The impact of flow PRA on outcome in pediatric heart recipients in modern era: An analysis of the Pediatric Heart Transplant Study database.

Data from patients in the Pediatric Heart Transplant Study (PHTS) registry transplanted between 2010 and 2014 were analyzed to determine the association between HLA antibody (PRA) determined by SPA using Luminex or flow cytometry with a positive retrospective cross-match and the post-transplant outcomes of acute rejection and graft survival. A total of 1459 of 1596 (91%) recipients had a PRA reported pretransplant; 26% had a PRA > 20%. Patients with a PRA > 20% were more likely to have CHD, prior cardiac surgery, ECMO support at listing, and waited longer for transplantation than patients with a PRA <20%. Patients with higher PRA% determined by SPA were predictive of a positive retrospective cross-match determined by flow cytometric method (P < .001). A PRA > 50% determined by SPA was independently associated with worse overall graft survival after first month of transplant in both unadjusted and adjusted for all other risk factors. In this large multicenter series of pediatric heart transplant recipients, an elevated PRA determined by SPA remains a significant risk factor in the modern era.

The caspase-independent algorithm of programmed cell death in Leishmania induced by baicalein: the role of LdEndoG, LdFEN-1 and LdTatD as a DNA 'degradesome'.

In the post-genomic perspective, the quest of programmed cell death (PCD) mechanisms in kinetoplastid parasites lies in the identification and characterization of cell death executer proteins. Here, we show that baicalein (BLN), a potent topoisomerase IB inhibitor, generates an oxidative stress in the parasites leading to altered physiological and morphological parameters, which are characteristic of PCD. For the first time we elucidate that, caspase-independent activation of a novel effector molecule, endonuclease G (LdEndoG), mediates BLN-induced cell death. Functional characterization of LdEndoG identifies Flap endonuclease-1 (LdFEN-1) and LdTatD-like nuclease as other effector molecules. BLN treatment translocates LdEndoG from mitochondria to nucleus, where it forms separate complexes with LdFEN-1 and LdTatD to constitute a DNA 'degradesome' unique to these parasites. Conditional antisense knockdown of LdEndoG provides protection against PCD. This knowledge paves the path toward a better understanding of the PCD pathway in simpler systems, which could be exploited in anti-leishmanial chemotherapy.

Apoptosis is induced in leishmanial cells by a novel protein kinase inhibitor withaferin A and is facilitated by apoptotic topoisomerase I-DNA complex.

Protein kinase C (PKC) is an important constituent of the signaling pathways involved in apoptosis. We report here that like staurosporine, withaferin A is a potent inhibitor of PKC. In Leishmania donovani, the inhibition of PKC by withaferin A causes depolarization of DeltaPsim and generates ROS inside cells. Loss of DeltaPsim leads to the release of cytochrome c into the cytosol and subsequently activates caspase-like proteases and oligonucleosomal DNA cleavage. Moreover, in treated cells, oxidative DNA lesions facilitate the stabilization of topoisomerase I-mediated cleavable complexes, which also contribute to DNA fragmentation. However, withaferin A and staurosporine cannot induce cleavable complex formation in vitro with recombinant topoisomerase I nor with nuclear extracts from control cells. Taken together, our results indicate that inhibition of PKC by withaferin A is a central event for the induction of apoptosis and that the stabilization of topoisomerase I-DNA complex is necessary to amplify apoptotic process.

Camptothecin induced mitochondrial dysfunction leading to programmed cell death in unicellular hemoflagellate Leishmania donovani.

The parasites of the order kinetoplastidae including Leishmania spp. emerge from most ancient phylogenic branches of unicellular eukaryotic lineages. In their life cycle, topoisomerase I plays a significant role in carrying out vital cellular processes. Camptothecin (CPT), an inhibitor of DNA topoisomerase I, induces programmed cell death (PCD) both in the amastigotes and promastigotes form of L. donovani parasites. CPT-induced cellular dysfunction in L. donovani promastigotes is characterized by several cytoplasmic and nuclear features of apoptosis. CPT inhibits cellular respiration that results in mitochondrial hyperpolarization taking place by oligomycin-sensitive F0-F1 ATPase-like protein in leishmanial cells. During the early phase of activation, there is an increase in reactive oxygen species (ROS) inside cells, which causes subsequent elevation in the level of lipid peroxidation and decrease in reducing equivalents like GSH. Endogenous ROS formation and lipid peroxidation cause eventual loss of mitochondrial membrane potential. Furthermore, cytochrome c is released into the cytosol in a manner independent of involvement of CED3/CPP32 group of proteases and unlike mammalian cells it is insensitive to cyclosporin A. These events are followed by activation of both CED3/CPP32 and ICE group of proteases in PCD of Leishmania. Taken together, our study indicates that different biochemical events leading to apoptosis in leishmanial cells provide information that could be exploited to develop newer potential therapeutic targets.

Raman, fluorescence, and time-resolved light scattering as optical diagnostic techniques to separate diseased and normal biomedical media.

Studies of Raman scattering, fluorescence and time-resolved light scattering were conducted on cancer and normal biomedical media. Fourier transform Raman spectroscopic measurements were performed on human normal, benign and cancerous tissues from gynecological (GYN) tracts. A comparison of the intensity differences between various Raman modes as well as the number of Raman lines, enables one to distinguish normal GYN tissues from diseased tissues. Fluorescence spectroscopic measurements on human breast tissues show that the ratio of fluorescence intensity at 340 nm to that at 440 nm can be used to distinguish between cancerous and non-cancerous tissues. Separate studies on normal and cancerous breast cell lines show spectral differences. The measurements of back-scattered ultrafast laser pulses from human breast tissues show differences in the scattered pulse profiles for different tissues. These studies show that various optical techniques have the potential to be used in medical diagnostic applications.

Outbreaks of egg drop syndrome due to EDS-76 virus in quail (Coturnix coturnix japonica).

Two outbreaks of the egg drop syndrome were observed in quail flocks maintained on a farm together with chickens. The decrease in egg production ranged from 10.6 per cent to 50.6 per cent, and the number of soft-shelled eggs increased with the decline in egg production. In both the outbreaks haemagglutination inhibiting antibodies to EDS-76 virus were detected. Fluorescent viral antigen specific to EDS-76 virus was also detected in the lining epithelial and glandular cells of the uterus.

Serum-ascites albumin gradient: a predictor of esophageal varices with ascites.

Other investigators have found that in adults the Serum-Ascites Albumin Gradient (SAAG) to be 1.1 g/dl or greater in the presence of portal hypertension (PTHN) and less than that in its absence. We sought to determine the correlation between the level of SAAG and the complications of PTHN, manifested by the presence of esophageal varices in children with ascites. Our study included 26 patients with cirrhosis, diagnosed by liver biopsy and 14 patients with nephrotic syndrome (NS) diagnosed by established criteria. The SAAG was measured in all patients. The patients with cirrhosis had upper gastrointestinal (GI) endoscopy for assessment of esophageal varices (EV). We found that 84.6% (22 of 26) patients with cirrhosis had High SAAG (> or = 1.1 g/dl) and 15.4% (4 of 26) had low SAAG (< 1.1 g/dl) (p < 0.001). EV was found in 91% (20 of 22) patients with high SAAG and in 50% (2 of 4) patients with low SAAG (p = 0.013). The SAAG differentiated cirrhosis with EV from those without EV (sensitivity = 91%, specificity = 50%, positive predictive value = 91%, negative predictive value = 50% and efficacy = 85%). The high SAAG is a useful means to predict the presence of EV in children with ascites.

Juvenile T waves (a study of 100 normal subjects).

"Juvenile pattern" of T wave inversion in the precordial leads of electrocardiogram was studied in 100 normal healthy Indians of 0 to 70 years of age in both sexes. Incidence of such pattern was observed in 25 subjects upto 19 years of age. Persons of weight/height ratio less than 30% had higher incidence of juvenile T waves. Maximum incidence (48%) of juvenile T waves was observed in the annual income group of Rs. 15000/-. Exercise ECC showed slight flattening of the inverted T waves in V1 to V3 leads in eight subjects only.

Optical biopsy of benign and malignant tissue by time resolved spectroscopy.

Pathological condition of malignant tissue could be analyzed by spectral domain or time domain spectroscopy, the two being the complementary to each other in optical biopsy (OB) of cancer. This paper reports results of time resolved emission spectroscopy (TRS) of 24 excised tissue samples of breast and prostate (normal control = 12; benign = 4; malignant = 8), employing a 390 nm, 100 fs, Ti-Sapphire laser pulses.The fluorescence decay times were measured using streak camera and the resultant data were fitted for single and bi-exponential decays with reliability of 97%. Our results show the distinct difference between normal, benign and malignant tissues mostly due to the emission spectra of Nicotinamide Adenine Dinucleotide (NADH), Flavin Mononucleotide (FAD) and also due to the heterogeneity of micro environments associated with the diseased tissues. In this short report, fit is also shown that TRS of breast tissues are similar to those of prostate tissues.

Time-resolved fluorescence polarization dynamics and optical imaging of Cytate: a prostate cancer receptor-targeted contrast agent.

Cypate-octreote peptide analogue conjugate (Cytate) was investigated as a prostate cancer receptor-targeted contrast agent. The absorption and fluorescence spectra of Cytate were ranged in the near-infrared "tissue optical window." Time-resolved investigation of polarization-dependent fluorescence emitted from Cytate in solution as well as in cancerous and normal prostate tissues was conducted. Polarization preservation characteristics of Cytate in solution and tissues were studied. Fluorescence intensity emitted from the Cytate-stained cancerous prostate tissue was found to be much stronger than that from the Cytate-stained normal prostate tissue, indicating more Cytate uptake in the former tissue type. The polarization anisotropy of Cytate contained in cancerous prostate tissue was found to be larger than that in the normal prostate tissue, indicating a larger degree of polarization preservation in Cytate-stained cancerous tissue. The temporal profiles of fluorescence from Cytate solution and from Cytate-stained prostate tissue were fitted using a time-dependent fluorescence depolarization model. The photoluminescence imaging of Cytate-stained cancerous and normal prostate tissues was accomplished, showing the potential of Cytate as a fluorescence marker for prostate cancer detection.

Left ventricular diastolic dysfunction in children and young adults with Marfan syndrome.

Adults with Marfan syndrome (MFS) demonstrate abnormal aortic elastic properties manifest by decreased aortic distensibility and increased aortic stiffness. Left ventricular (LV) diastolic dysfunction has been reported in adults with MFS. The objective of this study was to assess the frequency of LV diastolic dysfunction in a group of children and young adults with MFS and to determine whether diastolic dysfunction is associated with hemodynamic alterations of the aorta. Review of echocardiographic findings in 40 patients with MFS was performed to assess LV size, systolic function, isovolumic relaxation time (IVRT), mitral inflow velocities, deceleration time (DT) of mitral E wave, and aortic root dimension. No patient had significant valvar disease or was on any cardiac medication at the time of study. A group of 40 age and sex-matched healthy subjects undergoing echocardiography served as controls. Significant differences in LV diastolic function were found between MFS patients and controls. MFS patients had prolonged DT and IVRT and decreased mitral E/A ratio, suggesting impaired LV relaxation. No relationship between aortic root dimension and diastolic performance was identified. Left ventricular diastolic dysfunction may be an early marker of myocardial involvement in young MFS patients occurring independently of aortic root dilatation.

Exercise capacity in pediatric heart transplant candidates: is there any role for the 14 ml/kg/min guideline?

A peak oxygen consumption (VO2) of < 14 ml/kg/min has been identified as a predictor of l-year mortality in adults with congestive heart failure (CHF) and is used as a criterion for listing for cardiac transplantation (OHT). The role of VO2 measurement in children awaiting OHT has not been thoroughly evaluated. We sought to assess the degree of exercise impairment and the clinical applicability of the 14 ml/kg/min rule in children awaiting OHT. Cardiopulmonary exercise test (CPT) and cardiac catheterization data in all patients listed for OHT during the period of 1995-2003 were reviewed. Fourteen patients with a mean age of 15.5 +/- 2.9 years underwent CPT with no serious adverse events at an interval of 6.6 +/- 5.1 months prior to OHT. The etiology of CHF was multifactorial. Patients had impaired aerobic capacity with a mean peak VO2 of 20.4 +/- 6.8 ml/kg/min. Eleven of 14 patients (79%) had a peak VO2 higher than the adult cutoff value of 14 ml/kg/min. Pediatric ambulatory patients with CHF can safely undergo CPT. Because of age-related differences in oxygen consumption and varied etiologies of CHF a peak VO2 of < 14 ml/kg/min is not a useful criterion for listing for OHT in this population.

Amylopectinosis disease isolated to the heart with normal glycogen branching enzyme activity and gene sequence.

We report a 17-month-old female patient with a rare cause of cardiomyopathy secondary to accumulation of amylopectin-like material (fibrillar glycogen) isolated to the heart. Evidence of amylopectinosis isolated to cardiac myocytes in this patient was demonstrated by histology and electron microscopy. Glycogen content, glycogen branching enzyme (GBE) activity, as well as phosphofructokinase enzyme activities measured in liver, skeletal muscle, fibroblasts and ex-transplanted heart tissue were all in the normal to lower normal ranges. Normal skeletal muscle and liver tissue histology and GBE activity, normal GBE activity in skin fibroblasts, plus normal GBE gene sequence in this patient exclude the classical branching enzyme deficiency (type IV GSD). We believe that this is an as yet uncharacterized and novel phenotype of GSD associated with cardiomyopathy, in which there is an imbalance in the regulation of glycogen metabolism limited to the heart.

Minimum blood concentration--a universal concept of anaesthetic potency.

Although brain concentration of anaesthetic agents is attractive as a method of expressing anaesthetic potency, both theoretical and practical considerations limit the usefulness of this index. Therefore, the MAC concept has been utilized to develop another expression of potency: the minimum blood concentration. This takes into account the blood/gas solubility coefficient, and the fact that at equilibrium the amount of anaesthetic agent present in brain is reflected in the blood concentration, and it is applicable to both parenteral and to inhalation anaesthetic agents.

Repolarization abnormalities in children with a structurally normal heart and ventricular ectopy.

In adults, increased QT dispersion has been shown to predict arrhythmic risk as well as risk of sudden death in several clinical settings. It is controversial whether QT and JT dispersion are increased in children with ventricular ectopy and a structurally normal heart. We studied two groups of children: 25 patients with ventricular ectopy and 25 healthy children as controls. Standard electrocardiograms were reviewed and dispersions of both corrected QT (QTc) and JT (JTc) intervals were compared. We conclude that QTc and JTc dispersions are significantly increased in children with ventricular ectopy compared to control subjects.

Imaging of a translucent object hidden in a highly scattering medium from the early portion of the diffuse component of a transmitted ultrafast laser pulse.

A translucent object hidden in a highly scattering medium is shown to be visible when the early portion of the transmitted diffuse pulse (snake photons) is detected. The use of the snake scattered photons to image objects depends on the scattering characteristics along a quasi-straight-line path that the photons traverse across the medium. A translucent object with different scattering characteristics compared with its surrounding medium will change the intensity of the snake photons. By scanning the medium across the laser beam and detecting only the snake photons, a translucent object hidden in a highly scattering medium could be located.

Berry syndrome with trisomy 13.

We report on 2-day-old neonate with trisomy 13 with coexistent distal aortopulmonary septal defect, aortic origin of the right pulmonary artery, interrupted aortic arch, intact ventricular septum, and a patent ductus arteriosus diagnosed by two-dimensional and color Doppler echocardiography. Review of the literature reveals that this patient is the 24th reported case of Berry syndrome and the first case of this unusual combination of cardiovascular defects associated with trisomy 13 syndrome.

Neonatal aortic arch thrombosis masquerading as coarctation of the aorta.

We report two neonates with thrombosis of the aortic arch and isthmus diagnosed by echocardiography. Neonatal transient protein C deficiency was present in one patient and the other patient had severe perinatal asphyxia. Both patients presented with markedly reduced left ventricular function and severe aortic obstruction. In one infant, the thrombus was successfully removed surgically, and the second infant died due to serious complications of perinatal asphyxia.