High Frequency Oscillatory Ventilation (HFOV) and Inhaled Nitric Oxide (iNO) Use During Neonatal Emergency Transport - Feasibility and Efficacy in India.

A retrospective study of 24 neonates to evaluate the feasibility and efficacy of high frequency oscillatory ventilation (HFOV) and inhaled nitric oxide (iNO) for transferring critically ill neonates to tertiary neonatal intensive care, who were transported by road ambulance was done. Efficacy was measured by clinical improvement, patient safety was assessed by comparing cardiorespiratory indicators before and after transport, and adverse events during transport. Significant oxygenation improvement was observed in neonates transported with HFOV ± iNO compared to earlier ventilator settings. Pre- and post-transport vital signs were stable, and no transport-related deaths occurred. A substantial rise in median SpO2 was seen after transfer [86 (81, 91) vs. 93 (89, 97) before transport, p <0.001]. Twelve of twenty-one newborns who received nitric oxide demonstrated significant improvement in oxygenation index (a 10% decrease from prior value). Overall survival was 70.8%, however non-transfer or inadequate respiratory treatment may have exacerbated mortality.

Intervention with metabolites emulating endogenous cell transitions accelerates muscle regeneration in young and aged mice.

Tissue regeneration following an injury requires dynamic cell-state transitions that allow for establishing the cell identities required for the restoration of tissue homeostasis and function. Here, we present a biochemical intervention that induces an intermediate cell state mirroring a transition identified during normal differentiation of myoblasts and other multipotent and pluripotent cells to mature cells. When applied in somatic differentiated cells, the intervention, composed of one-carbon metabolites, reduces some dedifferentiation markers without losing the lineage identity, thus inducing limited reprogramming into a more flexible cell state. Moreover, the intervention enabled accelerated repair after muscle injury in young and aged mice. Overall, our study uncovers a conserved biochemical transitional phase that enhances cellular plasticity in vivo and hints at potential and scalable biochemical interventions of use in regenerative medicine and rejuvenation interventions that may be more tractable than genetic ones.

To Evaluate Platelet Indices, Platelet to Lymphocyte Ratio and Neutrophil to Lymphocyte Ratio as Prognostic and Risk Factors in Patients with Coronary Artery Disease.

To evaluate platelet indices, platelet to lymphocyte ratio and neutrophil to lymphocyte ratio as prognostic and risk factors in patients with coronary artery disease Introduction: cardiovascular diseases have 12 million deaths annually which is one of the commonest causes of death globally. Platelet parameters like Mean platelet volume (MPV), platelet distribution width (PDW) and WBC parameters like Neutrophil to lymphocyte ratio (NLR) and platelet to lymphocyte ratio(PLR) have recently been emerging as a new prognostic marker in number of coronary artery disease (CAD) with limited studies to explore their utility. AIMS AND OBJECTIVES: this study aimed to evaluate MPV, P-LCR, PDW, PLR, NLR and GRACE risk score in CAD patients. MATERIAL AND METHODS: this descriptive cross-sectional study was done in 330 cases of CAD and 200 healthy controls were studied.MPV, platelet-large cell ratio (P-LCR), PLR, NLR and PDW of patients were analyzed. RESULTS: In our study, 245 were males and 85 were females. Mean PDW, MPV, Mean PLCR, Mean NLR was significantly higher in CAD cases as compared to controls (p value < 0.05). Mean PLR was not significantly different in CAD cases and controls. MPV was more associated risk predictor of CAD (8.98 times) followed by NLR (2.79 times), PDW (1.53 times) and PLCR (1.02 times). DISCUSSION AND CONCLUSION: platelet indices, NLR and PLR are simple cost effective parameters and in future these might be useful adjuvant tests in conjunction with conventional biochemical cardiac markers in early prediction of risk of CAD in patients admitted to hospital and can guide clinicians in assessing the prognosis on short and long term follow up of these patients in terms of morbidity and mortality.

Overview of acute type A aortic dissection in India.

Acute type A aortic dissection (TAAD) is a life-threatening surgical emergency. Though the entity is associated with high mortality and morbidity, with well-executed and timely surgical intervention, mortality and morbidity could be reduced to a reasonable extent. Information about demographics, clinical pattern, and results of management of acute TAAD from the Indian subcontinent largely remains unpublished. There are only a few specialized centers performing aortic operations. Very often, the patients with acute TAAD are operated on by surgeons with limited experience and resources. The surgeon is operating like a "lone warrior" without the support of a specialized radiologist, interventionalist, and specialized anesthetist. In most of the hospitals, facilities for sophisticated monitoring, sealants, specialized grafts, and stent graft are not available. We follow a simple algorithm of diagnosis and surgical management. The goal of treatment is to save the life. We follow a conservative approach best suited to our circumstances. Mild hypothermia, carotid cannulation, and antegrade cerebral perfusion as cerebral protection strategies have yielded satisfactory results. In case of organ malperfusion, with some exceptions, we perform aortic repair first. Our policy, towards arch management is less aggressive. In high-risk cases, we perform an endovascular-compliant hemiarch or arch replacement, followed by stent grafting in the post-operative period.

Open arch replacement: my way.

With advancement of hybrid and endovascular techniques, there are very few indications for open arch replacement. Major advancements in open arch replacement include antegrade perfusion-based cerebral protection, and an endovascular compliant arch replacement. In the present article, we demonstrate and describe our technique of Bentall's procedure and endovascular compliant arch replacement in a young Marfan's patient with chronic type A dissection and root aneurysm. Supplementary Information: The online version contains supplementary material available at 10.1007/s12055-023-01560-1.

Ocular morbidity among children (aged 6-18 yr) of the tribal area of Melghat, India: A community-based study.

BACKGROUND OBJECTIVES: Most of the ocular morbidities among school children are preventable or treatable. Melghat, a difficult to access, hilly, forest, tribal area with poorly developed infrastructure in the Amravati district of Maharashtra. Scarcity of ophthalmologists and low health-seeking behaviour of tribal people contributes to the high burden of ocular morbidity. Given the lack of published studies on the ocular morbidity among children in Melghat, outreach programmes are essential to diagnose and treat visual impairments promptly. The objective was to determine the prevalence of ocular morbidity among children in the tribal area of Melghat. METHODS: A community-based observational study was carried out in the Chikhaldara and Dharni blocks of Melghat. Children from 15 tribal villages were screened for eye disorders by trained paramedics. Most of the children were examined by an ophthalmologist. We used Chi-square test for categorical variables. RESULTS: A total of 4357 children aged between 6 and 18 yr were examined. Of these 2336 (53.6%) were females and 2021 (46.4%) were males. Out of 4357 children, 507 (11.63%) had an ocular morbidity. The prevalence of ocular morbidity and refractive error increased in the age group of 8-10 yr (P<0.05 and <0.001, respectively). Refractive error was the most common ocular morbidity (n=339; 7.8%), followed by vitamin A deficiency (VAD) (n=120; 2.8%). INTERPRETATION CONCLUSIONS: The prevalence of refractive error and VAD in this study was significantly higher than the rest of India and the world. For the prevention of childhood blindness, immediate intervention programme, including eye screening by trained paramedics, treatment by an ophthalmologist and prophylaxis, is crucial.

Genome-wide CRISPR activation screening in senescent cells reveals SOX5 as a driver and therapeutic target of rejuvenation.

Our understanding of the molecular basis for cellular senescence remains incomplete, limiting the development of strategies to ameliorate age-related pathologies by preventing stem cell senescence. Here, we performed a genome-wide CRISPR activation (CRISPRa) screening using a human mesenchymal precursor cell (hMPC) model of the progeroid syndrome. We evaluated targets whose activation antagonizes cellular senescence, among which SOX5 outperformed as a top hit. Through decoding the epigenomic landscapes remodeled by overexpressing SOX5, we uncovered its role in resetting the transcription network for geroprotective genes, including HMGB2. Mechanistically, SOX5 binding elevated the enhancer activity of HMGB2 with increased levels of H3K27ac and H3K4me1, raising HMGB2 expression so as to promote rejuvenation. Furthermore, gene therapy with lentiviruses carrying SOX5 or HMGB2 rejuvenated cartilage and alleviated osteoarthritis in aged mice. Our study generated a comprehensive list of rejuvenators, pinpointing SOX5 as a potent driver for rejuvenation both in vitro and in vivo.

Intradermal but not intramuscular modified vaccinia Ankara immunizations protect against intravaginal tier2 simian-human immunodeficiency virus challenges in female macaques.

Route of immunization can markedly influence the quality of immune response. Here, we show that intradermal (ID) but not intramuscular (IM) modified vaccinia Ankara (MVA) vaccinations provide protection from acquisition of intravaginal tier2 simian-human immunodeficiency virus (SHIV) challenges in female macaques. Both routes of vaccination induce comparable levels of serum IgG with neutralizing and non-neutralizing activities. The protection in MVA-ID group correlates positively with serum neutralizing and antibody-dependent phagocytic activities, and envelope-specific vaginal IgA; while the limited protection in MVA-IM group correlates only with serum neutralizing activity. MVA-ID immunizations induce greater germinal center Tfh and B cell responses, reduced the ratio of Th1 to Tfh cells in blood and showed lower activation of intermediate monocytes and inflammasome compared to MVA-IM immunizations. This lower innate activation correlates negatively with induction of Tfh responses. These data demonstrate that the MVA-ID vaccinations protect against intravaginal SHIV challenges by modulating the innate and T helper responses.

Single-cell individual full-length mtDNA sequencing by iMiGseq uncovers unexpected heteroplasmy shifts in mtDNA editing.

The ontogeny and dynamics of mtDNA heteroplasmy remain unclear due to limitations of current mtDNA sequencing methods. We developed individual Mitochondrial Genome sequencing (iMiGseq) of full-length mtDNA for ultra-sensitive variant detection, complete haplotyping, and unbiased evaluation of heteroplasmy levels, all at the individual mtDNA molecule level. iMiGseq uncovered unappreciated levels of heteroplasmic variants in single cells well below the conventional NGS detection limit and provided accurate quantitation of heteroplasmy level. iMiGseq resolved the complete haplotype of individual mtDNA in single oocytes and revealed genetic linkage of de novo mutations. iMiGseq detected sequential acquisition of detrimental mutations, including large deletions, in defective mtDNA in NARP/Leigh syndrome patient-derived induced pluripotent stem cells. iMiGseq identified unintended heteroplasmy shifts in mitoTALEN editing, while showing no appreciable level of unintended mutations in DdCBE-mediated mtDNA base editing. Therefore, iMiGseq could not only help elucidate the mitochondrial etiology of diseases, but also evaluate the safety of various mtDNA editing strategies.

An unusually long intra-atrial course of the right coronary artery detected on CT angiography.

We hereby present an unusually long intra-atrial course of the right coronary artery incidentally detected on computed tomography angiography. Although usually asymptomatic, an intra-atrial right coronary artery may be injured during iatrogenic procedures which require right heart catheterisation.

V2 hotspot optimized MVA vaccine expressing stabilized HIV-1 Clade C envelope Gp140 delays acquisition of heterologous Clade C Tier 2 challenges in Mamu-A*01 negative Rhesus Macaques.

Stabilized HIV envelope (Env) trimeric protein immunogens have been shown to induce strong autologous neutralizing antibody response. However, there is limited data on the immunogenicity and efficacy of stabilized Env expressed by a viral vector-based immunogen. Here, we compared the immunogenicity and efficacy of two modified vaccinia Ankara (MVA) vaccines based on variable loop 2 hotspot (V2 HS) optimized C.1086 envelope (Env) sequences, one expressing the membrane anchored gp150 (MVA-150) and the other expressing soluble uncleaved pre-fusion optimized (UFO) gp140 trimer (MVA-UFO) in a DNA prime/MVA boost approach against heterologous tier 2 SHIV1157ipd3N4 intrarectal challenges in rhesus macaques (RMs). Both MVA vaccines also expressed SIVmac239 Gag and form virus-like particles. The DNA vaccine expressed SIVmac239 Gag, C.1086 gp160 Env and rhesus CD40L as a built-in adjuvant. Additionally, all immunizations were administered intradermally (ID) to reduce induction of vaccine-specific IFNγ+ CD4 T cell responses. Our results showed that both MVA-150 and MVA-UFO vaccines induce comparable Env specific IgG responses in serum and rectal secretions. The vaccine-induced serum antibody showed ADCC and ADCVI activities against the challenge virus. Comparison with a previous study that used similar immunogens via intramuscular route (IM) showed that ID immunizations induced markedly lower SHIV specific CD4 and CD8 T cell responses compared to IM immunizations. Following challenge, MVA-UFO vaccinated animals showed a significant delay in acquisition of SHIV1157ipd3N4 infection but only in Mamu-A*01 negative macaques with an estimated vaccine efficacy of 64% per exposure. The MVA-150 group also showed a trend (p=0.1) for delay in acquisition of SHIV infection with an estimated vaccine efficacy of 57%. The vaccine-induced IFNγ secreting CD8 T cell responses showed a direct association and CD4 T cells showed an inverse association with delay in acquisition of SHIV infection. These results demonstrated that both MVA-150 and MVA-UFO immunogens induce comparable humoral and cellular immunity and the latter provides marginally better protection against heterologous tier 2 SHIV infection. They also demonstrate that DNA/MVA vaccinations delivered by ID route induce better antibody and lower CD4 and CD8 T cell responses compared to IM.

LINE-1 RNA causes heterochromatin erosion and is a target for amelioration of senescent phenotypes in progeroid syndromes.

Constitutive heterochromatin is responsible for genome repression of DNA enriched in repetitive sequences, telomeres, and centromeres. During physiological and pathological premature aging, heterochromatin homeostasis is profoundly compromised. Here, we showed that LINE-1 (Long Interspersed Nuclear Element-1; L1) RNA accumulation was an early event in both typical and atypical human progeroid syndromes. L1 RNA negatively regulated the enzymatic activity of the histone-lysine N-methyltransferase SUV39H1 (suppression of variegation 3-9 homolog 1), resulting in heterochromatin loss and onset of senescent phenotypes in vitro. Depletion of L1 RNA in dermal fibroblast cells from patients with different progeroid syndromes using specific antisense oligonucleotides (ASOs) restored heterochromatin histone 3 lysine 9 and histone 3 lysine 27 trimethylation marks, reversed DNA methylation age, and counteracted the expression of senescence-associated secretory phenotype genes such as p16, p21, activating transcription factor 3 (ATF3), matrix metallopeptidase 13 (MMP13), interleukin 1a (IL1a), BTG anti-proliferation factor 2 (BTG2), and growth arrest and DNA damage inducible beta (GADD45b). Moreover, systemic delivery of ASOs rescued the histophysiology of tissues and increased the life span of a Hutchinson-Gilford progeria syndrome mouse model. Transcriptional profiling of human and mouse samples after L1 RNA depletion demonstrated that pathways associated with nuclear chromatin organization, cell proliferation, and transcription regulation were enriched. Similarly, pathways associated with aging, inflammatory response, innate immune response, and DNA damage were down-regulated. Our results highlight the role of L1 RNA in heterochromatin homeostasis in progeroid syndromes and identify a possible therapeutic approach to treat premature aging and related syndromes.

Cannulation strategies in aortic surgery: techniques and decision making.

Arterial cannulation for cardiopulmonary bypass (CPB) is an important determinant of outcome in aortic surgery. Unlike traditional cardiac operations, aortic pathology may preclude the cannulation of the distal ascending aorta. In other cases, special need of the pathology/operation may demand an alternative cannulation site. Choosing the right cannulation site, especially in type A aortic dissection, is the most crucial initial step. The decision about cannulation sites should be individualized and patient-specific. Various cannulation techniques include femoral, right axillary, innominate, carotid, central aortic, direct true lumen, transapical, and trans-atrial left ventricle cannulation. The ideal cannulation should be easy, quick, and suitable for all clinical scenarios. It should allow smooth conduct of CPB without malperfusion or cerebral embolization. The cannulation strategy should also provide an option for selective antegrade cerebral perfusion and it should be free from neurovascular and local site complications. There is no ideal cannulation technique. Each technique has its pros and cons. Excellent results and drawbacks have been reported with each technique. Final selection of the cannulation site is dependent upon several factors. However, a surgeon's familiarity with a particular technique plays a major role in selection. Despite this, there is a definite shift in surgeons' preference from femoral to central cannulation (axillary, carotid, innominate, aortic) over the last few decades. The aim of this review is to give a brief overview of the cannulation techniques in aortic surgery and discuss the decision-making process.

In vivo partial cellular reprogramming enhances liver plasticity and regeneration.

Mammals have limited regenerative capacity, whereas some vertebrates, like fish and salamanders, are able to regenerate their organs efficiently. The regeneration in these species depends on cell dedifferentiation followed by proliferation. We generate a mouse model that enables the inducible expression of the four Yamanaka factors (Oct-3/4, Sox2, Klf4, and c-Myc, or 4F) specifically in hepatocytes. Transient in vivo 4F expression induces partial reprogramming of adult hepatocytes to a progenitor state and concomitantly increases cell proliferation. This is indicated by reduced expression of differentiated hepatic-lineage markers, an increase in markers of proliferation and chromatin modifiers, global changes in DNA accessibility, and an acquisition of liver stem and progenitor cell markers. Functionally, short-term expression of 4F enhances liver regenerative capacity through topoisomerase2-mediated partial reprogramming. Our results reveal that liver-specific 4F expression in vivo induces cellular plasticity and counteracts liver failure, suggesting that partial reprogramming may represent an avenue for enhancing tissue regeneration.

Myc Supports Self-Renewal of Basal Cells in the Esophageal Epithelium.

It is widely believed that cellular senescence plays a critical role in both aging and cancer, and that senescence is a fundamental, permanent growth arrest that somatic cells cannot avoid. Here we show that Myc plays an important role in self-renewal of esophageal epithelial cells, contributing to their resistance to cellular senescence. Myc is homogeneously expressed in basal cells of the esophageal epithelium and Myc positively regulates their self-renewal by maintaining their undifferentiated state. Indeed, Myc knockout induced a loss of the undifferentiated state of esophageal epithelial cells resulting in cellular senescence while forced MYC expression promoted oncogenic cell proliferation. A superoxide scavenger counteracted Myc knockout-induced senescence, therefore suggesting that a mitochondrial superoxide takes part in inducing senescence. Taken together, these analyses reveal extremely low levels of cellular senescence and senescence-associated phenotypes in the esophageal epithelium, as well as a critical role for Myc in self-renewal of basal cells in this organ. This provides new avenues for studying and understanding the links between stemness and resistance to cellular senescence.

Greater Curvature, Centerline or Straightened Centerline Length - Which length is Most Accurate for Preprocedural Planning in Thoracic Endovascular Aortic Repair?

BACKGROUND: Automated centerline (CL) measurements have been conventionally used for stent-graft length estimation during thoracic endovascular aortic repair (TEVAR). The purpose of this study was to assess the accuracy of greater curvature length (GL), semiautomated CL and straightened centerline length (SCL) for preprocedural planning in TEVAR. METHODS: Immediate postprocedural CT Angiographies of 30 patients (22 males, age-49.2 ± 10.1years) who underwent TEVAR between 2015 and 2017 were retrospectively analyzed. CL, GL, SCL and the straightline length(SL) were measured between proximal and distal ends of the stent-graft and results were compared with the true length of the stent-graft (TL). Tortuosity index (TI = CL/SL) was calculated. RESULTS: GL (17.92 ± 4.78 cm) was the closest in predicting the TL (17.75 ± 4.29 cm) (P = 0.414) overall, as well as in both dissection and aneurysm subgroups (P= 0.9). There was a significant difference between CL (16.67 ± 4.07 cm) and TL (P< 0.0001) as well as between SCL (16.86 ± 4.16 cm) and TL (P= 0.001). These differences were greater in dissection subgroup than in the aneurysm group (P< 0.0001 and P= 0.03 for TL-CL and TL-SCL, respectively). The extent of mismatch between GL or CL and TL did not correlate with tortuosity, but the difference between TL and SCL had a significant positive correlation with tortuosity (r = 0.375, P= 0.04). TL-GL had a negative linear correlation with the stent-graft length (TL) in the dissection group (r = 0.50, P= 0.03). CONCLUSIONS: The greater curvature length predicts the actual total length of the deployed stent-graft more accurately than centerline or straightened centerline lengths. Hence, it should be used in planning for the length of stent-graft required for TEVAR.

Pasteurized Donor Human Milk Should Not Replace Mother's Own Milk in Preterm Neonates: A Quality Initiative Toward Decreasing the "PDHM Dependency".

Aim: Mother's own milk (MOM) is the preferred source of neonatal nutrition. Due to various challenges, mothers are often unable to provide exclusive MOM to neonates admitted in neonatal intensive care units (NICUs) and depend on pasteurized donor human milk (PDHM). The aim of this quality improvement (QI) initiative was to enable mothers to provide MOM and consequently decrease the "PDHM dependency." Methods: Neonates <32 weeks of gestation (n = 120) were included. A multidisciplinary team was formed, and a detailed root cause analysis was done to understand the cause of PDHM dependency during the observation phase (November 1 to December 15, 2019). Various evidence-based practices were planned, tested, and implemented through Plan-Do-Study-Act cycles during the intervention phase (December 16, 2019 to January 31, 2020). These were further strengthened and adopted as a unit culture during the maintenance phase (February 1 to July 31, 2020). Results from the observation and intervention phases were compared. Results: Within 6 weeks of QI interventions, the average proportion of MOM significantly increased from 74.4% to 93.5% (p = 0.0003), and the proportion of PDHM significantly decreased from 20.5% to 4.6% (p = 0.005). The proportion of MOM remained at 82.5% during the maintenance phase. There was a significant decrease in the number of days to reach full feeds and regain birth weight. Conclusions: Provision of PDHM from our newly functional milk bank led to a reduced drive to express MOM in mothers of NICU babies. Our QI project focused on various strategies to improve MOM feeding and reduce PDHM dependence.

In vivo partial reprogramming alters age-associated molecular changes during physiological aging in mice.

Partial reprogramming by expression of reprogramming factors (Oct4, Sox2, Klf4 and c-Myc) for short periods of time restores a youthful epigenetic signature to aging cells and extends the life span of a premature aging mouse model. However, the effects of longer-term partial reprogramming in physiologically aging wild-type mice are unknown. Here, we performed various long-term partial reprogramming regimens, including different onset timings, during physiological aging. Long-term partial reprogramming lead to rejuvenating effects in different tissues, such as the kidney and skin, and at the organismal level; duration of the treatment determined the extent of the beneficial effects. The rejuvenating effects were associated with a reversion of the epigenetic clock and metabolic and transcriptomic changes, including reduced expression of genes involved in the inflammation, senescence and stress response pathways. Overall, our observations indicate that partial reprogramming protocols can be designed to be safe and effective in preventing age-related physiological changes. We further conclude that longer-term partial reprogramming regimens are more effective in delaying aging phenotypes than short-term reprogramming.

In vivo partial reprogramming of myofibers promotes muscle regeneration by remodeling the stem cell niche.

Short-term, systemic expression of the Yamanaka reprogramming factors (Oct-3/4, Sox2, Klf4 and c-Myc [OSKM]) has been shown to rejuvenate aging cells and promote tissue regeneration in vivo. However, the mechanisms by which OSKM promotes tissue regeneration are unknown. In this work, we focus on a specific tissue and demonstrate that local expression of OSKM, specifically in myofibers, induces the activation of muscle stem cells or satellite cells (SCs), which accelerates muscle regeneration in young mice. In contrast, expressing OSKM directly in SCs does not improve muscle regeneration. Mechanistically, expressing OSKM in myofibers regulates the expression of genes important for the SC microenvironment, including upregulation of p21, which in turn downregulates Wnt4. This is critical because Wnt4 is secreted by myofibers to maintain SC quiescence. Thus, short-term induction of the Yamanaka factors in myofibers may promote tissue regeneration by modifying the stem cell niche.