Early role of CD4+ Th1 cells and antibodies in HER-2 adenovirus vaccine protection against autochthonous mammary carcinomas

HER-2 is an oncogenic tumor-associated Ag that is overexpressed in several human tumors including breast and ovarian cancer. The efficacy and mechanism of a HER-2-expressing recombinant adenoviral vaccine to protect against tumorigenesis was examined using HER-2 transgenic (BALB-neuT) mice, which develop spontaneous breast tumors in all 10 mammary glands, and also using a transplantable mouse tumor model. Vaccination beginning at 6-8 wk of age (through 19 wk of age) prevented development of spontaneous mammary tumors even after 50 wk, whereas the animals in the control groups had tumors in all mammary glands by 25 wk. Such long-term protection after the last boost has not been achieved previously in this transgenic mouse in which the oncogene is continuously spawning tumorigenesis. Using beta(2)-microglobulin-knockout, IFN-gamma-knockout, and B cell-deficient mice, CD4(+) and CD8(+) cell depletion, and Ab transfer studies, we show that induction of anti-HER-2/neu Abs are both necessary and sufficient for protection, and the IgG2a isotype is most effective. In contrast, CD8(+) T cells are not necessary at all, and CD4(+) T cells are necessary for only 36-48 h after immunization to provide help for B cells but not as effector cells. Equal protection in immunized mice deficient in FcgammaRI/III excluded an FcR-mediated mechanism. Anti-HER-2 serum not only inhibited growth of mammary tumor cell lines expressing HER-2 in vitro but also protected mice from tumors in vivo, suggesting a direct action of Ab on the tumor cells. Such a vaccine may provide Ab-mediated protection against HER-2-expressing breast cancers in humans.     

Effects of NaCl on 4-hydroxy-2-hexenal formation in yellowtail meat stored at 0 degrees C

Changes in the 4-hydroxy-2-hexenal (HHE) and malonaldehyde (MA) contents were investigated in the meat of the yellowtail Seriola quinqueradiate containing 0, 0.3, 0.6, and 0.9 M NaCl stored at 0 degrees C for 7 days. After 7 days of storage, the HHE content was significantly lower and the MA content significantly higher in the meat containing NaCl than in the control without NaCl.     

Casting the critical regions in nucleotide binding oligomerization domain 2 protein: a signature mediated structural dynamics approach

Nucleotide binding oligomerization domain 2 (NOD2), a protein involved in the first line defence mechanism has a pivotal role in innate immunity. Impaired function of this protein is implicated in disorders such as Blau syndrome and Crohn’s disease. Since an altered function is linked to protein’s structure, we framed a systematic strategy to interpret the structure–function relationship of the protein. Initiated with mutation-based pattern prediction and identified a distant ortholog (DO) of NOD2 from which the intra-residue interaction network was elucidated. The network was used to identify hotspots that serve as critical points to maintain the stable architecture of the protein. Structural comparison of NOD2 domains with a DO revealed the minimal number of intra-protein interactions required by the protein to maintain the structural fold. In addition, the conventional molecular dynamics simulation emphasized the conformational transitions at hot spot residues between native NOD2 domains and its respective mutants (G116R, R42W and R54A) structures. The analysis of intra-protein interactions globally and the displacement of residues locally around the mutational site revealed loss of several critical bonds and residues vital for the protein’s function. Conclusively we report, about 10 residues in leucine-rich repeat, 13 residues in NOD and 6 residues in CARD domain are required by the NOD2 to maintain its function. This protocol will help the researchers to achieve for more prospective studies to attest druggable site utility in discovering novel drug candidates.     

Digital analysis of changes by Plasmodium vivax malaria in erythrocytes

Blood samples of malaria patients (n = 30), selected based on the severity of parasitemia, were divided into low (LP), medium (MP) and high (HP) parasitemia, which represent increasing levels of the disease severity. Healthy subjects (n = 10) without any history of disease were selected as a control group. By processing of erythrocytes images their contours were obtained and from these the shape parameters area, perimeter and form factor were obtained. The gray level intensity was determined by scanning of erythrocyte along its largest diameter. A comparison of these with that of normal cells showed a significant change in shape parameters. The gray level intensity decreases with the increase of severity of the disease. The changes in shape parameters directly and gray level intensity variation inversely are correlated with the increase in parasite density due to the disease.     

Associations between low Apgar scores and mortality by race in the United States: A cohort study of 6,809,653 infants

BackgroundApgar scores measure newborn health and are strongly associated with infant outcomes, but their performance has largely been determined in primarily white populations. Given the majority of the global population is not white, we aim to assess whether the association between low Apgar score and mortality in infants varies across racial groups.Methods and findingsPopulation-based cohort study using 2016 to 2017 United States National Vital Statistics System data. The study included singleton infants born between 37⁺⁰ and 44⁺⁶ weeks to mothers over 15 years, without congenital abnormalities. We looked at 3 different mortality outcomes: (1) early neonatal mortality; (2) overall neonatal mortality; and (3) infant mortality. We used logistic regression to assess the association between Apgar score (categorized as low, intermediate, and normal) and each mortality outcome, and adjusted for gestational age, sex, maternal BMI, education, age, previous number of live births, and smoking status, and stratified these models by maternal race group (as self-reported on birth certificates). The cohort consisted of 6,809,653 infants (52.8% non-Hispanic white, 23.7% Hispanic, 13.8% non-Hispanic black, 6.6% non-Hispanic Asian, and 3.1% non-Hispanic other). A total of 6,728,829 (98.8%) infants had normal scores, 63,467 (0.9%) had intermediate scores, and 17,357 (0.3%) had low Apgar scores. Compared to infants with normal scores, low-scoring infants had increased odds of infant mortality. There was strong evidence that this association varied by race (p < 0.001) with adjusted odds ratios (AORs) of 54.4 (95% confidence interval [CI] 49.9 to 59.4) in non-Hispanic white, 70.02 (95% CI 60.8 to 80.7) in Hispanic, 23.3 (95% CI 20.3 to 26.8) in non-Hispanic black, 100.4 (95% CI 74.5 to 135.4) in non-Hispanic Asian, and 26.8 (95% CI 19.8 to 36.3) in non-Hispanic other infants. The main limitation was missing data for some variables, due to using routinely collected data.ConclusionsThe association between Apgar scores and mortality varies across racial groups. Low Apgar scores are associated with mortality across racial groups captured by United States (US) records, but are worse at discriminating infants at risk of mortality for black and non-Hispanic non-Asian infants than for white infants. Apgar scores are useful clinical indicators and epidemiological tools; caution is required regarding racial differences in their applicability.

Emma Gillette and co-authors assess the associations between low Apgar scores and mortality in infants by race in the United States.     

Simple and efficient genomic DNA extraction protocol for molecular characterisation of Phytophthora colocasiae causing taro leaf blight

Genomic DNA extraction protocol with relatively high quantity and purity is prerequisite for the successful molecular identification and characterisation of plant pathogens. Conventional DNA extraction methods are often time-consuming and yield only very poor quantity of genomic DNA for samples with higher mycelial age. In our laboratory, we have aimed at establishing an efficient DNA isolation procedure, exclusively for the oomycete pathogen Phytophthora colocasiae causing serious leaf blight disease in taro. For this a phenol free protocol was adopted, which involves SDS/Proteinase K-based inactivation of protein contaminants, extraction of nucleic acids using chloroform: isoamyl alcohol and later precipitation of genomic DNA using isopropanol and sodium acetate. The purity of the isolated DNA was analysed by A_260/280 and A_260/230 spectrophotometric readings and confirmed by restriction digestion with restriction enzyme Eco RI. In this study, a comparative assessment was done with CTAB method and the commercial genomic DNA purification kit (Thermo Fisher Scientific, Fermentas, EU). The extracted DNA was found to be suitable for further downstream applications like ITS amplification of the rDNA ITS region and PCR amplification with species-specific primers.     

Incidence and identification of Cassava tuber rot caused by Phytophthora palmivora

The occurrence of Cassava tuber rot in regions of Kolli hills, Kollam, and Kottayam of South India, causes major economic loss up to 70% in Cassava production. The disease tuber is characterised by brown watery lesions with foul smell, making it unfit for further use. The sporangia of the pathogen were oval and ellipsoid with a short pedicle. Identification of the isolate from these regions was also confirmed by ribosomal internal transcribed spacer (ITS) of rDNA region. The pathogen was highly aggressive when pathogenicity was tested. Based on morphological, pathogenicity and ITS sequences, the pathogen was identified as Phytophthora palmivora. Development of integrated disease management practices is essential to combat the disease. This is the first report recording the spread of Cassava tuber rot disease in regions of Kolli hills of Tamil Nadu and Kollam and Kottayam, of Kerala.     

Intra-abdominal fat burden discriminated in vivo using proton magnetic resonance spectroscopy

Objective: To assess proton magnetic resonance spectroscopy (¹H‐MRS) as a means to distinguish among mice with disparate intra‐abdominal body fat compositions, and to measure changes in intra‐abdominal fat burden during weight loss and regain. Research Methods and Procedures: Intra‐abdominal fat burden was analyzed as a ratio of integrated areas under the curves of fat to water ¹H‐MRS signals collected from a region of interest standardized across B6.V‐Lep^ob, C57BL/6, and A‐ZIP/F mice that exhibited various genotypically related body fat compositions, ranging from obese (B6.V‐Lep^ob) to minimal body fat (A‐ZIP/F). ¹H‐MRS analysis of fat burden was compared with intra‐abdominal fat volume and with a single cross‐sectional intra‐abdominal fat area calculated from segmented magnetic resonance images. Similar measurements were made from obese B6.V‐Lep^ob mice before, during, and after they were induced to lose weight by leptin administration. Results: Relative amounts of intra‐abdominal fat analyzed by ¹H‐MRS differed significantly according to body composition and genotype of the three strains of mice (p < 0.05). Intra‐abdominal fat assessed by ¹H‐MRS correlated with both intra‐abdominal fat volume (r = 0.88, p < 0.001) and body weight (r = 0.82, p < 0.001) among, but not within, all three genotypes. During weight loss and regain, there was a significant overall pattern of changes in intra‐abdominal fat quantity that occurred, which was reflected by ¹H‐MRS (p = 0.006). Discussion: Results support the use of localized ¹H‐MRS for assessing differences in intra‐abdominal fat. Refinements in ¹H‐MRS voxel region of interest size and location as well as instrument precision may result in improved correlations within certain body compositions.     

Secreted Protein Acidic and Rich in Cysteine (SPARC) in Human Skeletal Muscle

Secreted protein acidic and rich in cysteine (SPARC)/osteonectin is expressed in different tissues during remodeling and repair, suggesting a function in regeneration. Several gene expression studies indicated that SPARC was expressed in response to muscle damage. Studies on myoblasts further indicated a function of SPARC in skeletal muscle. We therefore found it of interest to study SPARC expression in human skeletal muscle during development and in biopsies from Duchenne and Becker muscular dystrophy and congenital muscular dystrophy, congenital myopathy, inclusion body myositis, and polymyositis patients to analyze SPARC expression in a selected range of inherited and idiopathic muscle wasting diseases. SPARC-positive cells were observed both in fetal and neonatal muscle, and in addition, fetal myofibers were observed to express SPARC at the age of 15–16 weeks. SPARC protein was detected in the majority of analyzed muscle biopsies (23 of 24), mainly in mononuclear cells of which few were pax7 positive. Myotubes and regenerating myofibers also expressed SPARC. The expression-degree seemed to reflect the severity of the lesion. In accordance with these in vivo findings, primary human-derived satellite cells were found to express SPARC both during proliferation and differentiation in vitro. In conclusion, this study shows SPARC expression both during muscle development and in regenerating muscle. The expression is detected both in satellite cells/myoblasts and in myotubes and muscle fibers, indicating a role for SPARC in the skeletal muscle compartment. (J Histochem Cytochem 57:29–39, 2009)     

Ab Initio Coordination Chemistry for Nickel Chelation Motifs

Chelation therapy is one of the most appreciated methods in the treatment of metal induced disease predisposition. Coordination chemistry provides a way to understand metal association in biological structures. In this work we have implemented coordination chemistry to study nickel coordination due to its high impact in industrial usage and thereby health consequences. This paper reports the analysis of nickel coordination from a large dataset of nickel bound structures and sequences. Coordination patterns predicted from the structures are reported in terms of donors, chelate length, coordination number, chelate geometry, structural fold and architecture. The analysis revealed histidine as the most favored residue in nickel coordination. The most common chelates identified were histidine based namely HHH, HDH, HEH and HH spaced at specific intervals. Though a maximum coordination number of 8 was observed, the presence of a single protein donor was noted to be mandatory in nickel coordination. The coordination pattern did not reveal any specific fold, nevertheless we report preferable residue spacing for specific structural architecture. In contrast, the analysis of nickel binding proteins from bacterial and archeal species revealed no common coordination patterns. Nickel binding sequence motifs were noted to be organism specific and protein class specific. As a result we identified about 13 signatures derived from 13 classes of nickel binding proteins. The specifications on nickel coordination presented in this paper will prove beneficial for developing better chelation strategies.