Different Treatments for Sugarcane Juice Preservation.

This investigation aimed to optimize the time, pH, pressure, and temperature of sugarcane juice pasteurization and to develop a "ready to serve" bottled sugarcane juice with a high preservation efficiency. Fresh sugarcane juice was extracted from sugarcane genotype Co 89003, and beverage samples were collected using three different treatments: sulphitation of juice with the addition of potassium metabisulphite (KMS-25, 50, 100, and 150 ppm), acidification of juice (addition of citric acid, to reduce the pH of the juice to 4.8, 4.5, and 4.25), and steam treatment of the canes (5 min, 10, and 15 min at 7 psi). In all treatments, the juice was pasteurized in glass bottles @ 65 °C for 25 min and stored at low temperature (5 °C) in pre-sterilized glass bottles. Juice properties such as the ˚Brix, total sugar, pH, and total phenolic content decreased with storage, whereas the microbial count, titrable acidity, and reducing sugar content significantly increased during storage. The addition of KMS, citric acid, and the steam treatment reduced the browning of juice and maintained the color of juice during storage, by inhibiting the polyphenol oxidase enzyme activity, from 0.571 unit/mL to 0.1 unit/mL. Among the selected treatments, sugarcane juice with KMS (100 and 150 ppm) and steam treatment of the canes for 5 and 10 min at 7 psi showed the minimum changes in physico-chemical properties, sensory qualities, and restricted microbial growth. Thesulphitation treatment with pasteurization proved best for increasing the shelf life of sugarcane juice upto 90 days with refrigeration. Similarly, the steam-subjected cane juice (10 and 15 min at 7 psi) could be effectively preserved for upto 30 days with refrigeration, without any preservative.

Sternocleidomastoid flap for pedicled reconstruction in head & neck surgery- revisiting the anatomy and technique.

BACKGROUND: Previous studies on sternocleidomastoid flaps, have defined the importance of preserving sternocleidomastoid (SCM) branch of superior thyroid artery (STA). This theory drew criticism, as this muscle is known to be a type II muscle, i.e., the muscle has one dominant pedicle (branches from the occipital artery at the superior pole) and smaller vascular pedicles entering the belly of muscle (branches from STA and thyrocervical trunk) at the middle and lower pole respectively. It was unlikely for the SCM branch of STA to supply the upper and lower thirds of the muscle. We undertook a cadaveric angiographic study to investigate distribution of STA supply to SCM muscle. METHODS: It is a cross-sectional descriptive study on 10 cadaveric SCM muscles along with ipsilateral STA which were evaluated with angiography using diatrizoate (urograffin) dye. Radiographic films were interpreted looking at the opacification of the muscle. Results were analyzed using frequency distribution and percentage. RESULTS: Out of ten specimens, near complete opacification was observed in eight SCM muscle specimens. While one showed poor uptake in the lower third of the muscle, the other showed poor uptake in the upper third segment of muscle. CONCLUSION: Based on the above findings we suggest to further investigate sternocleidomastoid muscle as a type III flap, as the STA branch also supplies the whole muscle along with previously described pedicle from occipital artery. However, this needs to be further corroborated intra-operatively using scanning laser doppler. This also explains better survival rates of superior thyroid artery based sternomastoid flaps.

Biomass potential of novel interspecific and intergeneric hybrids of Saccharum grown in sub-tropical climates.

Sugarcane-derived biomass is a promising source of renewable energy to meet the growing demands for biofuel. Currently, modern sugarcane cultivars are unable to provide enough biomass due to their narrow genetic base and susceptibility to abiotic and biotic stresses. We have evaluated total of 23 hybrids derived from diverse genetic backgrounds of different Saccharum spp. and allied genera, one inbred and compared with commercial checks. Intergeneric hybrids (IGHs) KGS 99-100 and GU 04-432, produced significantly higher biomass (43.37 t ha-1 and 35.24 t ha-1, respectively) than commercial sugarcane have genes derived from Erianthus arundinaceus. Interspecific hybrids (ISHs) GU 07-3704 and 99-489, also produced significantly higher amounts of biomass (37.24 t ha-1 and 33.25 t ha-1, respectively) than commercial checks have genes from S. officinarum and S. spontaneum backgrounds. ISHs recorded significantly higher biomass yield, number of stalks and total dry matter percentage whereas, IGH group recorded significantly higher fibre percent. Furthermore, the clones resistant to red rot and sugarcane borers were identified. The estimated energy value for seven hybrid clones was found to be very high. Cluster analysis of genetic traits revealed two major clusters in traits improving biomass. Our study has revealed that the genetic diversity present in these hybrids could be used for improving biomass production and tolerance to abiotic and biotic stresses in cultivated sugarcanes.

Association of functional genetic variants of transcription factor Forkhead Box P3 and Nuclear Factor-κB with end-stage renal disease and renal allograft outcome.

BACKGROUND: The transcription factor FOXP3 and NF-κB regulates the expression of various genes that play an important role in the regulation of renal inflammation. We investigated the association of FOXP3 (rs2232365, rs3761548, rs5902434 and rs2294021) and NF-κB1 (rs28362491 and rs696) gene variants in susceptibility and prognosis of end stage renal disease (ESRD) and renal allograft outcome. METHODS: We genotyped four common polymorphisms of FOXP3 and two-tag SNPs of NF-κB1 genes in 350 ESRD cases and 350 controls. Single marker analysis and SNP-SNP interaction model (one to six way combinations) was used for determination of clinical outcome of ESRD and acute rejection episode (ARE). RESULTS: We observed significantly higher occurrence of mutant genotypes of tag-SNPs of FOXP3 namely; rs2232365 and rs3761548 along with NF-kB1 namely; rs28362491 and rs696 in ESRD and ARE cases, suggested a risk association for ESRD and ARE. Interestingly, multifactor dimension reduction analysis suggested an increased risks of nearly 6-folds for ESRD and 23-folds for ARE cases under the six factors model which consists of tag-SNPs of FOXP3 (rs2232365, rs3761548, rs5902434 and rs2294021) and NF-kB1 (rs28362491 and rs696). Kaplan-Meier survival analysis showed the lowest overall survival for mutant genotypes compared with wild and heterozygous genotypes of rs2232365 and rs3761548 tag SNPs of FOXP3 as well as NF-kB1 tag-SNPs rs28362491 and rs696 in renal allograft recipients. The crude and adjusted hazard ratios in univariate and multivariate Cox regression models showed almost 2-folds to 3-folds risk for overall survival against mutant genotypes of tag-SNPs of FOXP3 (rs2232365 and rs3761548) and NF-kB1 (rs28362491 and rs696) genes. CONCLUSIONS: These results suggest that variants of transcription factor FOXP3 and NF-kB1 might be associated with increased risk to the clinical outcome of ESRD and renal allograft survival.

Genetic variation in Micro-RNA genes of host genome affects clinical manifestation of symptomatic Human Cytomegalovirus infection.

BACKGROUND: Micro-RNAs are implicated in various physiological and pathologic processes. In this study, we tested whether Micro-RNA gene variants of host-genome affect clinical manifestation of symptomatic HCMV infection. METHODOLOGY: HCMV infection was detected by fluorescent PCR and immuno-histochemistry. The detection of genetic variants of four studied Micro-RNA tag-SNPs was done through PCR-RFLP assay and validated with DNA sequencing. RESULTS: We observed an increased risk ranged from 3-folds to 5-folds among symptomatic HCMV cases for mutant genotype of rs2910164 (crude OR=3.11, p=0.009 and adjusted OR=3.25, p=0.007), rs11614913 (crude OR=3.20, p=0.006 and adjusted OR=3.48, p=0.004) and rs3746444 (crude OR=4.91, p=0.002 and adjusted OR=5.28, p=0.002) tag-SNPs. Interestingly, all the tag-SNPs that were significant after multiple comparisons at a FDR of 5% in symptomatic HCMV cases remained significant even after bootstrap analysis, providing internal validation to these results. Multifactor Dimensionality Reduction (MDR) analysis revealed 5-folds increased risk for symptomatic HCMV cases under the four-factor model (rs2910164, rs2292832, rs11614913 and rs3746444). CONCLUSIONS: These results suggest that Micro-RNA gene variants of host-genome may affect clinical manifestation of symptomatic HCMV infection.

HLA-G gene expression influenced at allelic level in association with end stage renal disease and acute allograft rejection.

BACKGROUND: Human leukocyte antigen (HLA)-G is a non-classical major-histocompatibility complex class-I molecule associated with immunosuppressive function. We have evaluated the impact of HLA-G allele associated with untranslated-region (UTR)-haplotype in end stage renal disease (ESRD) and acute allograft rejection (AR) cases. The mRNA levels of different HLA-G isoforms were evaluated in ESRD and AR cases. Subsequently, the total HLA-G mRNA levels and protein concentration were evaluated against its UTR-haplotype among ESRD and AR cases. METHODOLOGY: Sequence based typing of the promoter region was carried-out to evaluate the impact of HLA-G haplotype in 350 ESRD cases and 300 controls. HLA-G gene expression was evaluated at the transcriptional level using semi-quantitative and quantitative PCR, whereas protein concentration was determined by ELISA among both cases and control. RESULTS: Increased risk was observed for G*01:01:01:03, G*01:01:02, G*01:06 and G*01:05:N haplotypes while G*01:01:01:01 and G*01:04:01 haplotypes showed a protective effect in ESRD and AR cases. Higher level of soluble HLA-G isoforms (G5 and G6) was observed among ESRD cases. Reduced levels of soluble isoform (G5) and increased levels of membrane bound (G1 and G3) isoforms were found among AR cases, revealing risk association. Decreased HLA-G expression was observed at both mRNA and protein level for G*01:01:01:03 and G*01:05:N haplotypes in ESRD and AR cases. CONCLUSIONS: These results suggest that the variation in the expression profile of membrane bound and soluble isoforms may modulate the risk for ESRD and AR. UTR-haplotypes appear to be involved in different HLA-G expression patterns at transcriptional and translational levels.

Association of CTLA-4 gene polymorphism with end-stage renal disease and renal allograft outcome.

Cytotoxic T-lymphocyte antigen 4 (CTLA-4) is upregulated in effector-T-cells after activation that may alter signal transduction and subsequently cytokine production. The present study was designed to investigate the impact of CTLA-4+49 A>G (rs231775), -318 C>T (rs5742909), -658 C>T (rs11571317), -1147 C>T (rs16840252), -1661 A>G (rs4553808), +6230 A>G (rs3087243) SNPs, and microsatellite (AT)n repeat polymorphism among end-stage renal disease (ESRD), acute allograft rejection (AR), and delayed graft function (DGF) cases. In this regard, 350 ESRD patients and 350 controls were included. Polymerase chain reaction (PCR)-restriction fragment length polymorphism analysis method was used for genotyping of CTLA-4 SNPs, while PCR-polyacrylamide gel electrophoresis method was adopted for studying CTLA4 (AT)n polymorphism. The mutant genotype GG of CTLA-4+49A>G,+6230 A>G, and longer alleles of (AT)n repeats polymorphisms were risk associated with ESRD, AR, and DGF cases. The distribution of haplotype+49G:+6230G and GCTTGG (constructed by using 6 studied SNPs) showed risk association for ESRD, DGF, and AR cases. Further, linkage analysis demonstrated strong to moderate linkage disequilibrium in our study populations. The meta-analysis also revealed risk associations for AR cases against GG genotype of CTLA-4+49A>G SNP, while CTLA-4 -318C>T polymorphism showed no correlation against TT genotype among AR cases. Subsequently, no correlation was established against the CTLA-4 -318C>T, -658 C>T, -1147 C>T, and -1661 A>G SNPs in the promoter region. Survival analysis revealed risk associations against GG genotype of CTLA-4+49A>G, +6230 A>G SNP's with overall survival (OS), and higher hazard for the OS. These results suggested that CTLA-4 variants might be involved in susceptibility to ESRD, AR, and DGF.

Teratogenic effects of 5-fluorouracil on rat brain.

Nine female pregnant rats were treated with single dose (30mg/kg) of 5-fluorouracil on 12th day of gestation. Gross defects in developing brain of rat fetuses were observed. 5-Fluorouracil is an antineoplastic drug, which has not been adequately studied. 5-FU induced about 5% mortality with significant reduction in body weight and various dimensions of the developing brain (p<0.001). Macroscopic findings of the developing brain revealed microcephaly, regression or absence of olfactory lobe and obliteration of the various fissures on the dorsal and ventral surfaces of the brain. Neuroembryopathic effects of 5-FU is more marked when given in late phase of gestation. So, it is advisable that the drug should be avoided during this period of pregnancy.

Neuroembryopathic effect of clobazam in rat: a histological study.

Clobazam is a derivative of the benzodiazepines used as an anti-epileptics drug. The pregnant rat had received, orally, 125 mg/kg of clobazam daily from 1st-7th day of gestation and fetuses were collected on 20th day of pregnancy. Brain of the clobazam treated fetuses showed no significant change in weight and size in comparision to that of the controls. Other gross abnormalities were also not found in the brains of treated group. Histological examination revealed the alteration in cytoarchitecture of the cerebral and the cerebellar cortex. Strips of focal coagulate necrosis extended through most of the layers of the cerebral cortex. There were paucity of neurons and neurological elements in the cortex and the central grey area of spinal cord. Many of these neurons showed pyknotic nuclei. The intercellular spaces increased either because of decrease in cell number or due to shrinkage and clumping of degenerated neurons. Subcortical (white matter) zone showed the paucity and derangement of the oligodendrocytes and the astrocytes, which revealed vigorous proliferation in the treated group. These findings showed that the benzodiazepines and their derivatives have teratogenic effect in the developing mammalian central nervous system.

Neuroembryotoxic effect of 5- Fluorouracil in rat: injecting in late phase of gestation.

Macro and microscopic findings in developing brain of rat fetuses were observed after intraperitoneal injection with a single dose (30mg/kg) of 5-Fluorouracil (5-FU) in late phase of gestation. 5-FU induced more than 60.0% lethality with significant reduction (p<0.001) in weight and various dimensions of the developing brain. Macroscopic findings of the developing brain revealed microcephaly, regression or absence of the olfactory lobe and obliteration of the various fissures on the dorsal surface. Microscopic examination of the olfactory lobe of treated brain showed the obliteration of the olfactory ventricle, distortion of the cellular arrangement of various layers of the olfactory cortex with clumping of degenerated neurons and glial cells. Cerebral cortex of the treated brain revealed the distortion of normal cytoarchitecture of the various cortical layers. The neurons of the treated brain revealed the degeneration, deeply stained eccentric nucleus with loss of mitotic figures and pyknotic changes. Subcortical zone of the treated cerebrum showed the degenerative changes in the fibrous structure along with paucity of the glial cells. The hippocampus of the treated brain revealed the loss of normal cytoarchitecture and shrinkage of all the layers. Neuroembyopathic effect of 5-FU is severe, when given in late phase of pregnancy, so it is advisable that the drug should be avoid during the late period of pregnancy.

Anatomical variation in origin and course of the thoracoacromial trunk and its branches.

The variations in origin and course of the thoracoacromial trunk (TAT) and its branches were studied in 178 cadavers during the routine dissection from the year 1982 to 2002. The TAT originated from the first part of the axillary artery (AA) in 13.4% cases of the right and 10.6% of the left axilla. The variations in origin of the branches of TAT were divided into three groups. First variation group showed deltoacromial (DA) and clavipectoral (CP) subtrunks of the TAT originating directly from the AA in majority of cases. Second group revealed clavicular branch of the TAT originating from the AA, whereas in the third group all classical branches originated directly from the AA and there was no existence of the TAT. The superior thoracic artery (STA) originated from the TAT in 16.8% (Confidence Interval, CI: 11.12-21.89) cases of the right and 6.1% (CI: 2.59-9.53) of the left axilla and the lateral thoracic artery in 39.8% (CI: 32.01-46.10) cases of the right and 29.3% (CI: 15.01-26.80) of the left axilla. The incidence of variations in origin of the TAT and its branches was found higher on right side. The knowledge of these variations is of anatomical and surgical interest. This information is useful for the surgeons dealing with the axillary region especially in case of reconstructive surgery.

Growth retardation and lethality induced by adriblastina in chick embryo.

The low (1.2 mg/kg) and high (2.4 mg/kg) therapeutic dose of adriblastina, dissolved in 0.04 ml of distilled water was injected into the chick embryo at different duration of the incubation. The control chick embryo received equal volume of distilled water at the same duration. All groups of embryo were collected on day 19 of gestation. The treated embryo showed growth retardation and lethality in a dose dependence response. The lethality and growth retardation in both treated groups were found significantly different (p<0.001) as compared with the control chick embryo. Similarly the groups treated on different days showed a time sequential effect on the developing chick embryo. Our observation had revealed that the drug is teratogenic to the chick embryo.

Articular branch of the axillary artery and its clinical implication.

Origin and course of the articular branch of the axillary artery was recorded in 151 (43.9%) out of 344 axilla during the routine dissection. The branch was observed in 60.5% right side and 39.4% left side of male axilla, whereas in 55.5% right and 44.4% left side of female axilla and the difference was found significant (P < 0.001) among the gender. The orientation of the articular branch on right and left side was of similar proportion in both the sexes. The articular branch entered into the shoulder joint either upper, middle or lower part of the capsule of joint. This articular branch originated mostly from the lateral aspect of second part of the axillary artery below to the origin of the lateral thoracic artery. This is a rare observation and is of importance to clinicians dealing with the shoulder joint.