Antimicrobial activity of Citrus spp. and Anethum graveolens components against Candida metapsilosis in ranch sauce.

In this paper, a study was carried out to test the inhibitory effect of a natural food compound (NFC), based on flavonoids (naringenin, hesperetin, tangeritin, luteolin, apigenin and kaempferol) from citrus and dill, in ranch sauce. A strain of C. metapsilosis, isolated from a spoiled sample of ranch sauce, was used as target pathogen microorganism. The inhibitory effect of NFC was compared with a common mixture of chemical preservatives used in this type of sauces: potassium sorbate and sodium benzoate (S/B). An in vitro test was performed by the microtiter plate assay at 10, 25 and 37 °C for 24 h in modified Tryptic Soy Broth. An additive antimicrobial effect had been observed in the combination of acetic acid and NFC. The results of the microtiter assay were validated in a challenge test in ranch sauce at 5, 25 and 37 °C for 10 weeks. NFC showed partial fungicidal effect against C. metapsilosis, reducing two logarithmic units at 5 °C for 10 weeks. At 5 °C, the traditional doses of S/B used in ranch sauce decreased viable cells to non-detectable counts from the second week of the experiment. At 25 and 37 °C, the use of S/B mixture or the use of NFC showed the same fungicidal effect. The incorporation of NFC, alone or in combination with acetic acid, opens the possibility of formulating clean label sauces with good protection against the development of the acid resistant yeast C. metapsilosis.

[SIXTH JESUS CULEBRAS' LECTURE: GLUTAMINE AND THE CRITICAL PATIENT: THE END OF AN AGE?]. / SEXTA LECCIÓN JESÚS CULEBRAS: GLUTAMINA Y PACIENTE CRÍTICO ¿EL FIN DE UNA ERA?

In the last few years, glutamine has changed its status from a "non-essential" amino acid to "almost essential or indispensable" in the critical patient. This has occurred thanks to a series of studies and meta-analysis highlighting the beneficial effects on nosocomial infection, stay in ICU and hospital stay and mortality. After two multicentre studies (REDOXS and MetaPlus) which reviewed the effects of glutamine on critically ill patients, comments changed to: "we do strongly recommend that glutamine is not used in critically ill patients in shock or multiple organ failure" and: "there is an important questioning about the safety of this approach (combination of high- dose enteral and parenteral glutamine) which should not be ignored" and, therefore: "the committee decides to decrease the degree of recommendation for endovenous glutamine"; it currently states that glutamine "should be considered". According to another multicentre study with severe trauma patients our group (a group which in theory was much benefitted from glutamine actions), and 143 patients, did not experience any observable benefit at the usual parenteral doses. We do agree with previous studies on the prognostic value of low levels of glutamine at admittance, which was confirmed if those levels were not back to normal after its administration, although there are no readily available analytic trials for this. This divergence about the usefulness of glutamine grows up as more and more multicentre studies in critical patients show there should be a change of attitude, and probably the clinical guidelines that welcomed its use should now be amended.

La glutamina es un aminoácido que en pocos años ha pasado de "no esencial" a "casi imprescindible en el enfermo crítico", gracias a una serie de estudios y metaanálisis en los que destacaban sus beneficiosos efectos sobre la infección nosocomial, estancias en UCI y hospitalarias y mortalidad, sobre todo tras dos estudios multicéntricos (REDOXS y MetaPlus) que revisaban los efectos de la glutamina en pacientes críticos, los comentarios pasaban a: "recomendamos fuertemente que la glutamina no sea utilizada en pacientes críticos en shock o fallo multiorgánico" a través de un "importante cuestionamiento sobre la seguridad de esta estrategia (combinación de altas dosis de glutamina enteral y parenteral) que no debe ser ignorada" y, por tanto, "el comité decide disminuir el grado de recomendación para la glutamina endovenosa"; y actualmente destaca que la misma "debería ser considerada". Nuestro grupo, también según otro estudio multicéntrico en enfermos traumáticos graves, un grupo teóricamente más beneficiado de la acción de la glutamina, y en 143 pacientes, a las dosis parenterales habituales, no observamos ningún beneficio. Sí que coincidimos con anteriores estudios en el valor pronóstico de valores bajos de glutaminemia al ingreso, que se veía confirmado si no se normalizaban tras su administración, aunque su determinación no es una prueba analítica asequible. Esta divergencia sobre la utilidad de la glutamina aparece con la proliferación de estudios multicéntricos en pacientes críticos que obligan a un cambio de actitud y, probablemente también, en las guías clínicas que tan favorablemente acogieron su uso.

A practical guide for the diagnosis of primary enteric nervous system disorders.

OBJECTIVE: Primary gastrointestinal neuropathies are a heterogeneous group of enteric nervous system (ENS) disorders that continue to cause difficulties in diagnosis and histological interpretation. Recently, an international working group published guidelines for histological techniques and reporting, along with a classification of gastrointestinal neuromuscular pathology. The aim of this article was to review and summarize the key issues for pediatric gastroenterologists on the diagnostic workup of congenital ENS disorders. In addition, we provide further commentary on the continuing controversies in the field. RESULTS: Although the diagnostic criteria for Hirschsprung disease are well established, those for other forms of dysganglionosis remain ill-defined. Appropriate tissue sampling, handling, and expert interpretation are crucial to maximize diagnostic accuracy and reduce interobserver variability. The absence of validated age-related normal values for neuronal density, along with the lack of correlation between clinical and histological findings, result in significant diagnostic uncertainties while diagnosing quantitative aberrations such as hypoganglionosis or ultrashort Hirschsprung disease. Intestinal neuronal dysplasia remains a histological description of unclear significance. CONCLUSIONS: The evaluation of cellular quantitative or qualitative abnormalities of the ENS for clinical diagnosis remains complex. Such analysis should be carried out in laboratories that have the necessary expertise and access to their own validated reference values.

Muscle atrophy and functional deficits of knee extensors and flexors in people with chronic stroke.

BACKGROUND: Further clarification is needed with regard to the degree of atrophy in individual muscle groups and its possible relationship to joint torque deficit poststroke. OBJECTIVE: The purpose of this study was to investigate quadriceps and hamstring muscle volume and strength deficits of the knee extensors and flexors in people with chronic hemiparesis compared with a control group. DESIGN: This was a cross-sectional study. METHODS: Thirteen individuals with hemiparesis due to chronic stroke (hemiparetic group) and 13 individuals who were healthy (control group) participated in this study. Motor function, quadriceps and hamstring muscle volume, and maximal concentric and eccentric contractions of the knee extensors and flexors were assessed. RESULTS: Only the quadriceps muscle of the paretic limb showed reduced muscle volume (24%) compared with the contralateral (nonparetic) limb. There were no differences in muscle volume between the hemiparetic and control groups. The peak torque of the paretic-limb knee extensors and flexors was reduced in both contraction modes and velocities compared with the nonparetic limb (36%-67%) and with the control group (49%-75%). The nonparetic limb also showed decreased extensor and flexor peak torque compared with the control group (17%-23%). Power showed similar deficits in strength (12%-78%). There were significant correlations between motor function and strength deficits (.54-.67). LIMITATIONS: Magnetic resonance imaging coil length did not allow measurement of the proximal region of the thigh. CONCLUSIONS: There were different responses between quadriceps and hamstring muscle volumes in the paretic limb that had quadriceps muscle atrophy only. However, both paretic and nonparetic limbs showed knee extensor and flexor torque and power reduction.

Endoscopic transesophageal vs. thoracoscopic removal of mediastinal lymph nodes: a prospective randomized trial in a long term animal survival model.

BACKGROUND AND STUDY AIMS: In cases where biopsies remain inconclusive, removal of mediastinal lymph nodes for further analysis requires surgical means. Natural orifice transluminal endoscopic surgery (NOTES) procedures allow incision/closure of the gut wall, which might enable endoscopic excision of pre-marked nodes. The aims of the current study were to investigate the feasibility, safety, and reproducibility of lymph node generation in an animal model to enable endoscopic ultrasound-guided (EUS) lymph node removal (ELR) using transesophageal NOTES access/closure and to compare this procedure with thoracoscopic lymph node removal (TLR) in a randomized long term survival animal study. PATIENTS AND METHODS: Lymph node creation using graphite injection was performed in 12 pigs. After randomization into ELR and TLR groups, lymph nodes were marked with newly developed anchors under EUS guidance and removed using either ELR or TLR. ELR included incision of the esophageal wall and closure after lymph node removal. The main outcome measures were success in lymph node generation, technical success of lymph node removal, complications, and comparability of ELR and TLR. RESULTS: Generation of lymph nodes proved successful in all animals in 46/48 sites injected (96 %). Anchors were placed through the selected nodes in a mean of 9.4 minutes. TLR and ELR were successful in all cases. One bleeding occurred during esophageal incision in ELR, which was stopped endoscopically. After lymph node removal, endoscopic suturing of the incision took a mean of 18 minutes. Procedure time was longer for ELR than TLR (mean 48 vs. 42 minutes). All animals survived the procedures. Autopsy after 4 weeks showed two thoracic wall abscesses in the TLR group and none in the ELR group.  Microscopic analysis revealed well healed esophageal scars. CONCLUSION: ELR proved to be feasible in this limited sample size and complications were not observed more frequently in this group than in the TLR group.

Quantitation of cellular components of the enteric nervous system in the normal human gastrointestinal tract--report on behalf of the Gastro 2009 International Working Group.

BACKGROUND: Patients with gastrointestinal neuromuscular diseases may undergo operative procedures that yield tissue appropriate to diagnosis of underlying neuromuscular pathology. Critical to accurate diagnosis is the determination of limits of normality based on the study of control human tissues. Although robust diagnostic criteria exist for many qualitative alterations in the neuromuscular apparatus, these do not include quantitative values due to lack of adequate control data. PURPOSE: The aim of this report was to summarize all relevant available published quantitative data for elements of the human enteric nervous system (neuronal cell bodies, glial cells, and nerve fibers) from the perspective of the practicing pathologist. Forty studies meeting inclusion criteria were systematically reviewed with data tabulated in detail and discussed in the context of methodological variations and limitations. The results reveal a lack of concordance between observations of different investigators resulting in data insufficient to produce robust normal ranges. This diversity highlights the need to standardize the way pathologists collect, process, and quantitate neuronal and glial elements in enteric neuropathologic samples, as suggested by recent international guidelines on gastrointestinal neuromuscular pathology.

The feasibility of wireless capsule endoscopy in detecting small intestinal pathology in children under the age of 8 years: a multicentre European study.

OBJECTIVE: To systematically evaluate the feasibility and methodology to carry out wireless capsule endoscopy (WCE) in children <8 years to define small intestinal pathology. DESIGN: Prospective European multicentre study with negative prior investigation. PATIENTS AND INTERVENTIONS: 83 children aged 1.5-7.9 years were recruited. Initially, all were offered "swallowing" (Group 1) for capsule introduction. If this failed endoscopic placement (Group 2) was used and the Roth net, Advance or custom-made introducers were compared. OUTCOME MEASURES: Primary endpoint: to determine pathology; secondary endpoint: comparison of capsule introduction methods. RESULTS: Capsule introduction: 20 (24%) children aged 4.0-7.9 years (mean, 6.9 years; 14 male) comprising Group 1 were older (p<0.025) than 63 (76%) aged 1.5-7.9 years (mean, 5.25 years; 30 male) forming Group 2. COMPLICATIONS: Roth net mucosal trauma in 50%; no others occurred. The available recording apparatus was inappropriate for those <3 years. INDICATIONS: gastrointestinal bleeding: n = 30 (16 positive findings: four ulcerative jejunitis, four polyps, two angiodysplasia, two blue rubber blebs, two Meckel's diverticula, one anastomotic ulcer, one reduplication); suspected Crohn's disease: n = 20 (11 had Crohn's disease); abdominal pain: n = 12 (six positive findings: three Crohn's disease, two lymphonodular hyperplasia, one blue rubber bleb); protein loss: n = 9 (four lymphangectasia); malabsorption: n = 12 (seven positive findings: six enteropathy, one ascaris). No abnormalities overall: 45%. CONCLUSION: WCE is feasible and safe down to the age of 1.5 years. 20 children >4 years swallowed the capsule. The Advance introducer proved superior for endoscopic placement. The pathologies encountered showed age specificity and, unlike in adolescents, obscure gastrointestinal bleeding was the commonest indication.

Which parameters might predict complications after natural orifice endoluminal surgery (NOTES)? Results from a randomized comparison with open surgical access in pigs.

BACKGROUND AND STUDY AIMS: Natural orifice transluminal endoscopic surgery (NOTES) is currently developed and assessed mainly in pig experiments. The vast majority of studies show a good outcome in short-term follow-up. The current study aims at comparing various parameters of postinterventional assessment and surveillance in relation to clinical behavior and autopsy results to find suitable control parameters and also to assess the pig as suitable model for NOTES compared with open surgery. METHODS: Within the framework of a randomized prospective study of 20 pigs with iatrogenic colonic perforation comparing endoscopic with open surgical closure, clinical examination, including observation of behavior, food intake, and body temperature, was carried out daily. Laboratory parameters (white blood cells [WBC], granulocytes) were measured in 14 animals. Weight was measured preoperatively and on days 2 and 7 postoperatively. Results were matched with complications found during/after 2 weeks' survival. Pre-autopsy sterile cultures were taken from the peritoneal cavities to determine possible bacterial contamination. RESULTS: Three animals from the surgical group were sacrificed on days 4, 8, and 12 because they became severely ill, with autopsy revealing intussusception from adhesions, peritoneal abscess, and peritonitis, in one pig each; another animal had culture positive for ESCHERICHIA COLI. Three minor complications (2 cough, 1 continuing fever with adhesions to the bladder found on autopsy) occurred in the endoscopic group without compromised recovery. WBC were measured in 14 animals, and found to be elevated (8 - 36 x 10 (9)/l) in six on day 2 including the two animals with severe complications. Between pre- and post-procedure, WBC increased about twofold in the uneventful cases but fourfold in the two animals with severe complications. Cultures from the abdominal cavity before autopsy were negative in all but one animal. CONCLUSION: Animal behavior was a reliable indicator of severe complications. Fever, body weight, and the results of in vitro cultures of the peritoneal fluid did not indicate complications. WBC proved not to be specific but showed a larger increase in pigs with severe complications.

Successful resection of a congenital superior mesenteric arteriovenous malformation.

A 15-month-old male presented with severe gastrointestinal bleeding and heart failure. Imaging revealed a superior mesenteric artery arteriovenous malformation, associated with a congenital portosystemic shunt. The heart failure was cured by resection of the arteriovenous malformation.

Pharmacokinetic evaluation of gemcitabine and 2',2'-difluorodeoxycytidine-5'-triphosphate after prolonged infusion in patients affected by different solid tumors.

BACKGROUND: The study determined pharmacokinetic parameters, toxicity profile and preliminary clinical activity of gemcitabine administered i.v. at different infusion rates in patients with a range of solid tumors. PATIENTS AND METHODS: Twenty patients were enrolled for both pharmacokinetic and clinical studies. Gemcitabine 300 mg/m(2) was administered during 1 h, 2 h or 3 h, and as a conventional dose of 1000 mg/m(2) during 30 min infusion. Administration was on days 1, 8 and 15 every 4 weeks. RESULTS: Patients were randomly assigned to one of the four arms. After 30 min infusion of 1000 mg/m(2) gemcitabine the plasma concentration remained above the saturation level of 10-20 microM, whereas after 1, 2 or 3 h infusion 300 mg/m(2) gemcitabine it remained below the saturation level for most of the time (being in the range 2.5-10 microM). Gemcitabine triphosphate was determined in the four arms in white blood cells; for infusion times from 0.5 to 3 h there was a progressive enhancement of gemcitabine triphosphate levels. In all evaluable patients the toxicity was mild, myelosuppression being the main toxicity. No grade 3 or 4 toxicities occurred. Clinical response was similar in patients receiving 300 mg/m(2) gemcitabine in 2 and 3 h and in the 1000 mg/m(2) arm. CONCLUSIONS: 300 mg/m(2) gemcitabine during 3 h infusion produced the highest accumulation of gemcitabine triphosphate. Thus, to achieve the highest possible gemcitabine triphosphate level, prolonged infusion time would appear to be more important than a high dose administered as a short infusion. However, there was no substantial difference in toxicity or antitumoral activity in the all different patient groups.

Systemic vasculitis: a cause of indeterminate intestinal inflammation.

OBJECTIVES: Indeterminate intestinal inflammation may result from a variety of inflammatory conditions in addition to ulcerative colitis and Crohn disease. The primary systemic vasculitides may present with intestinal inflammation and an indeterminate colitis. We set out to describe a series of children with primary systemic vasculitis who initially presented with clinical features suggestive of inflammatory bowel disease (IBD) to establish criteria that might help discriminate between IBD and primary systemic vasculitis. METHODS: Ten children (6 boys, median age at presentation 8.9 years, range 0.9-14.5 years) satisfied inclusion criteria. RESULTS: All had abdominal pain, weight loss, diarrhea (6 of 10 bloody) and laboratory evidence of a severe acute phase response. Extraintestinal clinical features included vasculitic rash, renal impairment, myalgia, testicular pain and polyarthritis. Endoscopy showed vascular changes or other macroscopic findings suggestive of vasculitis in 5 of 10 patients. Gut histology revealed indeterminate chronic inflammatory mucosal changes and one patient with small artery fibrinoid necrosis in the submucosal vessels. Extraintestinal biopsy was performed in 6 patients and had a higher yield for the demonstration of vasculitis than intestinal biopsy. The results of selective visceral angiography was suggestive of vasculitis in all patients, but was normal in 7 cases of treatment-unresponsive classic IBD. Treatment comprised corticosteroid and azathioprine in all patients. Cyclophosphamide was given to 7 of 10 patients. CONCLUSIONS: Extraintestinal manifestations and inflammatory responses that may be disproportionate to the degree of intestinal inflammation provide clues to the presence of an underlying primary systemic vasculitis, and these data suggest that selective visceral angiography plays a key role in the diagnosis of vasculitis in this context. It is important to identify and treat any vasculitic component because failure to do so may result in consequential morbidity or mortality.

CathCam guide wire-directed colonoscopy: first pilot study in patients with a previous incomplete colonoscopy.

BACKGROUND AND STUDY AIMS: Conventional colonoscopy as the gold standard for large-bowel diagnostics and therapy may fail in 5 %-20 % of cases, depending on the experience of the examiner. Colonoscopy is regarded as difficult and painful by many patients. In an attempt to overcome the limitations of conventional colonoscopes, a guide wire-directed, thin, flexible diagnostic colonoscope, the CathCam was developed. In this prospective pilot study, we report its use in patients in whom conventional colonoscopy had failed. PATIENTS AND METHODS: 49 patients with a previous or current failure of complete colonoscopy were invited to participate in a trial using the new CathCam system, and 14 (nine men; mean age 59 years) accepted. The CathCam is an 11-mm diameter disposable, multilumen catheter, with visualization by a 3-mm camera with six light-emitting diodes. In the first five patients, the CathCam was inserted over a newly developed 0.024-inch, hinged, lumen-seeking guide wire. Subsequently, a modified combined approach was used: a conventional colonoscope was introduced into the sigmoid or left colon, then the guide wire was advanced as far as possible, followed by CathCam insertion over it. Caecal intubation rate, insertion times and patient discomfort were recorded; patients received low-dose midazolam sedation (2-5 mg). RESULTS: One patient was excluded during colonoscopy. The caecum could be eventually reached in 12 of 13 patients; in the remaining patient a significant sigmoid stricture could be passed, but further advancement appeared too risky. The mean caecal intubation time was 24 minutes (range 3-105 min). Only two patients experienced pain and discomfort during the procedure (one immediate assessment and one case reported at later telephone interview). No complications occurred, and previously undiagnosed important findings were obtained in 9 cases. CONCLUSIONS: A combined approach, consisting of guide wire insertion via a partially introduced colonoscope followed by CathCam or colonoscope insertion into the caecum was successful in over 90 % of patients with previous failure of complete colonoscopy. Further improvements may make this system suitable for use as a standard diagnostic colonoscope, either as a single unit (CathCam plus guide wire) or using the guide wire alone with a standard colonoscope in difficult cases.

Management of fulminating ulcerative colitis in childhood with chimeric anti-CD25 antibody.

Fulminating acute ulcerative colitis (UC) is a potentially life threatening medical emergency. Up to 30% of individuals respond poorly to corticosteroids alone and second line medical or surgical therapies are indicated. We describe the successful use of chimeric anti-CD25 therapy in 4 such children poorly responsive to combined therapy with intravenous steroids and calcineurin inhibitors with a pretreatment predictive risk of colectomy of 85-100%. Clinical disease activity scores normalized within 72 hours of anti-CD25 administration and colonic histology provided evidence of mucosal healing within 10-14 days. None required emergency colectomy. Anti-CD25 is efficacious in fulminating UC and randomized placebo controlled trials appear indicated.

Consumerism in healthcare can be detrimental to child health: lessons from children with functional abdominal pain.

AIMS: To determine prognostic indicators in children with severe functional abdominal pain (FAP) and to test the hypothesis that "healthcare consumerism" in these families might be deleterious to the child. METHODS: Retrospective analysis of a cohort of 23 children aged <16 years fulfilling the Rome II diagnostic criteria for FAP during the period December 1997 to February 2001. Poor outcome was defined as continued pain and failure to return to normal functioning >12 months after onset. RESULTS: Poor outcome was associated with refusal to engage with psychological services, involvement of more than three consultants, lodging of a manipulative complaint with hospital management by the child's family, and lack of development of insight into psychosocial influences on symptoms. Three of four adverse prognostic indicators reflected healthcare consumerism by the families. CONCLUSIONS: Actions of families who lack insight into their child's illness may perpetuate FAP in childhood. A culture of parental consumerism in healthcare, however well intentioned, needs to be accompanied by robust systems to protect the interests of the child.

Hypertrophic eosinophilic gastroenteropathy is associated with reduced enterocyte apoptosis.

AIMS: To investigate the cause of grossly elongated villi in four children presenting with obstruction due to a novel form of eosinophilic gastroenteropathy in which there was profound hyperplasia of the intestinal villi with grossly increased villous/crypt ratio and prominent mucosal eosinophilia. Increased eosinophils were also present in the muscularis propria and submucosa. All had intermittent diarrhoea and signs of a protein-losing enteropathy. METHODS AND RESULTS: The cause of the grossly elongated villi was investigated by studying enterocyte proliferation (Ki67), survival factors (bcl-2) and apoptosis (TUNEL) in these patients (n = 4) and normal (jejunum n = 6, ileum n = 6) and disease (n = 6) controls. The most remarkable finding was that apoptotic enterocytes were undetectable in the elongated villi. CONCLUSIONS: It seems likely that a defect in the regulation of apoptosis of the epithelium occurs which could explain the remarkable hyperplasia of the villi seen.

The influence of Hox genes and three intercellular signalling pathways on enteric neuromuscular development.

Normal intestinal motility requires orderly development of the complex nerve plexuses and smooth muscular layers in the gut wall. Organization of these structures results, in part, from cell autonomous programmes directed by transcription factors, which orchestrate appropriate temporal and spatial expression of specific target genes. Hox proteins appear to function in combination to dictate regional codes that establish major structural landmarks in the gut such as sphincters and muscle layers. These codes are translated in part by intercellular signals, which allow populations of cells in the embryonic gut wall to alter the developmental fate of their neighbours. Some of the best characterized intercellular signalling pathways involved in enteric neurodevelopment are mediated by GDNF/GFRa1/RET, EDN3/ENDRB, and NETRINS/DCC. These signals affect enteric neural precursors as they colonize the gut, and perturbations of these molecules are associated with various types of intestinal neuropathology.

[Indirect calorimetry in critical ill patients: validity of measurement for ten minutes]. / Calorimetría indirecta en el enfermo crítico: validez de la medición durante 10 minutos.

GOAL: There are no gold standards on the duration and frequency of the measurement of indirect calorimetry, a fact of importance in daily clinical practice. An assessment of is made of the degree of concordance between energy expenditure at rest (EER) measured over a short interval (10 minutes) versus another prolonged measurement (1 hour). PATIENTS: Sixty critically-ill patients, under sedation and analgesia with connection to mechanical ventilation, were studied. INTERVENTIONS: EER values were determined by means of a metabolic computer analysis (Engström Eliza) at rest. The reproducibility and the degree of concordance were assessed in the measurements made with both periods. RESULTS: The mean values of the EER determinations at 10 and 60 minutes were 1,818 +/- 319 kilocalories/day and 1,815 +/- 318 Kcal/day. The limits of the concordance between both times were -101 and +117 kilocalories/day and the correlation was significant (r = 0.98, p < 0.0001). CONCLUSIONS: In critically-ill patients under sedation and with mechanical ventilation, the measurement of EER may be taken over short periods of time (10 minutes) providing that baseline examination conditions are met, thus giving greater availability of the resources used to study indirect calorimetry.

In yeast sterol biosynthesis the 3-keto reductase protein (Erg27p) is required for oxidosqualene cyclase (Erg7p) activity.

In Saccharomyces cerevisiae, the 3-keto reductase (Erg27p) encoded by ERG27 gene is one of the key enzymes involved in the C-4 demethylation of the sterol intermediate, 4,4-dimethylzymosterol. The oxidosqualene cyclase (Erg7p) encoded by the ERG7 gene converts oxidosqualene to lanosterol, the first cyclic component of sterol biosynthesis. In a previous study, we found that erg27 strains grown on cholesterol- or ergosterol-supplemented media did not accumulate lanosterol or 3-ketosterols but rather squalene, oxidosqualene, and dioxidosqualene intermediates normally observed in ERG7 (oxidosqualene cyclase) mutants. These results suggested a possible interaction between these two enzymes. In this study, we present evidence that Erg27p interacts with Erg7p, facilitating the association of Erg7p with lipid particles (LPs) and preventing digestion of Erg7p both in the endoplasmic reticulum (ER) and LPs. We demonstrate that Erg27p is required for oxidosqualene cyclase (Erg7p) activity in LPs, and that Erg27p co-immunoprecipitates with Erg7p in LPs but not in microsomal fractions. While Erg27p is essentially a component of the ER, it can also be detected in LPs. In erg27 strains, a truncated Erg7p mislocalizes to microsomes. Restoration of Erg7p enzyme activity and LPs localization was achieved in an erg27 strain transformed with a plasmid containing a wild-type ERG27 allele. We suggest that the physical interaction of Erg27p with Erg7p is an essential regulatory tool in yeast sterol biosynthesis.

Eosinophilic myenteric ganglionitis is associated with functional intestinal obstruction.

The diagnostic features and clinical course of three children (aged 1 month to 15 years) with severe functional intestinal obstruction and inflammation of the colonic lamina propria and myenteric plexus are described. The myenteric inflammatory infiltrate was eosinophil predominant with none of the immunological characteristics of lymphocytic ganglionitis. Neurones in the myenteric ganglia expressed the potent eosinophil chemoattractant interleukin 5. None responded to dietary exclusion but all three responded symptomatically to immunosuppression/anti-inflammatory treatments. Eosinophilic ganglionitis is associated with a pseudo-obstructive syndrome which is amenable to anti-inflammatory treatment.

The histological appearances of Nissen-type fundoplication in the ferret.

Nissen fundoplication is of proven effectiveness in the surgical control of gastro-oesophageal reflux. However, our understanding of the effects of fundoplication upon foregut physiology is incomplete and post-operative symptoms are often poorly understood. This experimental study aimed systematically to characterize the tissue response to fundoplication in an animal model, to improve understanding of the effects of anti-reflux surgery upon foregut physiology. Nissen-type fundoplication was performed in the ferret, and the tissue response at 3 months examined histologically. Sham-operated animals that underwent laparotomy but no dissection or wrap, acted as controls. In fundoplicated animals, serosal fibrosis was observed in the gut wall, with patchy replacement of muscle by fibrous tissue. The ventral and dorsal vagal nerve trunks were identified intact within the wrap. In cases where the wrap had spontaneously disrupted, fibrosis was more extensive and there was evidence of nerve damage. This is the first systematic description of the histopathological response to Nissen fundoplication. In the intact wrap, the vagal trunks appear spared, but there is fibrosis in the serosa, extending into the muscularis of the distal oesophagus and region of the cardia. These findings are discussed in relation to the effects of Nissen fundoplication upon gastric physiology and postoperative symptoms.