Abstract

Background and Objective: Assessment of alimentary tract helminthes of rodents has a great zoonotic importance.  This study aimed at determining the prevalence of helminth infections in rodents in Hamadan.

Material and Methods: a total of 100 rodents trapped from different parts of city were transported to laboratory. After anesthetizing by chloroform, the animals were undergone an operation to isolates the helminthes. The isolates were stained by Carmine and identified at the genus and species levels. Furthermore, age, sex, weight of rodent and size of various organs of body were determined.

Results: Totally, 62% of the rodents were infected to intestinal helminthes. All trapped rodents were Rattus norvegicus. Six species of helminthes, including three Nematode (45%), 3 Cestode (51%) and no Trematode were isolated from rodents. The infection rate for different helminthes was as follow: Hymenolepis nana 21%, H. diminuta 29%, Heterakis spomosa 43%, Strongyloides sp. 1% Trichuris muris 1% and Cysticercus fasciolaris 1%.

Conclusion: in this area, infection rate of alimentary tract helminthes in the Rattus norvegicus, especially zoonotic helminthes, is relatively high, and the rate of Cestodes is higher than those of Nematodes and Trematodes.

Key words: Prevalence, Helminthes, Alimentary tract, Rodents

Abstract

Background and Objective: Ophthalmic pterygium is a potentially vision-threatening lesion of unknown etiology that often extends on the corneal surface and has a worldwide distribution. Despite various studies, the pathogenesis of pterygium remains unclear and the involvement of human papillomavirus is controversial. We aimed to investigate the involvement of papillomavirus in pterygium formation.

Material and Methods: This case-control study was conducted on 50 tissue specimens of pterygium from the patients who had pterygium surgery as the case group and 10 conjunctival biopsy specimens of individuals without pterygium including the patients with  cataract surgery, as controls. The evidence of papillomavirus infection was tested by polymerase chain reaction (PCR).

Results: All samples, case and control, were not positive for papillomavirus. Both groups were positive for beta-globulin gene used to check the quality of extracted DNA.

Conclusion: In this study, due to the absence of papillomavirus in the context of Pterygium it seems that other factors are involved in causing the disease.

Keywords: Pterygium; Human Papilloma Virus; PCR.

Abstract

       Background and Objective: Various cellular factors affect the process of HIV activity. One of these cellular factors are structures known as microRN that are expected to be involved in controlling HIV replication and infectivity. The expression of one or a set of them may represent the patient's clinical conditions. In this study,  the expression of miR-29a and miR-29b involved in regulating viral genes’ expression was evaluated in three HIV-positive groups and a healthy control group. Later,  the expression level of these microRNAs  was compared between the cases and controls.

      Methods: Total RNA extraction was performed on the collected samples using RNx-plus kit and then the microRNA expression levels were evaluated using Relative Real-time PCR. The obtained data was entered into SPSS 22 and Graphpad softwares and analyzed using Kruskal-Wallis and Man-Whitney tests. P-value of less than 0.05 was considered as statistical significance level.

     Results: The expression level of miR-29a  was reduced in patients under treatment and drug-resistant patients ( P ≤ 0.05)  . All three HIV-positive groups including people without drug treatment, patients under treatment and drug-resistant patients showed reduced miR-29b expression level compared to  control group (P ≤ 0.05).

     Conclusion: the decreased expression of miR-29a and miR-29b in patients under treatment and drug-resistant patients indicates an  increased viral replication and reduced CD4 cell count. It may be possible to predict the progression of the disease by miRNA measurement or control viral replication using these mir-RNAs that requires further studies.

        Keywords: HIV, expression, mir-29a, mir-29b.

Abstract

      Background and Objective: Agr is the most important regulatory system for the expression of Staphylococcus aureus virulence factors in different conditions. Agr acts as a quorum sensing system in this bacterium which is activated by increased cell concentration during the transition from logarithmic growth phase to stationary phase. Its role is to upregulate the secretory virulence factors such as alpha-hemolysin and inhibit the transcription of surface proteins including protein A-encoding gene. The aim of this study was to assess the relationship between the agr system expression and some virulence factors of Staphylococcus aureus in Brain-heart infusion (BHI) culture medium.

     Methods: The expression level of agrA and RNAIII genes from the agr locus along with the expression of hla, spa and mecA genes in BHI broth were assessed in different growth phases using Real time-PCR. Also, gyrB was used as an internal control in this study.

     Results: The growth curve of the five tested isolates in BHI broth at 24 hours showed that all the isolates had relatively similar growth patterns. AgrA gene expression in the stationary phase was decreased by 0.89-fold compared with the logarithmic phase. Although the expression of RNAIII gene increased by 3-fold, hla expression decreased by 0.47-fold.

     Conclusion: An inactive agr system is observed in the BHI broth medium. BHI broth medium contains high amounts of suitable nutrients for the growth of Staphylococcus aureus, thus the bacteria do not require the activity of the agr system for the regulation of the virulence genes in these conditions.

    

Abstract

      Background and objective: Hepatitis B virus (HBV) is a DNA virus with high tendency toward hepatic tissue. There are currently about 3 million HBV-infected people and 350 to 400 million chronic carriers of this virus in the world. X protein plays a role in the over-expression of oncogenes, carcinogenicity of liver cells and overlaps with the basal core promoter of the virus. Mutations at specific nucleotides of this region increase viral replication and liver disease progression. The aim of this study was to investigate the frequency of mutations at nucleotides 1762, 1764 and 1766 of HBV X gene in patients with chronic hepatitis B and hepatitis B-related cirrhosis.

      Methods: In this study, 102 patients including 68 chronic hepatitis patients and 34 patients with hepatitis B-related cirrhosis were enrolled. After DNA extraction, HBV X gene was amplified and sequenced using Semi Nested-PCR. Obtained gene sequences were compared with the standard sequence of HBV virus X gene available in the gene bank (Okamoto AB033559). Then, the mutations in the gene X of HBV were identified.

      Results: Comparison of the standard sequence with sequences obtained from patients showed the presence of A1762T / G1764A mutation in 12 chronic (17.64%) and 13 cirrhotic (38.23%) patients. Also, C1766G / G1764T mutations were found in 8.23% of chronic patients and 17.64% of cirrhotic patients.

      Conclusion: A1762T / G1764A mutations in the overlapping region of the basal core promoter with gene X C-terminal may lead to liver disease progression from chronic hepatitis to cirrhosis, by changing the amino acid sequence of the X protein.

    

ABSTRACT

       Background and Objective: The association of Triglyceride/High Density Lipoprotein-Cholesterol (TG/HDL-C) ratio with fasting serum insulin, which is an alternative method of insulin resistance (IR) measurement, is well-recognized. Thus, the measurement of TG/HDL-C ratio is useful to determine both IR and dyslipidemia, which itself is a characteristic of individuals with IR. Therefore, this study aimed to investigate the relationship between TG/HDL ratio as an indicator of IR, with different fasting blood glucose levels.

      Methods: This case-control study was performed on 343 volunteers with no history of diabetes or use of blood glucose-lowering medications and fasting blood sugar (FBS) levels of less than 126 mg/dl. After sampling, the subjects were divided into three groups based on their FBS level. First group included healthy subjects with FBS of less than 100 mg/dl. Second group consisted of subjects with impaired fasting glucose (IFG) and FBS of 110-100 mg/dl and a third group including those with impaired glucose tolerance (IGT) and FBS of 110-125 mg/dl.

       Results: The amount of TG/HDL-C ratio was 3.8 ± 2.8, 4.0 ± 2.1 and 5.4 ± 3.8 for the healthy group, individuals with IFG and IGT, respectively. The TG/HDL index was significantly different among the tested groups with no significant difference between healthy subjects and subjects with IFG. Moreover, there was a statistically significant difference between the IGT and IFG groups with healthy individuals.

       Conclusion: Considering the significant increase of the TG/HDL ratio in groups with impaired glucose, using this index can be helpful in evaluation of glycemic disorder.

  

ABSTRACT

      Background and Objective: Glycyrrhizaglabra L. is one of the most widely used medicinal herbs in Golestan province that is known for its anti-inflammatory, carminative, antiviral, anti-infection and anti-ulcer properties in Iranian traditional medicine. This study aimed to assess the anti-bacterial and anti-oxidative activity of G. glabrafrom the Golestan province.

        Methods:  The rip root of the plant was collected in autumn 2013. The ethanolic extract of the plant was prepared by maceration method. The anti-oxidative property of the plant was assessed by total antioxidant capacity (TAC), reducing power (RP) and 1, 1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging capacity assays. The anti-bacterial activity was assessed using agar-well diffusion method and the minimum inhibitory concentration (MIC) assay.

        Results: The ethanolic extract of G. glabrahad relatively high anti-oxidative activity with IC50 value of 130 µg/ml, especially in the DPPH method. The extract also exhibited high anti-bacterial activity against the following Gram-positive bacteria: Staphylococcus aureus (21.1±0.7 mm), Staphylococcus epidermidis (19.6±0.2 mm), Bacillus subtilus (19.3±0.6 mm), followed by Escherichia coli (12.1±0.8 mm), Enterococcus faecalis (13.2±0.1 mm) and Kelebsiellapneumoniae (11.5±0.4 mm) with MIC values in the range of 31 - 132 mg. mL^-1.

         Conclusion: According to results, the root extract of G.glabrais a good source of antioxidant compounds with suitable anti-bacterial activity, which can be used as natural anti-infection and anti-inflammatory agent for treatment of many diseases.

      

ABSTRACT

         Background and objectives: Globally, about one third of the population has been infected with Hepatitis B virus (HBV) and more than 400 million people have become chronically infected. Nearly, 20-25% of all carriers develop serious liver diseases such as cirrhosis, chronic hepatitis and hepatocellular carcinoma (HCC). According to the World Health Organization, HBV infection causes more than one million deaths every year. Co-infection with Human Immunodeficiency virus (HIV) and HBV is common, since both viruses have the same routes of transmission. Approximately 10 -15% of HIV-infected individuals develop chronic hepatitis B. The risk of liver diseases-related deaths is also higher in the co-infected patients. According to previous studies, mutation of the pre-core (PC) and basal-core promoter (BCP) regions may play an important role in development of HBV-related HCC and severe liver disease. The aim of this study was to investigate mutations in the BCP, PC and core regions of HBV in HIV-positive patients.

          Methods: DNA was extracted using commercial kits to determine the BCP, PC/core mutations in 124 HIV/HBV co-infected patients (32.4% female and 67.6% male). Polymerase chain reaction (PCR) was performed using specific primers. The positive PCR products were subjected to automated sequencing. Then, nucleotide sequences were aligned with the standard hepatitis B sequence [Gene bank, accession number: AB033559] for mutation detection and analysis.

          Results: In this study, three patients (8.1%) were HBeAg-positive and all of them were HBsAg-positive. The mean of CD4 cell count was 120 cells/mL. The mean age of the patients was 36.16 years. The important pathological mutations in HBV patients including 1752A (73%), 1773C (70.3%), 1753C (10.8%), 1896A (8.1%) and 1762T/1764A (2.7%) were detected in this study.

         Conclusion: Identification of mutations in co-infected patients is of greater importance compared to mono-infected patients, because it can be useful for prediction of HCC-related mutations. Co-infection with HIV has important effects on the natural history of HBV infection, and creates different mutational patterns compared to mono-infected patients.

           Keywords: HBV, HIV, Mutation.

ABSTRACT

         Cardiovascular disease (CVD) is one of the most important causes of mortality worldwide. Several risk factors have been proposed to be associated with CVD. The main risk factors include family history, smoking, hypertension, history of diabetes, hyperlipidemia and obesity. However, new factors have been recently proposed to be associated with CVD. These include blood homocysteine, fibrinogen, plasma factor VII activity, lipoprotein (a) and several other factors. Nowadays, the role of homocysteine in the development and exacerbation of several diseases has been demonstrated. Extensive studies have been performed on the role of homocysteine in diabetes, preeclampsia, depression, Alzheimer's disease and multiple sclerosis. Homocysteine level increases with age. Moreover, absorption of micronutrients, especially vitamin B12, decreases in old age that could cause a further increase in homocysteine level. On the other hand, elasticity of blood vessels decline with aging, and increased levels of homocysteine accelerate atherosclerosis. The sum of these processes highlights the role of homocysteine in the development and exacerbation of several diseases.

         Keywords: Homocysteine, Cardiovascular Disease, Diabetes, Preeclampsia, Depression, Alzheimer's Disease.

ABSTRACT

          Background and Objective: Several studies have

shown the antimicrobial activity of lactobacilli against Salmonella enterica (serotype typhimurium). The aim of this study was to evaluate the inhibitory potential of metabolites produced by probiotic culture of Lactobacillus casei against S. typhimurium and its impact on S. typhimurium motility and biofilm formation.

         Methods: In order to evaluate the impact of the metabolites, L. casei cell-free culture supernatant (CFCS) was collected by centrifugation of L. casei secondary cultures. Effectiveness of the CFCS against Salmonella was evaluated by the well-diffusion method. In addition, in vitro effect of this treatment on motility in Swarm agar and biofilm formation by the bacteria was investigated.

         Results: Inhibition zone diameters of S. typhimurium were 0.83 and 12.1 mm at concentrations of 50 and 100 μl of Lactobacillus CFCS against the log4 of S. typhimurium, respectively. Moreover, CFCS treatment inhibited the motility and biofilm formation by Salmonella. Concentrations of 5% and 10% were determined as the minimum inhibitory concentrations for motility and biofilm formation by S. typhimurium. Furthermore, effectiveness of the CFCS against Salmonella was dose-dependent (P<0.05).

         Conclusion: L. casei CFCS is able to inhibit the growth, motility and biofilm formation in S. typhimurium.

         Keywords: Anti-Bacterial, Lactobacillus Casei Metabolites, Phenotypic Characteristics of S. typhimurium

ABSTRACT
        Background and Objectives: The unfavorable lipid profile in obese individuals is associated with high incidence of various diseases including cardiovascular disease, hypertension, etc. Dieting for weight loss and physical activity are among the most important factors affecting the serum lipid profile. The aim of this study was to compare the effect of diet with and without physical activity on body mass index and serum lipid profile of obese women.
      Methods: This clinical trial was performed in 2011 on 39 obese women referred to a weight loss and nutrition counseling center. Subjects were randomly divided into an active group (diet and exercise) and inactive group (diet without exercise). Blood sampling was done before the intervention and two months after the intervention. Data was analyzed using SPSS-16 and t-test.
       Results: Mean level of triglyceride reduced significantly in both groups. Mean level of cholesterol and low-density lipoprotein decreased significantly only in the active group. The Mean level of high-density lipoprotein in the two groups had no significant difference.
Conclusion: The results indicate that dieting for weight loss along with short-term physical activity improves serum levels of cholesterol and low-density lipoprotein, but does not affect serum HDL level.
        Keywords: Diet, Exercise, Triglyceride, Cholesterol, LDL, HDL.

ABSTRACT
            Background and Objectives: Diagnosis of hepatitis E virus (HEV) infection could be missed in some cases if serological tests are used solely. Molecular characterization of HEV is essential for diagnosis of acute and chronic HEV infections, and evaluating the chronic HEV infection status in immunocompromised patients. The aim of this study was to prepare a suitable HEV positive control, determine the limit of detection (LOD) of HEV RNA for a specific molecular test, and evaluate the efficiency and precision of the test.
           Methods: Genomic region of HEV NCBI reference sequence was constructed. LOD, intra-assay precision, and inter-assay precision were calculated to evaluate the efficiency and precision of the test. Then, tenfold serial dilutions of the HEV positive control were prepared. Real time PCR was performed three times for each dilution. Mean, standard deviation, and coefficient of variation of cycle thresholds obtained in three independent and simultaneous tests were calculated, and the results were analyzed.
          Results: The LOD of this test was determined as 1.4×104 copy/ml or 42 copy/reaction or 14 copy/µl. Intra-assay precision and inter-assay precision for all assays were lower than 2.5% and 10%, respectively.
          Conclusion: We propose that the real time PCR assay targeting the ORF2/3 overlapping conserved region is suitable for detection of a wide range of different HEV genotypes found in acute and chronic HEV infections. However, the precision of the test should be improved for detecting HEV RNA lower than 103 copy/ml.
          Keywords: Hepatitis E virus, Limit of Detection, Real Time PCR.

ABSTRACT
        Background and objectives: Synthesis and characterization of several metal complexes (metal ions: Co2+, Ti4+ and Ce3+) of macroacyclic Schiff base ligand have been reported. The Schiff base ligand is prepared from the condensation reaction of 1, 4-di-(4-fluoro-2-aminophenoxy) butane with salicylaldehyde in ethanol. In addition, antibacterial activity of metal ions, Schiff base ligand and their complexes have been investigated. Recent studies show that many Schiff base complexes have antibacterial activity against Gram-positive and Gram-negative bacteria. Therefore, we aimed to synthesize new Schiff base complexes and evaluate their antibacterial activity against a number of Gram-positive and Gram-negative bacteria.
         Methods: Schiff base ligand and their complexes were characterized by mass spectrometry and IR, H-NMR and C-NMR spectroscopy. The in vitro antibacterial activity of the Schiff base ligand, metal ions and their complexes were evaluated against some Gram-positive and Gram-negative bacteria by disk diffusion method and determining minimum inhibitory concentration.
          Results: In this study, the Schiff base complexes had good antibacterial activity, but the Schiff base ligand and metal ions did not show any antibacterial effect. In some cases, the antibacterial effect of the complexes was higher than that of the standard antibiotics tetracycline and gentamycin. The titanium complex showed the highest antibacterial activity in both methods. This complex created the largest growth inhibition zone (diameter: 100mm) against Staphylococcus aureus, and had the lowest minimum inhibitory concentration against Bacillus subtilis (6.75 mg/ml).
          Conclusion: The compounds synthesized in our study have strong antibacterial activity.
          Keywords: Schiff base complex, Schiff base ligand, antibacterial activity, Staphylococcus aureus, Bacillus subtilis.

ABSTRACT
       Background and Objective: MiRNAs are small RNAs that are expressed in most eukaryotes, and can regulate gene expression by attaching to the 3’ end of target mRNA. MicroRNA-101 (miR-101) post-transcriptional regulation is important for host-virus interactions. In addition, miR-101 has a tumor suppressive role in liver cancer and metastasis, and induces apoptosis in tumor cells. We examined miR-101 expression in patients with chronic hepatitis B, hepatitis B virus (HBV)-associated cirrhosis and healthy individuals.
       Methods: The study was performed on 108 whole blood samples (36 samples from each group) collected in EDTA tubes. RNA was extraction by RNX-plus kit according to the manufacturer’s protocol. Finally, miRNA expression was evaluated using relative real time PCR.
         Results: A 2.4-fold increase was observed in miR-101 expression in patients with chronic hepatitis B, while there was a 3.5-fold increase in miR-101 expression in patients with HBV-associated cirrhosis compared with healthy controls (P=0.003). MiR-101 overexpression in patients with HBV-associated cirrhosis was more notable that in patients with chronic hepatitis B.
         Conclusion: According to the results, evaluating miR-101 expression may predict disease progression from chronic hepatitis B to HBV-associated cirrhosis.
         Keywords: MicroRNAs, Chronic Hepatitis B, Liver Cirrhosis, MiR-101.

ABSTRACT
         Background and Objectives: Capparis spinosa has been used in traditional medicine for various applications including treatment of liver disorders and cancer. We studied the effects of this plant on cell proliferation and morphological characteristics of PLC/PRF/5 liver cancer cell line.
        Methods: After preparing ethanolic extract of the plant, the inhibitory effect of the extract was assessed using MTT assay, and morphological changes were assessed by an inverted microscope.
       Results: C. spinosa ethanolic extract exhibited anti-cancer effects in a concentration-dependent manner. Half-maximal inhibitory concentration of the extract was 1051±4.21 mg/mL. Morphological changes including cell shrinkage, reduction of cell volume and nuclear condensation confirmed the inhibitory effect of C. spinosa on PLC/PRF/5 cells.
        Conclusion: According to the results of this study, extract of C. spinosa seems to be suitable for prevention and treatment of liver cancer. Further studies on animal models could verify the efficiency of the extract against cancer cells.

KEYWORDS: Plants, Medicinal, Capparis spinosa, Liver Neoplasms.       

ABSTRACT
          Background and Objectives: C-C chemokine receptor type 5 (CCR5) is a chemokine receptor expressed at high levels on the surface of T-cells. A 32-bp deletion in the coding region of the CCR5 (CCR5Δ32) leads to production of an incomplete protein that is not expressed on the cell surface. CCR5Δ32 may be involved in development of autoimmune disease, such as systemic lupus erythematosus. We investigated frequency of the CCR5Δ32 polymorphism in SLE patients and healthy controls, and evaluated the relationship between the CCR5Δ32 polymorphism and susceptibility to SLE in Golestan Province, Iran.
          Methods: Whole blood samples were taken from 80 SLE patients admitted to Shahid Sayyad Shirazi hospital and 80 healthy controls (from a blood bank) in the Golestan Province, in 2016. Baseline clinical and laboratorial characteristics were evaluated regarding the CCR5Δ32 genotypes. The CCR5Δ32 polymorphism was determined from genomic DNA by polymerase chain reaction.
          Result: Genotype frequencies of both groups were in the Hardy-Weinberg equilibrium. The frequencies of the CCR5 and the CCR5Δ32 alleles were 98.13% and 1.88% among the patients, and 98.75% and 1.25% among the controls, respectively. Homozygote CCR5Δ32 was not observed in the subjects. The frequency of heterozygous Δ32 was 3.8% and 2.5% among the SLE patients and controls, respectively (P-value>0.05). There was no significant association between the CCR5 status and clinical signs of SLE (P>0.05).
          Conclusion: Our data suggest that the CCR5Δ32 polymorphism has no correlation with SLE in our study population. In addition, the frequency of the Δ32 polymorphism in SLE patients and controls does not follow the Hardy-Weinberg equilibrium
          Keywords: CCR5, Homozygote CCR5Δ32, Heterozygote CCR5Δ32, CCR5Δ32 allele, SLE.

ABSTRACT
          Background and Objectives: Nonviral carriers including those based on synthetic cationic lipids, offer several advantages over the viral counterparts. These carriers are able to form complexes with nucleic acids and deliver genes into the cells via the cellular endocytosis pathway, without significant toxicity. The level of transgenes expression depends on some experimental variables including cell type and density, Lipofectamine and DNA concentrations and Lipofectamine-DNA complexing time. The main objective of this study was to optimize transfection of SW480 colon cancer cells with Lipofectamine 2000.
          Methods: In this study, SW480 cells were transfected with plasmid containing green fluorescent protein reporter gene using Lipofectamine 2000. Green fluorescent protein expression was studied under a reverse fluorescence microscope and the results were analyzed with the ImageJ software. Effect of different quantities of plasmid DNA and different Lipofectamine 2000 volumes on cell transfection efficiency was evaluated.
          Results: The optimal volume of Lipofectamine and quantity of plasmid was 2 µl and 1µg, respectively, which showed 59% efficiency for the transfection of SW480 cells at 24 hours post-transfection.
          Conclusion: This study shows that Lipofectamine 2000 is an efficient reagent for the delivery of genes into SW480 cells. According to the results, the quantity of DNA per transfection and reagent concentrations are essential factors for a successful transfection.
          Keywords: Optimization; pEGFP-NI; Lipofectamine; SW480.

ABSTRACT
             Background and Objectives: Cancer and its conventional treatments may bring about some psychological and health-related symptoms leading to unstable cortisol level. Since evidence has shown that mindfulness based approaches can be helpful to reduce cortisol level, we aimed at investigating the efficacy of   mindfulness-based cognitive therapy (MBCT) on decreasing salivary cortisol level in the women with breast cancer. 
             Methods: in this one- group pretest posttest study, the women with early stage breast cancer were assessed to select eligible ones (N = 15). They were first trained for collecting salivary cortisol sample (before, after and follow-up)   and then were asked to participate in MBCT, an eight-week session approach.  Participants provided salivary samples in the morning (7: 30- 8:30), afternoon (1-3) and night (9-11).  To measure cortisol, Cortisol Enzyme Immunoassay Kit was used. Analysis was performed on the data of 12 participants remained, using repeated measurement.
              Results: the results of repeated measure ANOVA revealed that cortisol level of posttest (after 8-weeks of MBCT and follow-up period) in comparison with pretest was significantly reduced for morning and afternoon cortisol while it was not true for night cortisol.  The post-treatment cortisol was not significantly correlated with age groups, education levels and socio-economic status using repeated measure ANCOVA.   
             Conclusion: Given that the women with breast cancer are at the high risk of psychological distresses and unstable cortisol level, we recommend mindfulness-based approaches esp. MBCT to health providers specifically those working with breast cancer patients.
             Keywords: Mindfulness, MBCT, Cortisol , Salivary Cortisol, Breast cancer.