially in this region, an observation that is consistent with recent findings in other regions.

Cytotoxicity Evaluation of Combination Irrigant Regimens with MTAD on Two Different Cell Lines.

BACKGROUND: Effective management of smear layer ensures adequate clinical success. Use of sodium hypochlorite (NaOCl)/ethylenediaminetetraacetic acid regimen has been the gold standard with limitations. Commercial irrigants incorporate surface modifiers to address these drawbacks. The aim of this study was to evaluate the cytotoxicity of combination regimens on target and nontarget cell lines by trypan blue assay. MATERIALS AND METHODS: Nonsurfactant combination regimen of chlorhexidine (CHX) and NaOCl (2% CHX + 2.5% NaOCl) and surfactant regimens of CHX with cetrimide (CTR) (2% CHX + 0.5% CTR) and CHX with sodium dodecyl sulfate (2% CHX + 1% SDS) were prepared. 0.9% normal saline (NS) and Biopure MTAD (100%) served as control. Cytotoxicity was evaluated on human gingival fibroblast (HGF) and Henrietta Lacks (HeLa) cell lines by trypan blue assay. Thirty microliter of the cell suspension was treated with 20 µl of irrigants. The cell suspension was loaded into Neubauer chamber after 5 min and cell count was performed under inverted microscope and expressed as viability percentage. RESULTS: Nonsurfactant combination comprising of 2% CHX + 2.5% NaOCl formed a brownish precipitate while surfactant combination regimes were stable without any precipitate formation. NS and 2% CHX + 0.5% CTR had greater viability scores on both cell lines. Two percent CHX + 1% SDS had better viability on HeLa but were severely cytotoxic on HGF. Two percent CHX + 2.5% NaOCl and MTAD were found to be severely cytotoxic on HeLa with limited viability on HGF. CONCLUSION: The variation in data obtained could be possibly attributed to the difference in the cellular membrane composition and mechanism of action of combination regimens. Experimental surfactant regimen 2% CHX + 0.5% CTR shows lower cytotoxicity than MTAD.