Targeting tumour markers in ovarian cancer treatment.

Ovarian cancers (OC) are the most common, lethal, and stage-dependent cancers at the global level, specifically in female patients. Targeted therapies involve the administration of drugs that specifically target the alterations in tumour cells responsible for their growth, proliferation, and metastasis, with the aim of treating particular patients. Presently, within the realm of gynaecological malignancies, specifically in breast and OCs, there exist various prospective therapeutic targets encompassing tumour-intrinsic signalling pathways, angiogenesis, homologous-recombination deficit, hormone receptors, and immunologic components. Breast cancers are often detected in advanced stages, primarily due to the lack of a reliable screening method. However, various tumour markers have been extensively researched and employed to evaluate the condition, progression, and effectiveness of medication treatments for this ailment. The emergence of recent technological advancements in the domains of bioinformatics, genomics, proteomics, and metabolomics has facilitated the exploration and identification of hitherto unknown biomarkers. The primary objective of this comprehensive review is to meticulously investigate and analyze both established and emerging methodologies employed in the identification of tumour markers associated with OC.

Small molecules that disrupt RAD54-BLM interaction hamper tumor proliferation in colon cancer chemoresistance models.

RAD54 and BLM helicase play pivotal roles during homologous recombination repair (HRR) to ensure genome maintenance. BLM amino acids (aa 181-212) interact with RAD54 and enhance its chromatin remodeling activity. Functionally, this interaction heightens HRR, leading to a decrease in residual DNA damage in colon cancer cells. This contributes to chemoresistance in colon cancer cells against cisplatin, camptothecin, and oxaliplatin, eventually promoting tumorigenesis in preclinical colon cancer mouse models. ChIP-Seq analysis and validation revealed increased BLM and RAD54 corecruitment on the MRP2 promoter in camptothecin-resistant colon cancer cells, leading to BLM-dependent enhancement of RAD54-mediated chromatin remodeling. We screened the Prestwick small-molecule library, with the intent to revert camptothecin- and oxaliplatin-induced chemoresistance by disrupting the RAD54-BLM interaction. Three FDA/European Medicines Agency-approved candidates were identified that could disrupt this interaction. These drugs bound to RAD54, altered its conformation, and abrogated RAD54-BLM-dependent chromatin remodeling on G5E4 and MRP2 arrays. Notably, the small molecules also reduced HRR efficiency in resistant lines, diminished anchorage-independent growth, and hampered the proliferation of tumors generated using camptothecin- and oxaliplatin-resistant colon cancer cells in both xenograft and syngeneic mouse models in BLM-dependent manner. Therefore, the 3 identified small molecules can serve as possible viable candidates for adjunct therapy in colon cancer treatment.

A Randomized Trial of Inpatient and Home-Based Maternal Oral Hydration Therapy in Isolated Oligohydramnios and Its Effect on Amniotic Fluid Index and Perinatal Outcome.

Background Isolated oligohydramnios, without any known fetal/maternal abnormality, may be associated with insufficient oral intake (such as water, glucose, and rehydration therapy). Therefore, the present study was conducted to assess the improvement following maternal hydration. Method A total of 50 cases of isolated oligohydramnios (other high-risk pregnancy conditions not present) were included in the study (25 in each group). Patients were encouraged for an additional 2 liters of oral rehydration solution intake daily along with regular diet. The fluid intake was unsupervised in the home group and supervised in the hospital group. Serial amniotic fluid index (AFI) measurements and fetal monitoring were performed. Birth weight and APGAR scores were recorded, and data were analyzed. Results The two groups were comparable in terms of demographics and baseline laboratory findings. AFI significantly improved in the hospital group compared to the home group (p-value: <0.001). Birth weight, placental weight, and APGAR scores were also significantly better in the hospital group than in the home group. Conclusion Maternal oral hydration therapy improves the amniotic fluid volume and subsequently improves the perinatal outcome. Due to poor compliance with home-based treatment, institution of supervised hydration therapy is recommended.

Menstrual Hygiene Practices in Indian Tribal Females: A Systematic Review and Meta-Analysis.

India is native to many tribal communities: Bharia (Madhya Pradesh), Bihl (Rajasthan), Santhal (Bihar, Jharkhand), Bodo (Assam, West Bengal), and many more. They reside in isolated geographical regions, which poses challenges in reaching out to them. In addition, they still have firm beliefs and taboos regarding menstruation. Knowledge about menstrual health and hygiene is one of the most important aspects of tribal health. Therefore, it is important to synthesize the results of menstrual hygiene data from the Indian tribal population. We have calculated the pooled prevalence of sanitary pad use, dustbin disposal, and hygienic reuse of menstrual materials. Online databases, namely PubMed, Cochrane Central, CINAHL, Pan African Journals, EBSCO, and Google Scholar, were searched. After the removal of duplicates, a quality check, and screening of cross-references, 19 articles were selected for final review. Statistical analysis was done by Revman 5.4 and STATA 17.0. A p-value of <0.05 was considered statistically significant. PRISMA guidelines were followed. The protocol registration number was CRD42022331376. This is a non-funded article. The pooled prevalence of sanitary pad use in Indian tribal females was 2% (95% CI 1 to 3). The pooled prevalence of dustbin disposal of menstrual material was 1% (95% CI: 0.00 to 0.02). The pooled prevalence of hygienic reuse of menstrual materials was 1%. Sanitary menstrual hygiene practices are very less prevalent in Indian tribal females. Awareness programs and tribal health policies need to be accelerated for the promotion of menstrual hygiene. Also, literature on the use, disposal, and storage of menstrual adsorbents is scarce in Indian tribes. Health research in this area needs to be emphasized.

Seed nutritional quality in lentil (Lens culinaris) under different moisture regimes.

The world's most challenging environmental issue is climate change. Agricultural productivity and nutritional quality are both substantially threatened by extreme and unpredicted climate events. To develop climate resilient cultivars, stress tolerance along with the grain quality needs to be prioritized. Present study was planned to assess the effect of water limitation on seed quality in lentil, a cool season legume crop. A pot experiment was carried out with 20 diverse lentil genotypes grown under normal (80% field capacity) and limited (25% field capacity) soil moisture. Seed protein, Fe, Zn, phytate, protein and yield were recorded in both the conditions. Seed yield and weight were reduced by 38.9 and 12.1%, respectively, in response to stress. Seed protein, Fe, Zn, its availability as well as antioxidant properties also reduced considerably, while genotype dependent variation was noted with respect to seed size traits. Positive correlation was observed between seed yield and antioxidant activity, seed weight and Zn content and availability in stress. Based on principal component analysis and clustering, IG129185, IC559845, IC599829, IC282863, IC361417, IG334, IC560037, P8114 and L5126 were promising genotypes for seed size, Fe and protein content, while, FLIP-96-51, P3211 and IC398019 were promising for yield, Zn and antioxidant capacity. Identified lentil genotypes can be utilized as trait donors for quality improvement in lentil breeding.

Hyperubiquitylation of DNA helicase RECQL4 by E3 ligase MITOL prevents mitochondrial entry and potentiates mitophagy.

Mutations in the DNA helicase RECQL4 lead to Rothmund-Thomson syndrome (RTS), a disorder characterized by mitochondrial dysfunctions, premature aging, and genomic instability. However, the mechanisms by which these mutations lead to pathology are unclear. Here we report that RECQL4 is ubiquitylated by a mitochondrial E3 ligase, MITOL, at two lysine residues (K1101, K1154) via K6 linkage. This ubiquitylation hampers the interaction of RECQL4 with mitochondrial importer Tom20, thereby restricting its own entry into mitochondria. We show the RECQL4 2K mutant (where both K1101 and K1154 are mutated) has increased entry into mitochondria and demonstrates enhanced mitochondrial DNA (mtDNA) replication. We observed that the three tested RTS patient mutants were unable to enter the mitochondria and showed decreased mtDNA replication. Furthermore, we found that RECQL4 in RTS patient mutants are hyperubiquitylated by MITOL and form insoluble aggregate-like structures on the outer mitochondrial surface. However, depletion of MITOL allows RECQL4 expressed in these RTS mutants to enter mitochondria and rescue mtDNA replication. Finally, we show increased accumulation of hyperubiquitylated RECQL4 outside the mitochondria leads to the cells being potentiated to increased mitophagy. Hence, we conclude regulating the turnover of RECQL4 by MITOL may have a therapeutic effect in patients with RTS.

Transglutaminase-Polyethyleneimine Nanoflowers Mediated Cellular Delivery of Anti-miR-210 for Effective Glioblastoma Therapy.

Glioblastoma (GBM) is a deadly tumor of the central nervous system (CNS) having a dismal prognosis. miRNA-based therapeutics hold immense potential for GBM therapy; however, its delivery remains a daunting challenge. MicroRNA-210 has been established as a critical oncomiR in GBM. Our group has developed novel, PEI-functionalized transglutaminase-based nanoflowers (TGNFs, ∼61 nm in diameter) for the efficient delivery of anti-miR-210 to glioblastoma cells in vitro. TGNFs show low cytotoxicity to normal human fibroblasts, do not affect the liver and kidney health of CD1 mice, and offer >95% anti-miR encapsulation efficiency, serum stability, and protection against polyanion moieties. Their synthesis is cost-effective and does not involve the application of harsh chemicals. TGNFs successfully delivered anti-miR-210 to glioblastoma cells, decreasing cellular proliferation and migration and increasing apoptosis. Overall, this research highlights the potential of TGNFs as delivery agents in miRNA inhibition therapy and encourages further preclinical studies to explore the potential of miR-210 as a therapeutic target in GBM and various other cancers where the oncogenic role of miR-210 has been well-established.

Monitoring effects of heavy metal stress on biochemical and spectral parameters of cotton using hyperspectral reflectance.

Monitoring heavy metal pollution in agricultural ecosystems is crucial to ensure environmental safety. Heavy metals interfere with plants' biochemical characteristics, such as chlorophyll content and photosynthesis, and also influence leaves' spectral properties. Spectral changes caused by heavy metal stress can easily be measured using proximal sensing or in-field spectroscopy. This research utilizes a combined approach of biochemical and spectral characteristics to evaluate cotton crops' performance under different heavy metal (Pb & Cd) stress after artificial contamination with the metal under study. A detailed study of spectroscopy and lab-based measurements for chlorophyll and heavy metal content during the crop's growth cycle revealed some significant findings. Results indicated that the chlorophyll pigments decreased significantly with increased heavy metal levels. Pb accumulation is high in cotton as compared to Cd. The most sensitive stage for the accumulation of Pb is the initial vegetative stage of cotton. The transfer factor from soil to plant was higher for Pb, indicating the feasibility of growing cotton in Pb-contaminated soil. The spectral measurement showed no characteristic changes in standard reflectance spectra due to heavy metal stress. Wavelet decomposition of reflectance spectra amplified the changes indicating Pb stress in cotton during the initial vegetative stage. The significant correlation of greater than - 0.70 between the reconstructed detail wavelet coefficients at the third level of the decomposition in the wavelength range of 651-742 nm suggested that Pb stress caused spectral changes in near-infrared and visible ranges in cotton plants. The effects of Cd stress on the cotton plant were negligible due to less absorption. Thus, detailed wavelet coefficients at the third level of decomposition in the range of 651-742 nm are a potential indicator of Pb stress. The results of this study can provide a basis for quantifying heavy metal stress in a particular region.

Correlating the patterns of diabetic macular edema, optical coherence tomography biomarkers and grade of diabetic retinopathy with stage of renal disease.

PURPOSE: To correlate optical coherence tomography (OCT)-based morphological patterns of diabetic macular edema (DME), biomarkers and grade of diabetic retinopathy (DR) in patients with various stages of chronic kidney disease (CKD) secondary to diabetes. DESIGN: Multicentric retrospective cross-sectional study was conducted at seven centers across India. METHODS: Data from medical records of patients with DME and CKD were entered in a common excel sheet across all seven centers. Staging of CKD was based on estimated glomerular filtration rate (eGFR). RESULTS: The most common morphological pattern of DME was cystoid pattern (42%) followed by the mixed pattern (31%). The proportion of different morphological patterns did not significantly vary across various CKD stages (p = 0.836). The presence of external limiting membrane-ellipsoid zone (ELM-EZ) defects (p < 0.001) and foveal sub-field thickness (p = 0.024) showed a direct correlation with the stage of CKD which was statistically significant. The presence of hyperreflective dots (HRD) and disorganization of inner retinal layers (DRIL) showed no significant correlation with the stage of CKD. Sight threatening DR was found to increase from 70% in CKD stage 3 to 82% in stages 4 and 5 of CKD, and this was statistically significant (p = 0.03). CONCLUSION: Cystoid morphological pattern followed by mixed type was the most common pattern of DME on OCT found in patients suffering from stage 3 to 5 of CKD. However, the morphological patterns of DME did not significantly vary across various CKD stages. ELM-EZ defects may be considered as an important OCT biomarker for advanced stage of CKD.

Root Trait Variation in Lentil (Lens culinaris Medikus) Germplasm under Drought Stress.

Drought is the most critical environmental factor across the continents affecting food security. Roots are the prime organs for water and nutrient uptake. Fine tuning between water uptake, efficient use and loss determines the genotypic response to water limitations. Targeted breeding for root system architecture needs to be explored to improve water use efficiency in legumes. Hence, the present study was designed to explore root system architecture in lentil germplasm in response to drought. A set of 119 lentil (Lens culinaris Medik.) genotypes was screened in controlled conditions to assess the variability in root traits in relation to drought tolerance at seedling stage. We reported significant variation for different root traits in lentil germplasm. Total root length, surface area, root volume and root diameter were correlated to the survival and growth under drought. Among the studied genotypes, the stress tolerance index varied 0.19-1.0 for survival and 0.09-0.90 for biomass. Based on seedling survival and biomass under control and drought conditions, 11 drought tolerant genotypes were identified, which may be investigated further at a physiological and molecular level for the identification of the genes involved in drought tolerance. Identified lines may also be utilised in a lentil breeding program.

CDX2 inducible microRNAs sustain colon cancer by targeting multiple DNA damage response pathway factors.

Meta-analysis of transcripts in colon adenocarcinoma patient tissues led to the identification of a DNA damage responsive miR signature called DNA damage sensitive miRs (DDSMs). DDSMs were experimentally validated in the cancerous colon tissues obtained from an independent cohort of colon cancer patients and in multiple cellular systems with high levels of endogenous DNA damage. All the tested DDSMs were transcriptionally upregulated by a common intestine-specific transcription factor, CDX2. Reciprocally, DDSMs were repressed via the recruitment of HDAC1/2-containing complexes onto the CDX2 promoter. These miRs downregulated multiple key targets in the DNA damage response (DDR) pathway, namely BRCA1, ATM, Chk1 (also known as CHEK1) and RNF8. CDX2 directly regulated the DDSMs, which led to increased tumor volume and metastasis in multiple preclinical models. In colon cancer patient tissues, the DDSMs negatively correlated with BRCA1 levels, were associated with decreased probability of survival and thereby could be used as a prognostic biomarker. This article has an associated First Person interview with the first author of the paper.

Growth and Antioxidant Responses in Iron-Biofortified Lentil under Cadmium Stress.

Cadmium (Cd) is a hazardous heavy metal, toxic to our ecosystem even at low concentrations. Cd stress negatively affects plant growth and development by triggering oxidative stress. Limited information is available on the role of iron (Fe) in ameliorating Cd stress tolerance in legumes. This study assessed the effect of Cd stress in two lentil (Lens culinaris Medik.) varieties differing in seed Fe concentration (L4717 (Fe-biofortified) and JL3) under controlled conditions. Six biochemical traits, five growth parameters, and Cd uptake were recorded at the seedling stage (21 days after sowing) in the studied genotypes grown under controlled conditions at two levels (100 µM and 200 µM) of cadmium chloride (CdCl2). The studied traits revealed significant genotype, treatment, and genotype × treatment interactions. Cd-induced oxidative damage led to the accumulation of hydrogen peroxide (H2O2) and malondialdehyde in both genotypes. JL3 accumulated 77.1% more H2O2 and 75% more lipid peroxidation products than L4717 at the high Cd level. Antioxidant enzyme activities increased in response to Cd stress, with significant genotype, treatment, and genotype × treatment interactions (p < 0.01). L4717 had remarkably higher catalase (40.5%), peroxidase (43.9%), superoxide dismutase (31.7%), and glutathione reductase (47.3%) activities than JL3 under high Cd conditions. In addition, L4717 sustained better growth in terms of fresh weight and dry weight than JL3 under stress. JL3 exhibited high Cd uptake (14.87 mg g-1 fresh weight) compared to L4717 (7.32 mg g-1 fresh weight). The study concluded that the Fe-biofortified lentil genotype L4717 exhibited Cd tolerance by inciting an efficient antioxidative response to Cd toxicity. Further studies are required to elucidate the possibility of seed Fe content as a surrogacy trait for Cd tolerance.

MITOL-dependent ubiquitylation negatively regulates the entry of PolγA into mitochondria.

Mutations in mitochondrial replicative polymerase PolγA lead to progressive external ophthalmoplegia (PEO). While PolγA is the known central player in mitochondrial DNA (mtDNA) replication, it is unknown whether a regulatory process exists on the mitochondrial outer membrane which controlled its entry into the mitochondria. We now demonstrate that PolγA is ubiquitylated by mitochondrial E3 ligase, MITOL (or MARCH5, RNF153). Ubiquitylation in wild-type (WT) PolγA occurs at Lysine 1060 residue via K6 linkage. Ubiquitylation of PolγA negatively regulates its binding to Tom20 and thereby its mitochondrial entry. While screening different PEO patients for mitochondrial entry, we found that a subset of the PolγA mutants is hyperubiquitylated by MITOL and interact less with Tom20. These PolγA variants cannot enter into mitochondria, instead becomes enriched in the insoluble fraction and undergo enhanced degradation. Hence, mtDNA replication, as observed via BrdU incorporation into the mtDNA, was compromised in these PEO mutants. However, by manipulating their ubiquitylation status by 2 independent techniques, these PEO mutants were reactivated, which allowed the incorporation of BrdU into mtDNA. Thus, regulated entry of non-ubiquitylated PolγA may have beneficial consequences for certain PEO patients.

Quantitative Evaluation of Apically Extruded Debris in Root Canals prepared by Single-file Reciprocating and Single File Rotary Instrumentation Systems: A Comparative In vitro Study.

BACKGROUND: Prevention of the debris from extruding to the periapical region is an important aspect to be taken care of as its extrusion can result in pain, periapical inflammation, and delayed healing owing to the presence of highly infective components in the debris. AIM: The present trial was aimed to compare and assess the amount of debris extruded apically using the single-file reciprocating system - WaveOne GOLD and single file rotary system - OneShape in root canal preparations in vitro. MATERIALS AND METHODS: Thirty mandibular premolars were divided into two groups (15 samples each) prepared with a single-file reciprocating system - WaveOne GOLD and single-file rotary system - OneShape. Debris extruded was collected and then stored in an incubator. The final weight was calculated after obtaining the mean of three consecutive weights obtained for each tooth. The data were compared using the ANOVA and Turkey's post hoc analysis. RESULTS: Debris extrusions were recorded in both groups and analysis revealed statistical differences in weight before and after the instrumentation procedures in both groups. When compared, reciprocating single file - WaveOne GOLD extruded more debris than rotary single file system - OneShape with a P < 0.001. CONCLUSIONS: The rotary instrumentation systems extrude lesser debris apically than the reciprocating instrumentation system. The difference found was statistically significant.

Reliability of C-reactive Protein as a Biomarker for Cardiovascular and Oral Diseases in Young and Old Subjects.

BACKGROUND: Increased C-reactive protein (CRP) levels are associated with various diseases such as cardiovascular diseases (CVDs), periodontal diseases, and other inflammatory diseases and can be an effective biomarker for predicting future risks of periodontal diseases. AIMS: The present trial was carried out to evaluate if CRP levels are lower in healthy and young individuals as compared to individuals with high body mass index (BMI), gingival index, and CVDs by assessing CRP levels in young and old individuals (more than 45 years). MATERIALS AND METHODS: A total of 174 participants were recruited for the study and were divided into three following groups: Less than 45 years healthy participants (n = 124), more than 45 years healthy participants, and more 45-year-old participants with CVDs (n = 25). The collected data were subjected to the statistical analysis. RESULTS: In Group I, females had a CRP level of 8476 ± 6.86 pg, females of Group II had a CRP value of 13,262 ± 3.76, and in females of Group III CRP value was 22,761 ± 6.24 pg. These values, respectively, for males were 8494 ± 7.68, 13,840 ± 4.82, and 13,839 ± 3.8. In smokers, the CRP values for Group I, II, and III were 29,012 ± 14.66, 12,568 ± 4.16, and 16,826 ± 3.42, respectively. CONCLUSION: CRP levels were higher in males as compared to females. Higher BMI was associated with higher values of CRP. The participants with higher BMI presented higher CRP values irrespective of age.

Evaluation of Remineralization Seen in Dentin Related to Ceramic Restorations.

BACKGROUND: Recently, the caries management had shifted from extension for prevention to early detection and remineralization of remaining tooth structure using noninvasive techniques. AIM: The present trial was carried out to assess the effect of adding mono-n-Dodecyl phosphate as a surfactant to ceramic on remineralization of dentin. MATERIALS AND METHODS: The 32 teeth were divided into the following four groups randomly (n = 8 each group) - Group I: cavities not restored, Group II: Restored with ceramic, Group III: restored with ceramic and 2% mono-n-Dodecyl phosphate, and Group IV: restored with ceramic and 5% mono-n-Dodecyl phosphate. Knoop indenter microhardness tests were done at 10, 20, and 40 days. The surface was evaluated under scanning electron microscope. The collected data were subjected to statistical evaluation, and the results were formulated. RESULTS: In ceramic restored (Group II), an increase in Knoop microhardness (KHN) values was seen at 10-, 20-, and 40-day assessment with the respective values of 102.2 ± 20.3, 118.7 ± 19.4, and 127 ± 15.1. Similarly, a constant increase at different assessments was observed for Groups III and IV with 2% and 5% of mono-n-Dodecyl phosphate with ceramic with a 40-day KHN value of 128.2 ± 21.4 for Group III and 113.5 ± 17.3 for Group IV. A statistically significant difference was seen between ceramic and 5% mono-n-Dodecyl phosphate (Group IV) with hardness values of 30.31%, 29.8%, and 21.78% for Groups II, III, and IV, respectively, at 20 days. CONCLUSION: The present study concludes that ceramic restorations are effective in promoting dentin remineralization. However, the addition of only 2% mono-n-Dodecyl phosphate was beneficial, whereas 5% mono-n-Dodecyl phosphate reduced the efficacy of ceramic on dentin remineralization.

BLM Potentiates c-Jun Degradation and Alters Its Function as an Oncogenic Transcription Factor.

Mutations in BLM helicase predispose Bloom syndrome (BS) patients to a wide spectrum of cancers. We demonstrate that MIB1-ubiquitylated BLM in G1 phase functions as an adaptor protein by enhancing the binding of transcription factor c-Jun and its E3 ligase, Fbw7α. BLM enhances the K48/K63-linked ubiquitylation on c-Jun, thereby enhancing the rate of its subsequent degradation. Functionally defective Fbw7α mutants prevalent in multiple human cancers are reactivated by BLM. However, BS patient-derived BLM mutants cannot potentiate Fbw7α-dependent c-Jun degradation. The decrease in the levels of c-Jun in cells expressing BLM prevents effective c-Jun binding to 2,584 gene promoters. This causes decreases in the transcript and protein levels of c-Jun targets in BLM-expressing cells, resulting in attenuated c-Jun-dependent effects during neoplastic transformation. Thus, BLM carries out its function as a tumor suppressor by enhancing c-Jun turnover and thereby preventing its activity as a proto-oncogene.

MRN complex-dependent recruitment of ubiquitylated BLM helicase to DSBs negatively regulates DNA repair pathways.

Mutations in BLM in Bloom Syndrome patients predispose them to multiple types of cancers. Here we report that BLM is recruited in a biphasic manner to annotated DSBs. BLM recruitment is dependent on the presence of NBS1, MRE11 and ATM. While ATM activity is essential for BLM recruitment in early phase, it is dispensable in late phase when MRE11 exonuclease activity and RNF8-mediated ubiquitylation of BLM are the key determinants. Interaction between polyubiquitylated BLM and NBS1 is essential for the helicase to be retained at the DSBs. The helicase activity of BLM is required for the recruitment of HR and c-NHEJ factors onto the chromatin in S- and G1-phase, respectively. During the repair phase, BLM inhibits HR in S-phase and c-NHEJ in G1-phase. Consequently, inhibition of helicase activity of BLM enhances the rate of DNA alterations. Thus BLM utilizes its pro- and anti-repair functions to maintain genome stability.