Experiences and perception towards reproductive health education among secondary school teachers in South India.

BACKGROUND: Reproductive health education (RHE) is an important component of school curricula. It helps students in the decision-making process regarding several issues concerning reproductive health. However delivering RHE at schools is a difficult task for the teachers. METHODS: This study was conducted to assess the experiences and perceptions towards reproductive health education (RHE) among 236 secondary school teachers in January 2019. Data were collected using a self-administered questionnaire. RESULTS: Only 21 (8.9%) were trained in RHE. Majority [179 (75.8%)] identified cultural barriers as the major challenge involved in its implementation. 95 (40.3%) teachers felt that the provision of sexual education as a part of RHE will promote pre-marital sexual activity among the students. Of the total, 185 (78.4%) had average while 51 (21.6%) participants had a good perception towards RHE. It was taught in only 3 (16.7%) out of the 18 schools surveyed. Only 11 (4.7%) participants felt that the availability of teaching aids to conduct RHE classes at their schools was adequate. Hardly 14 (5.9%) teachers had taken RHE classes for students. Among the rest, 135 (60.8%) expressed their willingness to take RHE classes with appropriate training. In multi variable analysis, participants aged ≤ 40 years (p = 0.031), those belonging to nuclear families (p = 0.013), and those who had taken classes in RHE (p = 0.037) had significantly good perception level towards RHE. CONCLUSIONS: Teachers therefore need to be trained and given more opportunities to take RHE sessions which will help improve their perception towards RHE. Schools need to be better equipped with resources and various perceived barriers need to be overcome before RHE can be successfully implemented.

This study was conducted to assess the experiences and perceptions towards reproductive health education (RHE) secondary school teachers. The participants provided the required information by filling a questionnaire. Hardly one in ten of them had prior training in RHE and one in twenty had taken RHE classes at schools. More than three-fourth of them felt that cultural barriers could pose problems in its implementation at schools. One in four teachers had good perception towards RHE. Two in three among teachers, who had not taken RHE classes before, expressed their willingness to take RHE classes with appropriate training. Favourable perception towards RHE were expressed by teachers who were young, from small families and those who had taken RHE classes before.

Zebrafish model of amyloid light chain cardiotoxicity: regeneration versus degeneration.

Cardiac dysfunction is the most frequent cause of morbidity and mortality in amyloid light chain (AL) amyloidosis caused by a clonal immunoglobulin light chain (LC). Previously published transgenic animal models of AL amyloidosis have not recapitulated the key phenotype of cardiac dysfunction seen in AL amyloidosis, which has limited our understanding of the disease mechanisms in vivo, as well as the development of targeted AL therapeutics. We have developed a transgenic zebrafish model in which a λ LC derived from a patient with AL amyloidosis is conditionally expressed in the liver under the control of the Gal4 upstream activation sequence enhancer system. Circulating LC levels of 125 µg/ml in these transgenic zebrafish are comparable to median pathological serum LC levels. Functional analysis links abnormal contractile function with evidence of cellular and molecular proteotoxicity in the heart, including increased cell death and autophagy. However, despite pathological and functional phenotypes analogous to human AL, the lifespan of the transgenic fish is comparable to control fish without the expressed AL-LC transgene. Nuclear labeling experiments suggest increased cardiac proliferation in the transgenic fish, which can be counteracted by treatment with a small molecule proliferation inhibitor leading to increased zebrafish mortality because of cardiac apoptosis and functional deterioration. This transgenic zebrafish model provides a platform to study underlying AL disease mechanisms in vivo further. NEW & NOTEWORTHY Heart failure is a major cause of mortality in amyloid light (AL) amyloidosis, yet it has been difficult to model in animals. We report the generation of a transgenic zebrafish model for AL amyloidosis with pathological concentration of circulating human light chain protein that results in cardiac dysfunction. The light chain toxicity triggers regeneration in the zebrafish heart resulting in functional compensation early in life, but with age develops into cardiac dysfunction.

Assessing Inflammatory Status of Pulp in Irreversible Pulpitis Cases with Pulse oximeter and Dental Hemogram.

Objective: This study evaluated pulse oximetry and dental hemogram in teeth with the clinical diagnosis of irreversible pulpitis (IP) to assess the inflammatory status of the pulp. Study design: The study and control groups (30n each) had teeth with IP and sound teeth respectively. Patients in the study group had night pain with or without pain on mastication (NM, N). Blood oxygen saturation (%SpO2) was recorded with a custom made pulse oximeter (CPO). For dental and peripheral hemogram, smears were made for each patient from the first drop of blood while entering the pulp and finger blood respectively. Results: Control group had mean %SpO2 in finger 91% (86-97); and in teeth 84% (80-91), while the study group had mean %SpO2 in finger 92% (88-98) and in teeth 83% (71-94). Fifty percent of IP cases were vital while no tooth showed necrosis according to CPO which was further confirmed by bleeding status from the pulp. Based on the findings of the clinical diagnosis, %SpO2 and bleeding status of IP and normal cases, the terminology as coronal or total pulpitis seems more appropriate. The statistical difference was significant in fingers while non-significant in teeth of IP and normal pulp cases. Dental hemogram of IP cases showed an overall significant fall of neutrophil, lymphocyte, eosinophil and monocyte counts compared to normal. Conclusion: Pulse oximetry was the most accurate pulp test to diagnose vitality in normal as well as inflamed pulps while hemogram was inconclusive for the same.

In Vivo Evaluation of Customized Pulse Oximeter and Sensitivity Pulp Tests for Assessment Of Pulp Vitality.

OBJECTIVE: The purpose of this study was to test a customized pulse oximeter (CPO) for evaluation of pulp vitality in primary and permanent teeth against clinical diagnosis (vital and untreated non-vital) in order to expand its clinical use for pulp preservation. STUDY DESIGN: CPO was evaluated on intact primary and permanent central or lateral incisor (CI, LI) teeth-vital (group 1, 20n each); untreated non-vital (group 2, 10n each) and; root filled non-vital (group 3, 10n each) of children 4-12 years according to inclusion/ exclusion criteria. For each patient CPO was first applied on finger followed by vitality tests in following sequence-electrical, pulse oximetry and thermal tests. RESULTS: Mean oxygen saturation (%SpO2) in permanent and primary-vital teeth was 88.78% & 87.77% respectively; non-vital teeth was 74.67% & 75.00% respectively; and in all root filled teeth was 0%. Tooth and finger oxygen saturation values showed strong positive relationship in vital primary or permanent teeth and; no correlation in untreated non-vital primary or permanent teeth. The accuracy rate of thermal pulp test and pulse oximetry was 100% and for electrical pulp test it was 90% for permanent and 86.67% for primary teeth. CONCLUSION: The CPO tested in this study proved to be a valuable adjunct for diagnosing pulp vitality by objective means.

Histologic validation of ICDAS-II and polarization sensitive optical coherence tomography to detect smooth surface early carious lesions.

AIM: This in vitro study aimed to histologically validate and compare the methods for detection of smooth surface early carious lesions (ECLs) that is, International caries detection and assessment system for the smooth surface (ICDAS-II-SSC), Polarization sensitive optical coherence tomography (PS-OCT), and radiography. METHODOLOGY: PS-OCT images for scores 0-3 of ICDAS-II-SSC were standardized according to ECLs' depth. Preliminary PS-OCT images for ICDAS-II-SSC score-2 of pigmented ECLs showed reduced lesion depth and therefore were dichotomized into scores 2 and 2p for white and pigmented lesions (ICDAS-II-SSCm). ECLs on one hundred freshly extracted teeth were scored by three examiners for ICDAS-II-SSCm, PS-OCT, radiography, and histology. RESULTS: Compared to histology, ICDAS-II-SSCm showed a strong positive correlation followed by PS-OCT and radiographic evaluation. ICDAS-II-SSCm had a strong positive correlation with PS-OCT, while both variables had a weak positive correlation with radiography. PS-OCT detected the activity of ECLs by directly relating the image depth of ECLs to their mineral volume content. CONCLUSION: The current scope of ICDAS-II should be reviewed since the pigmentation can be misinterpreted as an active lesion. Till then, ICDAS-II-SSC is an effective visual method for early caries detection. PS-OCT has the potential to become a probe with the proper algorithm for diagnostic purposes.

CaMKIIδ subtypes differentially regulate infarct formation following ex vivo myocardial ischemia/reperfusion through NF-κB and TNF-α.

Deletion of Ca2+/calmodulin-dependent protein kinase II delta (CaMKIIδ) has been shown to protect against in vivo ischemia/reperfusion (I/R) injury. It remains unclear which CaMKIIδ isoforms and downstream mechanisms are responsible for the salutary effects of CaMKIIδ gene deletion. In this study we sought to compare the roles of the CaMKIIδB and CaMKIIδC subtypes and the mechanisms by which they contribute to ex vivo I/R damage. WT, CaMKIIδKO, and mice expressing only CaMKIIδB or δC were subjected to ex vivo global ischemia for 25min followed by reperfusion. Infarct formation was assessed at 60min reperfusion by triphenyl tetrazolium chloride (TTC) staining. Deletion of CaMKIIδ conferred significant protection from ex vivo I/R. Re-expression of CaMKIIδC in the CaMKIIδKO background reversed this effect and exacerbated myocardial damage and dysfunction following I/R, while re-expression of CaMKIIδB was protective. Selective activation of CaMKIIδC in response to I/R was evident in a subcellular fraction enriched for cytosolic/membrane proteins. Further studies demonstrated differential regulation of nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) signaling and tumor necrosis factor alpha (TNF-α) expression by CaMKIIδB and CaMKIIδC. Selective activation of CaMKIIδC was also observed and associated with NF-κB activation in neonatal rat ventricular myocytes (NRVMs) subjected to oxidative stress. Pharmacological inhibition of NF-κB or TNF-α significantly ameliorated infarct formation in WT mice and those that re-express CaMKIIδC, demonstrating distinct roles for CaMKIIδ subtypes in I/R and implicating acute activation of CaMKIIδC and NF-κB in the pathogenesis of reperfusion injury.

Identification of Novel Inhibitors of Leishmania donovani γ-Glutamylcysteine Synthetase Using Structure-Based Virtual Screening, Docking, Molecular Dynamics Simulation, and in Vitro Studies.

Trypansomatids maintain their redox balance by the trypanothione-based redox system, enzymes of which exhibit differences from mammalian homologues. γ-Glutamylcysteine synthetase (Gcs) is an essential enzyme in this pathway that performs the first and rate-limiting step. l-Buthionine-(S,R)-sulfoximine (BSO), a specific inhibitor of Gcs, induces toxicity in hosts infected with Trypanosoma brucei, underlining the need for novel Gcs inhibitors. The present study reports identification of Leishmania donovani Gcs (LdGcs) inhibitors using computational approaches and their experimental validation. Analysis of inhibitor-LdGcs complexes shows modifications that could result in increased efficacy of these compounds.

Discovery of novel inhibitors for Leishmania nucleoside diphosphatase kinase (NDK) based on its structural and functional characterization.

Nucleoside diphosphate kinases (NDKs) are ubiquitous enzymes that catalyze the transfer of the γ-phosphate moiety from an NTP donor to an NDP acceptor, crucial for maintaining the cellular level of nucleoside triphosphates (NTPs). The inability of trypanosomatids to synthesize purines de novo and their dependence on the salvage pathway makes NDK an attractive target to develop drugs for the diseases they cause. Here we report the discovery of novel inhibitors for Leishmania NDK based on the structural and functional characterization of purified recombinant NDK from Leishmania amazonensis. Recombinant LaNDK possesses auto-phosphorylation, phosphotransferase and kinase activities with Histidine 117 playing an essential role. LaNDK crystals were grown by hanging drop vapour diffusion method in a solution containing 18% PEG-MME 500, 100 mM Bis-Tris propane pH 6.0 and 50 mM MgCl2. It belongs to the hexagonal space group P6322 with unit cell parameters a = b = 115.18, c = 62.18 Å and α = ß = 90°, γ = 120°. The structure solved by molecular replacement methods was refined to crystallographic R-factor and Rfree values of 22.54 and 26.52%, respectively. Molecular docking and dynamics simulation-based virtual screening identified putative binding compounds. Protein inhibition studies of selected hits identified five inhibitors effective at micromolar concentrations. One of the compounds showed ~45% inhibition of Leishmania promastigotes proliferation. Analysis of inhibitor-NDK complexes reveals the mode of their binding, facilitating design of new compounds for optimization of activities as drugs against leishmaniasis.

Biotransformation of lignan glycoside to its aglycone by Woodfordia fruticosa flowers: quantification of compounds using a validated HPTLC method.

CONTEXT: Saraca asoca Linn. (Caesalpiniaceae) is an important traditional remedy for gynaecological disorders and it contains lyoniside, an aryl tetralin lignan glycoside. The aglycone of lyoniside, lyoniresinol possesses structural similarity to enterolignan precursors which are established phytoestrogens. OBJECTIVES: This work illustrates biotransformation of lyoniside to lyoniresinol using Woodfordia fruticosa Kurz. (Lythraceae) flowers and simultaneous quantification of lyoniside and lyoniresinol using a validated HPTLC method. MATERIALS AND METHODS: The aqueous extract prepared from S. asoca bark was fermented using W. fruticosa flowers. The substrate and fermented product both were simultaneously analyzed using solvent system:toluene:ethyl acetate:formic acid (4:3:0.4) at 254 nm. The method was validated for specificity, accuracy, precision, linearity, sensitivity and robustness as per ICH guidelines. RESULTS: The substrate showed the presence of lyoniside, however, it decreased as the fermentation proceeded. On 3rd day, lyoniresinol starts appearing in the medium. In 8 days duration most of the lyoniside converted to lyoniresinol. The developed method was specific for lyoniside and lyoniresinol. Lyoniside and lyoniresinol showed linearity in the range of 250-3000 and 500-2500 ng. The method was accurate as resulted in 99.84% and 99.83% recovery, respectively, for lyoniside and lyoniresinol. CONCLUSION: Aryl tetralin lignan glycoside, lyoniside was successfully transformed into lyoniresinol using W. fruticosa flowers and their contents were simultaneously analyzed using developed validated HPTLC method.

Optimization of microwave-assisted extraction conditions for preparing lignan-rich extract from Saraca asoca bark using Box-Behnken design.

CONTEXT: Lyoniside is the major constituent of Saraca asoca Linn. (Caesalpiniaceae) bark. There is an immediate need to develop an efficient method to isolate its chemical constituents, since it is a therapeutically important plant. OBJECTIVE: A rapid extraction method for lyoniside based on microwave-assisted extraction of S. asoca bark was developed and optimized using response surface methodology (RSM). MATERIALS AND METHODS: Lyoniside was analyzed and quantified by high-performance liquid chromatography coupled with ultraviolet detection (HPLC-UV). The extraction solvent ratio (%), material solvent ratio (g/ml) and extraction time (min) were optimized using Box-Behnken design (BBD) to obtain the highest extraction efficiency. The optimal conditions were the use of 1:30 material solvent ratio with 70:30 mixture of methanol:water for 10 min duration. RESULTS: The optimized microwave-assisted extraction yielded 9.4 mg/g of lyoniside content in comparison to reflux extraction under identical conditions which yielded 4.2 mg/g of lyoniside content. Under optimum conditions, the experimental values agreed closely with the predicted values. The analysis of variance (ANOVA) indicated a high goodness-of-fit model and the success of the RSM method for optimizing lyoniside extraction from the bark of S. asoca. DISCUSSION: All the three variables significantly affected the lyoniside content. Increased polarity of solvent medium enhances the lyoniside yield. CONCLUSION: The present study shows the applicability of microwave-assisted extraction in extraction of lyoniside from S. asoca bark.

Formulation and evaluation of guggul lipid nanovesicles for transdermal delivery of aceclofenac.

CONTEXT: Most new drugs have low water solubility and liposome is an important formulation to administer such drugs; however, it is quite unstable and has negligible systemic absorption. OBJECTIVE: Aceclofenac nanovesicles were made using guggul lipid for formulating stable transdermal formulation. MATERIALS AND METHODS: Guggul lipid was formulated into vesicles along with cholesterol and dicetyl phosphate using film hydration method. The formulations were analyzed for physicochemical properties and stability. Then its skin permeation and anti-inflammatory activity were determined. RESULTS: Both categories of vesicles (PC and GL) showed optimum physicochemical properties; however, accelerated stability study showed considerable differences. GL-1 was appreciably stable for over 6 months at 4 °C. Corresponding gels (PCG-1 and GLG-1) showed C max values at 4.98 and 7.32 µg/mL along with the Tmax values at 4 and 8 hours, respectively. GLG-1 inhibited edema production by 90.81% in 6 hours. Discussion. PC liposomes are unstable at higher temperature and upon longer storage. The formulation with higher lipid content (GL-1) showed good drug retention after 24 hours and appreciable stability both at higher temperature and for longer duration. Guggul lipid being a planar molecule might be stacked in vesicle wall with cholesterol. CONCLUSION: The composition of the nanovesicle played an important role in stability and drug permeation. Guggul lipid is suitable for producing stable vesicles.

Ceramide-2 nanovesicles for effective transdermal delivery: development, characterization and pharmacokinetic evaluation.

UNLABELLED: Abstract Context: The vesicles based on skin lipid have a drug localization effect and its main lipid, ceramide provides protective and regenerative effects while oleic acid (OA) is a penetration enhancer, however, it causes slight irritation, so we have formulated formulation incorporating both of these to develop a transdermal formulation for better permeation. OBJECTIVE: Present study investigated the preparation and characterization of physicochemical properties and permeation of nanovesicles of ceramide-2 containing OA and palmitic acid (PA) respectively and a commercial gel. MATERIALS AND METHODS: The vesicles were made using ceramide 2, cholesterol (Chol), cholesteryl sulfate (CS) and OA or PA, respectively, using film hydration method. The vesicles were characterized for physicochemical properties, ex vivo permeation using human skin and pharmacokinetic parameters and anti-inflammatory activity in rats. RESULTS: The vesicles showed size at 102-125 nm while PDI was 0.11-0.13 and negative zeta potential. OV-3 showed highest entrapment efficiency. The drug fluxes were 92.02 and 8.920 µg/cm(2)/h, respectively, for OV-3 and PV-1. The Cmax were 7.91 and 4.01 µg/ml at 4 and 6 h for OV-3 (2.5 mg) and PV-1 (10 mg), respectively. OV-3 and PV-1 showed 98.8% and 77.36% edema inhibition, respectively, at 3 h. DISCUSSION: Both formulations showed similar physical parameters and different permeation since OA get incorporated in vesicles and increases its permeability and ceramide makes sure that vesicles can rapidly traverse the stratum corneum. CONCLUSION: OV-3 containing 3% OA showed optimum physical parameters and good permeation with maximum anti-inflammatory activity.

Preparation, characterization and permeation studies of a nanovesicular system containing diclofenac for transdermal delivery.

CONTEXT: Transdermal formulations contain permeation enhancer which causes skin damage. Ceramide 2 is natural lipid found in stratum corneum (SC). OBJECTIVE: Drug-loaded nanovesicles of ceramide-2, cholesterol, palmitic acid, cholesteryl sulfate were formulated and analyzed for physical and biological properties. Diclofenac was used as a model drug. MATERIALS AND METHOD: The vesicles were prepared using the film hydration method and characterized for physical parameters, in vitro drug release, accelerated stability studies and formulated into gel. Respective gels were compared with a commercial formulation (CEG) and plain carbopol gel (CG) containing drug for ex vivo, in vivo drug permeation and anti-inflammatory activity. RESULTS: The vesicles were stable with optimum physical parameters. DCG-1 showed 92.89% in vitro drug release. Ceramide vesicles showed drug release between 18 and 25 µg/cm(2) whereas CG and CEG released 0.33 and 1.35 µg/cm(2) drug, respectively. DCG-1 and CEG showed corresponding Cmax at 6 and 4 h, respectively. DCG-1 showed six times AUC than CEG. DCG-1 inhibited edema by 86.37% by 4th hour of application. DISCUSSION: The presence of ceramide 2 specifically promotes the drug permeation through SC and dermis and also contribute towards stability and non-irritancy. CONCLUSION: The composition of the nanovesicle played an important role in physical properties and drug permeation.

A Rare Complication of Proteus Brain Abscess Due to Coccidioidomycosis Causing Hydrocephalus.

Proteus mirabilis is a very common gram-negative facultative anaerobe seen in urinary tract infections. This rod-shaped bacterium tends to cause urolithiasis via its ability to alkalinize the urine. However, in some cases, this bacterium has been shown to cause bacteremia as well as other complicated infections. Here we would like to present a rare case of Proteus mirabilis that has invaded the brain in a patient that has a ventriculoperitoneal (VP) shunt placed due to coccidioidal meningitis causing hydrocephalus. We would also like to discuss the importance of the monitoring of VP shunt and discuss their likelihood of infections and the medical as well as surgical management.

Human amyloidogenic light chain proteins result in cardiac dysfunction, cell death, and early mortality in zebrafish.

Systemic amyloid light-chain (AL) amyloidosis is associated with rapidly progressive and fatal cardiomyopathy resulting from the direct cardiotoxic effects of circulating AL light chain (AL-LC) proteins and the indirect effects of AL fibril tissue infiltration. Cardiac amyloidosis is resistant to standard heart failure therapies, and, to date, there are limited treatment options for these patients. The mechanisms underlying the development of cardiac amyloidosis and AL-LC cardiotoxicity are largely unknown, and their study has been limited by the lack of a suitable in vivo model system. Here, we establish an in vivo zebrafish model of human AL-LC-induced cardiotoxicity. AL-LC isolated from AL cardiomyopathy patients or control nonamyloidogenic LC protein isolated from multiple myeloma patients (Con-LC) was directly injected into the circulation of zebrafish at 48 h postfertilization. AL-LC injection resulted in impaired cardiac function, pericardial edema, and increased cell death relative to Con-LC, culminating in compromised survival with 100% mortality within 2 wk, independent of AL fibril deposition. Prior work has implicated noncanonical p38 MAPK activation in the pathogenesis of AL-LC-induced cardiotoxicity, and p38 MAPK inhibition via SB-203580 rescued AL-LC-induced cardiac dysfunction and cell death and attenuated mortality in zebrafish. This in vivo zebrafish model of AL-LC cardiotoxicity demonstrates that antagonism of p38 MAPK within the AL-LC cardiotoxic signaling response may serve to improve cardiac function and mortality in AL cardiomyopathy. Furthermore, this in vivo model system will allow for further study of the molecular underpinnings of AL cardiotoxicity and identification of novel therapeutic strategies.

Stanniocalcin1 is a key mediator of amyloidogenic light chain induced cardiotoxicity.

Immunoglobulin light chain (LC) amyloidosis (AL) results from overproduction of circulating amyloidogenic LC proteins and subsequent amyloid fibril deposition in organs. Mortality in AL amyloidosis patients is highly associated with a rapidly progressive AL cardiomyopathy, marked by profound impairment of diastolic and systolic cardiac function and significant early mortality. While myocardial fibril deposition contributes to the severe diastolic dysfunction seen in AL cardiomyopathy patients, the degree of fibril deposition has not been found to correlate with prognosis. Previously, we and others showed a direct cardiotoxic effect of amyloidogenic LC proteins (AL-LC), which may contribute to the pathophysiology and mortality observed in AL cardiomyopathy patients. However, the mechanisms underlying AL-LC related cardiotoxicity remain unknown. Mammalian stanniocalcin1 (STC1) is associated with a number of cellular processes including oxidative stress and cell death. Herein, we find that STC1 expression is elevated in cardiac tissue from AL cardiomyopathy patients, and is induced in isolated cardiomyocytes in response to AL-LC, but not non-amyloidogenic LC. STC1 overexpression in vitro recapitulates the pathophysiology of AL-LC mediated cardiotoxicity, with increased ROS production, contractile dysfunction and cell death. Overexpression of STC1 in vivo results in significant cardiac dysfunction and cell death. Genetic silencing of STC1 prevents AL-LC induced cardiotoxicity in cardiomyocytes and protects against AL-LC induced cell death and early mortality in zebrafish. The cardiotoxic effects of STC1 appears to be mediated via mitochondrial dysfunction as indicated by loss of mitochondrial membrane potential, ROS production and increased mitochondrial calcium levels. Collectively, this work identifies STC1 as a critical determinant of AL-LC cardiotoxicity.

Location matters: clarifying the concept of nuclear and cytosolic CaMKII subtypes.

RATIONALE: Differential effects of δ(B) and δ(C) subtypes of Ca²âº/calmodulin-dependent protein kinase (CaMKII) on cardiomyocyte Ca²âº handling and survival have been suggested to result from their respective nuclear versus cytosolic localizations. CaMKIIδ subtype localization and its relationship to enzyme activation and target phosphorylation have not, however, been systematically evaluated. OBJECTIVE: To determine whether CaMKIIδ subtypes are restricted to a particular subcellular location and assess the relationship of localization to enzyme activation and function. METHODS AND RESULTS: CaMKIIδ is highly expressed in mouse heart and cardiomyocytes and concentrated in sarcoplasmic reticulum (SR)/membrane and nuclear fractions. CaMKIIδ(B) and δ(C) subtypes differ by a nuclear localization sequence, but both are present in nuclear and SR/membrane fractions. Nonselective subtype distribution is also seen in mice overexpressing CaMKIIδ(B) or δ(C), even in a CaMKIIδ null background. Fluorescently tagged CaMKIIδ(B) expressed in cardiomyocytes concentrates in nuclei whereas δ(C) concentrates in cytosol, but neither localization is exclusive. Mouse hearts exposed to phenylephrine show selective CaMKIIδ activation in the nuclear (versus SR) compartment, whereas caffeine selectively activates CaMKIIδ in SR (versus nuclei), independent of subtype. Compartmentalized activation extends to functional differences in target phosphorylation at CaMKII sites: phenylephrine increases histone deacetylase 5 phosphorylation (Ser498) but not phospholamban (Thr17), whereas the converse holds for caffeine. CONCLUSIONS: These studies demonstrate that CaMKIIδ(B) and δ(C) are not exclusively restricted to the nucleus and cytosol and that spatial and functional specificity in CaMKIIδ activation is elicited by mobilization of different Ca²âº stores rather than by compartmentalized subtype localization.

Malposition of the central venous catheter secondary to accessory hemiazygos vein variant.

A 36-year-old female with sickle cell disease presented with sickle cell pain crisis. After failure to establish peripheral venous access, an internal jugular central venous catheter (CVC) was placed. Confirmation of internal jugular cannulation was performed with bedside ultrasound. A confirmatory chest X-ray revealed an unusual position of the catheter, taking a course inferiorly, making a loop and remaining on the left side of the mediastinum. A lateral view was done and revealed that the catheter passed inferiorly through the internal jugular vein then posteriorly and inferiorly giving the looped appearance. This is better delineated on a sagittal view CT scan showing the tip of the catheter terminating in the accessory hemiazygos vein. This unusual course is due to a variant of the accessory hemiazygos vein which is connected to the left superior intercostal vein. This creates a lower resistance pathway for the CVC which passes from the internal jugular vein, down the left superior intercostal vein (instead of the left brachiocephalic vein) and into the accessory hemiazygos vein. Discussion: The correct tip placement of an internal jugular CVC terminates in the superior vena cava just above the cardiac silhouette. In 1%-2% of individuals, a connection between the accessory hemiazygos and the left superior intercostal vein is present. Rare cases are discovered incidentally during CVC placement. The diameter of the accessory hemiazygos vein is less than half of that of the superior vena cava. The catheter should not be used as central venous access and removal is recommended. Malpositioning of central catheters is unpredictable but can be easily avoided by using intraprocedural methods to confirm tip position. Such modalities include intracavitary ECG or ultrasound with agitated saline injection as described in the SIC (Safe Insertion of Centrally Inserted Central Catheters) protocol.

A Unique Case of COVID-19 Presenting as Focal Seizures With Impaired Awareness.

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has very rapidly become a global pandemic with millions of confirmed cases worldwide. In early 2021, viral encephalitis was the first neurological complication associated with COVID-19 and since then rise in cases has been reported with this association. A review highlighting 3 potential mechanisms linking a correlation between seizures and COVID-19 was previously reported. Herein described is a unique case of SARS-CoV2 infection that presented with focal seizure with impaired awareness.