Immune checkpoint inhibitors for POLE or POLD1 proofreading-deficient metastatic colorectal cancer.

BACKGROUND: POLE and POLD1 proofreading deficiency (POLE/D1pd) define a rare subtype of ultramutated metastatic colorectal cancer (mCRC; over 100 mut/Mb). Disease-specific data about the activity and efficacy of immune checkpoint inhibitors (ICIs) in POLE/D1pd mCRC are lacking and it is unknown whether outcomes may be different from mismatch repair-deficient (dMMR)/microsatellite instability-high (MSI-H) mCRCs treated with ICIs. PATIENTS AND METHODS: In this global study, we collected 27 patients with mCRC harboring POLE/D1 mutations leading to proofreading deficiency and treated with anti-programmed cell death-ligand 1 alone +/- anti-cytotoxic T-lymphocyte antigen-4 agents. We collected clinicopathological and genomic characteristics, response, and survival outcomes after ICIs of POLE/D1pd mCRC and compared them with a cohort of 610 dMMR/MSI-H mCRC patients treated with ICIs. Further genomic analyses were carried out in an independent cohort of 7241 CRCs to define POLE and POLD1pd molecular profiles and mutational signatures. RESULTS: POLE/D1pd was associated with younger age, male sex, fewer RAS/BRAF driver mutations, and predominance of right-sided colon cancers. Patients with POLE/D1pd mCRC showed a significantly higher overall response rate (ORR) compared to dMMR/MSI-H mCRC (89% versus 54%; P = 0.01). After a median follow-up of 24.9 months (interquartile range: 11.3-43.0 months), patients with POLE/D1pd showed a significantly superior progression-free survival (PFS) compared to dMMR/MSI-H mCRC [hazard ratio (HR) = 0.24, 95% confidence interval (CI) 0.08-0.74, P = 0.01] and superior overall survival (OS) (HR = 0.38, 95% CI 0.12-1.18, P = 0.09). In multivariable analyses including the type of DNA repair defect, POLE/D1pd was associated with significantly improved PFS (HR = 0.17, 95% CI 0.04-0.69, P = 0.013) and OS (HR = 0.24, 95% CI 0.06-0.98, P = 0.047). Molecular profiling showed that POLE/D1pd tumors have higher tumor mutational burden (TMB). Responses were observed in both subtypes and were associated with the intensity of POLE/D1pd signature. CONCLUSIONS: Patients with POLE/D1pd mCRC showed more favorable outcomes compared to dMMR/MSI-H mCRC to treatment with ICIs in terms of tumor response and survival.

Exploring bacterial diversity in Arctic fjord sediments: a 16S rRNA-based metabarcoding portrait.

The frosty polar environment houses diverse habitats mostly driven by psychrophilic and psychrotolerant microbes. Along with traditional cultivation methods, next-generation sequencing technologies have become common for exploring microbial communities from various extreme environments. Investigations on glaciers, ice sheets, ponds, lakes, etc. have revealed the existence of numerous microorganisms while details of microbial communities in the Arctic fjords remain incomplete. The current study focuses on understanding the bacterial diversity in two Arctic fjord sediments employing the 16S rRNA gene metabarcoding and its comparison with previous studies from various Arctic habitats. The study revealed that Proteobacteria was the dominant phylum from both the fjord samples followed by Bacteroidetes, Planctomycetes, Firmicutes, Actinobacteria, Cyanobacteria, Chloroflexi and Chlamydiae. A significant proportion of unclassified reads derived from bacteria was also detected. Psychrobacter, Pseudomonas, Acinetobacter, Aeromonas, Photobacterium, Flavobacterium, Gramella and Shewanella were the major genera in both the fjord sediments. The above findings were confirmed by the comparative analysis of fjord metadata with the previously reported (secondary metadata) Arctic samples. This study demonstrated the potential of 16S rRNA gene metabarcoding in resolving bacterial composition and diversity thereby providing new in situ insights into Arctic fjord systems.

The seasonal distribution and pollution potential of dissolved heavy metals and nutrients in subterranean estuaries in southern India.

In order to better understand the distribution pattern, pollution degree and the submarine groundwater discharge (SGD) of dissolved heavy metals, 15 subterranean estuaries (STEs) along southwest Indian coast were sampled over three contrasting seasons. The average concentration of metals were ranked as, pre-monsoon > monsoon > post-monsoon with 3 to 12-fold higher groundwater metal concentrations than the adjacent seawater. Average SGD derived essential metal fluxes were five times higher than the toxic metal fluxes of which Fe and Zn together contributed >90 %. Using the Single Factor Contamination Index, the majority of sites were minimally contaminated with only two sites indicating moderate ecological risk due to As. Higher fluxes of Fe, Cu and Zn were likely a result of rising anthropogenic activities. The SGD derived nutrient fluxes were an important source of DIP for primary production in coastal waters and represented 30 % and 44 % of the DIN and DIP inputs respectively.

High Counts in Hematologic Malignancies Predict Low Metaphase Yield for Cytogenetic Analysis.

OBJECTIVES: This study examined the impact of various preanalytical variables on metaphase yield in hematologic malignancies. METHODS: Marrow samples from patients with hematologic malignancies that were subjected to cytogenetic analysis were categorized into two groups: one with samples that yielded an adequate number of metaphases, defined as at least 20, and a second with a low number of metaphases (LNM), having fewer than 20 metaphases. Age, sex, bone marrow nucleated cell (MNC) count, and peripheral blood counts (hemoglobin, total WBC count, and platelet count) were analyzed for an association with LNM. RESULTS: Of 455 samples, 17% (79/455) belonged to the LNM group, including 6% (27/455) that yielded no metaphases. MNCs and WBCs were higher in the LNM group (P < .001 for both). MNCs were higher in LNM groups in both acute myeloid leukemia (P = .008) and acute lymphoblastic leukemia (P = .001). Receiver operating characteristic curves showed moderate prediction of MNC and WBC counts for LNM with areas under the curves of 0.7. Other analyzed parameters showed no significant associations with LNM. CONCLUSIONS: Low metaphase yields occur frequently in hematologic malignancies with high counts. This could reflect biological characteristics of these malignancies that merit further investigation.

A new Tardigrade species, Stygarctus keralensis sp. nov. (Arthrotardigrada: Stygarctidae) from the intertidal zone of Southwest coast of India.

Stygarctus keralensis sp. nov. (Arthrotardigrada: Stygarctidae), is described from the intertidal sandy sediments of Vadakara beach, Kerala, Southwest coast of India. To a certain extent, this new species shows morphological similarity with Stygarctus gourbaultae Renaud-Mornant, 1981, however it can be differentiated by some significant distinguishable characters like double looped seminal receptacle ducts and the presence of a bow shaped internal thickening in between the opening of seminal receptacles situated below the level of gonopore and above the anus; unsculptured body plates I-III; paired sub-cephalic pore/muscle attachment; shape and structure of caudal processes. Presence of minute spikes on third lateral processes in the new species is another unique character by which it can be easily differentiated from S. gourbaultae.

Assessment of benthic macroinvertebrate response to anthropogenic and natural disturbances in the Kodungallur-Azhikode estuary, southwest coast of India.

Benthic biotic indices are important ecological tools extensively used to understand the ecological quality of coastal wetlands. The present study aimed to assess the ecological status of Kodungallur-Azhikode estuary for the first time by using widely used benthic indices such as species richness (S), Shannon diversity index (H'log2), BENTIX, benthic opportunistic polychaetes amphipods (BOPA), AZTI's Marine Biotic Index (AMBI) and multivariate AMBI (M-AMBI). In the canonical correspondence analysis, salinity, dissolved oxygen, organic matter, sediment Eh, sediment pH and sand were identified as important variance descriptors. A single species of an opportunist, Americorophium triaeonyx, an amphipod belonging to the ecological group (EG) III, significantly contributed to the total macrofaunal density. Other dominant opportunistic species included Obelia bidentata (EGII), Arcuatula senhousia (EGIII), Cirolana fluviatilis (EGII), Prionospio cirrifera (EGIV) and Capitella sp. (EGV). The overall assessment indicated a 'good to moderate' condition in AMBI, 'good to poor' condition in M-AMBI, 'high to moderate' condition in BENTIX, 'high to poor' condition in BOPA and 'moderate to poor' condition in univariate Shannon diversity index. All the multivariate indices tested in the study were correlated with each other except BOPA and M-AMBI. The group of stations dominated with a sandy substrate and a moderate level of organic content indicated high to good conditions while other stations demonstrated moderate to poor conditions. However, no significant variation in indices tested between seasons was observed. The present study recommends long-term monitoring of benthic macroinvertebrate assemblages with proper taxonomic identification and functional trait analysis for better calibration of indices, which is the key factor for getting better results.

Evaluation of Cytogenetic Abnormalities in Patients with Acute Lymphoblastic Leukemia.

Acute lymphoblastic leukemia (ALL) accounts for 20% of all adult leukemias and is the most common leukemia during childhood (80%). We present data on cytogenetics of ALL from a tertiary centre in India correlating it with clinical factors. Karyotyping of bone marrow samples of 204 patients with newly diagnosed ALL was performed with standard G-banding technique. Clinical data of patients was obtained from case records. Survival was estimated using Kaplan-Meir curves and compared by the log-rank test. Univariate and multivariate analysis was done for survival with age, sex, immunophenotype, hyperleukocytosis, risk type, remission status and cytogenetics. The most common karyotypes observed were normal in 39.7% (N = 81), hyperdiploidy in 12.7% (N = 26), t(9;22) in 4.4% (N = 9), t(1;19) in 3.9% (N = 8). Adults with ALL had worse survival compared with pediatric patients (HR 3.62; 2.03-6.45 95% CI, p < 0.001). Patients not in morphologic remission after induction chemotherapy fared poorly (HR 4.86; 2.67-8.84 95% CI, p < 0.001). Patients with favourable cytogenetics had better overall survival (HR 0.36; 0.12-1.05 95% CI, p < 0.05). On multivariate analysis, achievement of morphologic remission emerged as single most significant predictor of survival (p < 0.001). MLL gene rearrangement and t(12;21) were seen less commonly as compared to Western data. However, incidence rates of various cytogenetic abnormalities were similar to that reported from other centres from India. Age, morphologic remission at end of induction chemotherapy and favourable cytogenetics correlated significantly with survival.

Chronic effects of copper and zinc on the fish, Etroplus suratensis (Bloch, 1790) by continuous flow through (CFT) bioassay.

Copper (Cu) and zinc (Zn) play a vital role in the growth and development, however increased uptake causes deleterious effects in normal functioning of organisms. We have demonstrated in this contribution the tolerance limit of Cu and Zn on Etroplus suratensis (pearl spot) by Continuous Flow Through (CFT) bioassay and the biomarker responses. The accumulation for Cu, Zn and selected trace metals (Cr, Cd, Ni and Pb) from field conditions, as well Geo-accumulation index (I geo) and Contamination factor (C.F) suggested moderate to heavy pollution in the Cochin estuarine system. The 96 h LC50 values for Cu was 1.74 ±â€¯0.04 mg L-1and that for Zn was 24.36 ±â€¯0.58 mg L-1 at 95% confidence interval. No observed effect concentration (NOEC) and low observed effect concentration (LOEC) for Cu and Zn were derived based on the survival rates. Chronic toxicity values for Cu and Zn were 0.23 mg L-1 and 2.005 mg L-1 respectively for 30 days period. The histological, biochemical, hematological and behavioral parameters showed significant variations at sublethal concentrations. Lamellar hyperplasia in gills, vacuolation combined with necrosis in liver, increased occurrence of melanomacrophage centres in spleen were noticed at chronic levels for both Cu and Zn. Tissue specific bioconcentration was observed for zinc and copper in gill and liver respectively, with least rate of bioconcentration observed in muscle tissues. Malaonate Dehydrogenase (MDH), Super oxide dismutase (SOD), Nonspecific esterase (EST) activity significantly varied compared to control at NOEC and LOEC values in both the metals. The hematological and genotoxic alterations as decrease in erythrocyte count, lymphocytes, hemoglobin concentration and hematocrit percentage were significantly reduced (p < 0.05) and increased thrombocytes and neutrophils, increased frequency of micronuclei, lobed, blebbed and notched nuclei and binucleate cells were characteristic for the metals at the sublethal concentrations. The frequency of behavioral changes remained significantly higher at chronic level than the control group. Thus such CFT based studies are important for precisely mapping the toxicity changes in organisms and also to develop suitable water quality guidelines.

Taxonomy of myid bivalves from fragmented brackish-water habitats in India, with a description of a new genus Indosphenia (Myidae, Myoidea, Myidae).

A group of small bivalves inhabiting Indian brackish-water estuaries and lagoons (known locally as backwaters), variously assigned to Corbula, Cuspidaria, and Sphenia, are reviewed and, based on shell characters, shown to be congeneric. Molecular (COI) and morphological data indicate that this group belongs to the family Myidae. Furthermore, the combined data suggest that these Indian myids are a sister taxon of the genus Sphenia. The Indian material studied herein exhibits a functional morphology typical of infaunal bivalves, whereas typical Sphenia are nestling and epibyssate. A new genus, Indosphenia, is thus erected for the Indian group and includes five species, one of which is named in this study. Indospheniakayalum Oliver, Hallan & Jayachandran, gen. et sp. n. is described from the Cochin Backwater on the western coast of India. Cuneocorbulacochinensis (Preston, 1916) is transferred to Indosphenia. Additionally, the west coast taxa I.abbreviata (Preston, 1907), I.abbreviatachilkaensis (Preston, 1911) and I.sowerbyi (EA Smith, 1893) are recognised herein. Corbulaalcocki Preston, 1907, Corbulagracilis Preston, 1907, Corbulacalcaria Preston, 1907 and Corbulapfefferi Preston, 1907 are placed in synonymy with I.abbreviata, and Cuspidariaannandalei Preston, 1915 is synonymised with I.abbreviatachilkaensis.

Toxicity effects of zinc on two marine diatoms, under varying macronutrient environment.

Diatoms constitute one of the fundaments in the trophic food chains and are responsible for much of the bioaccumulation of heavy metals, in particular zinc, throughout the web of food chains. Chaetoceros calcitrans and Thalassiosira weissflogii are widely present as a dominant marine diatom in estuarine and coastal waters. The zinc uptake, its fractionation in subcellular structures and macromolecular compartments in two marine diatoms, C. calcitrans and T. weissflogii and trophic transfer under different macronutrient concentrations were investigated. The study reveals a significant interaction between macronutrients and metal uptake by the two diatom species. Increased cellular accumulation of zinc and N, P-limited condition were found to affect N, P and Si assimilation by diatoms negatively. Conversely, nitrogen (N) limitation inhibited the intracellular uptake of zinc. However, at higher concentration of zinc, the difference in zinc uptake between nutrient enriched condition (+NP) and nutrient limited (N or P) condition become smaller, indicating that the zinc uptake by diatoms is less dependent on N containing protein ligands at high Zn concentration. Nitrogen concentration in the medium was also found to affect the relative distribution of zinc in subcellular structures and macromolecular components. However, major portion of zinc was distributed in soluble substance and in the protein of the algal cells. The N limited condition facilitates the accumulation of zinc in cell organelles (insoluble substance) leading to increased toxicity. Trophic transfer of zinc was also measured by calculating percentage of metal retained in mussel, Perna viridis over the experiment period. Regardless of the algal species, the percentage of accumulation of zinc was found to be high in mussels fed with algal cells acclimated to nutrient enriched condition (31 and 38%). Our study therefore suggests that N enrichment may lead to an increase in Zn uptake and transfer in marine plankton.

New species of Victoriopisa Karaman Barnard, 1979 (Crustacea: Amphipoda: Eriopisidae) from Vembanad backwaters, Southwest coast of India.

The new species is the third of genus Victoriopisa from Indian waters.The species is described from the Valanthakad mangrove area of Vembanad backwater. Victoriopisa cusatensis sp. nov differs from other species of Victoriopisa by presenting: (1) a characteristic projection at dorsolateral margin of peduncle article 2 of antenna 1in male while smooth in female; (2) broad, triangular lateral cephalic lobe; (3) posteroventral tooth in epimeral plates; (4) presence of lateral and subapical spines in telson (5) smooth palm of gnathopod 2 without any excavations.

A Qualitative Analysis of Periodontal Pathogens in Chronic Periodontitis Patients after Nonsurgical Periodontal Therapy with and without Diode Laser Disinfection Using Benzoyl-DL Arginine-2-Naphthylamide Test: A Randomized Clinical Trial.

INTRODUCTION: Periodontal diseases are caused by pathogenic bacteria locally colonized in the dental biofilm creating infection; the main etiological factor is represented by dental plaque and in particular by anaerobic Gram-negative bacilli. For that reason, the first phase of periodontal treatment is always represented by the initial preparation which primarily aims at the elimination or reduction of bacterial infection and the control of periodontal plaque-associated inflammation. Yet, another innovative causal therapy is represented by the irradiation of periodontal pockets with LASER. The aim of this randomized clinical study is to compare and to detect the presence of periodontal pathogens in chronic periodontitis patients after nonsurgical periodontal therapy with and without diode LASER disinfection using BANA test. MATERIALS AND METHODS: This randomized clinical trial includes 20 patients having chronic periodontitis. From each patient, one test site and one control site were selected and assessed for gingival index (GI), oral hygiene index (OHI), pocket probing depth and clinical attachment level (CAL), and presence of BANA pathogens. The test site underwent scaling and root planning along with diode LASER therapy as an adjuvant while the control site received scaling and root planning alone. Patients were recalled for review after 2 weeks and 2 months where periodontal parameters were assessed and plaque samples were collected and analyzed for BANA pathogens. RESULTS: The test site where LASER was used as an adjuvant showed significant reduction in pocket probing depth, CAL, OHI, GI, and periodontal pathogens which shows that the amount of recolonization of microbes is less when LASER is used as an adjuvant to conventional therapy. CONCLUSION: Diode LASER as an adjuvant to SRP has shown additional benefits over conventional therapy in all the clinical parameters evaluated and this can be associated in the treatment of periodontal therapy. BANA-enzymatic kit is a simple chair side kit which can be reliable indicator of BANA positive species in dental plaque.

Tri-Layered Nanocomposite Hydrogel Scaffold for the Concurrent Regeneration of Cementum, Periodontal Ligament, and Alveolar Bone.

A tri-layered scaffolding approach is adopted for the complete and concurrent regeneration of hard tissues-cementum and alveolar bone-and soft tissue-the periodontal ligament (PDL)-at a periodontal defect site. The porous tri-layered nanocomposite hydrogel scaffold is composed of chitin-poly(lactic-co-glycolic acid) (PLGA)/nanobioactive glass ceramic (nBGC)/cementum protein 1 as the cementum layer, chitin-PLGA/fibroblast growth factor 2 as the PDL layer, and chitin-PLGA/nBGC/platelet-rich plasma derived growth factors as the alveolar bone layer. The tri-layered nanocomposite hydrogel scaffold is cytocompatible and favored cementogenic, fibrogenic, and osteogenic differentiation of human dental follicle stem cells. In vivo, tri-layered nanocomposite hydrogel scaffold with/without growth factors is implanted into rabbit maxillary periodontal defects and compared with the controls at 1 and 3 months postoperatively. The tri-layered nanocomposite hydrogel scaffold with growth factors demonstrates complete defect closure and healing with new cancellous-like tissue formation on microcomputed tomography analysis. Histological and immunohistochemical analyses further confirm the formation of new cementum, fibrous PDL, and alveolar bone with well-defined bony trabeculae in comparison to the other three groups. In conclusion, the tri-layered nanocomposite hydrogel scaffold with growth factors can serve as an alternative regenerative approach to achieve simultaneous and complete periodontal regeneration.

Periodontal Specific Differentiation of Dental Follicle Stem Cells into Osteoblast, Fibroblast, and Cementoblast.

The dental follicle is a source of dental follicle stem cells (DFCs), which have the potential to differentiate into the periodontal lineage. DFCs therefore are of value in dental tissue engineering. The purpose of this study was to evaluate the effect of growth factor type and concentration on DFC differentiation into periodontal specific lineages. DFCs were isolated from the human dental follicle and characterized for the expression of mesenchymal markers. The cells were positive for CD-73, CD-44, and CD-90; and negative for CD-33, CD-34, and CD-45. The expression of CD-29 and CD-31 was almost negligible. The cells also expressed periodontal ligament and cementum markers such as periodontal ligament-associated protein-1 (PLAP-1), fibroblast growth factor-2 (FGF-2), and cementum protein-1 (CEMP-1), however, the expression of osteoblast markers was absent. Further, the DFCs were cultured in three different induction medium to analyze the osteoblastic, fibroblastic, and cementoblastic differentiation. Runt-related transcription factor 2 (RUNX-2), alkaline phosphatase (ALP) activity, alizarin staining, calcium quantification, collagen type-1 (Col-1), and osteopontin (OPN) expression confirmed the osteoblastic differentiation of DFCs. DFCs cultured in recombinant human FGF-2 (rhFGF-2) containing medium showed enhanced PLAP-1, FGF-2, and COL-1 expression with increasing concentration of rhFGF-2 which thereby confirmed periodontal ligament fibroblastic differentiation. Similarly, DFCs cultured in recombinant human cementum protein-1 (rhCEMP-1) containing medium showed enhanced bone sialoprotein-2 (BSP-2), CEMP-1, and COL-1 expression with respect to rhCEMP-1 which confirmed cementoblastic differentiation. The expression of osteoblast, fibroblast, and cementoblast-related genes of DFCs cultured in induction medium was enhanced in comparison to DFCs cultured in noninduction medium. Thus, growth factor-dependent differentiation of DFCs into periodontal specific lineages was proved by quantitative analysis.

Evaluation of antimicrobial efficacy and release pattern of tetracycline and metronidazole using a local delivery system.

The present study compared the effectiveness of two antimicrobials, tetracycline and metronidazole, in reducing subgingival microorganisms in periodontal pockets using an ethyl cellulose strip as the delivery medium. The study involved 30 patients, with a minimum of 3 periodontal pockets with probing depths > 6 mm throughout the oral cavity. Patients were given supragingival scaling and then divided into 5 groups, depending on the length of time the medication was in place. Sites were marked for tetracycline, metronidazole, and placebo. Sites were wiped and isolated, and baseline microbiology samples were taken for Gram staining and culture methods. After treatment, subgingival microbiological samples were taken again. The ethyl cellulose strips were removed and analyzed for any remaining drug. Results showed that tetracycline and metronidazole can both be applied locally to periodontal sites using ethyl cellulose strips and markedly suppress the subgingival bacteria over a period of several days. The tetracycline showed a faster release; however, the metronidazole required a lesser concentration to achieve complete reduction of the subgingival flora.

Long stay patients in a state mental hospital.

A study of the long stay patients in a state mental hospital on 30th June 1980 revealed that 70 per cent of the total 1,434, i.e. 1015 had spent more than two years continuously; most of them are chronic schizophrenics, admitted by the police, lacking social support. The need for rehabilitating them both within and outside the hospital is stressed.