Plant-microbe interactions ameliorate phosphate-mediated responses in the rhizosphere: a review.

Phosphorus (P) is one of the essential minerals for many biochemical and physiological responses in all biota, especially in plants. P deficiency negatively affects plant performance such as root growth and metabolism and plant yield. Mutualistic interactions with the rhizosphere microbiome can assist plants in accessing the available P in soil and its uptake. Here, we provide a comprehensive overview of plant-microbe interactions that facilitate P uptake by the plant. We focus on the role of soil biodiversity in improved P uptake by the plant, especially under drought conditions. P-dependent responses are regulated by phosphate starvation response (PSR). PSR not only modulates the plant responses to P deficiency in abiotic stresses but also activates valuable soil microbes which provide accessible P. The drought-tolerant P-solubilizing bacteria are appropriate for P mobilization, which would be an eco-friendly manner to promote plant growth and tolerance, especially in extreme environments. This review summarizes plant-microbe interactions that improve P uptake by the plant and brings important insights into the ways to improve P cycling in arid and semi-arid ecosystems.

The role of hand fingerprints on predisposition of cancer development.

Fingerprints or dermatoglyphics contain patterns that were formed by parallel ridges on the bare skin of fingertips. This property on the skin, especially on the finger, makes it possible to hold objects with our fingers, and this feature can also be used to determine identity. After cardiovascular diseases, cancer is the second cause of death worldwide. In this paper, we reviewed the associations reported between fingerprint patterns (dermatoglyphics) and cancer types. In this review, we focused on six types of cancer, including gynecological cancers, oral cancer, prostate cancer, gastric cancer, leukemia, and pituitary tumors, and their connection with fingerprints. The dermatoglyphic could be a potentially useful tool for early diagnosis of predisposition in developing some diseases. As some patterns inform us about leading to deadly diseases, such as cancer, which could be prevented, or at least by early diagnosis and taking proper care, the mortality rate could decline. Thus, the fingerprints that have been primarily observed in particular cancers require more research.

Streptomyces strains can improve the quality properties and antifungal bioactivities of tomato fruits by impacting WRKY70 transcription factor gene and nitrate accumulation.

The current study evaluated the effect of plant growth-promoting (PGP) strains of Streptomyces on yield, quality, and nitrate content of fruits, plant-microbe responses, and antifungal effect against blight disease caused by fungus pathogen Alternaria solani on tomato fruits in commercial greenhouse conditions. Greenhouse trials were done with four treatments including strains Y28, IC10, IT25, and commercial bio-fertilizer (Barvar NPK®) on tomato plants. In PGP treatments, the number of infected fruits significantly reduced (60%) compared to Barvar and control. Strain Y28 improved the quality of tomatoes more than other treatments. All three PGP treatments contained a higher level of total sugar concentration and antioxidant enzyme activities than Barvar and control. In contrast, PGP strains, especially Y28, significantly reduced nitrate accumulation (25%) compared to Barvar and control tomatoes. Streptomyces treatments induced more than a 20-fold increase in UDP and WRKY70 transcription factor gene expression relative to the control (P < 0.01). Based on the results, microbe-dependent plant defense induced by these strains is positively correlated to WRKY70 expression and nitrate reduction in commercial greenhouse conditions. These findings suggest that the commercial application of specific strains not only can illustrate an eco-friendly solution to induce resistance against fungal pathogens but also improve the quality properties of food plants with lower nitrate content.

Overexpression of long intergenic noncoding RNAs in bladder cancer: A new insight to cancer diagnosis.

There is increasing evidence that show long noncoding RNAs including long intergenic noncoding RNAs (lincRNA) play a pivotal regulatory role in the biological processes. Differential expression of lincRNAs can be utilized for cancer diagnosis, prognosis, and targeted therapy. Little is known about their expressions in urothelial tumors. Concerning the potential role of lincRNAs in cancer development, we aimed to investigate the expression levels of LINC00958 and DNM3OS in bladder cancer. Fifty tumor and 50 adjacent non-tumor tissue samples along with their clinicopathological parameters were obtained from bladder cancer patients. Expressions of LINC00958 and DNM3OS were analyzed by Real-time PCR. ROC curve analysis was used to evaluate the diagnostic power of LINC0095 and DNM3OS for BC. Expression level of LINC00958 was considerably increased in cancerous tissues (P < 0.001) and in correlation with cigarette smoking (P = 0.043). DNM3OS expression was higher in the tumor tissues than normal tissues (P < 0.001) and showed a significant association with age (P = 0.038). By using the ROC curve, the diagnostic power of LINC00958 and DNM3OS transcript levels in bladder cancer were estimated to be 87% and 75%, respectively. Our findings offer some important intuitions into the oncogenic role of LINC00958 and DNM3OS in bladder cancer and suggest that they can be candidate biomarkers and may provide new approaches for the diagnosis and therapy if being validated in a larger sample size of clinical samples as well as functional studies.

Responsiveness of Elite Cultivars vs. Ancestral Genotypes of Barley to Beneficial Rhizosphere Microbiome, Supporting Plant Defense Against Root-Lesion Nematodes.

Harnessing plant-microbe interactions to advance crop resistance to pathogens could be a keystone in sustainable agriculture. The breeding of crops to maximize yield in intensive agriculture might have led to the loss of traits that are necessary for beneficial plant-soil feedback. In this study, we tested whether the soil microbiome can induce a stronger plant defense against root-lesion nematodes in ancestral genotypes of barley than in elite cultivars. Plants were grown in a sterile substrate with or without the inoculation of rhizosphere microbiomes, and Pratylenchus neglectus was inoculated to the roots. Unexpectedly, elite cultivars profited significantly more from the microbiome than ancestral genotypes, by the reduction of nematodes in roots and the increased shoot weight relative to control plants. The elite cultivars had higher microbial densities in the rhizosphere, which were correlated with root weight. The structure of the bacterial and fungal community of elite and ancestral genotypes differed, as compared by 16S rDNA or internal transcribed spacer amplicon profiles in denaturing gradient gel electrophoresis. The elite cultivars differed in responsiveness to the microbiome. For the most responsive cultivars Beysehir and Jolgeh, the strong microbe-induced suppression of nematodes coincided with the strongest microbe-dependent increase in transcripts of salicylic acid-regulated defense genes after nematode invasion, while the jasmonate-regulated genes LOX2 and AOS were downregulated in roots with the inoculated microbiome. The microbe-triggered modulation of defense gene expression differed significantly between elite and ancestral genotypes of barley. Soil microbiomes conditioned by maize roots suppressed the nematodes in elite cultivars, while the corresponding bulk soil microbiome did not. In conclusion, cultivars Beysehir and Jolgeh harbor the genetic background for a positive plant-microbiome feedback. Exploiting these traits in breeding for responsiveness to beneficial soil microbiomes, accompanied by soil biome management for compatible plant-microbe interactions, will support low-input agriculture and sustainability.

Streptomyces strains modulate dynamics of soil bacterial communities and their efficacy in disease suppression caused by Phytophthora capsici.

The responses of rhizosphere bacterial communities of Streptomyces (SS14 and IT20 stains) treated-pepper plants following inoculation by Phytophthora capsici (PC) was investigated using Illumina MiSeq sequencing. Distinct modulation of the bacteriome composition was found for PC samples with the highest relative abundance (RA) of Chitinophaga (22 ± 0.03%). The RA of several bacterial operational taxonomic units (OTUs) was affected and caused changes in alpha and beta-diversity measures. In IT20, the RA of Cyanobacteria was enriched compared to SS14 (72%) and control samples (47%). Phylotypes belonging to Devosia, Promicromonospora, Kribbella, Microbacterium, Amylocolatopsis, and Pseudomonas genera in the rhizosphere were positively responding against the pathogen. Our findings show that the phosphate solubilizing strain IT20 has higher microbial community responders than the melanin-producing strain SS14. Also, positive interactions were identified by comparing bacterial community profiles between treatments that might allow designing synthetic bio-inoculants to solve agronomic problems in an eco-friendly way.

Tissue-specific synergistic bio-priming of pepper by two Streptomyces species against Phytophthora capsici.

Among several studied strains, Streptomyces rochei IT20 and S. vinaceusdrappus SS14 showed a high level of inhibitory effect against Phytophthora capsici, the causal agent of pepper blight. The effect of two mentioned superior antagonists, as single or combination treatments, on suppression of stem and fruit blight diseases and reproductive growth promotion was investigated in pepper. To explore the induced plant defense reactions, ROS generation and transcriptional changes of selected genes in leaf and fruit tissues of the plant were evaluated. The plants exposed to the combination of two species responded differently in terms of H2O2 accumulation and expression ratio of GST gene compared to single treatments upon pathogen inoculation. Besides, the increment of shoot length, flowering, and fruit weight were observed in healthy plants compared to control. Likely, these changes depended on the coordinated relationships between PR1, ACCO genes and transcription factors WRKY40 enhanced after pathogen challenge. Our findings indicate that appropriate tissue of the host plant is required for inducing Streptomyces-based priming and relied on the up-regulation of SUS and differential regulation of ethylene-dependent genes.

Streptomyces Strains Induce Resistance to Fusarium oxysporum f. sp. lycopersici Race 3 in Tomato Through Different Molecular Mechanisms.

Plant growth promoting rhizobacteria (PGPR) are potential natural alternatives to chemical fungicides in greenhouse production via inducing plant immune system against biotic stresses. In this research, 126 Streptomyces isolates were recovered from rhizosphere soils of 13 different commercial vegetable greenhouses in Iran. Streptomyces isolates were screened for in vitro Plant growth promoting (PGP) traits and ability to antagonize Fusarium oxysporum f. sp. lycopersici race 3 (FOL), the causal agent of Fusarium wilt of tomato (FWT). Six isolates with the highest antagonistic activity and at least three PGP traits were selected and compared with chemical fungicide Carbendazim® in a greenhouse experiment. All bacterial treatments mitigated FWT disease symptoms like chlorosis, stunting and wilting at the same level or better than Carbendazim®. Strains IC10 and Y28 increased shoot length and shoot fresh and dry weight compared to not inoculated control plants. Phenotypic characterization and 16S rRNA gene sequencing showed, strains IC10 and Y28 were closely related to S. enissocaesilis and S. rochei, respectively. The ability of the superior biocontrol strains to induce antioxidant enzymes activity and systemic resistance (ISR) was investigated. Increased activity of catalase (CAT) in plant treated with both strains as well as an increase in peroxidase (POX) activity in plants treated with Y28 pointed to a strain specific-induced systemic resistance (ss-ISR) in tomato against FOL. The differential induced expression of WRKY70 and ERF1 (two transcription factors involved in plant defense) and LOX and TPX by the analyzed Streptomyces strains, especially after inoculation with FOL, suggests that ss-ISR is triggered at the molecular level.

Radioprotective effects of vitamin A against gamma radiation in mouse bone marrow cells.

Radioprotectors by neutralizing the effects of free radicals, reduce the destructive effects of radiation. In this protocol article, the radioprotectory effect of vitamin A on micronuclei induced by gamma radiation was evaluated using micronucleus test. Vitamin A was injected intraperitoneally at 100 and 400 mg/kg two hours before 2 Gray (Gy) of gamma radiation. Animals were sacrificed after 24 h, and then specimens of the bone marrow were smeared and stained. The number of micronuclei were counted in polychromatic cells. Both dosage of vitamin A reduced the micronucleus in bone marrow polychromatic erythrocytes (MnPCE) level, which is statistically significant. The appropriate amount of vitamin A for protection in mice is 100 mg/kg, which protect the bone marrow of mice against clastogenic effects of radiation. The results of the study showed that vitamin A, possibly with an antioxidant mechanism, eliminates the effects of free radicals from ionizing radiation on bone marrow cells and reduces genetic damage. •The data of radioprotective effects of vitamin A showed that administration of 100 mg/kg vitamin A to mice prior to 2 Gy of gamma radiation has reduced the micronucleus levels in PCE cells by a factor of 2.62.•Administration of 100 mg/kg vitamin A, which is much smaller than LD50 of vitamin A (LD50 for intraperitoneal injection = 1510 ± 240 mg/kg) can protect mice.•Vitamin A reduces the harmful effects of ionizing radiation on DNA, due to the antioxidant activity and the trapping of free radicals produced by radiation, and diminish the genetic damage caused by radiation.•Vitamin A has no effect on the proliferation and differentiation rate of bone marrow cells.

Measurement bone mineral density (BMD) of patients with beta thalassemia.

The aim of the study is determining bone mineral density (BMD) of Patients with beta thalassemia in order to find the prevalence and related factors on the conditions. Z-Score of femoral neck and lumbar vertebrae were reported comparing normal matched subjects. Age and bone mineral density were significantly correlated. Moreover, the disease had significantly higher severity in men than in women. A negative significant correlation was detected between BMD and the mean of hematocrit in the last 5 years. There was significant differences between sex hormone and bone density. A significant correlation between hydroxy urea and BMD were found. A significant relationship between the use of bisphosphonates and bone density were found. Osteopenia and osteoporosis were highly prevalent in our participants. Therefore, regular tests are required to examine bone mineral density in these patients. Furthermore, the exact effect of age on bone mineral density need to be clarified.

Dermatoglyphic patterns on fingers and gynecological cancers.

OBJECTIVES: Fingerprints have so far been used for determining the basis of certain malignant diseases, with positive outcomes. Considering the high rates of cancer-related mortality in Iran, this study was conducted for the purpose of examining the dermatoglyphic pattern of fingers in patients with gynecological cancers as compared to healthy people. STUDY DESIGN: The present study was conducted on 151 women with gynecological cancers as the case group and 152 healthy women with no history of such cancers as control group. The dematographic details of participants from both control and case groups were collected using a checklist, and the pattern of their fingerprints was prepared and examined. The data were analyzed for their significance using chi-square test and t- test. Odds ratio with 95% confidence intervals were calculated. RESULTS: Dermatoglyphic analysis showed that arch and loop patterns significantly changed in cases group as compared to control. However, the odds ratio suggested that loop pattern in 6 or more fingers might be a risk factor for developing gynecological cancers. CONCLUSION: Our results showed that there is an association between fingerprint patterns and gynecological cancers and so, dermatoglyphic analysis may aid in the early diagnosis of these cancers.

The Effects of Sex Protein Receptors and Sex Steroid Hormone Gene Polymorphisms on Breast Cancer Risk.

Breast cancer is a common disease and a major cause of death among women throughout the world. Various genes are believed to be involved in the initiation and progression of the disease. Some polymorphisms of these genes increase susceptibility to breast cancer in particular ethnicities. This study used electronic literature search to review the effects of different sex steroid hormone gene polymorphisms on breast cancer risk. Our findings indicated that some polymorphisms in estrogen receptor alpha (ER-α), ER-ß, progesterone receptor (PGR), pregnane X receptor (PXR), and cytochrome P450 enzymes (CYPs) affected breast cancer susceptibility, especially in African American women.

A rare FANCA gene variation as a breast cancer susceptibility allele in an Iranian population.

Fanconi Anemia (FA) is an autosomal recessive syndrome characterized by congenital abnormalities, progressive bone marrow failure and Fanconi anemia complementation group A (FANCA) is also a potential breast and ovarian cancer susceptibility gene. A novel allele with tandem duplication of 13 base pair sequence in promoter region was identified. To investigate whether the 13 base pair sequence of tandem duplication in promoter region of the FANCA gene is of high penetrance in patients with breast cancer and to determine if the presence of the duplicated allele was associated with an altered risk of breast cancer, the present study screened DNA in blood samples from 304 breast cancer patients and 295 normal individuals as controls. The duplication allele had a frequency of 35.4 and 21.2% in patients with breast cancer and normal controls, respectively. There was a significant increase in the frequency of the duplication allele in patients with familial breast cancer compared with controls (45.1%, P=0.001). Furthermore, the estimated risk of breast cancer in individuals with a homozygote [odds ratio (OR), 4.093; 95% confidence intervals (CI), 1.957­8.561] or heterozygote duplicated genotype (OR, 3.315; 95% CI, 1.996­5.506) was higher compared with the corresponding normal homozygote genotype. In conclusion, the present study indicated that the higher the frequency of the duplicated allele, the higher the risk of breast cancer. To the best of our knowledge, the present study is the first to report FANCA gene duplication in patients with breast cancer.

Modulation of Radiation-induced Base Excision Repair Pathway Gene Expression by Melatonin.

OBJECTIVE: Approximately 70% of all cancer patients receive radiotherapy. Although radiotherapy is effective in killing cancer cells, it has adverse effects on normal cells as well. Melatonin (MLT) as a potent antioxidant and anti-inflammatory agent has been proposed to stimulate DNA repair capacity. We investigated the capability of MLT in the modification of radiation-induced DNA damage in rat peripheral blood cells. MATERIALS AND METHODS: In this experimental study, male rats (n = 162) were divided into 27 groups (n = 6 in each group) including: irradiation only, vehicle only, vehicle with irradiation, 100 mg/kg MLT alone, 100 mg/kg MLT plus irradiation in 3 different time points, and control. Subsequently, they were irradiated with a single whole-body X-ray radiation dose of 2 and 8 Gy at a dose rate of 200 MU/min. Rats were given an intraperitoneal injection of MLT or the same volume of vehicle alone 1 h prior to irradiation. Blood samples were also taken 8, 24, and 48 h postirradiation, in order to measure the 8-oxoguanine glycosylase1 (Ogg1), Apex1, and Xrcc1 expression using quantitative real-time-polymerase chain reaction. RESULTS: Exposing to the ionizing radiation resulted in downregulation of Ogg1, Apex1, and Xrcc1 gene expression. The most obvious suppression was observed in 8 h after exposure. Pretreatments with MLT were able to upregulate these genes when compared to the irradiation-only and vehicle plus irradiation groups (P < 0.05) in all time points. CONCLUSION: Our results suggested that MLT in mentioned dose may result in modulation of Ogg1, Apex1, and Xrcc1 gene expression in peripheral blood cells to reduce X-ray irradiation-induced DNA damage. Therefore, administration of MLT may increase the normal tissue tolerance to radiation through enhancing the cell DNA repair capacity. We believed that MLT could play a radiation toxicity reduction role in patients who have undergone radiation treatment as a part of cancer radiotherapy.

Biocontrol Activities of Gamma Induced Mutants of Trichoderma harzianum against some Soilborne Fungal Pathogens and their DNA Fingerprinting.

BACKGROUND: Random induced mutation by gamma radiation is one of the genetic manipulation strategies to improve the antagonistic ability of biocontrol agents. OBJECTIVES: This study aimed to induce mutants with more sporulation, colonization rate leading to enhanced antagonistic ability (in vitro assay) comparing to wild type (WT) and the assessment of genetic differences (in situ evaluation) using molecular markers. The superior mutants could be appropriate biocontrol agents against soil borne fungal diseases. MATERIALS AND METHODS: In this research sampling and isolation of Trichoderma isolates were performed from soils with low incidence of soil borne disease. T. harzianum 65 was selected and irradiation was conducted with gammacell at optimal dose 250 Gray/s. Mutants (115) were obtained from the WT. The antagonistic abilities of twenty-four mutants were evaluated using dual culture and culture filtrate tests. RESULTS: The results of in vitro assays revealed that Th15, Th11 and Th1 mutants exhibited stronger growth inhibition (GI) and colonization rate on Macrophomina phaseolina and Rhizoctonia solani AG4 compared to the wild type. Th15 and Th11 mutants exhibited stronger GI and colonization rate on Sclerotinia sclerotiorum in dual culture and culture filtrate tests and Th1 and Th11 mutants exhibited stronger GI on Fusarium grminearum in culture filtrate test. The DNA fingerprinting was carried out using RAPD and rep-PCR markers. Two (Th9 and Th17) out of the 24 mutants categorized distantly from the rest based on different polymorphism obtained by molecular markers. However, Th9 was different in GI% from Th17. RAPD analysis separated WT from mutants, Th9 from Th17 and also phenotypically superior mutants from other mutants. Meanwhile, rep-PCR analysis categorized WT isolate and mutants according to their antagonistic properties. CONCLUSIONS: The latter marker (rep-PCR) appeared to be reproducible and simple to distinguish mutants from a single isolate of T. harzianum. Mutants (3 isolates) were phenotypically and genotypically distinct from WT. These mutants demonstrated a pronounced biocontrol activities against soilborne fungal phytopathogens.

The L-cell isolation from heterogonous population of intestinal cell line using antibiotic selection method.

Enteroendocrine cells are the largest population of hormone-producing cells in the body and play important roles in many aspects of body functions. The enteroendocrine cell population is divided into different subpopulations that secrete different hormones and peptides. Characterization of each subpopulation is particularly useful for analyzing the cellular mechanisms responsible for specific cell types. Therefore, the necessity of a pure cell line for a specific study purpose was the important motivation for the separation of cell lines for each subpopulation of enteroendocrine cells. The present research introduces a method for the isolation of L-cells, one of the important subpopulations of enteroendocrine cells. The antibiotic selection method was conducted in order to isolate the L-cells from a heterogonous population of intestinal cell line. In this method, a neomycin resistance gene (as selected marker) was expressed under the control of a specific promoter of L-cells. After transfection of manipulated plasmid, only the cells which determine the specific promoter and express neomycin resistance protein would be able to survive under Geneticin antibiotic treatment condition. In order to confirm that the isolated cells were L-cells, reverse transcriptase polymerase chain reaction (PCR) and quantitative PCR assays were performed. Based on the results, the isolated cells were pure L-cells that could be able to express specific mRNA of L-cells efficiently. This technique provides a unique method for the isolation and purification of any cell line. The purified isolated L-cells by this method can be used for future studies and for analyzing cellular mechanisms that involve L-cells' functions.

Association of estrogen receptor-α A908G (K303R) mutation with breast cancer risk.

Genetic mutations in premalignant breast lesions may have a role in malignancy progression or influence the behavior of subsequent disease. A point mutation in estrogen receptor-α (ER-α) as A908G (Lys303→Arg) was originally involved to hypersensitive to estrogen breast hyperplasia. We detected this mutation among Iranian women with invasive breast cancer. A population-based case-control study was conducted in 150 newly diagnosed invasive breast cancer and 147 healthy control individuals controls to screen for presence of the ER-α A908G mutation by using single-strand conformation polymorphism (SSCP) analysis and 33Pcycle DNA sequencing. We detected the 10.7% ER-α A908G mutation in the form of heterozygote genotype only among cancer patients (χ(2)=22.752, P=0.00). The allelic frequency of mutant allele AGG in codon 303 was significantly (χ(2)=29.709, P=0.001) higher in patients with the family history of breast cancer (28.9%) than those without the family history of breast cancer (1.9%). Our data suggest that ER-α codon 303 mutation is correlated with various aspects of breast cancer in Iran. ER-α genotype might represent a surrogate marker for predicting breast cancer developing later in life.

Estrogen receptor genes variations and breast cancer risk in Iran.

Evidence suggests that alterations in estrogen signaling pathways, including estrogen receptor α (ER-α) and estrogen receptor ß (ER-ß) occur during breast cancer development. ER-α and ER-ß genes polymorphisms have been found to be associated with breast cancer and clinical features of the disease in the western countries. In the current study, we evaluated the hypothesis that certain sequence variants of the ER-α and ER-ß genes are associated with an additively increased risk for breast cancer in Iranian women breast cancer patients. The genes were scanned in 150 Iranian patients with newly diagnosed invasive breast tumors and in healthy control individuals by PCR single-strand conformation polymorphism (SSCP) method. Three single nucleotide polymorphisms (SNPs) in codon10 (TCT→TCC), codon 352 (CCG→CCC) and codon 594 (ACG→ACA) in ER-α gene and one SNP codon 392 (CTC→CTG) in ER-ß were revealed have additive effects in developing breast cancer and LN metastases. Also, SNP in codon 392 of estrogen receptor-ß gene is more effective (threefold) than those SNPs in codons 10, 325, 594 of estrogen receptor-α gene in developing LN metastases in breast cancer patients. SNPs in estrogen receptor α and ß have additive effects in increasing risk for developing breast cancer with LN metastases among Iranian women breast cancer patients.

Estrogen receptor-beta gene polymorphism in women with breast cancer at the Imam Khomeini Hospital Complex, Iran.

ER-alpha and ER-beta genes have been proven to play a significant role in breast cancer. Epidemiologic studies have revealed that age-incidence patterns of breast cancer in Middle East differ from those in the Western countries. Two selected coding regions in the ER-beta gene (exons 3 and 7) were scanned in Iranian women with breast cancer (150) and in healthy individuals (147). PCR single-strand conformation polymorphism was performed. A site of silent single nucleotide polymorphism was found only on exon 7. The SNP was found only in breast cancer patients (5.7%) (chi2 = 17.122, P = 0.01). Codon 392 (C1176G) of allele 1 was found to have direct association with the occurrence of lymph node metastasis. Our data suggest that ER-beta polymorphism in exon 7 codon 392 (C1176G) is correlated with various aspects of breast cancer and lymph node metastasis in our group of patients.