Different roles of heat shock proteins (70 kDa) during abiotic stresses in barley (Hordeum vulgare) genotypes

In this work, the involvement of heat shock proteins (HSP70) in barley (Hordeum vulgare) has been studied in response to drought and salinity. Thus, 3 barley genotypes usually cultivated and/or selected in Italy, 3 Middle East/North Africa landraces and genotypes and 1 improved genotype from ICARDA have been studied to identify those varieties showing the best stress response. Preliminarily, a bioinformatic characterization of the HSP70s protein family in barley has been made by using annotated Arabidopsis protein sequences. This study identified 20 putative HSP70s orthologs in the barley genome. The construction of un-rooted phylogenetic trees showed the partition into four main branches, and multiple subcellular localizations. The enhanced HSP70s presence upon salt and drought stress was investigated by both immunoblotting and expression analyses. It is worth noting the Northern Africa landraces showed peculiar tolerance behavior versus drought and salt stresses. The drought and salinity conditions indicated the involvement of specific HSP70s to counteract abiotic stress. Particularly, the expression of cytosolic MLOC_67581, mitochondrial MLOC_50972, and encoding for HSP70 isoforms showed different expressions and occurrence upon stress. Therefore, genotypes originated in the semi-arid area of the Mediterranean area can represent an important genetic source for the improvement of commonly cultivated high-yielding varieties

Physiological and molecular osmotic stress responses in three durum wheat (Triticum turgidum ssp durum) genotypes

This study aims to investigate the activities and expression of enzymes of primary metabolism and relate these data with the growth performance of three dierent durum wheat genotypes (Maali; YT13; and ON66) under osmotic stress. Growth traits—including plant height, dry weight (DW) and relative water content (RWC)—were measured to classify genotypes depending on their tolerance to stress. Several enzymes were investigated: Ascorbate peroxidase (APX), Glutamine Synthetase (GS), Glutamine dehydrogenase (GDH), Glutamate synthase (GOGAT), Glucose 6-phosphate dehydrogenase (G6PDH), and Phosphoenolpyruvate Carboxylase (PEPC). The expression of the cytosolic and plastidic glutamine synthetase (TaGS1 and TaGS2), high anity nitrate transporters (TaNRT2.3) and Glutamate dehydrogenase (TaGDH) were also detected by qRT-PCR. The results indicated dierent growth performances among genotypes, indicating Maali and YT13 as tolerant genotypes and ON66 as a drought-susceptible variety. Data showed a decrease in PEPC and increase in APX activities under osmotic stress; a slight decrease in GS activity was observed, together with an increase in G6PDH in all genotypes; GS and NRT2 expressions changed in a similar pattern in the dierent genotypes. Interestingly, Maali and YT13 showed higher transcript abundance for GDH under stress compared to ON66, suggesting the implication of GDH in protective phenomena upon osmotic stress

Clinical Significance of Epigenetic Inactivation of hMLH1 and BRCA1 in Tunisian Patients with Invasive Breast Carcinoma

Aberrant hypermethylation of gene promoter regions is one of the mechanisms for inactivation of tumour suppressor genes in many human cancers including breast carcinoma. In the current study, we aimed to assess by MSP, the methylation pattern of two cancer-related genes involved in DNA repair: hMLH1 (mutL homolog 1, colon cancer, nonpolyposis type 2 (E. coli) and BRCA1 (breast cancer 1, early onset) in 78 primary breast cancers from Tunisian patients. The methylation frequencies were 24.36% for hMLH1 and 46% for BRCA1. BRCA1 methylation correlated with age at diagnosis (P = .015) and 5-years disease free survival (P = .016) while hMLH1 methylation was more frequent in larger tumors (P = .002) and in presence of distant metastasis (P = .004). Furthermore, methylation of hMLH1 significantly correlated with high level of P53 expression (P = .006) and with overall survival (P = .015) suggesting that silencing of hMLH1 through aberrant promoter methylation could be used as a poor prognosis indicator in breast cancer

Salt Stress Induces Differentiated Nitrogen Uptake and Antioxidant Responses in Two Contrasting Barley Landraces from MENA Region

The interaction between salinity and nitrogen metabolism has been investigated in two barley landraces, one tolerant (“100/1B”) and one susceptible to salinity (“Barley medenine”) from the Middle East and North Africa (MENA) region. Barley plants were exposed to 50 mM NaCl for 7 days; then, salinity was increased to 150 mM NaCl in the presence (10 mM) or limitation (1 mM) of ammonium as a nitrogen source. Upon salinity, “100/1B” was shown to support N assimilation by enhancing the glutamine synthetase (GS) and glutamine oxoglutarate aminotransferase (GOGAT) cycle under high N, and the stimulation of the glutamate dehydrogenase (GDH) pathway under low N treatment. In “Barley medenine”, salinity reduced the GS/GOGAT cycle, and increased GDH activity. Upon salinity, Heat Shock Proteins 70 and PEPC remained unchanged in “100/1B”, while they decreased in “Barley medenine”. The tolerance degree is a determining factor in enzymes’ occurrence and regulation: exposed to salinity, “100/1B” rapidly increased APX and PEPC activities, while this was delayed in “Barley medenine”. Salinity increased cyt-G6PDH levels in “100/1B”, while “Barley medenine” showed a decrease in G6PDH isoforms. Correlation analyses confirm GOGAT was related to G6PDH; GDH and APX with PEPC in “100/1B” under moderate salinity; severe salinity correlated GDH with G6PDH and PEPC. In “Barley medenine” under salinity, GOGAT was correlated with G6PDH, while APX showed a relation with PEPC. Therefore, specific enzymatic activities and occurrence can be used to determine stress responsiveness of different landraces. We suggest that the rapid increase in G6PDH, APX, and nitrogen assimilation enzymes activities represents an index of tolerance in “100/1B” and a stress symptom in “Barley medenine”

Pre-Field Screening of Potential Barley and Durum Wheat Mutants for Drought Tolerance

Drought is the most important stress reducing drastically crop production in Tunisia. Therefore, it is imperative to develop drought tolerant barley and durum wheat variety. This study aims to screen for water stress tolerance among generated variability of barley (Hordeum vulgare L.) and durum wheat (Triticum turgidum ssp. durum). Two commercial genotypes of barley "Rihane" and durum wheat "Maali" were exposed to three levels of γ-irradiation; 100 Gy, 150 Gy and 250 Gy. Selection was set up on M2 using early vigor (Z14) and stay green phenotype (Z59) parameters which allowed to identify 70 and 167 potential mutants of barley and durum wheat, respectively. All selected mutants were assessed for morphological, physiological and yield variability compared to control. Selected barley mutants showed higher yield components than the average non-irradiated controls. Durum wheat M2 selected mutants showed higher agro-morphological parameters under all gamma irradiations. Higher number and fertile tillers were observed under low irradiation levels of 100 and 150 Gy, respectively. Photosynthesis activity evaluated by Fv/Fm and Fv/F0 was improved under 100 Gy. Besides, yield components were also improved mostly under 250 Gy. Furthermore, results showed that δ15N (‰) and δ13C (‰) measured on M3 kernels were not able to discriminate tolerant mutant. All measured parameters will be used in further generations to ass the variability and the efficiency of mutation breeding process on barley and durum wheat

Gastrointestinal parasites of canids, a latent risk to human health in Tunisia

Abstract Background Although data on the parasite environmental contamination are crucial to implement strategies for control and treatment, information about zoonotic helminths is very limited in Tunisia. Contamination of areas with canid faeces harboring infective parasite elements represents a relevant health-risk impact for humans. The aim of this study was to assess the environmental contamination with eggs and oocysts of gastrointestinal parasites of dogs and wild canids in Tunisia with special attention to those that can be transmitted to humans. Results One thousand two hundred and seventy faecal samples from stray dogs and 104 from wild canids (red foxes and golden jackals) were collected from different geographical regions throughout Tunisia. The helminth eggs and protozoan oocysts were concentrated by sucrose flotation and identified by microscopic examination. The most frequently observed parasites in dog samples were Toxocara spp. (27.2%), E. granulosus (25.8%), and Coccidia (13.1%). For wild canid faeces, the most commonly encountered parasites were Toxocara spp. (16.3%) followed by Capillaria spp. (9.6%). The parasite contamination of dog faeces varied significantly from one region to another in function of the climate. Conclusion To our knowledge, the study highlights for the first time in Tunisia a serious environmental contamination by numerous parasitic stages infective to humans. Efforts should be made to increase the awareness of the contamination risk of such parasites in the environment and implement a targeted educational program

Different Roles of Heat Shock Proteins (70 kDa) During Abiotic Stresses in Barley (Hordeum vulgare) Genotypes

In this work, the involvement of heat shock proteins (HSP70) in barley (Hordeum vulgare) has been studied in response to drought and salinity. Thus, 3 barley genotypes usually cultivated and/or selected in Italy, 3 Middle East/North Africa landraces and genotypes and 1 improved genotype from ICARDA have been studied to identify those varieties showing the best stress response. Preliminarily, a bioinformatic characterization of the HSP70s protein family in barley has been made by using annotated Arabidopsis protein sequences. This study identified 20 putative HSP70s orthologs in the barley genome. The construction of un-rooted phylogenetic trees showed the partition into four main branches, and multiple subcellular localizations. The enhanced HSP70s presence upon salt and drought stress was investigated by both immunoblotting and expression analyses. It is worth noting the Northern Africa landraces showed peculiar tolerance behavior versus drought and salt stresses. The drought and salinity conditions indicated the involvement of specific HSP70s to counteract abiotic stress. Particularly, the expression of cytosolic MLOC_67581, mitochondrial MLOC_50972, and encoding for HSP70 isoforms showed different expressions and occurrence upon stress. Therefore, genotypes originated in the semi-arid area of the Mediterranean area can represent an important genetic source for the improvement of commonly cultivated high-yielding varieties.</jats:p

Negative/Low HER2 expression alone or combined with E-cadherin positivity is predictive of better prognosis in patients with breast carcinoma

The loss of E-cadherin expression leads to absence of tissue integrity, an essential step in tumor progression. Methylation of CpG islands in the promoter region of the CDH1 gene coding E-cadherin might be an alternative for gene silencing. In the present study, we investigate the expression of E-cadherin and hormone receptors in invasive ductal breast carcinoma (IDCs). Protein expression was analysed immunohistochemically in 87 cases, including 26 familial tumors. The most interesting results revealed a significantly reduced E-cadherin expression in cases with familial history compared to sporadic tumors (p=0.009), as well as with tumors ≤5cm (p=0.022). Moreover, HER2 over-expression was associated with distant metastasis (p=0.011) and overall survival (p log rank=0.028). Tumors displaying negative/low HER2 expression combined with E-cadherin positivity confer better patient survival (p=0.052). Triple Negative tumors (TN) were more frequently found in patients with advanced grade (GIII) (p=0.001) and TNM (III+IV) (p=0.018) which supports the aggressive behavior of TN tumors. On the other hand, hypermethylation of CDH1 gene promoter was observed in 46% of hereditary cases and strongly associated with loss of E-cadherin expression (p=0.002). Furthermore, patients with unmethylated CDH1 pattern have a better 5-year disease free survival (p=0.021). In conclusion, in patients with hereditary breast cancer, the CpG methylation event contributes to the loss of E-cadherin expression. On the other hand, HER2 over-expression is predictive of worse prognosis, either alone or combined with loss of E-cadherin expression in Tunisian patients with breast cancer