Biofluid infrared spectro-diagnostics: pre-analytical considerations for clinical applications.

Several proof-of-concept studies on the vibrational spectroscopy of biofluids have demonstrated that the methodology has promising potential as a clinical diagnostic tool. However, these studies also show that there is a lack of a standardised protocol in sample handling and preparation prior to spectroscopic analysis. One of the most important sources of analytical errors is the pre-analytical phase. For the technique to be translated into clinics, it is clear that a very strict protocol needs to be established for such biological samples. This study focuses on some of the aspects of the pre-analytical phase in the development of the high-throughput Fourier Transform Infrared (FTIR) spectroscopy of some of the most common biofluids such as serum, plasma and bile. Pre-analytical considerations that can impact either the samples (solvents, anti-coagulants, freeze-thaw cycles…) and/or spectroscopic analysis (sample preparation such as drying, deposit methods, volumes, substrates, operators dependence…) and consequently the quality and the reproducibility of spectral data will be discussed in this report.

Diagnosis of hepatocellular carcinoma in cirrhotic patients: a proof-of-concept study using serum micro-Raman spectroscopy.

Hepatocellular carcinoma (HCC) is the third most common cause of cancer death worldwide. The development of novel diagnostic methods is needed to detect tumours at an early stage when patients are eligible for curative treatments. The purpose of this proof-of-concept study was to determine if micro-Raman spectroscopy applied to the sera of cirrhotic patients may be an alternative method for rapidly discriminating patients with and without HCC. Serum samples were collected from 2 groups of patients: cirrhotic patients with HCC (n = 37) and without HCC (n = 34). Two different approaches were used, dried serum drops and freeze-dried serum, and micro-Raman spectra were acquired in the point-mode with a 785 nm laser excitation in the spectral range of 600-1800 cm(-1). Spectra were quality-tested and pre-processed (smoothing, baseline subtraction, vector normalization). Using principal component analysis, the 2 classes, corresponding to cirrhotic patients with and without HCC, could not be differentiated. In contrast, the support vector machine method using the leave-one-out cross validation procedure was able to correctly classify the two groups of patients with an overall rate of accuracy of 84.5% to 90.2% for dried serum drops and 86% to 91.5% for freeze-dried serum. These results are promising and support the concept that serum micro-Raman spectroscopy may become a useful diagnostic tool to detect biomarkers in the field of cancer, as described here for distinguishing between cirrhotic patients with and without HCC.

Increased risk of colonic neoplasia in patients with sporadic duodenal adenoma.

BACKGROUND: Recent studies have shown an increased risk of colorectal neoplasia in patients with duodenal neoplasia. The aim of this retrospective case-control study was to confirm this risk. PATIENTS AND METHODS: Rate of colorectal neoplasia in 29 patients with one or more duodenal adenomas were compared with controls matched for gender and age, but without duodenal adenomas (one case to two controls). Patients with neoplasia of the ampulla, familial adenomatous polyposis or other known hereditary conditions of the digestive tract were excluded. Indications for upper and lower gastrointestinal endoscopy in controls were abdominal pain or changes in bowel habits. Controls with anemia or digestive bleeding were not included. Neoplastic lesions found at colonoscopy were classified as adenomas, advanced adenomas (size > or =10 mm, villous component, high-grade dysplasia), cancers and advanced neoplasia (cancers and advanced adenomas). Comparison between groups was by Fisher's exact test or Student's t test. Odds-ratios (OR) and 95% confidence intervals were calculated, if the difference was significant. RESULTS: Mean age of the 29 cases (seven women, 22 men) was 63.2 years and that of the 58 controls (14 women, 44 men) was 62.5 years. First-degree family history of colorectal cancer was present in four cases (13.8%) and eight controls (13.8%) (NS). Colonoscopy showed at least one adenoma in 15 cases (51.7%) and 11 controls (19%) (P=0.0027; OR 1.87, 1.0-3.49), advanced adenomas in four cases (13.8%) and three controls (5.2%) (NS), and colonic adenocarcinoma in three cases (10.3%) and no controls (0%) (P=0.03). Advanced neoplasia was present in seven cases (24.1%) and three controls (5.2%) (P=0.014; OR 2.86, 0.96-8.52). Results were not significantly modified after the exclusion of patients with a family history of colorectal cancer. CONCLUSION: Although lacking in statistical power, these results confirm that patients with sporadic duodenal adenoma are at high risk of colonic adenoma and advanced neoplasia, warranting systematic colonoscopy.

Prevention by aluminium phosphate of gastric lesions induced by ethanol in the rat: role of endogenous prostaglandins and sulfhydryls.

This study was designed to demonstrate the cytoprotective effect of an antacid containing aluminium phosphate (Phosphalugel) against ethanol-induced gastric injury in the rat and to determine whether this cytoprotective effect is mediated by endogenous prostaglandins and sulfhydryls. We have quantitatively evaluated gastric mucosal lesions using macroscopic and histological techniques one hour after ethanol administration. Two ml of aluminium phosphate given orally one hour before administration of 2 ml of 100% ethanol significantly (p less than 0.01) reduced the area of macroscopic lesions induced by ethanol (3.3 +/- 0.9%) when compared to distilled water (20 +/- 4.8%). The histological study showed that aluminium phosphate prevented deep tissue necrosis. However, it did not protect surface epithelial cells against ethanol injury. Pretreatment with indomethacin, 5 mg/kg sc one hour before aluminium phosphate, slightly but significantly (p less than 0.05) reduced the cytoprotective effect of aluminium phosphate. Macroscopic lesions occupied 4.3 +/- 0.94% and 1.88 +/- 0.41% of total mucosal area in indomethacin group and in vehicle group, respectively. On the other hand, the sulfhydryl blocker, N-ethyl-maleimide, 10 mg/kg sc, given one hour before aluminium phosphate, completely abolished the cytoprotective effect of aluminium phosphate (32.92 +/- 4.85% in N-ethyl-maleimide group versus 3.78 +/- 1.41% in vehicle group; p less than 0.01). These results show that aluminium phosphate has a cytoprotective effect against ethanol injury in the rat. This property appears to be mediated by both endogenous prostaglandins and sulfhydryls.

Gastric emptying is accelerated in obese type 2 diabetic patients without autonomic neuropathy.

OBJECTIVE: To clarify the impact of type 2 diabetes mellitus on the gastric emptying rate. MATERIAL AND METHODS: Using a double-isotope scintigraphic technique, we assessed the gastric emptying of a standard liquid-solid meal in 13 obese type 2 diabetic patients without autonomic neuropathy (age: 47.4 +/- 8.6 yr, body mass index: 33.9 +/- 4.8 kg/m(2), glycaemia: 9.1 +/- 2.6 mmol/l) and in 7 controls with similar sex ratio, age, BMI and body fat distribution. RESULTS: The half gastric emptying time for the liquid phase was not significantly different between diabetic patients and controls (respectively: 52.7 +/- 14.5 min and 63.1 +/- 15.2 min). However, the half gastric emptying time for the solid phase was significantly shortened in diabetic patients versus controls (respectively 88.8 +/- 23.2 min in diabetic patients and 113.6 +/- 26.9 min in controls; p<0.04). Furthermore, a negative relationship was highlighted between the half gastric emptying time for the solid phase and basal glycaemia (r=-0.65, p<0.02) in diabetic patients. No significant relationship was found between gastric emptying parameters and cardiac autonomic nerve function, insulin or gastrin levels. CONCLUSION: Solid gastric emptying is accelerated in obese type 2 diabetic patients without patent autonomic neuropathy when compared to obese non diabetic patients.

Treatment of mild chronic hepatitis C with interferon alpha-2b: results of a multi-centre randomized study in 80 patients.

OBJECTIVE: The natural history of mild chronic hepatitis C is not well-known and the benefit of treating this form of the disease is not well-defined. We conducted a pilot study to answer this question. DESIGN: Mild chronic hepatitis C was defined by positivity for anti-HCV antibodies, detectable serum HCV RNA by PCR, and a Knodell score < or = 5 on a liver biopsy performed within the previous 6 months. Eighty patients from six centres were randomized into two groups receiving interferon alpha-2b, 3 MU three times a week for 6 months (group 1, n = 39) or no treatment (group 2, n = 41). Sustained response was defined by the loss of detectable serum HCV RNA at 6 months after therapy. RESULTS: The two groups were not different at entry with respect to age, sex ratio, source of infection, disease duration, genotype, viral load and Knodell score. One patient (group 1) was excluded from the study, while two patients in group 1 (5%) and seven in group 2 (17.1 %) did not complete the trial. A sustained response was observed in seven patients (18%) in group 1 versus none in group 2 (P < 0.01). The difference in mean Knodell score remained non-statistically significant between the two groups at the end of the study. Reduction or interruption of interferon was necessary in eight patients (24.2%). CONCLUSIONS: This first randomized controlled study in mild chronic hepatitis C shows a proportion of sustained responders to interferon alpha-2b similar to that observed in active chronic hepatitis C.

Vagal afferent innervation and regulation of gastric function.

In this article, we have presented evidence that vagal capsaicin-sensitive afferent fibers are involved in the regulation of gastric mucosal and motor function. Gastric acid secretion stimulated by gastric distension, histamine and central injection of TRH analog are all partly dependent on vagal capsaicin-sensitive afferent mechanisms. It is possible that as vagal efferent activity releases histamine, the common final pathway is the reduction in the response to histamine. At present, it is unclear as to the mechanism by which capsaicin-sensitive afferents are involved in the secretory response to histamine. With regard to the gastric acid and mucosal blood flow responses to TRH, it is not clear whether the sensory neurons represent a component of the efferent pathway that is activated by TRH or whether their role is to set the sensitivity of, or exert feedback control on this efferent pathway. As perineural capsaicin application decreases peptide content in the peripheral terminal fields of sensory neurons and these peptides may produce local effector functions within the tissue, it is possible that alterations in the gastric responses to TRH result from a decrease in the local effector functions of vagal neurons. From the experiments on electrical stimulation of the vagus nerve, it is evident that antidromic stimulation of vagal afferents can stimulate gastric mucosal blood flow, although under these experimental conditions there was no evidence for a capsaicin-sensitive stimulation of gastric acid secretion. The physiological relevance of this stimulation of gastric mucosal blood flow is at present unclear, but it is possible that physiological stimuli, such as distension or nutrients, may stimulate afferents and signal for an increase in gastric mucosal blood flow. In addition, pathophysiological or noxious stimulation of vagal afferents may also signal for an increase in gastric mucosal blood flow and may play a role in the response of the mucosa to injury.

[Colonic perforations complicating colonoscopy. Apropos of 15 consecutive cases observed over 16 years]. / Les perforations coliques compliquant la coloscopie. A propos de 15 cas consécutifs observés en 16 ans.

OBJECTIVES: Our study was undertaken to better assess the circumstances of perforation during colonoscopy. METHODS: All cases observed in our department between 1978 and 1994 were computer recorded. During this period, 5,822 colonoscopies were performed, including 872 with at least one polypectomy. RESULTS: All perforations occurred during colonoscopies performed under anaesthesia (n = 3,373 vs 2,449 without anaesthesia). There were a total of 15 observations of colonic perforations, 1 perforation in 388 colonoscopies. Ten perforations occurred during diagnostic colonoscopies (0.20%), 5 after polypectomy (0.57%). Eight of 10 perforations complicating diagnostic colonoscopies occurred below an impassable stricture. Two patients died from this complication (0.03% of all colonoscopies). CONCLUSIONS: Our study suggests that endoscopic exploration of colonic stricture, in addition to anaesthesia, increases the risk of colonic perforation.

[Gastroprotective effect of an antacid coating agent in rats. Role of endogenous prostaglandins, capsaicin-sensitive afferents neurons and nitric oxide]. / Etude de l'effect gastroprotecteur d'un pansement antiacide chez le rat. Rôle des prostaglandines endogènes, des neurones afférents sensibles à la capsaïcine et du monoxyde d'azote.

UNLABELLED: The aims of this study were: a) to demonstrate the gastroprotective effect of the antacid and mucosal coating agent Gastralgine (aluminium hydroxide and glycinate, magnesium trisilicate and simeticone) against ethanol- and indomethacin-induced gastric injury in the rat; b) to investigate whether gastroprotection elicited by this agent involves stimulation of capsaicin sensitive afferent neurons and activation of the nitric oxide system. METHODS: Rats received intragastrically 2 mL of the antacid or distilled water followed 1 hr later by 2 mL of 100% ethanol ig or 30 mg of indomethacin sc. The surface of ethanol-induced lesions and the length of indomethacin-induced lesions were measured. The role of afferent neurons and endogenous nitric oxide in the prevention of ethanol-induced gastric damage was determined using respectively capsaicin and the inhibitor of nitric oxide biosynthesis, NG-nitro-L-arginine. RESULTS: The antacid and mucosal coating agent significantly reduced the area of macroscopic lesions induced by ethanol and the length of lesions induced by indomethacin. Both functional ablation of afferent nerves and inhibition of nitric oxide synthesis significantly increased ethanol-induced gastric injury but failed to reverse the gastroprotective effect of the antacid against 100% ethanol. CONCLUSIONS: The antacid and mucosal coating agent Gastralgine has a gastroprotective effect against ethanol- and indomethacin-induced injury in the rat. This property does not involve stimulation of capsaicin sensitive afferent neurons or synthesis of endogenous nitric oxide.

[Partial regression of Barret esophagus with high grade dysplasia and adenocarcinoma after photocoagulation and endocurietherapy under antisecretory treatment]. / Régression partielle d'un endobrachyoesophage en dysplasie de haut grade avec adénocarcinome après photocoagulation et endocuriethérapie sous traitement antisécrétoire.

Barrett's oesophagus is a premalignant condition. The possibility of eradicating at least partially the metaplastic epithelium has been reported recently. In this case report, a patient with Barrett's oesophagus complicated by high grade dysplasia and focal adenocarcinoma was treated by Nd:Yag laser then high dose rate intraluminal irradiation while on omeprazole 40 mg/day. A partial eradication of Barrett's oesophagus and a transient tumoural regression were obtained. Histologically, residual specialized-type glandular tissue was observed beneath regenerative squamous epithelium. Four months after intraluminal irradiation, a local tumoural recurrence was detected while the area of restored squamous epithelium was unchanged on omeprazole 40 mg/day. This indicates that physical destruction of Barrett's oesophagus associated with potent antisecretory treatment can induce a regression of the metaplastic epithelium, even in presence of high grade dysplasia. The persistence of specialized-type glands beneath the squamous epithelium raises important issues about its potential malignant degeneration.

[Hepatocellular carcinoma invading the right atrium: clinical, echographic and angiographic study]. / Carcinome hépatocellulaire envahissant l'oreillette droite: étude clinique, échographique et angiographique.

The authors report the case of a 54 year old woman suffering from hepatocellular carcinoma with tumor growth into right hepatic vein, inferior vena cava and right atrium. On cardiac examination, a pansystolic bruit and a diastolic rumble were audible at the tricuspid focus. Diagnosis was confirmed by inferior vena cavography and two-dimensional echocardiography, which demonstrated a large mobile mass in the right atrium moving to and fro through the tricuspid valve. This case report emphasizes the value of routine cardiac examination during the course of hepatocellular carcinoma.

[Lymphocytic ascites revealing systemic mastocytosis]. / Ascite lymphocytaire révélatrice d'une mastocytose systémique.

Systemic mastocytosis is a chronic disease with proliferation of mastocytes in organs separate from skin. We report the case of a systemic mastocytosis revealed by a lymphocytic ascites in the absence of cutaneous involvement. The diagnosis was established by the ultrastructural examination of the bone marrow. Identification of few mastocytes in ascites suggested a mast cell infiltration of the peritoneum. A treatment by alpha-2b interferon was unsuccessful, and death rapidly occurred.

[Primary leiomyosarcoma of the liver 37 years after successful treatment of hereditary retinoblastoma]. / Léiomyosarcome primitif du foie survenu 37 ans après traitement curatif d'un rétinoblastome héréditaire.

In patients successfully treated for hereditary retinoblastoma, the risk of developing a second non-ocular tumor has been reported. We report the first case of primary hepatic leiomyosarcoma in a 39 year-old woman who has been treated 37 years before for hereditary retinoblastoma of the left eye. The patient presented with right upper quadrant abdominal pain and fever. Histological diagnosis was made by liver biopsy. As surgical resection was impossible, chemotherapy with epirubicin, then ifosfamide, etoposide and cisplatin was performed. The patient died 22 months after diagnosis. Genetic abnormalities observed in hereditary retinoblastoma, which probably resulted in a predisposition to the development of hepatic cancer in this patient, were not investigated.

[Subcutaneous seeding on the tract of percutaneous cytologic puncture with a fine needle of a hepatic metastasis from colonic adenocarcinoma]. / Essaimage sous-cutané sur le trajet d'une ponction cytologique percutanée à l'aiguille fine d'une métastase hépatique d'un adénocarcinome colique.

Subcutaneous seeding after percutaneous ultrasonically guided fine needle aspiration of liver tumours is a rare complication and only a few cases have been reported. We report a new case of cutaneous implantation following a fine needle (21.5 gauge of external diameter) aspiration biopsy of a superficial colonic liver metastasis, as shown by a subcutaneous nodule and increased serum carcinoembryonnary-antigen. We discuss the risk factors of this rare complication, especially the thickness of the liver parenchyma along the needle tract and precautions which can be taken for prevention.

[Diclofenac-induced colitis complicated by Klebsiella oxytoca infection]. / Colite induite par le diclofénac et compliquée d'une infection à Klebsiella oxytoca.

A 25-year-old man presented with abdominal pain and bloody diarrhea. Colonoscopy showed hemorrhagic proctocolitis with superficial erosions. Histology was consistent with the diagnosis of ulcerative colitis and biopsy cultures were negative. Despite treatment with prednisolone (40 mg/day), his clinical condition deteriorated and he was referred to our institution. On repeated questioning, the patient reported self-medication with diclofenac (200 mg/day) for 6 weeks to treat tendinitis prior to the beginning of digestive symptoms. Rectosigmoidoscopy confirmed bleeding colitis and repeated biopsy cultures showed Klebsiella oxytoca. Corticosteroids were stopped and ofloxacin (400 mg/day) was prescribed for 14 days. Diarrhea quickly resolved. Colonoscopy 8 weeks later showed only patchy erythematous mucosa without bleeding or erosions. Two years later, the patient remains asymptomatic with normal total colonoscopy. The definitive diagnosis was de novo NSAID-induced colitis subsequently complicated by Klebsiella oxytoca infection.

[Protective effect of appendectomy on the development of ulcerative colitis. A case-control study]. / Effet protecteur de l'appendicectomie contre la rectocolite hémorragique.

AIMS: To examine by a case-control study the relationship between appendectomy and subsequent ulcerative colitis development in a French population. METHODS: A total of 150 patients with ulcerative colitis were matched for age (+/- 5 years) and sex, with 150 controls recruited in a preventive medicine center. The following data were collected from medical records and by standardised questionnaire in consultation or by phone: appendectomy and tonsillectomy before the onset of ulcerative colitis, smoking habits and area of residence. RESULTS: The rate of previous appendectomy in patients with ulcerative colitis was 8% (12/150) compared with 30.6% (46/150) in the control group (P=0.001). There was no significant association between ulcerative colitis and tonsillectomy (25.3 and 27.3% in the control and the ulcerative colitis groups, respectively). Smoking was more frequent in the control group (36%) than in the ulcerative colitis group (25.3%) but the difference was not significant (P=0.07). In multivariate analysis, the risk of developing ulcerative colitis was significantly lower after previous appendectomy (odds ratio=0.26; 95% confidence interval: 0.13-0.55; P=7 x 10(-4)). CONCLUSION: Our study confirms the inverse association between appendectomy and subsequent ulcerative colitis, in a French population, after adjusting on smoking.

[Epidemiology and treatment of gastroduodenal lesions caused by non-steroidal anti-inflammatory agents]. / Epidémiologie et traitement des lésions gastroduodénales provoquées par les médicaments anti-inflammatoires non stéroïdiens.

The most common serious adverse reactions to nonsteroidal anti-inflammatory drugs (NSAIDs) are those affecting the gastro-intestinal tract. There is epidemiologic evidence that NSAIDs use is associated with the development of gastric ulcer and with upper gastrointestinal bleeding and perforation of both duodenal and gastric ulcers. The individual risk is low but given the widespread use of NSAIDs, the number of cases is large with appreciable morbidity and mortality. The main risk factors are age above 65, previous ulcer history and treatment with several NSAIDs. Prophylactic therapy is justified in high risk patients. Synthetic prostaglandin misoprostol has been shown to reduce significantly the frequency of gastric ulcer in patients on NSAIDs. By contrast, H2 receptor blockade with ranitidine has been demonstrated to prevent duodenal but not gastric ulcers. Gastric and duodenal ulcers associated with NSAIDs appear to heal on H2 receptors antagonists and prostaglandins even if NSAIDs are continued. However, large gastric ulcer may heal slowly over 8 to 12 weeks. The place of omeprazole remains to be determined.

[Comments on emergency treatment of biliary lithiasis in patients over 75 years of age. Apropos of 157 cases]. / Réflexions sur le traitement des urgences de la maladie biliaire lithiasique chez les sujets de plus de 75 ans. A propos de 157 observations.

Surgery remains the ideal emergency treatment for biliary lithiasis in elderly subjects despite perioperative morbidity and mortality. Minimally invasive techniques appear promising but require assessment. The aim of this work was to determine the usefulness of these techniques and evaluate outcome in a series of 157 patients over 75 years of age who were hospitalized in an emergency setting of complicated biliary lithiasis from January 1990 to December 1996. There were 103 women and 54 men, mean age 82 years. The patients' general status was evaluated according to the ASA classification; 66% of the patients were ASA III, IV or V. Diagnoses at admission were acute cholecystitis (n = 71, 45%), angiocholitis (n = 50, 31%) subintrant hepatic colic (n = 17, 10.8%), pancreatitis (n = 10, 6%), isolated jaundice (n = 2), peritonitis (n = 2) and occlusion (n = 5). Within 24 hours of admission, 7 patients underwent emergency surgery, and the 150 others were given medical treatment. Among these 150 patients, cure was considered to have been achieved with medical treatment alone in 41 (subsequent surgery being required in only one 6 months later), semi-emergency was performed in 17, and a minimally invasive procedure was performed in the 92 others (echo-guided percutaneous cholecystostomy in 42, endoscopic sphincterotomy in 50) followed by a subsequent operation in 29. In the 103 patients (65.5%) in this series who did not undergo surgery, mortality was 3.8% and in the 54 patients (34.5%) who did, mortality was 15%, but this rate was only 6.9% when the open procedure followed a minimally invasive technique. Surgical treatment of complicated biliary disease remains the ideal therapy but indications should be carefully weighed in these elderly fragilized subjects. Under surgical observation, abstention from surgery or use of minimally invasive techniques can play an important role in the therapeutic strategy aimed at lowering perioperative mortality.

Computed tomography follow-up of acute portal vein thrombosis.

PURPOSE: To assess the evolution of acute portal vein thrombosis by computed tomography (CT). PATIENTS AND METHODS: Retrospective single-centre study (2005-2011) including 23 patients who had an initial CT scan and a CT scan during the first year. The analysis compared the last CT scan available with that of the initial CT scan. Neoplastic thrombosis, extrinsic compressions and cavernomas were excluded. All patients received anticoagulant treatment. RESULTS: The causes included: cirrhoses (n = 6), blood disorders (n = 4), locoregional inflammations and infections (n = 8), abdominal surgery (n = 1). The thrombosis was idiopathic in 4 cases. After a mean follow-up of 7.7 months, 7 patients (30%) benefited from a restitutio ad integrum of the portal system, a stable or partially regressive thrombosis was noted in 12 patients (52%) and an aggravation of the thrombosis was noted in 4 patients (18%). In the sub-group of portal vein thrombosis, repermeabilisation was noted in 37.5% of the patients (6/16) and 6 cavernomas developed. CONCLUSION: CT monitoring helps follow the evolution of an acute portal vein thrombosis and demonstrates complete repermeabilisation of the portal vein in 30% of the patients.