kDa Hsp Research Articles

Esculetin Attenuates Doxorubicin-Induced Kidney Damage By Reducing Heat Shock Proteins Expression Levels

The effectiveness of Doxorobucin (DOX), a commonly used anti-cancer and immunosuppressive medication, is hindered by its potential for organ toxicity. Prolonged use of DOX is associated with severe hepatocellular toxicity. This study reveals fresh insights into the therapeutic impact of esculetin (E) on DOX-induced kidney cell damage. Esculetin demonstrates its remedial effects by modulating heat shock protein signaling pathways. In our research, we explored the impact of DOX and E on the expression of the 70 kDa HSP gene family, including Hspa1a, Hspa4, and Hspa5, which are small stress proteins in Rattus norvegicus. The study involved the assignment of five different groups (Control, DOX, E50 mg/kg, E100 mg/kg, DOX+ E50 mg/kg, and DOX+ E100 mg/kg). Subsequently, kidney tissues were collected from rats, and cDNA libraries were generated at the conclusion of the application process. The Real-Time PCR method was employed using these libraries to detect HSP70 genes. Analyses conducted on Hspa1a, Hspa4 and Hspa5 expression revealed a statistically significant increase in the DOX group compared to the control group. Additionally, the combination of DOX and esculetin demonstrates a reduction in the increase caused by DOX alone. The study suggests that esculetin could serve as a potential protective agent for shielding kidney tissue from oxidative damage and apoptosis.

Assessment of the therapeutic potential of Hsp70 activator against prion diseases using in vitro and in vivo models.

Prion diseases are deadly neurodegenerative disorders in both animals and humans, causing the destruction of neural tissue and inducing behavioral manifestations. Heat shock proteins (Hsps), act as molecular chaperones by supporting the appropriate folding of proteins and eliminating the misfolded proteins as well as playing a vital role in cell signaling transduction, cell cycle, and apoptosis control. SW02 is a potent activator of Hsp 70kDa (Hsp70). In the current study, the protective effects of SW02 against prion protein 106-126 (PrP106-126)-induced neurotoxicity in human neuroblastoma cells (SH-SY5Y) were investigated. In addition, the therapeutic effects of SW02 in ME7 scrapie-infected mice were evaluated. The results showed that SW02 treatment significantly increased Hsp70 mRNA expression levels and Hsp70 ATPase activity (p < 0.01). SW02 also significantly inhibited cytotoxicity and apoptosis induced by PrP106-126 (p < 0.01) and promoted neurite extension. In vivo, intraperitoneal administration of SW02 did not show a statistically significant difference in survival time (p = 0.16); however, the SW02-treated group exhibited a longer survival time of 223.6 ± 6.0days compared with the untreated control group survival time of 217.6 ± 5.4days. In addition, SW02 reduced the PrPSc accumulation in ME7 scrapie-infected mice at 5months post-injection (p < 0.05). A significant difference was not observed in GFAP expression, an astrocyte marker, between the treated and untreated groups. In conclusion, the potential therapeutic role of the Hsp70 activator SW02 was determined in the present study and may be a novel and effective drug to mitigate the pathologies of prion diseases and other neurodegenerative diseases. Further studies using a combination of two pharmacological activators of Hsp70 are required to maximize the effectiveness of each intervention.

Effects of heat shock protein 70 knockdown on the tolerance of the brine shrimp Artemia franciscana to aquaculture-related stressors: Implications for aquatic animal health and production

Temperature, pH, salinity and ammonia stress adversely affect the performance and well-being of aquatic organisms, especially those used in aquaculture, a major food production sector. Management strategy to mitigate stress caused by abiotic factors in farmed aquatic organisms is scarce but is highly warranted considering the negative impact of these factors on the welfare and economic production of the farmed animals. The induction of heat shock proteins, mainly 70 kDa Hsp (Hsp70), in farmed fish and shellfish, including the live feed brine shrimp Artemia franciscana, by exposure to a non-lethal heat shock (NLHS) of 37 °C for 30 min followed by 6 h recovery period caused a significant improvement in the tolerance of the animals towards subsequent challenge with an abiotic stress. Although the protective effect of NLHS was associated with enhanced Hsp70 accumulation in these species, the induction of other members of Hsp and their collective involvement in the stress tolerance could not be ignored. This triggered an argument as to whether only Hsp70 or its combination with other Hsps was responsible for inducing stress tolerance phenotype in these species. In this study, we employed RNAi technology in the biological model organism Artemia to gain insight into the functional role of Hsp70 in stress tolerance. The results showed that upon exposure to a NLHS, the wild-type Artemia nauplii capable of producing Hsp70 exhibited significantly higher survival than the unexposed control in all the abiotic stress tolerance tests performed in this study. However, the Hsp70-deficient mutant nauplii did not exhibit increased stress-tolerant phenotypes as the survival percentage of the nauplii decreased by 29, 19, and 15%, respectively, upon challenge with a median lethal concentration of salinity, pH, and ammonia. These results suggest that Hsp70 was involved in inducing tolerance against stress caused by the abiotic factors, and hence can be a potential target for developing strategies to protect economically important aquaculture species from environmental disturbances.

Southern introgression increases adaptive immune gene variability in northern range margin populations of Fire-bellied toad.

Northern range margin populations of the European fire‐bellied toad (Bombina bombina) have rapidly declined during recent decades. Extensive agricultural land use has fragmented the landscape, leading to habitat disruption and loss, as well as eutrophication of ponds. In Northern Germany (Schleswig‐Holstein) and Southern Sweden (Skåne), this population decline resulted in decreased gene flow from surrounding populations, low genetic diversity, and a putative reduction in adaptive potential, leaving populations vulnerable to future environmental and climatic changes. Previous studies using mitochondrial control region and nuclear transcriptome‐wide SNP data detected introgressive hybridization in multiple northern B. bombina populations after unreported release of toads from Austria. Here, we determine the impact of this introgression by comparing the body conditions (proxy for fitness) of introgressed and nonintrogressed populations and the genetic consequences in two candidate genes for putative local adaptation (the MHC II gene as part of the adaptive immune system and the stress response gene HSP70 kDa). We detected regional differences in body condition and observed significantly elevated levels of within individual MHC allele counts in introgressed Swedish populations, associated with a tendency toward higher body weight, relative to regional nonintrogressed populations. These differences were not observed among introgressed and nonintrogressed German populations. Genetic diversity in both MHC and HSP was generally lower in northern than Austrian populations. Our study sheds light on the potential benefits of translocations of more distantly related conspecifics as a means to increase adaptive genetic variability and fitness of genetically depauperate range margin populations without distortion of local adaptation.

Exogenous cysteine alleviates chromium stress via reducing its uptake and regulating proteome in roots of Brassica napus L. seedlings

Sulfur compounds are known to play crucial roles in plants facilitating tolerance to heavy metal stress. Previous studies have shown the protective role of exogenous cysteine (Cys) to improve chromium (Cr) tolerance. In the present study, we aimed to evaluate the capability of Cys (0.5 mM) to reduce chromium toxicity (100 μM) in rapeseed by physiological and proteomic analyses. Seedlings treated with Cr showed reduced growth and activities of antioxidant enzymes, and increased malondialdehyde (MDA) content. Exogenous application of Cys reduced Cr accumulation and MDA content, restored catalase (CAT) and peroxidase (POD) activities, and caused significant enhancements in seedling growth. 2-DE gel electrophoresis and mass spectrometry were performed to investigate changes in the root proteome in response to Cys and Cr. The Cr-induced down-regulations of glycine-rich RNA-binding protein, lactoylglutathione lyase, fructose-bisphosphate aldolase, and glutamine synthetase were restored with Cys. Additionally, Cys removed the Cr-induced up-regulation of some proteins including pathogenesis-related protein, major latex protein (MLP) like proteins, 22.0 kDa HSP, triosephosphate isomerase, glyceraldehyde-3-phosphate dehydrogenase, glutamine amidotransferase, cysteine synthase, and L-lactate dehydrogenase. Taken together, proteomic analysis revealed that exogenous Cys helps plants to overcome Cr stress by modulating energy metabolism, amino acid metabolism, and defense mechanism.

In Silico Analysis of HSP70 Gene Family in Bovine Genome

Heat shock proteins (HSPs), members of molecular chaperones families fulfill essential roles under normal conditions and provide protection and adaptation during and after stress. Among different HSPs, HSP70 kDa family of proteins is most abundant and well-studied in human and mouse but has not yet been characterized in bovines. In silico analysis was performed to characterize members of HSP70 gene family in bovine genome and a total of 17 genes of bovine HSP70 gene family were identified. The members of HSP70 family were distributed over 12 chromosomes with gene size ranging from 1911 (HSPA2) to 54,017bp (HSPA4). Five genes were intronless, while rest of 12 genes were multiexonic. Phylogenetic analysis of HSP70 gene family distinguished them into eight major evolutionary groups wherein members of group 1 were most divergent and quite dissimilar than from rest of the HSP70 sequences. Domain structure of all bovine HSP70 genes was conserved and three signature patterns HSP70_1, HSP70_2, and HSP70_3 were identified. HSPA8, HSP9, and HSPA1A showed comparatively higher expression in majority of tissues. Like humans, bovine HSP70 family was characterized by remarkable evolutionary diversity. The analysis also suggested resemblance of bovine HSP70 family to that of human compared to mouse. Overall, the study indicates the presence of diversity for structure, function, localization, and expression in the bovine HSP70 family chaperons which could form the basis to understand thermotolerance/adaptive changes in the bovines.

A label-free shotgun proteomics analysis of macadamia nut.

In this study, we present a systematic proteomic overview of macadamia nut using a label-free shotgun proteomic approach. We identified 947 proteins in 723 clusters and gene ontology analysis revealed proteins across 46 functional categories including carbohydrate metabolism (10%), protein metabolic processes (5%), amino acid metabolism (4%), transport (4%), stress response (3%), lipid metabolism (3%), protein folding (3%) and defense response (1.4%). The defense response proteins accounted for 24% of the total peptide abundance. The vicilin-like macadamia antimicrobial peptides 2-3 (MiAMP2) was the most abundant protein, followed by glyceraldehyde-3-phosphate dehydrogenase 3, 11S legumin-like protein, 2-phospho-D-glycerate hydrolase and heat shock 70kDa protein among others. The cascading of amino acid and carbohydrate metabolic pathways in macadamia nut were constructed against reference maps from KEGG and proposed for the first time. Results were also indicative of useful protein candidates with possible allergenic potential and cross-reactivity in macadamia nut. The in-silico analysis revealed homology and linear epitope similarities to known allergens such as conglutin β allergen from lupin, Jug r2 vicilin allergens from walnut, Ara h3 11S globulin from peanut, small rubber particle protein Hev b3, hevein, enolase 2, HSP 70kDa Cla h4, Der f28 allergen, and methylglyoxalases. Label-free shotgun proteomics reveal valuable insights into the genetic and biological makeup of macadamia nut proteome and provide guidance on protein candidates with allergenic potential for further immunological investigation. Data are available via ProteomeXchange with identifier PXD015364.

Influence Of Total Body Controlled Hyperthermic Perfusion on Levels of Heat Shock Proteins 70 kDa in Patients with Active Infective Endocarditis

Development of infective endocarditis initiates a complex immunological response of the organism changing over time: the prevalence of pro-and anti-inflammatory mechanisms at the beginning is replaced by immunosuppression. A significant role in antigen presentation, cross-presentation, activation of macrophages and lymphocytes play the heat shock proteins 70 kDa (HSP).&#x0D; The aim of the study was to investigate the relationship between systemic inflammatory response and HSP 70 kDa values in patients operated on under the total body controlled hyper-thermic perfusion (TBCHP).&#x0D; Materials and methods.The study included 18 patients with active infective valve endocarditis operated from 01.01.2016 to 01.01.2017 with the use of TBCHP. Assessment of the relationship between clinical characteristics of patients with infective endocarditis and the level of heat shock protein 70 kDa was performed preoperatively, 2 hours after TBCHP, 8 hours after TBCHP, 20 hours after TBCHP.&#x0D; Results.The presence of high levels of heat shock proteins 70 kDa (5.6 ± 3.3 ng/ml) pre-operatively indicated the depression of cellular and humoral immunity. Reduction in total blood plasma protein level as an indicator of catabolism was also combined with the registration of higher values of HSP70 kDa. After 2 hours after TBCHP a wide range of HSP 70 kDa levels was observed. A blood test performed after 2 hours after TBCHP showed HSP70 kDa – average level of 6.06 ± 3.8 ng/ml. Values range between 0.96 to 9.08 ng/ml. According to these data it was possible to distinguish two subgroups of patients who differ significantly in the average values of HSP70 kDa after TBCHP. There was a general tendency to reduce the values of HSP 70kDa from the initial preoperative level to the end of 20 hour after TBCHP – 5.6 ± 3.3 ng/ml and 1.6 ± 0.68 ng/ml correspondingly (p = 0.116).&#x0D; Conclusion. Taking into account the previously identified preoperative relationships between HSP70 kDa level and clinical parameters of patients we may suggest that reduction of HSP 70 kDa corresponds with elimination of signs of the syndrome of persistent inflammation, immunosuppression and catabolism.

The Autoantigenic Proinsulin B-Chain Peptide B11-23 Synergises with the 70 kDa Heat Shock Protein DnaK in Macrophage Stimulation.

Background Heat shock proteins (Hsp) act as intracellular chaperones and in addition are used as adjuvant in vaccines of peptides complexed with recombinant Hsp. By interacting with autologous peptides, Hsp may promote the induction of autoimmune reactivity. Objective Here, we analysed whether the effect of Hsp on macrophages is modulated by insulin peptides known to interact with Hsp. Results Combinations of the 70 kDa Hsp DnaK with peptide B11-23 from the core region of the proinsulin B-chain induced the release of the inflammatory mediators interleukin-6, tumor necrosis factor α, and interleukin-1β from cells of human and murine macrophage lines. In parallel, there was high-affinity binding of B11-23 to DnaK. DnaK mixed with peptides from other regions of the insulin molecule did not stimulate cytokine secretion. DnaK alone induced little cytokine production, and peptides alone induced none. Conclusion The macrophage-stimulating potential of Hsp70 family proteins when combined with the proinsulin B-chain peptide B11-23 may contribute to the immunodominance of this peptide in the development of beta cell-directed autoimmunity in type 1 diabetes.

Assessment of some genetic attributes in wheat (Triticum aestivum L.) using gene-specific molecular markers

Twenty-four wheat genotypes were characterized for the presence of three stress related genetic attributes using gene-specific molecular markers. Polymerase chain reaction (PCR) products of approximately 110 bp of 1RS rye chromosome fragment from 16 genotypes were obtained using specific primer pairs indicating the presence of translocation in these lines. The same genotypes were screened for the presence of dwarfing genes where 19 genotypes showed the presence of either the Rht-B1b or Rht-D1b allele having a semi-dwarf phenotype. Two of the genotypes (Kheri and Sufi) had wild type alleles in both the loci and two other genotypes showed the presence of double dwarf alleles (Rht-B1b + Rht-D1b). A 16.9 kDa HSP gene was characterized by validating a single-nucleotide polymorphism (SNP) linked with thermo tolerance in wheat. Thirteen of the 24 genotypes, which failed to amplify the specific PCR product due to the presence of an SNP, are expected to show tolerance for heat stress. Among the genotypes, Sonora-64, Balaka, Barkat, Aghrani and BARI Gom-24 (Prodip) tested positive for all the three markers evaluated (rye translocation, dwarfing genes and heat tolerance). These genotypes could be used to improve various stress tolerance attributes in wheat for future breeding programs.

The Reaction of Placental GRP78 Protein with Sera from Women with Multiple Sclerosis.

Multiple Sclerosis (MS) with four different types is one of the well studied autoimmune diseases of the central nervous system. Generally, two-thirds of MS patients are females who are at risk of pregnancy-related complications. Inappropriate responses of mother's immune system, such as antibody production against placental proteins, may lead to pregnancy-related disorders. The association between pregnancy complications and some autoantibodies including anti-phospholipid and anti-angiotensin II type-1 receptor antibodies are clear examples in this regard. To investigate the probable placental antigens that might be targeted by the antibodies in the sera of MS patients. Total placental proteins were extracted from normal fresh placentas and were separated using two-dimensional gel electrophoresis (2-DE) technique. The separated proteins were transferred onto a Polyvinylidene Fluoride (PVDF) membrane and blotted with the pooled sera of MS women or healthy controls (20 individuals in each group). The differentially blotted spot was identified by mass spectrometry and confirmed by western blot technique. The results indicated that the women afflicted with MS had an antibody against placental HSP 70kDa protein 5 (GRP78). In the present study, a new placental autoantigen candidate, which was targeted by antibody present in MS women sera, was found. The clinical importance of this finding regarding pregnancy complications in MS patients should be investigated by further experiments.

Functional characterization of the transcriptional regulatory elements of three highly expressed constitutive genes in the jewel wasp, Nasonia vitripennis.

The jewel wasp, Nasonia vitripennis Ashmead (Hymenoptera: Pteromalidae), is an easily reared parasitoid that is providing an ever increasingly malleable model for examining the biology and genetics of Hymenoptera. Utilizing genomic and transcriptome resources, 5' upstream transcriptional regulatory sequences (TREs) from three highly expressed genes were identified and cloned. Criteria for TRE selection included the presence of an adjacent gene 5' of the translation initiation site. One gene was methylated whereas the other two were nonmethylated. Each TRE, heat-shock protein 70 (hsp70), activator of 90 kDa hsp ATPase protein 1 (hsp90A), and lipid storage droplet surface-binding protein 1 (lsdp) was linked with enhanced green fluorescent protein (EGFP) coding sequence and cloned into both pDP9e somatic and piggyBac germline transformation vectors. EGFP expression patterns under control of each TRE were compared with patterns of DsRed fluorescence produced from the transformation vector cassette. Functional activity of each TRE was observed in cultured Spodoptera frugiperda 9 (Sf9) cells and Drosophila melanogaster as well as in N.vitripennis embryos demonstrating that all three sequences had functional transcriptional regulatory activity in three different insect orders. Identification and functional characterization of these three TREs will provide critical and necessary resources for further genetic analyses of N. vitripennis, Hymenoptera and other insects.

Nanogapped impedimetric immunosensor for the detection of 16 kDa heat shock protein against Mycobacterium tuberculosis

The 16 kDa heat shock protein (16 kDa HSP) against Mycobacterium tuberculosis (MT), expressed during the growth phase of MT, is a potential target in diagnostic tests for tuberculosis (TB). We describe here a method for impedimetric determination of the antigen by using a nanogapped dielectric surface consisting of a silver support coated with a thin finger-shaped coating made from zinc oxide and gold and patterned through a lift-off process. The electrode was characterized by scanning electron microscopy, field emission scanning electron microscopy, atomic force microscopy, and energy-dispersive X-ray spectroscopy. Surface chemical functionalization and immobilization of antibody against the 16 kDa HSP was evidenced by FTIR. In order to improve the detection limit, the antigen was conjugated to 10 nm gold nanoparticles. The resulting biosensor is capable of detecting the 16 kDa HSP in concentrations as low as 100 fM. The method covers a wide analytical range that extends from 100 fM to 1 nM.

Molecular cloning, characterization and expression analysis of a heat shock protein 10 (Hsp10) from Pennisetum glaucum (L.), a C4 cereal plant from the semi-arid tropics.

Heat shock proteins (Hsp10) belong to the ubiquitous family of heat-shock molecular chaperones found in the organelles of both prokaryotes and eukaryotes. Chaperonins assist the folding of nascent and stress-destabilized proteins. A cDNA clone encoding a 10 kDa Hsp was isolated from pearl millet, Pennisetum glaucum (L.) by screening a heat stress cDNA library. The fulllength PgHsp10 cDNA consisted of 297 bp open reading frame (ORF) encoding a 98 amino acid polypeptide with a predicted molecular mass of 10.61 kDa and an estimated isoelectric point (pI) of 7.95. PgHsp10 shares 70-98 % sequence identity with other plant homologs. Phylogenetic analysis revealed that PgHsp10 is evolutionarily close to the maize Hsp10 homolog. The predicted 3D model confirmed a conserved eight-stranded ß-barrel with active site between the ß-barrel comprising of eight-strands, with conserved domain VLLPEYGG sandwiched between two ß-sheets. The gene consisted of 3 exons and 2 introns, while the position and phasing of these introns were conserved similar to other plant Hsp10 family genes. In silico analysis of the promoter region of PgHsp10 presented several distinct set of cis-elements and transcription factor binding sites. Quantitative RT-PCR analysis showed that PgHsp10 gene was differentially expressed in response to abiotic stresses with the highest level of expression under heat stress conditions. Results of this study provide useful information regarding the role of chaperonins in stress regulation and generated leads for further elucidation of their function in plant stress tolerance.

Pretreatment Effect of Running Exercise on HSP70and DOX-Induced Cardiotoxicity

The purpose of this study was to determine pretreatment effects of moderate-term endurance training before the various dosages (10 and 20 mg.kg(-1)) of DOX on a heat shock protein (HSP70 kda) and cardiotoxicity in heart tissue. Forty-eight male rats were randomly assigned to nontraining (NT) and training (T) groups and three subgroups; DOX10 mg.kg(-1) and DOX20 mg.kg(-1) and saline treatment. The training program included treadmill running between 25-39 min/day and 15-17 m/min, 5 days/wk for 3 wk. DOX administration, in particularly with 20 mg.kg(-1), caused up-regulation of oxidants and cardiac damage (MDA, CK, CPK-MB and CK/ CPK-MB) and down-regulation of cardioprotection (HSP70, SOD) markers, as compared to NT+saline group. Pretreatment effect of treadmill running endurance exercise in the presence of DOX with 10 mg.kg(-1) caused a significant increase in HSP70, SOD and a significant decrease in MDA and insignificant decrease in CK, CPK-MB and CK/CPK-MB, in comparisonT+DOX10 with NT+DOX10 group. However, there was no significant difference between T+DOX10 mg.kg(-1) and T+DOX20 mg.kg(-1) in the aforesaid markers. Dox-induced cardiotoxicity is related to oxidative stress. Our study suggests that pretreatment with endurance exercise may be considered as a potentially useful strategy to improve myocardial tolerance against single dose DOX-induced oxidative damage.

Molecular mimicry between HSP 65 of Mycobacterium leprae and cytokeratin 10 of the host keratin; Role in pathogenesis of leprosy

Mycobacteria are known to induce autoimmune response in the host. Anti-host keratrin antibodies (AkAbs) might be responsible for the autoimmune phenomena in leprosy patients as majority of leprosy lesions are manifested in the skin and occurrence of keratosis is not an uncommon feature. The aim of this study was to find out the level of AkAbs in leprosy patients across the spectrum and to explore its correlation with the clinical manifestation of the disease. Further, mimicking epitopes of keratin and Mycobacterium leprae components were characterized. We screened 140 leprosy patients (27 BT, 28 BL, 41 LL, 25 T1R, 19 ENL), 74 healthy controls (HC) and 3 psoriasis patients as positive control. Highest AkAbs level was observed in the psoriasis patients followed by T1R, LL, BL, ENL, TT/BT. AkAbs level was significantly (p<0.05) higher in all the groups of leprosy patients except TT/BT in comparison to HC. Significant positive correlation was found between number of lesions and level of AkAbs in leprosy patients. Highest lympho-proliferation for keratin protein was observed in T1R, followed by BL/LL, TT/BT, ENL. Lympho-proliferation was significantly (p<0.05) higher in all groups of leprosy patients except ENL in comparison to HC.Interestingly, it was noted that hyperimmunization of inbred strains of female BALB/c mice and rabbit with M. leprae soluble antigen (MLSA) induce higher level of AkAbs. The percentage of FoxP3+ expressing Treg cells (total CD4+CD25+FoxP3+ andCD4+CD25+hiFoxP3+) in splenocytes and lymph nodes of hyperimmunized mice were declined in comparison to control mice. Further, it was found that this autoimmune response can be adoptively transferred in naïve mice by splenocytes and lymph node cells as well as T cells. Comparative molecular characterization between keratin and MLSA noted a cross-reactivity/similarity between these two antigens. The cross-reactive protein of keratin was found to be in molecular weight range ≈74–51kDa and at pI 4.5 while the cross-reactive protein of MLSA was found to be in molecular weight ≈65kDa and at pI 4–4.5. Cross-reactive protein of keratin and MLSA was identified and characterized by MALDI-TOF/TOF analysis and Mascot software. It was found that the keratin (host protein) which reacted with anti-M. leprae sera is cytokeratin-10 and MLSA which reacted with anti-keratin sera is heat shock protein 65 (HSP 65). Seven B-cell epitopes of cytokeratin-10 and HSP 65 was found to be similar by multiple sequence alignment using ClustalW server and out of which 6 B-cell epitopes were found to be on the surface of HSP 65.In conclusion, our study provides evidence for the existence of molecular mimicry between cytokeratin-10 of keratin (host protein) and 65kDa HSP (groEL2) of M. leprae. Presence of heightened CMI response of leprosy patients to keratin and positive correlation of AkAbs level with number of lesions of leprosy patients showed the clinical evidence for its role in the pathogenesis in leprosy.

A new cell line from Spodoptera exigua (Lepidoptera: Noctuidae) and its differentially expressed genes

AbstractA new cell line, designated IOZCAS‐Spex XI, was established from the pupal ovaries of Spodoptera exigua (Lepidoptera: Noctuidae) in TNM‐FH medium containing 10% foetal bovine serum. The spherical cells were predominant among the various cell types. The population‐doubling time during the logarithmic phase of growth was 81.7 h. It was confirmed that the cell line originated from S. exigua by DAF‐PCR technique. Analysis of susceptibility to baculovirus showed that the new cell line was susceptible to S. exigua nucleopolyhedrovirus (SeNPV), Autographa californica multiple NPV (AcMNPV) and slightly susceptible to S. litura NPV (SpltNPV), while not permissive to Helicoverpa armigera NPV and Hyphantria cunea NPV (HcNPV). Real‐Time PCR analysis was carried out to compare some differentially expressed genes between the cell line and the primary culture. The result showed that marked significant differences were observed in the expression of the genes of SUMO‐1 activating enzyme, BCCIP‐like protein, 10 kDa HSP, CypA, receptor for activated PKC, PDI‐like protein ERp57, ALDH, DEAD box ATP‐dependent RNA helicase‐like protein (P &lt; 0.01), while a significant difference was obtained in the expression of GST gene between the cell line and the primary culture (P &lt; 0.05).

Proteomic study identified HSP 70kDa protein 1A as a possible therapeutic target, in combination with histone deacetylase inhibitors, for lymphoid neoplasms

Histone deacetylase inhibitors (HDACi) demonstrate possible anticancer activities in various malignancies including lymphoid neoplasms. However, the anticancer effects of HDACi are often limited, and combination therapy with other drugs has been undertaken to improve the outcome of patients. Here we conducted proteomic investigation of 33 lymphoid cell lines to identify novel therapeutic targets for enhancing the effects of HDACi. Using the proteomic data in our published 2D-DIGE database, we examined the proteins associated with resistance to valproic acid (VPA). The lymphoid neoplasm cell lines in the database were grouped according to their sensitivity to VPA treatment. A comparative proteomic study of the cell line groups resulted in the identification of 10 protein spots, whose intensity was associated with chemosensitivity. Among the identified proteins, HSPA1A showed higher expression in cell lines with resistance to VPA, and the results were validated by Western blotting. In vitro experiments demonstrated that treatment with KNK-437, an inhibitor of HSPA1A, enhanced the cytotoxic effects of VPA, as well as vorinostat, in the lymphoid neoplasm cell line. Treatment with KNK-437 facilitated the apoptotic effects of VPA. In conclusion, we identified HSPA1A as a possible therapeutic target, in combination with HDACi, for lymphoid neoplasms.

Disseminated Mycobacterium abscessus Infection and Showerheads, Taiwan

Disseminated Mycobacterium abscessus Infection and Showerheads, Taiwan

Mycobacterium mageritensePulmonary Disease in Patient with Compromised Immune System

<i>Mycobacterium mageritense</i>Pulmonary Disease in Patient with Compromised Immune System