Absar-ul Hasnain

Operculina turpethum attenuates N-nitrosodimethylamine induced toxic liver injury and clastogenicity in rats.

The root extract of Operculina turpethum (OTE) has been used as an anti-inflammatory, purgative, and hepato-protective agent. N-Nitrosodimethylamine (NDMA) is a potent hepatotoxin that induces fibrosis of the liver. In the present study, we examined the therapeutic effects of OTE root extract against NDMA-induced hepatotoxicity and clastogenicity in rats. Hepatic fibrosis was induced in adult male albino rats through serial intraperitoneal administrations of NDMA at a concentration of 10mg/kg body weight on three consecutive days of each week over a period of three weeks. A group of rats received OTE orally in doses of 75, 150 and 200mg/kg body weight at 5h after the administration of NDMA. The controls and treated animals were sacrificed on days-7, 14 and 21 after the start of the administration of NDMA. The progression of hepatic fibrosis as well as the amelioration effect of OTE was evaluated through histopathologically as well as by immunohistochemical staining for the activation of hepatic stellate cells. Alterations in serum and liver biochemical parameters and LDH isoenzymes were also studied. Serial administration of NDMA resulted in well formed fibrosis in the liver and induction of micronuclei in the bone marrow cells. Staining of alpha-SMA demonstrated activated stellate cells from day-7 onwards which was dramatically increased on day-21. An elevation of micronuclei count, liver function enzymes, serum hydroxyproline levels and LDH isoenzymes 4 and 5 were also observed. All these changes were remarkably reduced in OTE administered animals and fibrogenesis was completely absent. Our results suggest that OTE has hepatoprotective and anti-clastogenic effects against NDMA-induced hepatic fibrosis. Therefore OTE may be used as a hepatoprotective agent against various liver diseases including toxic liver injury.

Changes in Some Biochemical Indices of Stability of Broiler Chicken Actomyosin at Different Levels of Sodium Bicarbonate in Presence and Absence of Sodium Chloride

This study examined some physicochemical properties of actomyosin isolated from pre-rigor chicken breast muscle (Pectoralis major) to gain insight on its interaction with 0.2–1.0M sodium bicarbonate (NaHCO3) either with or without 3% sodium chloride (NaCl). NaHCO3 has been a frequent quality raiser with respect to water holding capacity of meat, which is adversely affected at low pH. This study document a remarkable inactivation of ATPases and the weakening of actin-myosin interaction with increasing molarity of NaHCO3, irrespective of the presence or absence of NaCl. Similarly, there was a concomitant increase in turbidity of soluble fraction, increase in total protein solubility and reactive SH groups. The observations on protein solubility/insolubility are supported by SDS-PAGE profiles. Therefore, the conformational changes induced by NaHCO3 are concentration dependent and appear different from those during thermal transitions. The findings suggest that low concentration of NaHCO3 may be more suitable component of marinades, since it affects the main myofibrillar contractile complex less adversely.

Effects of an aqueous extract of Croton bonplandianum Baill in rats.

We have investigated the cytotoxic and biochemical effects of injecting aqueous phytoextract of Croton bonplandianum (Baill) leaves in male rats. Subchronic dosages of 3.25, 4.65 and 6.97mgphytoextractg(-1)b.wt.week(-1) were administered to rats. To test ameliorative effects, rats were injected with phytoextract mixed with 0.2mgg(-1)b.wt. of cyclophosphamide. Positive controls received only cyclophosphamide, while negative control groups were kept on normal diet and water. Our results demonstrate that phytoextract did not induce micronuclei formation in rats and shows insignificant amelioration (P<0.05). However, differences in serum LDH isoenzymes, ALP, SGOT, SGPT activities and bilirubin were remarkable and displayed dose as well as duration dependent variations. The most outstanding observation of this study was the release of cardiac TnI in sera of rats injected with 6.97mgg(-1)b.wt. of phytoextract for 21 days. Our findings suggest that at the highest concentrations used here phytoextract of C. bonplandianum is not clastogenic; instead it is cardio- and hepatotoxic.

Observation on the Changes in Lactate Dehydrogenase Isoenzymes in Post-Burn Patients: Significance in Relation to Creatine Kinase

Observation on the Changes in Lactate Dehydrogenase Isoenzymes in Post-Burn Patients: Significance in Relation to Creatine Kinase The present study deals with the quantitative assessment of lactate dehydrogenase isoenzymes in the sera of burn subjects. Efforts are also made here to show better predictive marker value of sera LDH, as a few other known protein markers like creatine kinase and myoglobin have limited analytical value in the management of thermal burns. Blood was initially collected at day-1 of admission from 29 burn and 10 healthy subjects. Further, the sampling was carried out at 2, 5, 10, 20 and 30 days of wound healing (recovery). Plasma and sera LDH isoenzymes were monitored on 7.5% polyacrylamide gel electrophoresis. Quantitative assessment of LDH isoenzymes was done from gelscans using GelPro and Scion Imaging softwares. Sera CK levels were estimated colorimetrically using reagent kits. Our results show that quantitative changes in LDH isoenzyme were more convincing and interpretable in the sera than plasma. Sera LDH-5 isoenzyme was detected as the major contributor of total sera LDH activity, which follows a change parallel to sera CK in burn subjects. Sera LDH-5 activity also remains significantly high for up to 10 days while sera CK levels were detected elevated up to 5 days (P<0.05) during the recovery of patients. Therefore, the present findings strongly recommended the use of sera to assess the LDH activity and indicate better stability of sera LDH-5 than sera CK during post burn wound healing. Opservacije O Promenama U Izoenzimima Laktat Dehidrogenaze Kod Pacijenata SA Opekotinama: Značaj U Odnosu NA Kreatin Kinazu Predmet studije je kvantitativna procena enzima laktat dehidrogenaze u serumu pacijenata sa opekotinama. Namera je bila da se prikaže bolja prediktivna vrednost markera LDH iz seruma, pošto nekoliko ostalih poznatih proteinskih markera poput kreatin kinaze i mioglobina imaju ograničenu analitičku vrednost u zbrinjavanju termičkih opekotina. Krv je uzeta prvog dana po prijemu od 29 pacijenata sa opekotinama i 10 zdravih subjekata. Zatim su uzimani uzorci posle 2, 5, 10, 20 i 30 dana zaceljivanja (oporavka). Izoenzimi LDH u plazmi i serumu praćeni su tokom elektroforeze na 7,5% poliakrilamidnom gelu. Kvantitativna procena izoenzima LDH izvršena je putem gel-skenova pomoću softvera GelPro i Scion Imaging. Nivoi CK u serumu određeni su kolorimetrijski korišćenjem komercijalnih reagenasa. Rezultati pokazuju da su kvantitativne promene u izoenzimima LDH bile ubedljivije i lakše za tumačenje u serumu nego u plazmi. Uočeno je da je izoenzim LDH-5 iz seruma glavni činilac ukupne aktivnosti LDH u serumu, koji prati paralelnu promenu CK u serumu pacijenata sa opekotinama. Takođe, aktivnost LDH-5 u serumu ostaje značajno povišena i do 10. dana, dok su nivoi CK u serumu povišeni do 5. dana (P<0,05) oporavka. Stoga naši rezultati ukazuju na bolju stabilnost LDH-5 u serumu nego CK u serumu tokom procesa zaceljivanja opekotina, te se preporučuje upotreba seruma za procenjivanje aktivnosti LDH.

Solubilisation of muscle proteins from chicken breast muscle by ultrasonic radiations in physiological ionic medium

Abstract Solubilisation of myofibrillar proteins in physiological or low ionic strength solutions is essential for their utilisation as supplementary protein food. In order to achieve low ionic strength solubility, ultrasonication as a physical force has been introduced as an effective method to shift the solubility range of myofibrillar proteins from high to low ionic medium. In this study, change in the solubility behaviour of extracted actomyosin by ultrasonication in tris-maleate (with/without 0.1 M NaCl) and water is studied. Our results demonstrate that ultrasonication solubilises actomyosin in all the three investigated systems i.e. tris-maleate (with 0.1 M NaCl), tris-maleate only (without 0.1 M NaCl) and water. A decreasing trend in the investigated biochemical parameters such as ATPases (Ca2+, Mg2+) and turbidity was observed as a result of ultrasonic exposure. Analysis of SDS-PAGE profiles showed least solubility of myosin heavy chain in water compared to tris-maleate (with/without 0.1 M NaCl), while results of electron micrographs reveal change in the degree of dissociation or disruption of actomyosin aggregates according to the time of sonication and the suspension-media type. In conclusion, the results of our study suggest that ultrasonication plays a significant role in solubilisation of major myofibrillar proteins most probably by altering the conformation of actomyosin complex.

Biochemical characteristics of myofibrillar proteins of the fish parasite Isoparorchis hypselobagri (Trematoda) as survival traits in an oxygen-rich environment

Abstract We have investigated biochemical properties of myofibrillar proteins of the digenetic trematode Isoparorchis hypselobagri, which correlate with its survival in the oxygen-rich swim bladder of its host catfish (Wallago attu). The polypeptide composition of the trematode’s natural actomyosin (NAM) was striated-muscle-like, with the exception that a 98-kD polypeptide corresponding to paramyosin also existed in its sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) profiles. The profiles of immunoprecipitated NAM of the trematode support these inferences. Ca2+-sensitivity of myofibrillar contractility and Mg2+-ATPase activity of I. hypselobagri resembled troponin-linked calcium regulation of the host striated muscle. Myofibrillar permeability to water influx was insensitive to calcium chelation at neutral pH. However, the host swim bladder myofibrils displayed smooth-muscle-like polypeptide composition, pH dependence of contractility, Ca2+-sensitivity, ATPase activities, and inactivation kinetics. We propose 2 survival strategies that I. hypselobagri appears to have co-evolved: (i) fast-muscle-like musculature with exceptionally high contractility or ATPase activity; and (ii) type-II myosin resembling the host muscle in functional plasticity.

Genetic diversity of serum proteins in some muslim populations of India.

Abstract The Muslim population of India is known for its historical and socioreligious significance. Literature on the genetic structure of this segment of Indias population is scanty. Therefore we have investigated the allele frequency distribution of haptoglobin (HP) and transferrin (TF) phenotypes among the Muslims to explore the genetic diversity of the Muslim immigrant populations of Aligarh. Aligarh is a city in Uttar Pradesh, India (latitude 27°54′N, longitude 78°5′E), situated 130 km southeast of Delhi. The population is mainly represented by Muslim immigrants from the eastern, northern, southern, and western regions of India and from abroad. Differences in allele frequencies of both HP and TF were statistically significant for the population of immigrants from western India and insignificant for others. The alleles HP*2 and TF*C2 show maximum frequencies in the southern population (0.882 and 0.822, respectively) followed by the eastern population (0.862 and 0.807) and the northern population (0.806 and 0.650). In the northern population a third allele, TF*C3, is also detected, with a mean frequency of 0.044. The average heterozygosity (HL) values for HP and TF are 0.273 and 0.361, respectively, and the pooled values for gene diversity parameters for both loci are HT = 0.4294 ± 0.0351, HS = 0.4225 ± 0.0271, and DST = 0.0069 ± 0.0051. The pooled GST value is 0.0153 ± 0.0108. The magnitude of these values indicates genetic similarity among the investigated populations. Our AMOVA results also demonstrate similarity among populations of the same geographic region. However, we note substantial differentiation among different regions (ΦCT = 0.221). The UPGMA dendrogram shows a cluster between the eastern and southern populations, to which the northern population joins. Our results reveal genetic similarity among different immigrant populations, with the western population being the most distant. Therefore the present study on culturally, geographically, and linguistically different endogamous groups of Muslims may have significance in understanding their genetic relationship.

Biochemical Variations in Salt Soluble Fractions of Ultrasonicated Actomyosin Isolated from Broiler Breast-Muscle

Solubility and quality of actomyosin play crucial roles during storage and processing of a particular muscle type. We report here the effect of low intensity (20 kHz) ultrasonication on the solubility and some other biochemical properties of the actomyosin isolated from chicken breast muscle. Although there was an overall enhancement in the solubility of actomyosin during sonication, the major increase of ~61.55% occurred at 0.2M NaCl after 10–12 min of exposure. The distinctive feature of sodium dodecyl sulphate-polyacrylamide gel electrophoretic (SDS-PAGE) profiles of this fraction was the presence of free or non-interacting actin in considerable amount. Even at higher salt concentrations (0.4 and 0.5M NaCl), the protein solubility was almost twice as high as that of the corresponding controls. At low salt concentrations (<0.3 M NaCl or KCl) where the protein solubility was high, Ca2+-, K+(EDTA)- and Mg2+-ATPase activities were low. However, a remarkable increase in each of the enzymatic activities occurred at the salt concentrations higher than 0.3 M. More importantly, the high level of Mg2+-ATPase strongly indicated that actomyosin was reconstituted. The ultraviolet spectra also supported the reconstitution of actomyosin at high salt concentrations. These data suggested that ultrasonication caused conformational changes which modified properties of myosin and actin affecting the intermolecular relationships within actomyosin complex. The shift in the solubility and functional modifications of sonicated actomyosin provide some basis to explain the processing behavior of sonicated poultry meat. In addition, the low frequency sonication may be suggested as a non-destructive method to compare actomyosin extracted from various sources.

Effect of pre-stain methanol washing on the sensitivity of CBBR-250 staining in SDS-polyacrylamide gels

Abstract Coomassie Brilliant Blue R-250 (CBBR-250) is the most frequently used dye stain in protein research. However, relatively poor sensitivity and long application durations are the major limitations of this stain. In the present study, we have investigated the effect of pre-stain methanol washing on the sensitivity of the conventional CBB staining method. Concentrations of methanol ranging from 5 to 30% were prepared in 5% acetic acid. Pre-stain washing of SDS-gels using acetic acid (5%) and methanol were assessed at various timings and temperatures. Our results demonstrate that pre-stain washing by methanol greatly improves the sensitivity of the CBB stain and facilitates the visualization of low molecular weight components present in a complex muscle protein, actomyosin. The outcome of this study also revealed duration of pre-stain washing and temperature as other factors that determine the sensitivity of CBB staining method. Therefore, it is suggested that pre-stain washing with methanol may improve the sensitivity of CBB stain in SDS-PA gels.

Presence of Circulating Lactate Dehydrogenase-Immunoglobulin G (LDH-IgG) Complex in the Sera of Post-Burn Patient

Presence of Circulating Lactate Dehydrogenase-Immunoglobulin G (LDH-IgG) Complex in the Sera of Post-Burn Patient Lactate dehydrogenase (LDH) is an enzyme that belongs to the class oxidoreductase. It actively participates in the inter-conversion of lactate to pyruvate during anaerobic glycolysis in cell cytoplasm. Available literature suggests that there exist only five isoenzymes of LDH which on polyacrylamide gels (Native PAGE) resolve into five bands in normal healthy individuals. However, the presence of extra band in humans is either detected in reproductive tissues or during some pathological state. Here, we report a case in which an extra sixth band of LDH was visualized which forms a complex with IgG in the circulating sera of burn subject. The presence of LDH-IgG complex was confirmed in freshly collected sera using PAGE and immunological approaches. The complex get disappears from 2nd day onwards. The probable reason of the formation of this complex and its relation with the progression of the disease is discussed.