Acceleration of cutaneous wound healing by Lucilia sericata maggots in diabetic Wistar rats.

The study was aimed to evaluate the effectiveness of maggot therapy in healing of cutaneous infected wound in streptozotocin (STZ) induced diabetic Wistar rat. For live maggots, the sterilized eggs of Lucilia sericata were obtained from colonies established in laboratory. Diabetes model was established in 48 male Wister rat by intra-peritoneal injection of STZ at the dose of 60 mg/kg body-weight. Cutaneous wounds exposed with mixed colonies of bacteria like Staphylococcus aureus, E. coli and Pseudomonas aeruginosa were prepared in all rat. The animals equally divided in 4 groups with 12 rats each being presented as treatment group of control, antibiotic, maggot and maggot with antibiotic in combination. All treatments were done once and hold for 24 hours. Wound kinetics and bacterial bio burden were measured at weekly interval to till complete healing. Significant reduction in wound area with maximum contraction was found (>95%) in maggot treated group when compared to antibiotic treated (79%) and control (72%). In maggot as well as maggot and antibiotic in combination group showed early elimination of bacterial bio-burden 7.88±0.03log CFU/ml to 1.12±0.65log CFU/ml and 7.86±0.04) log CFU/ml to 1.54±0.52log CFU/ml respectively in three weeks of time. Early healing indication was also experienced on histomorphological examination of wounded tissue of maggot treated groups by early and better epithelialization, collagenation and neovascularization with complete healing of wound in three weeks in comparison to antibiotic and control respectively. However, the present study did not show any difference in healing of wound with use of maggot alone or in antibiotic combination. Live maggot of Lucilia sericata effectively lower bacterial bioburden and and accelerate healing of infected cutaneous wound in diabetic conditions.

Acceleration of cutaneous wound healing by Lucilia sericata maggots in diabetic Wistar rats

@#The study was aimed to evaluate the effectiveness of maggot therapy in healing of cutaneous infected wound in streptozotocin (STZ) induced diabetic Wistar rat. For live maggots, the sterilized eggs of Lucilia sericata were obtained from colonies established in laboratory. Diabetes model was established in 48 male Wister rat by intra-peritoneal injection of STZ at the dose of 60 mg/kg body-weight. Cutaneous wounds exposed with mixed colonies of bacteria like Staphylococcus aureus, E. coli and Pseudomonas aeruginosa were prepared in all rat. The animals equally divided in 4 groups with 12 rats each being presented as treatment group of control, antibiotic, maggot and maggot with antibiotic in combination. All treatments were done once and hold for 24 hours. Wound kinetics and bacterial bio burden were measured at weekly interval to till complete healing. Significant reduction in wound area with maximum contraction was found (>95%) in maggot treated group when compared to antibiotic treated (79%) and control (72%). In maggot as well as maggot and antibiotic in combination group showed early elimination of bacterial bio-burden 7.88±0.03log CFU/ml to 1.12±0.65log CFU/ml and 7.86±0.04) log CFU/ml to 1.54±0.52log CFU/ml respectively in three weeks of time. Early healing indication was also experienced on histomorphological examination of wounded tissue of maggot treated groups by early and better epithelialization, collagenation and neovascularization with complete healing of wound in three weeks in comparison to antibiotic and control respectively. However, the present study did not show any difference in healing of wound with use of maggot alone or in antibiotic combination. Live maggot of Lucilia sericata effectively lower bacterial bioburden and and accelerate healing of infected cutaneous wound in diabetic conditions.

Variant morphology and arterial supply of diaphragmatic crura with probe patent foramen ovale.

Crura are the posterior pillars of the tendino-muscular partition between thoracic and abdominal cavity. This study highlights bilaterally an unfamiliar morphology of diaphragmatic crura with uncommon origin of inferior phrenic artery and accompanying atrial septal defect in a 63-years-old female cadaver with scoliosis. An attempt is made to portray the clinical implications of these variations. In addition authors have discussed the embryological and genetic basis of these variations. In the recent few decades, surgical correction of scoliosis involving intervention with the crura has gained pace. Knowledge about the normal as well as variant crural morphology is imperative for the surgeons to decide the correct approach during spinal corrective surgeries and for radiologist to prevent any diagnostic pitfall. Familiarity about the variant origin of inferior phrenic artery may prevent complications during treatment of pathological conditions related to inferior phrenic artery. Preoperative awareness about the coexisting atrial septal defect is vital for anesthesiologists promoting them to administer cardioprotective anesthetic drugs. To the best of our knowledge, description of such a combination of variations in anatomical literature is rare and scarcely reported. Awareness of these anatomical variations is relevant for operating surgeons, radiologists, anesthesiologists and anatomists.

Therapeutic use of Lucilia sericata maggot in controlling bacterial bio-burden in Rat wound model.

Delayed wound healing due to extraneous bacterial contamination, antibacterial resistance and other associated factors are of great concern in dealing patients having chronically infected wound. Medicinal properties of certain maggots of Calliphoridae family are known for its effective wound debridement therapy. The objective of the study was to evaluate the wound healing potential of maggots of Lucilia sericata in an experimentally infected cutaneous wound model in Wistar rat. The study was carried out by using male Wistar rats (n=48) by creating excisional wounds and later contaminated with mixed population of gram positive and gram-negative bacteria. Animals were divided randomly in to four groups with 12 individuals each, being denominated as control, antibiotic treated, maggot treated, and antibiotic plus maggot combination treated group. Ten pre-sterilized maggots were applied per centimetre square wound bed for 24 hours. Different wound kinetics in L. sericata maggot treated wounds revealed significant reduction in wound area with maximum contraction, early elimination of bacterial bioburden as compared to group of infected control and group of rats receiving only antibiotic treatment. The histopathological examination of wounded tissue of maggot treated groups showed early and better epithelialization, collagenation and neovascularization with complete healing of wound in two weeks. The maggot effects on healing when used singly or in combination with antibiotic were recorded to be similar. The results of the present study clearly demonstrate that the maggots of L. sericata possesses a definite antibacterial action along with removal of dead tissues and effectively reduced the bacterial bio-burden in infected wound and induced wound healing quickly.

Therapeutic use of Lucilia sericata maggot in controlling bacterial bio-burden in Rat wound model

@#Delayed wound healing due to extraneous bacterial contamination, antibacterial resistance and other associated factors are of great concern in dealing patients having chronically infected wound. Medicinal properties of certain maggots of Calliphoridae family are known for its effective wound debridement therapy. The objective of the study was to evaluate the wound healing potential of maggots of Lucilia sericata in an experimentally infected cutaneous wound model in Wistar rat. The study was carried out by using male Wistar rats (n=48) by creating excisional wounds and later contaminated with mixed population of gram positive and gram-negative bacteria. Animals were divided randomly in to four groups with 12 individuals each, being denominated as control, antibiotic treated, maggot treated, and antibiotic plus maggot combination treated group. Ten pre-sterilized maggots were applied per centimetre square wound bed for 24 hours. Different wound kinetics in L. sericata maggot treated wounds revealed significant reduction in wound area with maximum contraction, early elimination of bacterial bioburden as compared to group of infected control and group of rats receiving only antibiotic treatment. The histopathological examination of wounded tissue of maggot treated groups showed early and better epithelialization, collagenation and neovascularization with complete healing of wound in two weeks. The maggot effects on healing when used singly or in combination with antibiotic were recorded to be similar. The results of the present study clearly demonstrate that the maggots of L. sericata possesses a definite antibacterial action along with removal of dead tissues and effectively reduced the bacterial bio-burden in infected wound and induced wound healing quickly.

Evidence for magnetic Weyl fermions in a correlated metal.

Weyl fermions have been observed as three-dimensional, gapless topological excitations in weakly correlated, inversion-symmetry-breaking semimetals. However, their realization in spontaneously time-reversal-symmetry-breaking phases of strongly correlated materials has so far remained hypothetical. Here, we report experimental evidence for magnetic Weyl fermions in Mn3Sn, a non-collinear antiferromagnet that exhibits a large anomalous Hall effect, even at room temperature. Detailed comparison between angle-resolved photoemission spectroscopy (ARPES) measurements and density functional theory (DFT) calculations reveals significant bandwidth renormalization and damping effects due to the strong correlation among Mn 3d electrons. Magnetotransport measurements provide strong evidence for the chiral anomaly of Weyl fermions-namely, the emergence of positive magnetoconductance only in the presence of parallel electric and magnetic fields. Since weak magnetic fields (approximately 10 mT) are adequate to control the distribution of Weyl points and the large fictitious fields (equivalent to approximately a few hundred T) produced by them in momentum space, our discovery lays the foundation for a new field of science and technology involving the magnetic Weyl excitations of strongly correlated electron systems such as Mn3Sn.

Recombinant 20.8-kDa protein of Mycobacterium avium subsp. paratuberculosis-based sero-diagnosis of paratuberculosis.

Johne's disease or paratuberculosis is a chronic infectious enteric disease of ruminants caused by the intracellular pathogen. The control of the Johne's disease is hampered by lack of specific diagnostic tests. In this study, we have cloned and expressed the N-terminal region of the locus tag Map 1637c encoding 20.8-kDa (r20.8) protein of Mycobacterium avium subsp. paratuberculosis. The recombinant protein r20.8 was expressed in high levels in Escherichia coli. The protein r20.8 was purified by single-step chromatography using Ni-NTA agarose. The protein r20.8 was reacted with anti-r20.8 antibodies as well as cattle sera infected with Map on Western blot. ELISA using well-characterized sera (both positive and negative; n = 60 each) Map-infected and non-infected cattle, respectively, yielded a sensitivity of 73.3% and a specificity of 98.3%. The 20.8 kDa protein expressed in the present study will prove useful as reagent in diagnostic test.

A double-stranded probe coupled with isothermal amplification for qualitative and quantitative detection of avian reovirus.

We applied a probe-based real-time loop-mediated isothermal amplification (Cy5-RTqLAMP) technique targeting the avian reovirus (ARV) S3 gene to develop a rapid, sensitive, and specific method for virus detection and quantification. This test specifically detected the presence of ARV, but not other viruses or bacteria present in clinical or artificially spiked samples, including Newcastle disease virus, infectious bursal disease virus, fowl adenovirus, Marek's disease virus, Escherichia coli, and Salmonella spp. This test can detect ARV in less than one hour with an analytical sensitivity of 10 viral gene copies and 1 fg of total cDNA. The Cy5-RTqLAMP does not yield false positive results and is 100 times more sensitive than conventional PCR. This test was shown to be able to detect the presence of ARV in clinical samples. A similar strategy may be used for detection of other important human and animal viral pathogens.

Interplanar coupling-dependent magnetoresistivity in high-purity layered metals.

The magnetic field-induced changes in the conductivity of metals are the subject of intense interest, both for revealing new phenomena and as a valuable tool for determining their Fermi surface. Here we report a hitherto unobserved magnetoresistive effect in ultra-clean layered metals, namely a negative longitudinal magnetoresistance that is capable of overcoming their very pronounced orbital one. This effect is correlated with the interlayer coupling disappearing for fields applied along the so-called Yamaji angles where the interlayer coupling vanishes. Therefore, it is intrinsically associated with the Fermi points in the field-induced quasi-one-dimensional electronic dispersion, implying that it results from the axial anomaly among these Fermi points. In its original formulation, the anomaly is predicted to violate separate number conservation laws for left- and right-handed chiral (for example, Weyl) fermions. Its observation in PdCoO2, PtCoO2 and Sr2RuO4 suggests that the anomaly affects the transport of clean conductors, in particular near the quantum limit.

Expression and purification of a gene encoding a 9.7 kDa PE protein of Mycobacterium avium subsp. paratuberculosis.

Mycobacterium avium subsp. paratuberculosis (Map) contains PE family antigens which are Proline and glutamic acid rich and may play important role as T-cell antigens. In the present study, the Map 1507 ORF encoding 9.7 kDa PE protein was amplified by polymerase chain reaction and cloned into E. coli vector pQE30 UA. The recombinant plasmid designated as pQ PE was transformed into E. coli M15 cells and induced with IPTG revealed the high level expression of 11.9 kDa His-fusion protein as estimated by migration in 15 % sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Recombinant PE protein was purified by Ni-NTA agarose chromatography. Polyclonal antibodies raised against purified recombinant PE protein reacted with expressed PE protein as well as with Map sonicate. The recombinant PE protein was also recognized by serum from goat with clinical paratuberculosis. The protein elicited significant delayed type hypersensitivity (DTH) skin reaction in mice sensitized with Map. The results indicated that the recombinant PE protein of Map was associated with T-cell response.

Incidence of Haemoproteus columbae in pigeons of Jammu district.

Haemoproteus columbae is the major protozoan infection reported in pigeon and appeared in the erythrocyte of the peripheral circulation. Incidence and parasitaemia of H. columbae in pigeon was studied in different localities of Jammu, India for a period from April to September 2010 using thin blood smear examination. Of the 150 pigeons (wild: 70, domestic: 80), 92 (61.33 %) were found to be infected with H. columbae. Domestic pigeon showed higher incidence rate (74.28 %) than the wild (50 %). Mature and immature gametocyte encircled the erythrocyte nucleus to form a halter shaped appearance were characteristic feature of the parasite. Pseudolynchia canariensis, the vector for H. columbae was also recovered from beneath the feathers of pigeons. No other haemoprotozoan parasite was observed in the blood smear of examined pigeon. Counting of infected erythrocyte revealed higher affection of 1-2 erythrocytes indicative of milder infection in the birds. A long term study within bird population is essential in order to disclose seasonal variation in parasite, vector density and age of infection such as nesting area.

[Increased manganese superoxide dismutase and cyclin B1 expression in carnosine-induced inhibition of glioblastoma cell proliferation].

Carnosine is an endogenous dipeptide with antiproliferative properties. Here we show that carnosine selectively inhibits proliferation of human glioblastoma cells (U-118-MG) compared to breast (MB231) and oral (Cal27 and FaDu) cancer cells. Carnosine-induced inhibition of U-118-MG proliferation is associated with a significant: decrease in cellular reactive oxygen species levels, increase in manganese superoxide dismutase (MnSOD) and increase in cyclin B1 expression resulting in G2-block. We conclude that the antiproliferative property of carnosine is due to its ability to enhance MnSOD and cyclin B1 expression. These results will be of significance to the potential application of carnosine in brain cancer therapy.

Rotenone-induced neurotoxicity in rat brain areas: a study on neuronal and neuronal supportive cells.

The present study was conducted to correlate rotenone-induced neurotoxicity with cellular and molecular modifications in neuronal and neuronal supportive cells in rat brain regions. Rotenone was administered (3, 6 and 12 µg/µl) intranigrally in adult male Sprague-Dawley rats. After the 7th day of rotenone treatment, specific protein markers for neuronal cells - tyrosine hydroxylase (TH), astroglial cells - glial fibrillary acidic protein (GFAP), microglial cells - CD11b/c, and Iba-1 were evaluated by immunoblotting and immunofluorescence in the striatum (STR) and mid brain (MB). Apoptotic cell death was assessed by caspase-3 gene expression. Higher doses of rotenone significantly lowered TH protein levels and elevated Iba-1 levels in MB. All the doses of rotenone significantly increased GFAP and CD11b/c protein in the MB. In STR, rotenone elevated GFAP levels but did not affect TH, CD11b/c and Iba-1 protein levels. Caspase-3 expression was increased significantly by all the doses of rotenone in MB but in STR only by higher doses (6 and 12 µg). It may be suggested that astroglial activation and apoptosis play an important role in rotenone-induced neurotoxicity. MB appeared as more sensitive than STR toward rotenone-induced cell toxicity. The astroglial cells emerged as more susceptible than neuronal and microglial cells to rotenone in STR.

Preferential selection of MnSOD transcripts in proliferating normal and cancer cells.

Manganese superoxide dismutase (MnSOD) is a nuclear encoded and mitochondrial matrix-localized redox enzyme that is known to regulate the cellular redox environment. Cellular redox environment changes during transitions between quiescent and proliferative cycles. Human MnSOD has two poly(A) sites resulting in two transcripts: 1.5 and 4.2 kb. The present study investigates if the 3'-untranslated region (UTR) of MnSOD regulates its expression during transitions between quiescent and proliferating cycles, and in response to radiation. A preferential increase in the levels of the 1.5-kb MnSOD transcript was observed in quiescent cells, whereas the abundance of the longer transcript showed a direct correlation with the percentage of S-phase cells. The log-transformed expression ratio of the longer to the shorter transcript was also higher in proliferating normal and cancer cells. Deletion and reporter assays showed a significant decrease in reporter activity in constructs carrying multiple AU-rich sequence that are present in the 3'-UTR of the longer MnSOD transcript. Overexpression of the MnSOD 3'-UTR representing the longer transcript enhanced endogenous MnSOD mRNA levels, which was associated with an increase in MnSOD protein levels and a decrease in the percentage of S-phase cells. Irradiation increases the mRNA levels of the 1.5-kb MnSOD transcript, which was consistent with a significant increase in the reporter activity of the construct carrying the 3'-UTR of the shorter transcript. We conclude that the 3'-UTR of MnSOD regulates MnSOD expression in response to different growth states and radiation.

Coexpression of PPE 34.9 Antigen of Mycobacterium avium subsp. Paratuberculosis with Murine Interferon Gamma in HeLa Cell Line and Study of Their Immunogenicity in Murine Model.

Mycobacterium avium subsp. paratuberculosis (Map) is the causative agent of johne's disease whose immunopathology mainly depends on cell mediated immuneresponse. Genome sequencing revealed various PPE (Proline-Proline-Glutamic acid) protein family of Map which are immunologically importance candidate genes In present study we have developed a bicistrionic construct pIR PPE/IFN containing a 34.9 kDa PPE protein (PPE 34.9) of Map along with a cytokine gene encoding murine gamma Interferon gene (IFNγ) and a monocistrionic construct pIR PPE using a mammalian vector system pIRES 6.1. The construct were transfected in HeLa cell line and expression were studied by Western blot as well as Immunefluroscent assay using recombinant sera. Further we have compared the immunereactivity of these two constructs in murine model by means of DTH study, LTT, NO assay and ELISA. DTH response was higher in pIR PPE/IFN than pIR PPE group of mice, similar finding also observed in case of LTT and NO production assay . ELISA titer of the pIR PPE/IFN was less than that with PPE only. These preliminary finding can revealed a CMI response of this PPE protein of Map and IFNγ having synergistic effect on this PPE protein to elicit a T cell based immunity in mice.

Lung cancer statistics in Luxembourg from 1981 to 2008.

Lung cancer is the leading cause of cancer-related death in the world and in Luxembourg. As a part of "The health science initiative focused on personalized medicine", Luxembourg aims to participate by developing diagnostics to improve the detection and treatment of lung cancer. In line with this objective, this study made a review of evolution of lung cancer in Luxembourg from 1981 to 2008 and compared this statistics to the situation in the Nordic countries, Europe in general and the World. Incidence data of the national morphological tumour registry and mortality data of the service of statistics of the national ministry of health is depicted in charts with trend lines, in the framework of a statistical evaluation of relevant parameters. The data indicate that while male lung cancer incidence decreased in Luxembourg, the incidence in women and its mortality have doubled over the 28-year span considered. Notwithstanding this increase, the female lung cancer incidence and mortality remain low compared to the Nordic countries and Europe. Interestingly, the study also potentially suggests that the lung cancer pattern follows the smoking pattern in incidence and mortality.

Expression of a Gene Encoding 34.9 kDa PPE Antigen of Mycobacterium avium subsp. paratuberculosis in E. coli.

Mycobacterium avium subsp. paratuberculosis (Map) contains PPE family antigens which are Proline and glutamic acid rich and may play important role as T cell antigens. Hence the identification and generation of antigens are necessary for immunological characterization. In the present study, the epitopic region of a unique PPE gene encoding 34.9 kDa protein from Map was amplified by polymerase chain reaction. The gene was cloned into Escherichia coli vector pQE30 UA. The recombinant plasmid designated as pQPPE was transformed into E. coli M15 and induced with IPTG revealed the high level expression of 37.1 kDa His-fusion protein (34.9 kDa PPE and 2.2 kDa His-tag), which was confirmed by immunoblotting. Recombinant PPE protein was then purified by Ni-NTA agarose chromatography. The polyclonal antiserum raised against purified recombinant PPE protein reacted with expressed 37.1 kDa His-fusion protein as well as with Map sonicate. The protein elicited significant delayed type hypersensitivity (DTH) skin reaction in mice sensitized with Map. The results indicated that the recombinant PPE protein of Map was associated with cellular immune response.

Crossed cerebellar atrophy. A case report.

Crossed cerebellar atrophy is an uncommon sequelae of contralateral cerebral hemispheric lesion. This condition has been described in patients with supratentorial infarct, tumor, arteriovenous malformation and hemorrhage. Crossed cerebellar atrophy possibly occurs due to interruption of the corticopontocerebellar pathway at supratentorial level.