The prokaryotic and eukaryotic microbiome of Pacific oyster spat is shaped by ocean warming but not acidification.

Pacific oysters (Magallana gigas, a.k.a. Crassostrea gigas), the most widely farmed oysters, are under threat from climate change and emerging pathogens. In part, their resilience may be affected by their microbiome, which, in turn, may be influenced by ocean warming and acidification. To understand these impacts, we exposed early-development Pacific oyster spat to different temperatures (18°C and 24°C) and pCO2 levels (800, 1,600, and 2,800 µatm) in a fully crossed design for 3 weeks. Under all conditions, the microbiome changed over time, with a large decrease in the relative abundance of potentially pathogenic ciliates (Uronema marinum) in all treatments with time. The microbiome composition differed significantly with temperature, but not acidification, indicating that Pacific oyster spat microbiomes can be altered by ocean warming but is resilient to ocean acidification in our experiments. Microbial taxa differed in relative abundance with temperature, implying different adaptive strategies and ecological specializations among microorganisms. Additionally, a small proportion (~0.2% of the total taxa) of the relatively abundant microbial taxa were core constituents (>50% occurrence among samples) across different temperatures, pCO2 levels, or time. Some taxa, including A4b bacteria and members of the family Saprospiraceae in the phyla Chloroflexi (syn. Chloroflexota) and Bacteroidetes (syn. Bacteroidota), respectively, as well as protists in the genera Labyrinthula and Aplanochytrium in the class Labyrinthulomycetes, and Pseudoperkinsus tapetis in the class Ichthyosporea were core constituents across temperatures, pCO2 levels, and time, suggesting that they play an important, albeit unknown, role in maintaining the structural and functional stability of the Pacific oyster spat microbiome in response to ocean warming and acidification. These findings highlight the flexibility of the spat microbiome to environmental changes.IMPORTANCEPacific oysters are the most economically important and widely farmed species of oyster, and their production depends on healthy oyster spat. In turn, spat health and productivity are affected by the associated microbiota; yet, studies have not scrutinized the effects of temperature and pCO2 on the prokaryotic and eukaryotic microbiomes of spat. Here, we show that both the prokaryotic and, for the first time, eukaryotic microbiome of Pacific oyster spat are surprisingly resilient to changes in acidification, but sensitive to ocean warming. The findings have potential implications for oyster survival amid climate change and underscore the need to understand temperature and pCO2 effects on the microbiome and the cascading effects on oyster health and productivity.

A 3D in vitro model for assessing the influence of intraocular lens: Posterior lens capsule interactions on lens epithelial cell responses.

Posterior Capsule Opacification (PCO), the most frequent complication of cataract surgery, is caused by the infiltration and proliferation of lens epithelial cells (LECs) at the interface between the intraocular lens (IOL) and posterior lens capsule (PLC). According to the "no space, no cells, no PCO" theory, high affinity (or adhesion force) between the IOL and PLC would decrease the IOL: PLC interface space, hinder LEC migration, and thus reduce PCO formation. To test this hypothesis, an in vitro hemisphere-shaped simulated PLC (sPLC) was made to mimic the human IOL: PLC physical interactions and to assess their influence on LEC responses. Three commercially available IOLs with different affinities/adhesion forces toward the sPLC, including Acrylic foldable IOL, Silicone IOL, and PMMA IOL, were used in this investigation. Using the system, the physical interactions between IOLs and sPLC were quantified by measuring the adhesion force and interface space using an adhesion force apparatus and Optical Coherence Tomography, respectively. Our data shows that high adhesion force and tight binding between IOL and sPLC contribute to a small interface space (or "no space"). By introducing LECs into the in vitro system, we found that, with small interface space, among all IOLs, acrylic foldable IOLs permitted the least extent of LEC infiltration, proliferation, and differentiation (or "no cells"). Further statistical analyses using clinical data revealed that weak LEC responses are associated with low clinical PCO incidence rates (or "no PCO"). The findings support that the in vitro system could simulate IOL: PLC interplays and predict IOLs' PCO potential in support of the "no space, no cells, no PCO" hypothesis.

Deciphering carbon dioxide fluxes and interactions in the Ganga river Basin of South Asia.

Anthropogenic influences significantly modify the hydrochemical properties and material flow in riverine ecosystems across Asia, potentially accounting for 40-50% of global emissions. Despite the pervasive impact on Asian rivers, there is a paucity of studies investigating their correlation with carbon dioxide (CO2) emissions. In this study, we computed the partial pressure of CO2 (pCO2) using the carbonate equilibria-based model (pCO2SYS) and examined its correlation with hydrochemical parameters from historical records at 91 stations spanning 2013-2021 in the Ganga River. The investigation unveiled substantial spatial heterogeneity in the pCO2 across the Ganga River. The pCO2 concentration varied from 1321.76 µatm, 1130.98 µatm, and 1174.33 µatm in the upper, middle, and lower stretch, respectively, with a mean of 1185.29 µatm. Interestingly, the upper stretch exhibited elevated mean pCO2 and FCO2 levels (fugacity of CO2: 3.63 gm2d-1) compared to the middle and lower stretch, underscoring the intricate interplay between hydrochemistry and CO2 dynamics. In the context of pCO2 fluctuations, nitrate concentrations in the upper segment and levels of biological oxygen demand (BOD) and dissolved oxygen (DO) in the middle and lower segments are emerging as crucial explanatory factors. Furthermore, regression tree (RT) and importance analyses pinpointed biochemical oxygen demand (BOD) as the paramount factor influencing pCO2 variations across the Ganga River (n = 91). A robust negative correlation between BOD and FCO2 was also observed. The distinct longitudinal patterns of both parameters may induce a negative correlation between BOD and pCO2. Therefore, comprehensive studies are necessitated to decipher the underlying mechanisms governing this relationship. The present insights are instrumental in comprehending the potential of CO2 emissions in the Ganga River and facilitating riverine restoration and management. Our findings underscore the significance of incorporating South Asian rivers in the evaluation of the global carbon budget.

Partial pressure of carbon dioxide/pH interaction and its association with mortality among patients mechanically ventilated in the emergency department.

OBJECTIVES: There is currently conflicting data as to the effects of hypercapnia on clinical outcomes among mechanically ventilated patients in the emergency department (ED). These conflicting results may be explained by the degree of acidosis. We sought to test the hypothesis that hypercapnia is associated with increased in-hospital mortality and decreased ventilator-free days at lower pH, but associated with decreased in-hospital mortality and increased ventilator-free days at higher pH, among patients requiring mechanical ventilation in the emergency department (ED). METHODS: Secondary analysis of patient level data from prior clinical trials and cohort studies that enrolled adult patients who required mechanical ventilation in the ED. Patients who had a documented blood gas while on mechanical ventilation in the ED were included in these analyses. The primary outcome was in-hospital mortality, and secondary outcome was ventilator-free days. Mixed-effects logistic, linear, and survival-time regression models were used to test if pH modified the association between partial pressure of carbon dioxide (pCO2) and outcome measures. RESULTS: Of the 2348 subjects included, the median [interquartile range (IQR)] pCO2 was 43 (35-54) and pH was 7.31 (7.22-7.39). Overall, in-hospital mortality was 27%. We found pH modified the association between pCO2 and outcomes, with higher pCO2 associated with increased probability of in-hospital mortality when pH is below 7.00, and decreased probability of in-hospital mortality when pH is above 7.10. These results remained consistent across multiple sensitivity and subgroup analyses. A similar relationship was found with ventilator-free days. CONCLUSIONS: Higher pCO2 is associated with decreased mortality and greater ventilator-free days when pH is >7.10; however, it is associated with increased mortality and fewer ventilator-free days when the pH is below 7.00. Targeting pCO2 based on pH in the ED may be a potential intervention target for future clinical trials to improve clinical outcomes.

Multiple blood gas variables predict AKI survival in an independent manner.

BACKGROUND AND AIM: Acute kidney injury (AKI) is becoming increasingly prevalent among hospitalized patients and carries a poor prognosis. While new biomarkers show promise in identifying early stages of AKI, accurately predicting severe outcomes such as the need for kidney replacement therapy (KRT) or death remains a challenge. However, blood gas analyses (BGA) can be used to diagnose life-threatening complications associated with AKI. The objective of this study was to assess the role of BGA as a biomarker panel in both emerging and established cases of AKI. METHODS: Retrospective observational study examining subjects with newly developed acute kidney injury (AKI). The study will document venous and arterial pH, pCO2, and actual bicarbonate levels upon hospital admission and at the onset of AKI. The primary endpoints include in-hospital mortality, the need for kidney replacement therapy (KRT), and the recovery of kidney function (ROKF). RESULTS: A total of 202 individuals were included in the study. Three variables were found to be independent predictors of in-hospital survival: admission arterial pH, arterial pH at acute kidney injury (AKI) onset, and arterial pCO2 at AKI onset. Additionally, venous pCO2 at AKI onset was identified as an independent predictor for the need of kidney replacement therapy (KRT). CONCLUSIONS: Our study suggests that blood gas analysis may have a potential role in predicting severe outcome variables in acute kidney injury (AKI). The associated costs are minimal.

Ponte osteotomies for treatment of spinal deformities: they are not all made equal.

PURPOSE: To compare surgical outcomes of Ponte's osteotomies for treatment of adolescent idiopathic scoliosis (AIS), Scheuermann's kyphosis (SK), and adult scoliosis (AdS). METHODS: We conducted a retrospective review of patients with AIS, SK, and AdIS who underwent posterior spinal instrumented fusion (PSIF) at our Institution from January 2019 to December 2022. Demographics, imaging, and intraoperative data (including number of osteotomies performed, blood losses, surgical timing, and complications) were extracted from patient charts. RESULTS: A total of 80 patients (62 AIS, 7 SK, and 11 AdS) were enrolled in the study. All patients were treated with a PSIF and a total of 506 Ponte osteotomies were performed (5.8 ± 4.1, 9.3 ± 2.4, and 7.5 ± 2.5 average osteotomies per patient in the AIS, SK, and AdS group, respectively; p = 0.045). Average time per osteotomy was 6.3 ± 1.5 min in the AIS group, and 5.8 ± 2.1 and 8.7 ± 4.0 in the SK and AdS group, respectively (p = 0.002). Blood loss was significantly smaller in the SK group (8.6 ± 9.6 ml per osteotomy) compared to AIS group (34.9 ± 23.7 ml) and AdS group (34.9 ± 32.7 ml) (p = 0.001). A total of 4 complications were observed in the AIS group (1.1%) and 2 complications in the AdS group (2.4%), but this was not statistically significant. CONCLUSIONS: Our study shows that Ponte's osteotomies are safe and effective in surgical treatment of AIS, SK, and AdS. Blood loss and execution time per osteotomy are significantly smaller in the SK group compared to AIS and AdS. No significant differences were noted in terms of complications between the three groups.

Development and validation of impact of early integration of palliative care and oncology(IEI PCO) questionnaire: a survey for medical oncologists and nurses.

OBJECTIVES: Many associations have recently recommended early integration of oncology and palliative care for more standard cancer care and better quality of life. We aimed to create a questionnaire to assess the opinion of medical oncologists and nurses about the clinical impact of the integrated palliative care and oncology (PCO) program. METHODS: A novel semi-structured questionnaire called Impact of Early Integration of Palliative Care Oncology (IEI PCO) questionnaire was developed and tested for validity and reliability then distributed to the oncologists and nurses working in Kuwait Cancer Control Center. RESULTS: After the pilot stage, testing the final questionnaire for validity and reliability was done with satisfactory results. Finally, the complete questionnaires were 170 out of 256 (response rate 66.41%). More awareness about the available palliative care services and the new available PCO services (p-value < 0.001 for all). Most of the oncologists and nurses agreed with the currently available structure of PCO, appreciated the patients' discharge plan and continuity of care of palliative medicine, admitted less work burden, a better attitude, and higher satisfaction (p-value for all < 0.001) toward palliative care. Significant improvements in symptoms were appreciated by oncologists and nurses after the integration of palliative care (p-value for all < 0.001. Oncologists and nurses valued repeated honest communication, discussion of the goals of care, dealing more effectively with ending active treatment, and higher acceptance of patients and families of PC policy of transfer, and significant progress in the care of end-of-life symptoms (p-value for all < 0.001). CONCLUSIONS: The IEI PCO questionnaire demonstrated the psychometric criteria for content, face, and construct validity and reliability. It provides a valuable tool to assess the impact of PCO integration. The opinion of medical oncologists and nurses was significantly positive toward the early integration of PCO in Kuwait in most aspects of care. This integration led to improved symptom control, end-of-life care, communication, and planned discharge and follow-up plans. Moreover, decreases the work burden, improves attitude, higher satisfaction of the oncology staff, and continuity of care.

Can perioperative pCO2 gaps predict complications in patients undergoing major elective abdominal surgery randomized to goal-directed therapy or standard care? A secondary analysis.

The difference between venous and arterial carbon dioxide pressure (pCO2 gap), has been used as a diagnostic and prognostic tool. We aimed to assess whether perioperative pCO2 gaps can predict postoperative complications. This was a secondary analysis of a multicenter RCT comparing goal-directed therapy (GDT) to standard care in which 464 patients undergoing high-risk elective abdominal surgery were included. Arterial and central venous blood samples were simultaneously obtained at four time points: after induction, at the end of surgery, at PACU/ICU admission, and PACU/ICU discharge. Complications within the first 30 days after surgery were recorded. Similar pCO2 gaps were found in patients with and without complications, except for the pCO2 gap at the end of surgery, which was higher in patients with complications (6.0 mmHg [5.0-8.0] vs. 6.0 mmHg [4.1-7.5], p = 0.005). The area under receiver operating characteristics curves for predicting complications from pCO2 gaps at all time points were between 0.5 and 0.6. A weak correlation between ScvO2 and pCO2 gaps was found for all timepoints (ρ was between - 0.40 and - 0.29 for all timepoints, p < 0.001). The pCO2 gap did not differ between GDT and standard care at any of the selected time points. In our study, pCO2 gap was a poor predictor of major postoperative complications at all selected time points. Our research does not support the use of pCO2 gap as a prognostic tool after high-risk abdominal surgery. pCO2 gaps were comparable between GDT and standard care. Clinical trial registration Netherlands Trial Registry NTR3380.

Elevated nitrogen loadings facilitate carbon dioxide emissions from urban inland waters.

Carbon dioxide (CO2) production and emissions from inland waters play considerable roles in global atmospheric CO2 sources, while there are still uncertainties regarding notable nutrient inputs and anthropogenic activities. Urban inland waters, with frequently anthropogenic modifications and severely nitrogen loadings, were hotspots for CO2 emissions. Here, we investigated the spatiotemporal patterns of partial pressure of CO2 (pCO2) and CO2 fluxes (FCO2) in typical urban inland waters in Tianjin, China. Our observation indicated that pCO2 values were oversaturated in highly polluted waters, particularly in sewage rivers and urban rivers, exhibiting approximately 9 times higher than the atmosphere equilibrium concentration during sampling campaigns. Obviously, the spatiotemporal distributions of pCO2 and FCO2 emphasized that the water environmental conditions and anthropogenic activities jointly adjusted primary productivity and biological respiration of inland waters. Meanwhile, statistically positive correlations between pCO2/FCO2 and NH4+-N/NO3--N (p < 0.05) suggested that nitrogen biogeochemical processes, especially the nitrification, played a dominant role in CO2 emissions attributing to the water acidification that stimulated CO2 production and emissions. Except for slight CO2 sinks in waters with low organic contents, the total CO2 emissions from the urban surface waters of Tianjin were remarkable (286.8 Gg yr-1). The results emphasized that the reductions of nitrogen loadings, sewage draining waters, and agricultural pollution could alleviate CO2 emissions from urban inland waters.

The Evolution of Central Venous-to-arterial Carbon Dioxide Difference (pCO2 Gap) during Resuscitation Affects ICU Outcomes: A Prospective Observational Study.

How to cite this article: Sundarsingh V, Kumar M, Rodrigues PR. The Evolution of Central Venous-to-arterial Carbon Dioxide Difference (pCO2 Gap) during Resuscitation Affects ICU Outcomes: A Prospective Observational Study. Indian J Crit Care Med 2024;28(7):709.

Author Response: The Evolution of Central Venous-to-arterial Carbon Dioxide Difference (PCO2 Gap) during Resuscitation Affects ICU Outcomes: A Prospective Observational Study.

How to cite this article: Tiwari AM, Zirpe KG, Kulkarni AP. Author Response: The Evolution of Central Venous-to-arterial Carbon Dioxide Difference (PCO2 Gap) During Resuscitation Affects ICU Outcomes: A Prospective Observational Study. Indian J Crit Care Med 2024;28(7):710.

The Evolution of Central Venous-to-arterial Carbon Dioxide Difference (PCO2 Gap) during Resuscitation Affects ICU Outcomes: A Prospective Observational Study.

Introduction: The usual methods of perfusion assessment in patients with shock, such as capillary refill time, skin mottling, and serial serum lactate measurements have many limitations. Veno-arterial difference in the partial pressure of carbon dioxide (PCO2 gap) is advocated being more reliable. We evaluated serial change in PCO2 gap during resuscitation in circulatory shock and its effect on ICU outcomes. Materials and methods: This prospective observational study included 110 adults with circulatory shock. Patients were resuscitated as per current standards of care. We recorded invasive arterial pressure, urine output, cardiac index (CI), PCO2 gap at ICU admission at 6, 12, and 24 hours, and various patient outcomes. Results: Significant decrease in PCO2 gap was observed at 6 h and was accompanied by improvement in serum lactate, mean arterial pressure, CI and urine output in (n = 61). We compared these patients with those in whom this decrease did not occur (n = 49). Mortality and ICU LOS was significantly lower in patients with low PCO2 gap, while more patients with high PCO2 gap required RRT. Conclusion: We found that a persistently high PCO2 gap at 6 and 12 h following resuscitation in patients with shock of various etiologies, was associated with increased mortality, need for RRT and increased ICU LOS. High PCO2 gap had a moderate discriminative ability to predict mortality. How to cite this article: Zirpe KG, Tiwari AM, Kulkarni AP, Vaidya HS, Gurav SK, Deshmukh AM, et al. The Evolution of Central Venous-to-arterial Carbon Dioxide Difference (PCO2 Gap) during Resuscitation Affects ICU Outcomes: A Prospective Observational Study. Indian J Crit Care Med 2024;28(4):349-354.

Arterial Blood Gas Analysis in Patients with Stridor, and the Impact of Emergency Tracheostomy on It: A Tertiary Care Center Experience.

Comparative evaluation of arterial blood gas in patients with stridor, before and after emergency tracheostomy. The present prospective study was conducted in tertiary care Centre from February 2022 to June 2023 on 42 patients who presented with stridor and underwent emergency tracheostomy in our department. After proper history taking and clinical examination, nonsurgical cause of stridor was ruled out. Patients were then classified on the basis of location of cause of stridor (whether oropharyngeal, hypo-pharyngeal, supra-glottic, glottic or sub-glottic). Immediately an arterial blood gas (ABG) analysis was done, and emergency tracheostomy was performed. Following tracheostomy, ABG analysis was done immediately, after 12 h and after 24 h. The mean age of presentation of stridor in our study was 65.02 ± 3.23 years, with male preponderance (Male: female ratio being 3.66:1). Most common etiology of stridor in our study was glottic carcinoma comprising 50%, and least common etiology of stridor was hypopharyngeal carcinoma, and subglottic stenosis comprising 2.4% each. There was statistically significant normalization of ABG in terms of pH, PO2, PCO2, HCO3. Mean pH, PO2, PCO2, and HCO3 before tracheostomy were 7.31, 74.8, 60.6, and 29.8 respectively. Mean pH, PO2, PCO2, HCO3, immediately after tracheostomy were7.38, 91.3, 48.4, and 27.4 respectively. After 12 h of tracheostomy, mean pH, PO2, PCO2, HCO3 were 7.41, 95.4, 42.7, 25.3 respectively. Mean pH, PO2, PCO2, HCO3 24 h after emergency tracheostomy were 7.441, 95.5, 42.8, 24.6 respectively. Emergency tracheostomy in stridor patients improves the acid base and ventilatory status, by relieving the obstruction as evidenced by statistically significant improvement in arterial blood gas values, and can be used as a diagnostic tool in upper airway obstruction.

Water column properties associated with massive algal bloom of green Noctiluca scintillans in the Arabian Sea.

An algal bloom of Noctiluca scintillans (NS) was monitored for 20 days in the Arabian Sea during February 2017. The stations under the influence of NS had low temperature and high salinity compared to outside indicating influence of convective mixing. The microscopic cell count of NS reached a value of 52,600 cells l-1. The surface variability in oxygen and pCO2 measured alongside showed a strong disconnect. Modest supersaturation of surface waters (ΔpCO2 = 3-75 µatm) was observed around the NS bloom compared to outside. However, as these stations were under the influence of convective mixing, the observed change in pCO2 due to subsurface ventilation cannot be ruled out. From the viewpoint of climate change and its influence on oceanic processes, constant monitoring of this bloom becomes essential due to its survival strategy in nutrient-depleted conditions and light of the present observations.

Seasonally varying biogeochemical regime around the coral habitats off central west coast of India.

The Western Indian Continental Shelf (WICS) experiences upwelling during the Southwest Monsoon (SWM), leading to deoxygenation and acidification of subsurface waters. The region has patchy growth of corals, e.g. in the Grande Island and Angria Bank. Measurements made during the late SWM of 2022 reveal that the shelf waters around the Grande Island were subject to varying environmental conditions, viz. lower temperature (21.3-26.1°C), oxygen (0-4.9 mL L-1) and pHT (7.506-7.927). Complete anoxia was associated with sulphide build-up to a maximum of 5.9 µmol L-1 at 17 m depth. An additional episodic condition (high temperature, low oxygen and pH) also occurred associated presumably with a plankton bloom in April 2017. Hence, unlike the offshore coral site Angria Bank, waters around the Grande Island experiences extreme changes in physico-chemical conditions (e.g. Ωarg ∼1.2-1.8 during October 2022) seasonally as reported here. The biogeochemical conditions are however not as intense (Ωarg = 0.6) as observed along the eastern boundary upwelling system of the Pacific Ocean.

A Case of Nemaline Myopathy With Sleep-Related Hypoventilation Diagnosed Using Polysomnography During Daytime Napping.

This is the case of a 49-year-old woman who was admitted to the hospital for a close examination of pulmonary hypertension; however, the next morning, she developed carbon dioxide (CO2) narcosis and was started on artificial ventilation. As pulmonary arterial hypertension was ruled out, the patient was extubated, and 24-hour transcutaneous partial pressure of carbon dioxide (PCO2)(transcutaneous carbon dioxide (TcPCO2)) monitoring was performed to diagnose sleep-related hypoventilation. Polysomnography (PSG) during daytime napping revealed markedly decreased chest motion and a "pseudo-central event," which was neither central nor obstructive hypopnea. Based on the PSG results and physical examination findings, a neuromuscular disorder was suspected, and a muscle biopsy was performed to diagnose nemaline myopathy. Neuromuscular diseases are widely recognized for their association with sleep-disordered breathing; thus, sleep-related hypoventilation should also be considered. Monitoring of TcPCO2 and PSG are useful tools in identifying the cause of hypoventilation; however, overnight PSG may cause CO2 narcosis in some diseases. In such cases, PSG may be beneficial during daytime napping.

Carbonate Chemistry and the Potential for Acidification in Georgia Coastal Marshes and the South Atlantic Bight, USA.

In coastal regions and marginal bodies of water, the increase in partial pressure of carbon dioxide (pCO2) in many instances is greater than that of the open ocean due to terrestrial (river, estuarine, and wetland) influences, decreasing buffering capacity and/or increasing water temperatures. Coastal oceans receive freshwater from rivers and groundwater as well as terrestrial-derived organic matter, both of which have a direct influence on coastal carbonate chemistry. The objective of this research is to determine if coastal marshes in Georgia, USA, may be "hot-spots" for acidification due to enhanced inorganic carbon sources and if there is terrestrial influence on offshore acidification in the South Atlantic Bight (SAB). The results of this study show that dissolved inorganic carbon (DIC) and total alkalinity (TA) are elevated in the marshes compared to predictions from conservative mixing of the freshwater and oceanic end-members, with accompanying pH around 7.2 to 7.6 within the marshes and aragonite saturation states (ΩAr) <1. In the marshes, there is a strong relationship between the terrestrial/estuarine-derived organic and inorganic carbon and acidification. Comparisons of pH, TA, and DIC to terrestrial organic material markers, however, show that there is little influence of terrestrial-derived organic matter on shelf acidification during this period in 2014. In addition, ΩAr increases rapidly offshore, especially in drier months (July). River stream flow during 2014 was anomalously low compared to climatological means; therefore, offshore influences from terrestrial carbon could also be decreased. The SAB shelf may not be strongly influenced by terrestrial inputs to acidification during drier than normal periods; conversely, shelf waters that are well-buffered against acidification may not play a significant role in mitigating acidification within the Georgia marshes. Supplementary Information: The online version contains supplementary material available at 10.1007/s12237-023-01261-3.

Circ_0000099/miR-223-3p/CTGF Regulates the Growth, Metastasis, and EMT Processes in TGF-ß2-Stimulated Human Lens Epithelial Cells.

PURPOSE: Posterior capsule opacification (PCO) is the major complication of visual impairment after cataract surgery. Circular RNAs (circRNAs) are involved in the development of many diseases. The purpose of this study was to explore the role and molecular mechanism of circ_0000099 in PCO. METHODS: SRA01/04 cells were treated with TGF-ß2 to establish a PCO cell model. The expression of circ_0000099, miR-223-3p, and connective tissue growth factor (CTGF) mRNA was determined by real-time quantitative polymerase chain reaction (qRT-PCR). Western blot assay was used to analyze the protein expression. Cell proliferation, migration, and invasion were analyzed by (4-5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), 5-ethynyl-2 '-Deoxyuridine (EdU), transwell, and wound healing tests. The circ_0000099/miR-223-3p/CTGF relationship was verified by dual luciferase reporter gene and RNA binding protein immunoprecipitation (RIP) assays. RESULTS: TGF-ß2 treatment promoted SRA01/04 cell proliferation invasion, migration, and EMT. Circ_0000099 expression was increased in POC patients and TGF-ß2-treated SRA01/04 cells.Knockdown of circ_0000099 suppressed TGF-ß2-induced proliferation, invasion, migration, and EMT in SRA01/04 cells. miR-223-3p was identified as the target of circ_0000099, and miR-223-3p inhibitor might partly abolish the repression of circ_0000099 silencing on TGF-ß2-triggered SRA01/04 cell disorders. MiR-223-3p directly targeted CTGF. Knockdown of CTGF suppressed TGF-ß2-induced SRA01/04 cell injury. Circ_0000099 can regulate CTGF expression by targeting miR-223-3p. CONCLUSIONS: Circ_0000099 silencing might relieve TGF-2-induced SRA01/04 cell injury by the miR-223-3p/CTGF axis, providing new avenues for the prevention and treatment of PCO.

Role of Menstrual Blood Stem Cell-Derived Secretome, Follicular Fluid, and Melatonin in Oocyte Maturation and Embryo Development in Polycystic Ovary Syndrome.

BACKGROUND: In vitro maturation has been considered an approach to mature oocytes derived from women with polycystic ovary syndrome (PCOS). It is suggested that the IVM of oocytes may benefit from mesenchymal stem cells derived conditioned medium (CM-MSC). OBJECTIVE: The purpose of this study was to determine the efficacy of a cocktail of menstrual blood stem cell (MenSCs)-derived secretome, along with follicular fluid and melatonin, in oocyte maturation and embryo development in PCOS. METHODS: Four hundred left germinal vesicle oocytes were collected from 100 PCOS patients and randomly divided into four treatment groups: 1) control, 2) secretome, 3) follicular fluid, and 4) melatonin. Oocyte maturation, fertilization rate, and embryo development were monitored, as well as the expression levels of oocyte-secreted factors (GDF9- BMP15), oocyte maturation (MPK3), and apoptosis (BAX- Bcl2). RESULTS: The rate of oocyte maturation increased in all test groups, but only the results for the SEC group were significant (P= 0.032). There were no significant differences in oocyte fertilization and embryo yield among groups. However, the quality of embryos significantly increased in the melatonin group compared to the control. Cytoplasmic maturation was confirmed by the expression of oocyte maturation-related genes using Real-time PCR. Additionally, the expression level of BCL-2 was significantly higher in the SEC-FF-MEL group than in the control group (p ≤ 0.01). CONCLUSION: Enrichment of IVM media using MenSCs-secretome, particularly along with melatonin, could be an effective strategy to improve oocyte maturation and embryo development in PCOS.

Knockout of TGF-ß receptor II by CRISPR/Cas9 delays mesenchymal transition of Lens epithelium and posterior capsule opacification.

Posterior capsule opacification (PCO) is the most common postoperative complication of cataract surgery. Transforming growth factor-ß (TGF-ß) is related to epithelial-mesenchymal transition (EMT) of lens epithelial cells (LECs) that is proven to induce PCO formation in clinical and experimental studies. In this study, CRISPR sequences targeting exon of TGF-ßRII were knocked out with lentiviral transfection in LECs. Rabbits' PCO model was established and recombinant adeno-associated virus (AAV) for transferring the gRNA of TGF ßRII were intravitreally injected. SgRNA inhibited TGF-ßRII expression and human LECs proliferation. In TGF-ßRII knockout group, LECs motility and migration were suppressed, N-cadherin and vimentin expressions were significantly decreased, whereas E-cadherin was increased. The animal model showed that TGF-ßRII knockout in vivo was effective in suppressing PCO. The current study suggested that the CRISPR/Cas9 endonuclease system could suppress TGF-ßRII secretion, which participates in the EMT procedure of LECs in vitro and PCO in vivo. These findings might provide a new gene-editing approach and insight into a novel therapeutic strategy for PCO.