Higher Expression Levels of SSX1 and SSX2 in Patients with Colon Cancer: Regulated In Vitro by the Inhibition of Methylation and Histone Deacetylation.

Background and Objectives: Colon cancer (CC) has a high mortality rate and is often diagnosed at an advanced stage in Saudi Arabia. Thus, the identification and characterization of potential new cancer-specific biomarkers are imperative for improving the diagnosis of CC by detecting it at an early stage. Cancer-testis (CT) genes have been identified as potential biomarkers for the early diagnosis of various cancers. Among the CT genes are those belonging to the SSX family. In order to assess the usefulness of SSX family genes as cancer biomarkers for the detection of early-stage CC, the goal of this research was to validate the expressions of these genes in patients with CC and in matched patients with normal colons (NCs). Materials and Methods: RT-PCR assays were used to analyze the SSX1, SSX2, and SSX3 family gene expression levels in 30 neighboring NC and CC tissue samples from male Saudi patients. Epigenetic alterations were also tested in vitro using qRT-PCR analysis to determine whether reduced DNA methyltransferase or histone deacetylation could stimulate SSX gene expression via 5-aza-2'-deoxycytidine and trichostatin treatments, respectively. Results: The RT-PCR results showed SSX1 and SSX2 gene expression in 10% and 20% of the CC tissue specimens, respectively, but not in any of the NC tissue specimens. However, no SSX3 expression was detected in any of the examined CC or NC tissue samples. In addition, the qRT-PCR results showed significantly higher SSX1 and SSX2 expression levels in the CC tissue samples than in the NC tissue samples. The 5-aza-2'-deoxycytidine and trichostatin treatments significantly induced the mRNA expression levels of the SSX1, SSX2, and SSX3 genes in the CC cells in vitro. Conclusions: These findings suggest that SSX1 and SSX2 are potentially suitable candidate biomarkers for CC. Their expressions can be regulated via hypomethylating and histone deacetylase treatments, subsequently providing a potential therapeutic target for CC.

HSPB6 Is Depleted in Colon Cancer Patients and Its Expression Is Induced by 5-aza-2'-Deoxycytidine In Vitro.

Background and Objectives: Colon cancer (CC) is the second most common cancer in Saudi Arabia, and the number of new cases is expected to increase by 40% by 2040. Sixty percent of patients with CC are diagnosed in the late stage, causing a reduced survival rate. Thus, identifying a new biomarker could contribute to diagnosing CC in the early stages, leading to delivering better therapy and increasing the survival rate. Materials and Methods: HSPB6 expression was investigated in extracted RNA taken from 10 patients with CC and their adjacent normal tissues, as well as in DMH-induced CC and a colon treated with saline taken from a male Wistar rat. Additionally, the DNA of the LoVo and Caco-2 cell lines was collected, and bisulfite was converted to measure the DNA methylation level. This was followed by applying 5-aza-2'-deoxycytidine (AZA) to the LoVo and Caco-2 cell lines for 72 h to see the effect of DNA methylation on HSPB6 expression. Finally, the GeneMANIA database was used to find the interacted genes at transcriptional and translational levels with HSPB6. Results: We found that the expression of HSPB6 was downregulated in 10 CC tissues compared to their adjacent normal colon tissues, as well as in the in vivo study, where its expression was lower in the colon treated with the DMH agent compared to the colon treated with saline. This suggests the possible role of HSPB6 in tumor progression. Moreover, HSPB6 was methylated in two CC cell lines (LoVo and Caco-2), and demethylation with AZA elevated its expression, implying a mechanistic association between DNA methylation and HSPB6 expression. Conclusions: Our findings indicate that HSPB6 is adversely expressed with tumor progression, and its expression may be controlled by DNA methylation. Thus, HSPB6 could be a good biomarker employed in the CC diagnostic process.

The Expression Patterns of Human Cancer-Testis Genes Are Induced through Epigenetic Drugs in Colon Cancer Cells.

BACKGROUND: The expression of human germline genes is restricted to the germ cells of the gonads, which produce sperm and eggs. The germline genes involved in testis development and potentially activated in cancer cells are known as cancer-testis (CT) genes. These genes are potential therapeutic targets and biomarkers, as well as drivers of the oncogenic process. CT genes can be reactivated by treatment with drugs that demethylate DNA. The majority of the existing literature on CT gene activation focuses on X-chromosome-produced CT genes. We tested the hypothesis that epigenetic landscape changes, such as DNA methylation, can alter several CT gene expression profiles in cancer and germ cells. METHODS: Colon cancer (CC) cell lines were treated with the DNA methyltransferase inhibitor (DNMTi) 5-aza-2'-deoxycytidine, or with the histone deacetylase inhibitor (HDACi) trichostatin A (TSA). The effects of these epigenetic treatments on the transcriptional activation of previously published CT genes (CTAG1A, SCP2D1, TKTL2, LYZL6, TEX33, and ACTRT1) and testis-specific genes (NUTM1, ASB17, ZSWIM2, ADAM2, and C10orf82) were investigated. RESULTS: We found that treatment of CC cell lines with 5-aza-2'-deoxycytidine or TSA correlated with activation of X-encoded CT genes and non-X-encoded CT genes in somatic (non-germline) cells. CONCLUSION: These findings confirm that a subset of CT genes can be regulated by hypomethylating drugs and subsequently provide a potential therapeutic target for cancer.

Cancer-Testis Gene Biomarkers Discovered in Colon Cancer Patients.

In Saudi Arabia, colon cancer (CC) is the most prevalent cancer in men and the third most common cancer in women. Rather than being detected through screening programs, most CC cases are diagnosed mainly during clinical exams. Because of the slow growth of CC and its ability to be treated at an early stage, screening for CC can reduce the incidence of death and mortality. Consequently, there is an urgent need to identify a potential new cancer-specific biomarker for detecting early illness. Much research has been conducted on distinct antigen classes as potential new cancer-specific biomarkers for the early identification of malignancy. The cancer-testis antigens (CTAs) are one such category of antigens, with protein presence largely normally confined to human germ line cells in the testis and aberrantly produced in some cancer cells. CTAs are potentially valuable for use as cancer biomarkers and in cancer therapeutics due to their distinctive expression pattern. The aim of this current study was to identify potential cancer-testis (CT) gene biomarkers in Saudi Arabian CC patients. In this study, a total of 20 matching CC and normal colon (NC) tissues were obtained from the Saudi population. Any genes that showed expression in CC tissues but not in matching NC tissues were subsequently verified for mRNA expression in eight breast and eight leukemia malignancies using RT-PCR to determine the specificity of any CC biomarkers. CTAG1A, SPZ1, LYZL6, SCP2D1, TEX33, and TKTL2 genes were expressed in varying numbers of CC tissues compared to no measurable expressions in all NC tissue specimens, making these genes suitable potential candidates for CC markers. The most frequently expressed CT genes in CC patients were CTAG1A (35%) and SCP2D1 (35%), followed by TKTL2 (25%), SPZ1 (20%), LYZL6 (15%), and TEX33 (5%). The LYZL6 gene shows a weak RT-PCR product in 25% of breast cancer (BC) patients but not in leukemia patients. The SCP2D1 gene appears to display expression in all leukemia patients but not in the BC patients. TKTL2 expression was also observed in 50% of leukemia samples but not in the BC samples. More experiments at the protein level and with a larger cohort of patients are required to evaluate this finding.

Cigarette Smoke Regulates the Expression of EYA4 via Alternation of DNA Methylation Status.

Cigarette SMOKE (CS) considerably contributes to causing some diseases such as cancer, and it has a role in the alternation of gene expression through several mechanisms including epigenetics modification, particularly DNA methylation. EYA4 is one of the genes, that whose expression has been dysregulated in lung, colon, bladder, and breast cancer, leading to tumor progression. The alternation of DNA methylation levels has been implicated in regulating the expression of the EYA4 gene. Thus, in this study, we have shown the effect of CS on the DNA methylation level of the EYA4 promoter region as well as the methylation level on EYA4 expression. To determine the level of DNA methylation on the promoter region of the EYA4 gene, we have employed the bisulfite conversion treatment followed by the Sanger Sequence for 100 DNA samples taken from Saudi people (50 smokers and 50 nonsmokers). We found that 26% of DNA extracted from smoker samples is methylated, while there was no methylation identified in nonsmoker samples. Also, using the demethylating agents such as AZA on LoVo and Caco-2 cancer cell lines causes induction of transcription level of EYA4, implying the possible mechanism of DNA methylation in the upregulation of EYA4. These findings suggest the possible mechanism of CS in controlling the expression of EYA4 via changing the status of DNA methylation.

Hydrotropism: Analysis of the Root Response to a Moisture Gradient.

Plant adaptation to environmental stress generated by low water availability requires continuous search for moisture niches in the soil. Thus, roots have evolved a hydrotropic response to sense differences in water potential of the soil, and through asymmetric growth, roots can bend to avoid lower water potential sites. Different experimental systems have been devised for hydrotropism assays, which usually rely on air moisture or split agar assays. This latter system uses plates containing an osmolyte only in a region of the plate in order to generate a water potential gradient. Seedlings are placed on the agar plate containing normal medium (NM) so that their root tips are near the junction between NM and the region supplemented with the osmolyte. As a result, a hydrotropic response is elicited to avoid the low water potential medium, which is reflected in the root curvature angle.

Affinity Purification of Ubiquitinated Proteins Using p62-Agarose to Assess Ubiquitination of Clade A PP2Cs.

Certain E3 ubiquitin ligases play a key role in the abscisic acid (ABA) pathway by targeting clade A type 2C protein phosphatases (PP2Cs) for degradation. At early stages of ABA signaling, degradation of PP2Cs is a complementary step to PP2Cs inhibition by ABA receptors. At later steps, protein levels of PP2Cs are increased as a negative feedback mechanism. Subsequently, E3 ligases targeting PP2Cs are critical to recover the basal PP2C levels and reset the ABA signaling. BTB/POZ AND MATH DOMAIN proteins (BPMs) are substrate adaptors of a multimeric cullin3-RING based E3 ligase and target for degradation clade A PP2Cs. In this chapter, we provide a detailed protocol to assess the ubiquitination of PP2CA, a clade A PP2C, mediated by BPMs using agarose-immobilised p62-derived ubiquitin-associated (UBA) domain, which efficiently binds ubiquitinated proteins.

Ubiquitylation of ABA Receptors and Protein Phosphatase 2C Coreceptors to Modulate ABA Signaling and Stress Response.

Post-translational modifications play a fundamental role in regulating protein function and stability. In particular, protein ubiquitylation is a multifaceted modification involved in numerous aspects of plant biology. Landmark studies connected the ATP-dependent ubiquitylation of substrates to their degradation by the 26S proteasome; however, nonproteolytic functions of the ubiquitin (Ub) code are also crucial to regulate protein interactions, activity, and localization. Regarding proteolytic functions of Ub, Lys-48-linked branched chains are the most common chain type for proteasomal degradation, whereas promotion of endocytosis and vacuolar degradation is triggered through monoubiquitylation or Lys63-linked chains introduced in integral or peripheral plasma membrane proteins. Hormone signaling relies on regulated protein turnover, and specifically the half-life of ABA signaling components is regulated both through the ubiquitin-26S proteasome system and the endocytic/vacuolar degradation pathway. E3 Ub ligases have been reported that target different ABA signaling core components, i.e., ABA receptors, PP2Cs, SnRK2s, and ABFs/ABI5 transcription factors. In this review, we focused specifically on the ubiquitylation of ABA receptors and PP2C coreceptors, as well as other post-translational modifications of ABA receptors (nitration and phosphorylation) that result in their ubiquitination and degradation.

The protective effects of the methylenetetrahydrofolate reductase rs1801131 variant among Saudi smokers.

Methylenetetrahydrofolate reductase (MTHFR) polymorphism plays a fundamental role in susceptibility to various diseases, including cancers and autoimmune diseases. In the current study, we aimed to compare genotype and allele frequency variations of rs1801131, one of the most common variants found in the MTHFR gene, among Saudi smokers and non-smokers. We hypothesized that genetic variations of this gene are responsible for many diseases, particularly those caused by cigarette smoking (CS) such as pulmonary diseases, oral cancer and lung cancer. We performed a case-control study on a sample of 235 healthy smokers and 239 healthy non-smokers in Saudi Arabia. The rs1801131 SNP genotypes were determined using a genotyping assay and multiple in silico algorithmic software programs were used to identify the effects and structural functions of the rs1801131 (Glu429Ala) mutation. Using chi-squared tests, we found that, among smokers, TG and GG genotype carriers had 0.209-fold (OR = 0.209, P < 0.005) and 0.427-fold (OR = 0.427, P = 0.003) lower risks of CS-related disease compared to TT reference genotypes. In addition, this protective effect was observed in Saudi smokers independent of age, gender, types of smoking, duration, and average daily smoking consumption. Filling a research gap by exploring this topic in the Saudi population, the current findings indicate that genotype and allele distributions of MTHFR rs1801131 polymorphism present fundamental protective effects against the risk of CS-related disease. These findings should be verified in future studies with larger sample sizes, different ethnicities, and patients suffering from CS-related diseases, such as oral cancer and lung cancer.

The correlation between single nucleotide polymorphisms of the thymic stromal lymphopoietin receptor and breast cancer in a cohort of female patients in Saudi Arabia.

The current study aimed to examine thymic stromal lymphopoietin receptor (TSLPR) genetic variation and breast cancer (BC) susceptibility in women in Saudi Arabia. Therefore, 127 blood samples from female patients diagnosed with BC and 116 blood samples from healthy female controls were studied using a genotyping assay to determine the association between three TSLPR single nucleotide polymorphisms (SNPs)-P196L, X201W, and A238V-and the risk of BC progression. In addition, gene expression was evaluated in 20 matching BC and normal tissues using immunohistochemistry. TSLPR protein levels were higher among BC patients than those with matching normal breast tissue. In addition, TSLPR SNP P196L was found to have a significant protective effect on BC progression (OR = 0.4427), although only the T allele for TSLPR P196L had this protective effect against BC progression in participants who were younger than 48 years old. In contrast, no association was found between the T allele and risk of BC in participants who were older than 48 years old, and the CT and TT genotypes were significantly associated with BC risk protection in the older group. The effects of the TT genotype and the T allele were closely associated with a decreased risk of BC in participants with estrogen receptors (ER+) and without them (ER-). Overall, the findings revealed a significant correlation between SNPs in the TSLPR genes and BC progression among women in Saudi Arabia.

Biological toxicity assessment of carbamate pesticides using bacterial and plant bioassays: An in-vitro approach.

In recent times, agricultural practices mainly rely on agrochemicals and pesticides to safe-guard edible crops against various pests and to ensure high yields. However, their indiscriminate use may cause severe environmental hazards that directly and negatively affect soil microorganisms and crop productivity. Considering these, present study was aimed to assess the toxicity of carbamate pesticides namely carbamoyl (CBL), methomyl (MML) and carbofuran (CBN) using bacterial and plant (Vigna mungo L.) bioassays. All pesticide doses (25-100 µg mL-1) showed negative effect on bacteria as well as plant. Growth, morphology, survival, cellular respiration and inner membrane permeability of Sinorhizobiumsaheli was hampered when exposed to pesticides. Pesticide induced morphological changes viz. aberrant margins; cellular cracking and distortion/damage in S. saheli were obvious under scanning electron microscope (SEM). The 100 µgCBNmL-1 had maximum inhibitory effect and it reduced survivability of S. saheli by 75%. In addition, biofilm formation ability of S. saheli was inhibited in a pesticides-dose dependent manner and it was statistically (p ≤ 0.05) significant. Pesticides indorsed significant changes in biomarker enzymatic assays and oxidative stress parameters towards S. saheli. Furthermore, at 100 µgCBNmL-1, germination efficiency, root, shoot length, plant survival and tolerance index of V. mungo were decrease by 50, 75, 65, 70 and 66%, respectively over control. Staining of pesticide treated roots with fluorescently labeled dyes propidium iodide (PI) and acridine orange (AO) showed increased oxidative stress, ROS generation and membrane permeability as revealed under confocal laser scanning microscope (CLSM). Furthermore, stressor metabolites and antioxidant enzymes in plant seedlings were progressively enhanced with increasing concentration of pesticides. Conclusively, present finding bestow an insights into a mechanistic approach of carbamate pesticide induced phyto, morpho and cellular toxic effects towards soil bacterium as well as plant with forthcoming implications for designing the pesticides to reduce their toxic/harmful effects.

PYL8 ABA receptors of Phoenix dactylifera play a crucial role in response to abiotic stress and are stabilized by ABA.

The identification of those prevalent abscisic acid (ABA) receptors and molecular mechanisms that trigger drought adaptation in crops well adapted to harsh conditions such as date palm (Phoenix dactylifera, Pd) sheds light on plant-environment interactions. We reveal that PdPYL8-like receptors are predominantly expressed under abiotic stress, with Pd27 being the most expressed receptor in date palm. Therefore, subfamily I PdPYL8-like receptors have been selected for ABA signaling during abiotic stress response in this crop. Biochemical characterization of PdPYL8-like and PdPYL1-like receptors revealed receptor- and ABA-dependent inhibition of PP2Cs, which triggers activation of the pRD29B-LUC reporter in response to ABA. PdPYLs efficiently abolish PP2C-mediated repression of ABA signaling, but loss of the Trp lock in the seed-specific AHG1-like phosphatase PdPP2C79 markedly impairs its inhibition by ABA receptors. Characterization of Arabidopsis transgenic plants that express PdPYLs shows enhanced ABA signaling in seed, root, and guard cells. Specifically, Pd27-overexpressing plants showed lower ABA content and were more efficient than the wild type in lowering transpiration at negative soil water potential, leading to enhanced drought tolerance. Finally, PdPYL8-like receptors accumulate after ABA treatment, which suggests that ABA-induced stabilization of these receptors operates in date palm for efficient boosting of ABA signaling in response to abiotic stress.

Association between tobacco substance usage and a missense mutation in the tumor suppressor gene P53 in the Saudi Arabian population.

The tumor suppressor gene TP53 and its downstream genes P21 and MDM2 play crucial roles in combating DNA damage at the G1/S cell cycle checkpoint. Polymorphisms in these genes can lead to the development of various diseases. This study was conducted to examine a potential association between tobacco substance usage (TSU) and single-nucleotide polymorphism (SNP) at the exon regions of the P53, P21, and MDM2 genes by comparing populations of smokers and non-smokers from Saudi Arabia. P53 rs1042522 (C/G), P21 rs1801270 (A/C), and MDM2 rs769412 (A/G) were investigated by genotyping 568 blood specimens: 283 from male/female smokers and 285 from male/female non-smokers. The results obtained from the smokers and their control non-smokers were compared according to age, sex, duration of smoking, and type of TSU. Heterozygous CG, homozygous GG, and CG+GG genotypes, as well as the G allele of rs1042522 were significantly associated with TSU in Saudi smokers compared with non-smokers. The C allele frequency of rs1801270 was also associated with TSU in smokers (OR = 1.33, p = 0.049) in comparison with non-smokers, in younger smokers (≤29 years) (OR = 1.556, p = 0.03280) in comparison with non-smokers of the same age, in smokers who had smoked cigarettes for seven years or less (OR = 1.596, p = 0.00882), and in smokers who had consumed shisha (OR = 1.608, p = 0.04104) in comparison with the controls. However, the genotypic and allelic frequencies for rs769412 did not show significant associations with TSU in Saudis. The selected SNP of P53 was strongly associated with TSU and may be linked to TSU-induced diseases in the Saudi Arabian population.

A facile and green synthesis of CuO/NiO nanoparticles and their removal activity of toxic nitro compounds in aqueous medium.

In this present work, we report the green synthesis of mixed bimetal oxides (CuO/NiO) for the efficient reduction of toxic nitrophenols (NP, DNP and TNP) in aqueous medium. The CuO/NiO NPs were synthesized by green hydrothermal method combined calcination process. The physiochemical properties of the synthesized CuO/NiO NPs were systematically characterized by using XRD, XPS, FTIR, SEM, and HR-TEM techniques. The calcinated CuO/NiO NPs XRD pattern and SEM morphology show the high crystalline nature than the non-calcinated. Whereas, the XPS and FTIR results confirmed the formation of the metal oxide bonding and the interaction of the bimetals. The HR-TEM images showed the spherical crystals with average particle size about 25 nm. In addition, the SAED pattern confirmed the polycrystalline nature of CuO/NiO NPs. The catalytic reduction of nitro compounds to amino derivative was studied with reducing agent (NaBH4). The CuO/NiO NPs showed the high catalytic activity and completed the reduction reaction of NP, DNP and TNP with in 2, 5 and 10 min respectively. In addition, CuO/NiO NPS exhibited the excellent kinetic rate constant k value about 1.519, 0.5102, 0.4601 min-1 for NP, DNP and TNP respectively. Furthermore, the conversion product aminophenol was observed for these three nitro compounds. The proposed CuO/NiO NPs showed excellent crystal stability after the nitrophenol reduction reactions. An inexpensive CuO/NiO NPs is a promising catalysts for reduction of toxic nitro compounds to useful products in aqueous or non-aqueous medium.

Batch adsorption studies on surface tailored chitosan/orange peel hydrogel composite for the removal of Cr(VI) and Cu(II) ions from synthetic wastewater.

Elimination of heavy metals from wastewater has been a significant process to improve the aquatic source's quality. Various materials act as very effective adsorbents to remove heavy metals, which cause toxicity to plants and all other living organisms. Thus, the present work focuses on removing heavy metals chromium (Cr) and copper (Cu) ions containing wastewater using biodegradable and cost-effective chitosan-based hydrogel composite. The composite was prepared via chemical cross-linking of radical chitosan with polyacrylamide and N,N'-Methylene bisacrylamide and blended with orange peel. The synthesis of the adsorbent has been confirmed by using Fourier-transform infrared spectroscopy (FT-IR), Scanning electron microscopy - Energy dispersive X-ray analysis (SEM-EDAX) and X-ray diffraction (XRD) studies. The adsorption power of the composite of metal ions at different time, pH, adsorbent dosages, different metal ion concentrations were analyzed by using Atomic Absorption Spectroscopy (AAS). The results concluded that the optimum pH for Cr(VI) and Cu (II) were 4 and 5, contact time: 360 min, adsorbent dosage: 4 g, and initial metal ion concentration: 100 mg/L for each metal ions. The adsorption isotherm models follow the Freundlich model and pseudo-second-order kinetics. From the results, the adsorption capacity was observed to be 80.43% for Cr(VI) and 82.47% for Cu(II) ions, respectively.

A strategy to enhance the photocatalytic efficiency of α-Fe2O3.

The photocatalytic dye degradation of pure α-Fe2O3 and different concentration of Co doped α-Fe2O3 is explored. Facile hydrothermal method were employed to prepare pristine, 2% and 4% Co-Fe2O3 nanoparticles. Further, synthesized product confirmation studies were employed from X-ray diffraction, UV-vis spectrometry, Fourier-transform infrared, Raman, scanning electron microscope and transmission electron microscope studies. The rhombohedral nanoparticles developed were enhanced photocatalytic action. Photocatalytic dye degradation studies were analyzed for prepared three samples and the photocatalytic efficacy of the obtained photocatalysts was compared experimentally. Methylene blue dye was degraded under UV-light irradiation with 364 nm. The results showed that 4% Co doped α-Fe2O3 sample exhibited better dye degradation with 92% efficiency. The 4% doping of cobalt ions enhanced the photocatalytic property of Fe2O3 and is a good candidate for methylene blue dye degradation above 90%. In addition, strategy for photocatalytic efficiency enhancement was proposed.

Nitrile-calixarene grafted magnetic graphene oxide for removal of arsenic from aqueous media: Isotherm, kinetic and thermodynamic studies.

A novel adsorbent was developed based on nitrile functionalized calix [4]arene grafted onto magnetic graphene oxide (N-Calix-MGO) for remediation of arsenic (III) ions from aqueous media. The nanocomposite was characterized using Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy (EDX). The effective parameters on adsorption efficiency such as pH, adsorbent dosage, contact time, initial concentration, and temperature were studied. The adsorption process was provided with a high removal efficiency up to (90%) at pH 6 which followed by IUPAC Type II pattern. The mathematical models of kinetics and isotherm validated the experimental process. The adsorption kinetic is followed pseudo-first-order model with R2 > 0.9. The adsorption equilibrium was well fitted on the Freundlich model (R2 ∼ 0.96) as compared Langmuir model (R2 ∼ 0.75). Hence, the Freundlich model suggested a multilayer sorption pattern with a physisorption mechanism for arsenic (III) uptake ono developed nanocomposite with a sorption capacity of 67 mg/g for arsenic. The Gibbs free energy (ΔG° < -20 kJ/mol) showed As(III) uptake ono N-Calix-MGO nanocomposite was the physical adsorption mechanism.

Plant Osmotic Stress Signaling: MAPKKKs Meet SnRK2s.

Osmotic stress signaling in higher plants is crucial to cope with abiotic stress. RAF-like MAPKKKs are activated by hyperosmotic stress and activate downstream ABA-unresponsive and ABA-activated SnRK2s, integrating early osmotic stress and ABA signaling cascades. The connection of B2/B3/B4 RAF-like MAPKKKs with SnRK2s is a new paradigm in signal transduction.

Isolation and Characterization of Novel Trichomonas gallinae Ribotypes Infecting Domestic and Wild Birds in Riyadh, Saudi Arabia.

Trichomonas gallinae, a single-celled protozoan parasite, is a causative agent of the disease trichomonosis, which is distributed worldwide and has recently been highlighted as a pandemic threat to several wild bird species. The aim of this study was to determine the prevalence and genotypic diversity of T. gallinae in Riyadh, Saudi Arabia. For this purpose, 273 oral swab samples from different bird species (feral pigeon Columba livia, common mynah Acridotheres tristis, chicken Gallus gallus domesticus, turkey Meleagris gallopavo, and ducks Anatidae) were collected and tested for T. gallinae infection with InPouch™ TV culture kits. The results showed that the overall prevalence of T. gallinae in these samples was 26.4% (n = 72). The PCRs were used to detect the internal transcribed spacer (ITS) region of T. gallinae, and the results of the sequence analysis indicated genetic variation. Among 48 sequences, we found 15 different ribotypes, of which 12 were novel. Three had been previously described as ribotypes A, C, and II. To our knowledge, this study demonstrated the presence of T. gallinae strain diversity in Saudi Arabian birds for the first time and revealed that ribotypes A and C are predominant among Riyadh birds.

Aislamiento y caracterización de nuevos ribotipos de Trichomonas gallinae que infectan aves domésticas y silvestres en Riad, Arabia Saudita. Trichomonas gallinae, un parásito protozoario unicelular, es un agente causal de la enfermedad tricomonasis, que se distribuye en todo el mundo y recientemente se ha destacado como una amenaza pandémica para varias especies de aves silvestres. El objetivo de este estudio fue determinar la prevalencia y la diversidad genotípica de T. gallinae en Riad, Arabia Saudita. Para este propósito, se recolectaron y analizaron 273 muestras de hisopos orales de diferentes especies de aves (paloma silvestre Columba livia, miná común Acridotheres tristis, pollos Gallus gallus domesticus, pavos Meleagris gallopavo y patos Anatidae) para detectar la infección por T. gallinae mediante cultivo con el estuche InPouch ™ TV. Los resultados mostraron que la prevalencia general de T. gallinae en estas muestras fue del 26.4% (n = 72). Se utilizó la técnica de PCR para detectar la región espaciadora transcrita interna (ITS) de T. gallinae, y los resultados del análisis de secuencias indicaron variación genética. Entre 48 secuencias, se encontraron 15 ribotipos diferentes, de los cuales 12 eran nuevos. Tres habían sido descritos previamente como ribotipos A, C y II. Hasta donde se conoce, este estudio demostró la presencia de diversidad entre cepas de T. gallinae en aves de Arabia Saudita por primera vez y reveló que los ribotipos A y C son los predominantes entre las aves de Riad.

Improved subtyping affords better discrimination of Trichomonas gallinae strains and suggests hybrid lineages.

Trichomonas gallinae is a protozoan pathogen that causes avian trichomonosis typically associated with columbids (canker) and birds of prey (frounce) that predate on them, and has recently emerged as an important cause of passerine disease. An archived panel of DNA from North American (USA) birds used initially to establish the ITS ribotypes was reanalysed using Iron hydrogenase (FeHyd) gene sequences to provide an alphanumeric subtyping scheme with improved resolution for strain discrimination. Thirteen novel subtypes of T. gallinae using FeHyd gene as the subtyping locus are described. Although the phylogenetic topologies derived from each single marker are complementary, they are not entirely congruent. This may reflect the complex genetic histories of the isolates analysed which appear to contain two major lineages and several that are hybrid. This new analysis consolidates much of the phylogenetic signal generated from the ITS ribotype and provides additional resolution for discrimination of T. gallinae strains. The single copy FeHyd gene provides higher resolution genotyping than ITS ribotype alone. It should be used where possible as an additional, single-marker subtyping tool for cultured isolates.