Efficacy of Topical Palmitoylethanolamide (Levagen+) for the Management of Eczema Symptoms: A Double-Blind, Comparator-Controlled, Randomized Clinical Trial.

INTRODUCTION: Eczema is a debilitating skin disorder clinically characterised by the development of itchy, dry, rough, and scaling skin caused by a series of rudimentary clinical phenotypes. METHODS: This double-blind, randomised, comparator-controlled trial evaluated the effectiveness of topical application of a novel palmitoylethanolamide formulation (Levagen+) compared with a standard moisturiser (comparator) to reduce eczema severity and improve patient outcomes. Seventy-two participants aged over 18 years old with atopic eczema (symptoms including redness, dry skin, scaling, and/or itchiness) on their hands or arm were recruited. Participants were randomly allocated to one of two treatment groups (Levagen + or comparator). Treatment was applied to the affected area twice daily for 4 weeks. Outcome measures included Self-Assessed Eczema Area Severity Index (SA-EASI) scoring and Patient-Oriented Eczema Measure (POEM) from baseline to week 4. RESULTS: Levagen+ was effective at alleviating symptom severity of eczema over 4 weeks. Levagen+ significantly reduced redness, dryness, and total POEM score compared to a comparator cream. CONCLUSION: Levagen+ can significantly reduce eczema symptom severity compared to a comparator product, supporting its use as a potential treatment for eczema. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT05003453.

Impact of lifestyle factors and nutrients intake on occurrence of gastrointestinal cancer in Tunisian population.

This study aims to show the relationship between lifestyle and risk of colorectal and gastric cancers in Tunisian population. The food frequency survey method was used to obtain information about the dietary intake and way of life. Nutrients intake was calculated according to the food composition database. According to our results, the consumption of vegetables, fruits, fish, as well as coffee seems to be protective against digestive cancer, while the consumption of citrus and olive oil is protective against gastric cancer. Tobacco, alcohol, and tea represent a risk against gastrointestinal cancer. Highly educated people are more conscious of the crucial role of prevention. In addition, nutrients were significantly associated with colorectal and gastric cancer. The findings suggest that lifestyle is associated with a risk of gastrointestinal cancer. Moreover, higher intake of nutrients from foods was observed more in cases with colorectal and gastric cancer than controls.

TP53 mutations in colorectal cancer from Tunisia: relationships with site of tumor origin, microsatellite instability and KRAS mutations.

Loss of TP53 function through gene mutation is a critical event in the development and progression of colorectal cancer (CRC). Here we examined 51 primary CRC tumors from Tunisia for mutations in TP53 exons 4-9 using PCR-direct sequencing. TP53 status and mutation site/type were than correlated with nuclear protein accumulation, familial and clinicopathologic variables and data on KRAS mutations and microsatellite instability (MSI-H). The TP53 mutation analysis was possible in the tumor of 47 patients and a deleterious somatic mutation has been detected in 59.6% of the patients (28/47) including 20 (71.4%) missense mutations, 7 nonsense mutations (25%) and 1 (3.6%) frameshift mutation. 89.3% (25/28) of the detected mutations were in exons 5-8, whereas 10.7% (3/28) were in exon 4. Among the 27 non frameshift mutations, 89% (24/27) were transitions and 11% (3/27) were transversions. 64.3% (18/27) of the altered amino acids corresponded to arginine. 74% (20/27) were G>C to A>T transitions, and more than half (14/27) occur at hotspots codons with CpG sites. TP53 mutations correlated closely with TP53 accumulation (p = 0.0090) and inversely with MSI phenotype (p = 0.0658). A KRAS somatic mutation was identified in 25% (7/28) of the TP53 mutated tumors. All these mutations were G>A transitions in codon 12 and all the tumors with combined alterations but one were distally located and MSS. In conclusion, frequency and types of TP53 mutations and correlations with TP53 protein accumulation, and MSI were as reported for non-Tunisian patients. However, no significant associations have been detected between TP53 mutations and clinicopathological data in Tunisian patients as previously reported.

KRAS mutations in colorectal cancer from Tunisia: relationships with clinicopathologic variables and data on TP53 mutations and microsatellite instability.

Mutations in KRAS gene are among the critical transforming alterations occurring during CRC tumorigenesis. Here we screened 51 primary CRC tumors from Tunisia for mutations in KRAS (codons 12 and 13) using PCR-direct sequencing. Our aim was to analyze tumor mutation frequencies and spectra in Tunisian patients with CRC. KRAS status and mutation site/type were than correlated with familial and clinicopathologic variables and data on TP53 mutations and nuclear protein accumulation and microsatellite instability (MSI). A KRAS somatic mutation has been detected in the CRC tumor of 31.5 % (16/51) of the patients. 81.2 % had a single mutation at codon 12 and 23 % had a single mutation at codon 13. The most common single mutation (50 %) was a G>A transition in codon 12 (c.35G>A; p.Gly12Asp). 81.25 % of the KRAS mutations were transitions and 23 % were transversions. All the mutations in codon 13 were a c.38G>A transition, whereas both G>A transitions and G>T and G>C transversions were found in codon 12. The mutation spectrum was different between MSS and MSI-H tumors and more varied mutations have been detected in MSS tumors. Some amino acid changes were detected only in MSS tumors, i.e. p.Gly12Ser, p.Gly12Cys and p.Gly12Ala. Whereas, the KRAS mutation p.Gly13Asp have been detected only in MSI-H. 43.75 % of the patients harboured combined mutations in KRAS and TP53 genes and the tumor of 71.42 % of them showed TP53 overexpression. In conclusion, the frequency and types of KRAS mutations were as reported for non-Tunisian patients. However, no significant associations have been detected between KRAS mutations and clinicopathologic variables and MSI in Tunisian patients as previously reported.

Impaired control of effector T cells by regulatory T cells: a clue to loss of oral tolerance and autoimmunity in celiac disease?

OBJECTIVES: Regulatory T cells (Tregs) are instrumental for tolerance to self-antigens and dietary proteins. We have previously shown that interleukin (IL)-15, a cytokine overexpressed in the intestine of patients with celiac disease (CD), does not impair the generation of functional Tregs but renders human T cells resistant to Treg suppression. Treg numbers and responses of intestinal and peripheral T lymphocytes to suppression by Tregs were therefore compared in CD patients and controls. METHODS: Intraepithelial lymphocytes (IELs) and lamina propria lymphocytes (LPLs) were isolated from duodenal biopsy specimens of CD patients and controls. Concomitantly, CD4+CD25+ T lymphocytes (Tregs) were purified from blood. Responses of IELs and of LPLs, and peripheral lymphocytes (PBLs) to suppression by Tregs were tested by analyzing anti-CD3-induced proliferation and interferon (IFN)-γ production in the presence or absence of peripheral Tregs. Lamina propria and peripheral CD4+CD25+FOXP3+ T cells were assessed by flow cytometry. RESULTS: Although percentages of CD4+CD25+FOXP3+ LPLs were significantly increased in patients with active CD, proliferation and IFN-γ production of intestinal T lymphocytes were significantly less inhibited by autologous or heterologous Tregs in CD patients than in controls (P < 0.01). In all tested CD patients, IEL were unable to respond to Tregs. Resistance of LPLs and PBLs to Treg suppression was observed in patients with villous atrophy who had significantly enhanced serum levels of IL-15 compared with patients without villous atrophy and controls. CONCLUSIONS: Our results indicate that effector T lymphocytes from active CD become resistant to suppression by Tregs. This resistance might cause loss of tolerance to gluten, but also to self-antigens.

Lynch syndrome in Tunisia: first description of clinical features and germline mutations.

PURPOSE: High rates of early colorectal cancers (CRC) are observed in Tunisia suggesting genetic susceptibility. Nevertheless, up to now, no molecular study has been performed in the Tunisian population. In our research, we evaluated the clinical characteristics of Tunisian families suspected of Lynch syndrome and the contribution of DNA mismatch repair (MMR) genes. METHODS: Thirty-one unrelated families suspected of Lynch syndrome were studied. Probands were tested for the presence of germline mutations in the MMR genes MLH1, MSH2, MSH6 and in MUTYH. Available tumours were analysed for microsatellite instability and expression of MMR proteins. Detailed family and medical histories were collected. RESULTS: A total of 134 cancers were noted in the 31 families, the most frequent type of cancer corresponding to CRC (69%), followed by uterine cancer (7.5%). Germline mutations were identified in 11 (35.5%) families (six MSH2, five MLH1, including seven novel mutations), seven of which fulfilled the Amsterdam criteria (sensitivity, 63.6%; positive predictive value, 58.3%). Noteworthy, germline mutations were detected in 52.6% of male patients tested, but in only 8.3% of females (p = 0.02). Moreover, CRC were essentially left sided in families without detected mutation (p = 0.017). Ages of onset of cancers and tumour spectrum were very similar in families with or without MMR germline mutation, contrasting with previous studies performed in other populations. CONCLUSIONS: MMR genes contribute significantly to CRC susceptibility in the Tunisian population. However, the cause of early CRC susceptibility remains unknown in most cases, especially in women and in patients with early left colon or rectal cancer.

Colonoscopy in elderly: feasibility, tolerance and indications: about 901 cases.

BACKGROUND: Colonoscopy is the standard investigation for colonic disease, but clinicians often are reluctant to refer elderly patients for colonoscopy because of a perception of higher risk and a high rate of incomplete examinations. AIMS: To evaluate feasibility and tolerance of this investigation in elderly and to review the most frequent indications of colonoscopy in these patients. METHODS: A pilot retrospective study including 901 patients from January 2005 to December 2009; divided into two groups. Group (I) included patients 75 years old and more, group (II) included patients 45 years old or less. All those patients underwent colonoscopy at the gastroenterology department of Charles Nicole hospital. RESULTS: The 1st group included 231 patients, and the 2nd group included 670 one. A past history of colorectal cancer was more frequent in the group I (33.3% versus 9.90%; p<0.05) however history of chronic inflammatory bowel disease was more frequent in group II (0 versus 40.6%; p<0.05). The main indication of colonoscopy was constipation in group II (6.1% versus 27%; p<0.05) and chronic diarrhoea in group I (42.9% versus 16.4%; p<0.05). Bowel preparation was poor in 30.4% cases of the group I and 12.9% of group II (p<0.05). The tolerance was similar in the two groups. The incomplete colonoscopy rate was higher in the group I (38.3% versus 23.4%; p<0.05). The most frequent cause of colonoscopy interruption was the poor preparation in group I and the bad tolerance in group II. Diverticular disease, polyps and colorectal cancers prevailed in group I, whereas inflammatory bowel disease was current in group II. CONCLUSION: In elderly patients, colonoscopy is safe, well tolerated and offers a good diagnostic yield. Its non completion was essentially due to the poor preparation. Sedation did not seem essential. The optimisation of results of colonoscopy requires an improvement of quality preparation.

[Prevalence and risk factors of thromboembolic complications in inflammatory bowel disease]. / Prévalence et facteurs de risque des complications thromboemboliques au cours des maladies inflammatoires chroniques de l'intestin.

BACKGROUND: Patients with inflammatory bowel disease (IBD) are at increased risk of thromboembolic complications (TEC), which represent an important cause of morbidity and mortality. AIM: To assess the prevalence and risk factors of TEC in patients with IBD. METHODS: We conducted a retrospective study including all the IBD patients in the gastroenterological department of Charles Nicolle hospital between 2000 and 2010. Only thromboembolic events that had been diagnosed by an imaging procedure were counted. RESULTS: A total of 266 patients with IBD were consecutively included. TE events occurred in nine patients (3.4%); six men and three women. Their mean age was 31 years [15-64 years]. Five patients had Crohn's disease and four had ulcerative colitis. The types of TEC were deep venous thrombosis of the leg in five cases with pulmonary embolism in one of them, cerebral venous thrombosis in two cases, portal thrombosis in one case and jugular vein thrombosis in one case. Active disease was present in all cases at the time TEC occurred. CONCLUSIONS: In our study, the prevalence of TEC is 3.4% in patients with IBD. Deep venous thromboses of the leg are the most common TEC and all our cases occurs during the active phase of IBD.

[Nephrogenic ascites: an exceptional diagnosis. About one case]. / L'ascite Nephrogenique, Un Diagnostic Rare Auquel Il Faut Penser. A Propos D'une Observation.

BACKGROUND: Nephrogenic ascites is a clinical entity that manifests as refractory and exsudative ascites with unknown etiology in patients with end stage renal disease and often undergoing hemodialysis. This entity presents in practice many diagnostic and therapeutic difficulties. AIM: The aim of this study is to focus on these difficulties through a new observation. CASE REPORT: We report one case of nephrogenic ascites in chronic renal failure related to an idiopathic neurologic bladder. It's a 38 years old patient on hemodialysis for three years who consulted for exsudative ascites with a low rate of leucocytes. CONCLUSION: We conclude that nephrogenic ascites is rare. Its diagnosis is an exclusion diagnosis based on exclusion of other causes of ascites, particularly exsudative forms with low leucocytes rate. Its management is based essentially on renal transplantation and its prognosis is very poor.

[Autoimmune hepatitis type 1 triggered by cytomegalovirus infection. One rare case report]. / Infection à cytomégalovirus révélatrice d'une hépatite auto immune de type 1.

BACKGROUND: Autoimmune hepatitis is chronic and uncommon disease. The pathogenesis is a complex process. Several triggers for autoimmune hepatitis particularly viral herpesviridae infection, which may induce the development of autoimmunity in predisposed individuals. AIM: Report a new case. CASE REPORT: We report a case of 17-year- woman presented with autoimmune triggered by cytomegalovirus infection. Cytomegalovirus induced autoimmune hepatitis has not been reported previously. Evolution was favourable under antiviral treatment, corticosteroid and azathioprine.

Correlation between liver biopsy and fibrotest in the evaluation of hepatic fibrosis in patients with chronic hepatitis C.

AIM: To assess the diagnostic value of Fibrotest in comparison with liver biopsy, for the evaluation of hepatic fibrosis in patients with chronic hepatitis C. METHODS: This prospective study included in 2 years (2006-2007), consecutive patients with chronic hepatitis C naive to treatment. Fibrotest and liver biopsy were performed. Receiver operating characteristics (ROC) curves , the sensitivity, specificity, positive and negative predictive values were used to assess the diagnostic value of Fibrotest in comparison with the METAVIR classification. RESULTS: We recruited a total of 65 patients: 28 males and 37 females (mean age: 50 years); 92% of the patients had genotype 1. The histological fibrosis results were: 3.1% F0; 24.6% F1; 32.3% F2; 29.2% F3 and 10.8 % F4. The diagnostic value of Fibrotest in the detection of significant fibrosis (F2-F4) was 0. 87. A score > 0.5 has a sensitivity of 85. 1%, a specificity of 72. 2%, a positive predictive value of 88. 9%,and a negative predictive value of 65%. The diagnostic value of Fibrotest in the detection of cirrhosis (F4) was 0. 85. There were 13/65 cases of discordance (20%) for fibrosis, 4 cases were attributable to biopsy and 6 cases to Fibrotest. The discordance was unexplained in 3 cases. the size of biopsy< 15 mm [OR=2. 82, 95% CI, 1. 3-6. 07; p = 0. 008] and the stage of fibrosis F0, F1, F2 [OR =3. 35 , 95% CI, 1. 1-10. 2; p=0. 03] were considered as risk factors of discordance in multivariate analysis. CONCLUSION: This prospective study confirmed the good diagnostic value of Fibrotest as compared with the histological analysis of liver biopsy.

Evaluation of nutritional status in patients with liver cirrhosis.

BACKGROUND: Malnutrition is a frequently reported complication in patients with liver cirrhosis. It has a high clinical and economic impact reflected by an increased morbidity and prolonged hospital stay. AIM: This preliminary prospective study aimed to determine the prevalence of malnutrition in hospitalized cirrhotic patients and to investigate whether biological and anthropometric parameters are a valuable tool for identifying malnutrition in these patients. METHODS: The nutritional status of 44 consecutive cirrhotic patients (21 men, 23 women) was assessed according to the anthropometric measurements and biochemical analysis. The diagnosis of malnutrition was based on diminished values of Mid arm muscle circumference (MAMC) and/or Triceps skinfold thikness (TST) below the 5th percentile or less than 60%. RESULTS: The aetiology of cirrhosis was viral hepatitis in 29 patients (66%). Cirrhosis was classified Child Pugh A, B or C in respectively 9, 26 and 9 patients; 37 patients (84%) have mild or tense ascite. In this study, malnutrition was found in 35 patients (79.5%), whereas 9 patients has a good nutritional status. TST and MAMC less than 60% was found in respectively 72% and 25% of patients. No significant statistical difference in epidemiological characteristics was found between malnourished and well-nourished patients. TST and MAMC decreased significantly according to the Child score (p = 0.014 and 0.032 respectively; a positive correlation was found between these two parameters and the severity of cirrhosis. CONCLUSION: In this study, the high prevalence of denutrition was associated with the severity of cirrhosis. Anthropometric parameters are valuable tools for malnutrition diagnosis.

First genetic analysis in Tunisian familial adenomatous polyposis probands.

Familial adenomatous polyposis (FAP) is an autosomal dominant inherited disease characterized by the development of hundreds to thousands of adenomatous polyps in colon and rectum. The APC gene (adenomatous polyposis coli) is considered as the major mutated gene in FAP. It has been shown that biallelic germline mutations in the base-excision-repair gene MYH can be responsible for a recessive inheritance of adenomatous polyposis (AP). This study is the first Tunisian genetic analysis on AP patients. Multiplex ligation-dependent probe amplification (MLPA) was used to screen the APC gene for large genomic rearrangements. The total APC and MYH exon sequences and exon-intron edges were sequenced in an effort to detect germline mutations, four were explored. Mutations were detected in four patients that fulfil the clinical criteria of AP. Three mutations were found in the APC gene, of which two were novel (c.1636_1639delAGTG and c.2514 G>T) and all gave rise to a truncated APC protein. The missense G382D mutation, already described in north and south European populations was found in the MYH gene at the homozygous state in the fourth patient with moderate AP. Our preliminary study provides a basis for implementation of genetic counselling for AP.

Endoscopic ultrasonography versus helical CT in diagnosis and staging of pancreatic cancer.

AIM: Compare the performances of EUS to helical CT in the diagnosis and staging of pancreatic adenocarcinoma. METHODS: Forty two consecutive patients (mean age 63 years; 25 men, 17 women) who had surgical exploration and histologically proved pancreatic cancer were retrospectively included. All our patients underwent with endoscopic ultrasonography (EUS) and helical computed tomography (helical CT). Data analysis compared helical CT, EUS with the surgical data with or without histological study in diagnosis, staging and resectability of pancreatic cancer. Surgical findings were used as gold standard. RESULTS: For positive diagnosis EUS was more sensitive 100% (CI:93-100) than helical CT 88% (CI:77-95). But helical CT was more specific 89% (CI:64-98) than EUS 83% (CI:58-96) for small tumors whose diameter is below 2.5 cm in witch EUS was more sensitive in their detection (100% versus 83%). In evaluating venous involvement EUS was more sensitive than helical CT (96% versus 50%; p<0.05), while CT was more specific (81% versus 75%; p<0.05). Regarding lymph nodes invasion, the two imaging technique had the same sensibility (56%) with better specificity for helical CT (83% versus 75%; p<0.05). The accuracy of EUS in identifying the T and N stages were 80% and 67% respectively, while helical CT have an accuracy of 50% and 71% respectively. EUS and helical CT correctly identified all resectable tumors while EUS was more accurate than helical CT in detecting non resectable tumors 94% versus 69%. CONCLUSION: EUS remains superior to helical CT in positive diagnosis of pancreatic adenocarcinoma especially for small tumors and also for the diagnosis of venous invasion and in identifying non resectable tumors. The two techniques have the same accuracy in the detection of lymph node involvement.

Do we know all there is to know about Familial Adenomatous Polyposis?

Familial Adenomatous Polyposis (FAP) and Attenuated FAP (AFAP) are caused by a germline mutation in the Adenomatous polyposis coli (APC) gene. Recently, a new pathway characterized by a biallelic mutation in the MYH gene, with a recessive model of inheritance was discovered for this inherited syndrome. This report describes a Tunisian patient with an attenuated FAP phenotype, presenting seven colon polyps and an adenocarcinoma but no detectable germline mutations in the FAP target genes. A well known somatic mutation was found in the APC mutation cluster region (MCR). This case shows that further studies are needed to fully understand all the pathways of the FAP syndrome.

[Usefulness of endoscopic ultrasonography for diagnosis between multivesicular hydatic cyst and serous cystadenoma of the pancreas. Case report]. / Apport de l'echoendoscopie dans le diagnostic differentiel entre un kyste hydatique multivesiculaire et un cystadenome sereux du pancreas. A propos d'un cas.

Hydatid cyst of the pancreas is very rare. However, it should be suspected in every case of pancreatic cyst, especially in hydatid countries where hydatic cyst disease is endemic. In the paper we report the first case of a multivesicular hydatid cyst of the pancreas explored by endoscopic ultrasonography. Endoscopic ultrasonography is very reliable in the investigation of pancreatic cystic tumors. However it did not allow us to define semiological criteria capable of permitting a differential diagnosis between a hydatid cyst and serous cystadenoma. Per-echoendoscopic transduodenal puncture might improve the efficiency of this procedure with of hydatid cysts of the head of the pancreas.

[Place of endoscopic variceal ligation for the prophylaxis of rebleeding from oesophageal varices. Prospective study about 102 cases]. / Place de la ligature elastique dans la prevention secondaire de l'hemorragie digestive par rupture de varices oesophagiennes. Etude prospective de 102 cas.

The aim of this study was to assess the efficacy and morbidity of endoscopic variceal ligation for the prophylaxis of rebleeding from oesophageal varices. This work is a prospective study including 102 patients having had at least one episode of variceal bleeding. The mean age is 52.5 years. Oesophageal varices were eradicated in 83.3% of cases after an average of 3.5 sessions. Recurrence of oesophageal varices after eradication was observed in 22% after an average follow-up of 7 months. 10 patients died before eradication of oesophageal varices from causes unrelated to the technique. The endoscopic variceal ligation is effective for eradication of oesophageal varices, with a low morbidity. This technique appears to be a method of choice in the prophylaxis of rebleeding from oesophageal varices.

[Bone mineral density in patients with Crohn's disease. Contribution of bone densitometry in 53 cases]. / La densite minerale osseuse au cours de la maladie de Crohn: apport de l'osteodensitometrie chez 53 patients.

Fifty-three patients with Crohn disease were included in a prospective study. Bone mineral density was measured at the femoral neck and lumbar spine by dual-energy X-ray absorptiometry. Ten variables were analyzed in search of an association with bone demineralization; Age, sex, nutritional state, smoking, duration of the disease, hormonal status, inflammatory syndrome, site of disease: ileal, colic or ileocolic, accumulated doses of corticoids and intestinal resection. A bone demineralization was observed in 58.5% of cases. The ileum location and the corticothérapie were predictives of bone demineralization. Crohn disease is a disease at high risk of bone demineralization. Predictors of the bone loss were smoking, long duration of the disease, ileal site and systematic corticotherapy.

Thymidylate synthase polymorphism in sporadic colorectal and gastric cancer in Tunisian population: a predictive role in 5-fluorouracil based chemotherapy treatment.

In our study, we investigate the possible association of thymidylate synthase polymorphism, 28 bp tandem repeat in 5'-UTR (transcription enhancer element) with susceptibility of colorectal and gastric cancer in Tunisian population. Because thymidylate synthase provides an effective prediction of chemotherapy treatment based on 5-fluorouracil, our interest in this study was focused on finding an eventual interaction between thymidylate synthase polymorphism and treatment of sporadic colorectal and gastric cancer. Whole blood was collected into EDTA tube, after centrifugation for 15 min, the buffy coat was isolated, and genotyping of TS 5'-UTR polymorphism was carried by polymerase chain reaction method using appropriate primers. Determination of the different genotypes was done directly on the stained agarose gel. Our finding showed that the 5'tandem repeat polymorphism of the thymidylate synthase gene is associated with risk of colorectal cancer; thus, LL (3R/3R) genotype is significantly high in patients with colorectal cancer compared to controls (P = 0.002; OR 2.7; 95 % CI 1.4-5.2). In addition, we found a positive association between SL (2R/3R) genotype in the thymidylate synthase 5'-UTR and gastric cancer risk (P = 0.015; OR 4.46; 95 % CI 1.08-19.64). Furthermore, we found a correlation of thymidylate synthase polymorphism with the fluorouracil-based therapy regimes and also with preoperatory radiation in patients with colorectal cancer. Thymidylate synthase is associated with risk of colorectal cancer but not with gastric cancer; however, heterozygous SL (2R/3R) polymorphism is associated with risk of gastric cancer; moreover, the 5' tandem repeat polymorphism of thymidylate synthase gene was an independent predictor of the clinical treatment.