Peritoneal fluid cytokines and matrix metalloproteinases as early markers of anastomotic leakage in colorectal anastomosis: a literature review and meta-analysis.

AIM: There has been recent interest in the measurement of peritoneal fluid cytokines in the early postoperative period to help diagnose anastomotic leakage at a preclinical stage. The currently available literature on the early diagnosis of colorectal anastomotic leakage by estimation of drain fluid cytokines or matrix metalloproteinases (MMP) is reviewed. METHOD: A literature search was performed in PubMed, EMBASE and the Cochrane library for all publications studying the feasibility of diagnosing colorectal anastomotic leakage earlier by estimation of peritoneal fluid cytokine or MMP levels. A meta-analysis of the most commonly measured cytokines was performed. RESULTS: Eight publications were included. Tumour necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were most frequently studied. Most studies found significantly higher levels of TNF-α and IL-6 in patients with anastomotic leakage during the first three postoperative days. In the meta-analysis IL-6 levels were significantly higher from day 1 and TNF-α from day 2. MMP-9 was most often significantly elevated in patients with anastomotic leakage. CONCLUSION: Measurement of drain fluid cytokines and MMP has the potential to diagnose colorectal anastomotic leakage at a preclinical stage, but is not yet ready for clinical use. Further research is needed, possibly using IL-6 in combination with other cytokines and MMP as markers.

Transanal single port access to facilitate distal rectal mobilization in laparoscopic rectal sleeve resection with hand-sewn coloanal anastomosis.

BACKGROUND: Laparoscopic rectal sleeve resection is challenging and technically demanding. Exposure and mobilization of the most distal part of the rectum can be especially hazardous. We propose the use of a single port access device, placed in the anal canal after incision of the sleeve at the appropriate level, to facilitate dissection without sphincter damage. The case of a 51-year-old woman suffering from a recurrent supralevator abscess is presented to illustrate the technique. METHODS: The procedure consisted of laparoscopic rectal pull-through with rectal sleeve resection and coloanal anastomosis. Incision of the endopelvic fascia and mobilization of the distal mesorectum was performed via the single port device under direct control. Medial-to-lateral mobilization of the colon was performed with a 3-port technique. RESULTS: Total operating time was 122 min: 50 min for rectal mobilization, 42 min for the laparoscopic part of the procedure and 30 min for the coloanal anastomosis. The patient's recovery was uneventful, and at 1-month follow-up, she was asymptomatic. CONCLUSIONS: Laparoscopic-assisted transanal single port rectal mobilization seems to be a promising addition to the armamentarium of minimally invasive surgery.

Redox proteomics in aging rat brain: involvement of mitochondrial reduced glutathione status and mitochondrial protein oxidation in the aging process.

Increasing evidence supports the notion that increased oxidative stress is a fundamental cause in the aging process and in neurodegenerative diseases. As a result, a decline in cognitive function is generally associated with brain aging. Reactive oxygen species (ROS) are highly reactive intermediates, which can modify proteins, nucleic acids, and polyunsaturated fatty acids, leading to neuronal damage. Because proteins are major components of biological systems and play key roles in a variety of cellular functions, oxidative damage to proteins represents a primary event observed in aging and age-related neurodegenerative disorders. In the present study, with a redox proteomics approach, we identified mitochondrial oxidatively modified proteins as a function of brain aging, specifically in those brain regions, such as cortex and hippocampus, that are commonly affected by the aging process. In all brain regions examined, many of the identified proteins were energy-related, such as pyruvate kinase, ATP synthase, aldolase, creatine kinase, and α-enolase. These alterations were associated with significant changes in both cytosolic and mitochondrial redox status in all brain regions analyzed. Our finding is in line with current literature postulating that free radical damage and decreased energy production are characteristic hallmarks of the aging process. In additon, our results further contribute to identifying common pathological pathways involved both in aging and in neurodegenerative disease development.

Protective effect of ferulic acid ethyl ester against oxidative stress mediated by UVB irradiation in human epidermal melanocytes.

UV solar radiation is the major environmental risk factor for malignant melanoma. A great effort is currently posed on the search of new compounds able to prevent or reduce UV-mediated cell damage. Ferulic acid is a natural compound recently included in the formulation of solar protecting dermatological products. The purpose of the present work was to assess whether its ethyl ester derivative, FAEE, could protect skin melanocytes from UV-induced oxidative stress and cell damage. Experiments on human melanocytes irradiated with UVB showed that FAEE treatment reduced the generation of ROS, with a net decrease of protein oxidation. FAEE treatment was accompanied by an induction of HSP70 and heme oxygenase, by a marked suppression of PARP activation and a significant suppression of apoptosis. Moreover FAEE prevented iNOS induction, thus suppressing the secondary generation of NO-derived oxidizing agents. FAEE may represent a potentially effective pharmacological approach to reduce UV radiation-induced skin damage.

The Hammersmith functional score correlates with the SMN2 copy number: a multicentric study.

Previous studies showed that SMN2 copy number correlates inversely with the disease severity. Our aim was to evaluate SMN2 copy numbers and the Hammersmith functional motor scale in 87 patients with SMA II in order to establish whether, within SMAII, the number of copies correlates with the severity of functional impairment. Our results showed a relative variability of functional scores, but a significant correlation between the number of SMN2 genes and the level of function.

Randomized, double-blind, placebo-controlled trial of phenylbutyrate in spinal muscular atrophy.

OBJECTIVE: To assess the efficacy of phenylbutyrate (PB) in patients with spinal muscular atrophy in a randomized, double-blind, placebo-controlled trial involving 10 Italian centers. METHODS: One hundred seven children were assigned to receive PB (500 mg/kg/day) or matching placebo on an intermittent regimen (7 days on/7 days off) for 13 weeks. The Hammersmith functional motor scale (primary outcome measure), myometry, and forced vital capacity were assessed at baseline and at weeks 5 and 13. RESULTS: Between January and September 2004, 107 patients aged 30 to 154 months were enrolled. PB was well tolerated, with only one child withdrawing because of adverse events. Mean improvement in functional score was 0.60 in the PB arm and 0.73 in placebo arm (p = 0.70). Changes in the secondary endpoints were also similar in the two study arms. CONCLUSIONS: Phenylbutyrate was not effective at the regimen, schedule, and duration used in this study.

Reliability of the Hammersmith functional motor scale for spinal muscular atrophy in a multicentric study.

The aim of this study was to validate the Hammersmith functional motor scale for children with spinal muscular atrophy in a large cohort of 90 non-ambulant children with spinal muscular atrophy type 2 or 3. All had a baseline assessment (T0) and were reassessed either at 3 months (T1) (n = 66) or at 6 months (T2) (n = 24). Inter-observer reliability, tested on 13 children among 3 examiners, was > 95%. Of the 66 children examined after 3 months 4 had adverse effects in between assessments and were excluded from the analysis. Forty-two (68%) of the remaining 62 reassessed had no variation in scores between T0 and T1 and 13 (21%) were within +/- 1 point. 9 (37.5%) of the 24 children reassessed after 6 months had no variation in scores between T0 and T2 and another 9 (37.5%) had variations within +/- 1 point. Our study confirms previous observations of the reliability of the scale and helps to establish a baseline for assessing changes of functional ability over 3 and 6 month intervals. This information can be valuable in view of therapeutic trials.

Congenital form of spinal muscular atrophy predominantly affecting the lower limbs: a clinical and muscle MRI study.

We describe clinical and muscle magnetic resonance imaging (MRI) findings in 11 cases (three familial and eight sporadic) with the form of spinal muscular atrophy characterised by predominant involvement of the lower limbs with weakness of the proximal and distal muscles and marked atrophy of the distal leg and foot muscles. All patients presented at birth with talipes, which were in extension in seven of the 11. Arm muscle and function were preserved and lower limbs appeared to be disproportionately shorter compared to trunk and upper limbs. Functional abilities were markedly affected and only one of the 11 is able to walk independently for long distances, while six require support of crutches and two use callipers for walking. One child lost ambulation following a fall. The course of the disease is relatively stable and the progression of disability appeared to be related mostly to increased contractures rather than to loss of muscle strength. Respiratory and cardiac function were well preserved. A neurogenic disorder was suggested by electromyography and/or muscle biopsy in all patients, while motor nerve conduction was consistently normal. Muscle MRI of the thighs revealed diffuse atrophic appearance with relative hypertrophy of the adductor longus and of the semitendinosus. Genetic studies excluded the involvement of the survival motor neuron gene but none of these families was sufficiently informative to study linkage to the locus on chromosome 12q23-q24 previously found to be involved in patients with similar phenotype. In our experience this form of spinal muscular atrophy affecting predominantly the lower limbs is a relatively common form and should be considered in the differential diagnosis of infants with talipes and weakness in the lower limbs. The identical clinical and imaging features of the sporadic and familial cases suggest that these cases are likely to be affected by the same condition.

Muscle magnetic resonance imaging in patients with congenital muscular dystrophy and Ullrich phenotype.

The aim of this study was to evaluate muscle magnetic resonance imaging findings in patients with congenital muscular dystrophy and Ullrich phenotype. Fifteen children with congenital muscular dystrophy and Ullrich phenotype were included in the study. All patients had collagen VI studies in muscle and, when family structure was informative, linkage studies to the collagen 6 loci. Three of the 15 patients had reduced collagen in muscle. One of the three was from an informative family and linked to one of the collagen 6 loci. Another patient was linked to one of the collagen 6 loci but had normal expression of collagen in muscle. The remaining 11 all had normal collagen expression in muscle. Only two of these 11 were from informative families and linkage to collagen 6 loci was excluded in them. All patients had muscle magnetic resonance imaging of their leg muscles using transverse T1 sequences. With the exception of the two patients in whom linkage to the collagen 6 loci was excluded, the other 13 patients showed the same pattern of selective involvement on magnetic resonance imaging of thigh muscles. This consisted of relative sparing of sartorius, gracilis, adductor longus and rectus. This pattern was also found in the case linked COL6A1/A2 locus but with normal collagen. This finding, and the striking clinical and magnetic resonance imaging concordance between patients with normal and reduced collagen VI in muscle suggest that collagen VI could still be the culprit in several cases with normal collagen expression, or alternatively a primary defect in a protein that closely interacts with collagen VI. Mutation analysis of the collagen 6 genes in cases with normal collagen VI expression is needed to resolve this issue.

Assessment of water pollution and suitability to fish life in six Italian rivers.

This paper reports the results of a monitoring programme of six rivers (Serchio, Cecina, Cascina, ERa, Elsa and Pavone) that drain north-west Tuscany (Italy). In this area, agricultural, industrial and urbanization pressures affect water quality, also in terms of suitability for the survival of aquatic organisms. The river waters were monitored for several physico-chemical parameters (temperature, dissolved oxygen, suspended solids, total ammonia, nitrites, Cu, Cd, Cr, Ni and Pb) in order to assess the anthropogenic impact on the environmental conditions of these watercourses and their suitability for fish life. Statistical data handling was performed for each river and the percentage of physico-chemical parameters that, according to the E.C. law, were over the limits that must not be exceeded to protect fish life, was calculated and a classification was made of the six rivers in this sense. Concerning the metals, inter-element relationships (between chromium and nichel, lead and copper, nichel and lead and copper, copper and lead) were found which allow some remarks on the origins of the anthropogenic impacts, and comparisons were made with two metal-polluted watercourses in northern Italy, the Po and Lambro rivers. From the comparison, the Tuscan rivers showed generally lower metal contamination, especially with respect to the Lambro.

Reproductive male-mediated risk: spontaneous abortion among wives of pesticide applicators.

This study was conducted among 32 pesticide applicators occupationally exposed to pesticides to determine whether paternal exposure is associated with an increased risk of spontaneous abortion. The ratio of abortion was compared between applicators and a group of 51 food retailers (control population). The ratio of abortions/pregnancies for applicators was 0.27 and for retailers 0.07. OR for spontaneous abortion adjusted for age of wife and smoking of parents is 3.8 times greater than for the control population in the multiple logistic regression model and 7.6 times with interaction effects model.

Cysteinyldopaenkephalins: synthesis, characterization and binding to bovine brain opioid receptors.

The reaction of opioid peptides with mushroom tyrosinase in the presence of an excess of a thiol compound gives rise to cysteinyldopaenkephalins (CDEnks). The major product is represented by the 5-S-CDEnk (80%) and the minor one by the isomer 2-S-CDEnk (20%). The adducts between leucine-enkephalin (Leu-enk) and cysteine have been isolated by high performance liquid chromatography (HPLC) and identified by amino acid analysis and electrospray ion mass spectrometry. 5-S-CDEnk is able to bind to opioid receptors in bovine brain membranes. Its binding affinity is higher for delta than for mu receptors and about 8-fold lesser than that exploited by Leu-enk. In the presence of the peroxidase/H(2)O(2) system, CDEnks can be converted into the corresponding pheo-opiomelanins.

Production of melanin pigments by cytochrome c/H2O2 system.

In the presence of hydrogen peroxide cytochrome c can perform the oxidation of catecholamines and their S-cysteinyl-derivatives yielding melanins as final products. The initial reaction rate is linearly dependent on cytochrome c and H2O2 concentration; the reaction follows the Michaelis and Menten kinetics both for H2O2 and hydrogen donors. Sulfhydryl compounds inhibit the formation of the pigment. The reported data indicate that a heme-containing protein belonging to the mitochondrial chain can accelerate the oxidation of catecholamines to eumelanins.

Catecholamines oxidation by xanthine oxidase.

Dopamine and structurally related catecholamines in the presence of hydrogen peroxide are oxidized in vitro by xanthine oxidase producing the corresponding melanin pigments. The kinetic parameters of the reaction, measured as aminochrome formation, have been calculated. The rate of peroxidation depends on enzyme and hydrogen peroxide concentration. The optimum pH for the peroxidative activity of the enzyme is around 8.5. Activation of the peroxidative reaction is also elicited by catechol compounds through a redox cycle mechanism. Implications about the possible biochemical relevance of xanthine oxidase activity on catecholamines oxidation are discussed.

Effects of prolonged autovehicle driving on male reproduction function: a study among taxi drivers.

This study had the purpose of exploring the possible association between the work exposures of professional drivers and their reproductive health, by studying a group of 201 taxi drivers in the city of Rome. Data on work and reproductive history were collected by interviews. Biological markers examined in 72 subjects included salivary testosterone levels, sperm quality (i.e., sperm concentration, sperm morphology, and motility), and fertility experience, including time to pregnancy. Their spermatologic profile was compared with that of a control group of 50 healthy subjects of similar age and smoking habits. The results showed that taxi drivers, compared to the controls, had a significantly lower prevalence of normal sperm forms (45.8% vs. 64.0%); this was particularly true for those with a longer time on this job. This result was confirmed by a multivariate analysis in which confounders such as age, smoking, and alcohol consumption were controlled. The other sperm parameters did not differ in the study and the control groups. Among the life-style factors, we found smoking to be associated with poorer sperm morphology. Moderate alcohol consumption was associated with a better seminologic profile, while the pattern in respect to coffee intake was inconclusive. Subjects with poor semen quality also more frequently exhibited longer time to pregnancy of their partner. The results suggest that prolonged urban automobile driving might be a risk factors for sperm quality, and particularly for sperm morphology, but the finding needs further confirmation.

Delta-aminolevulinic acid dehydratase and zinc protoporphyrin in very low lead-exposed pets: a community study.

Markers of lead intoxication have been developed based on their capacity to identify lead intoxication at the preclinical, ie biochemical stage of manifestation. However, little information on these markers is available under conditions of low lead exposure. This prompted us to conduct a community-based study to determine the usefulness of theta-aminolevulinic acid dehydratase (ALAD) and zinc protoporphyrin (ZnPP) in conditions of low lead environmental exposure by studying the relationships between low blood lead levels, ALAD and ZnPP in a large group of healthy dogs living in an Italian urban area. The study population consisted of 79 dogs. Each sample was tested for ALAD, lead and ZnPP and for complete blood count, hemoglobin, AST, ALT, and urea values. A weak inverse relationship between ALAD and ZnPP was found. An inverse relationship between ALAD and lead concentrations was found in the whole group (p < 0.0005). This relationship remained significant when selecting the values falling between 2 standard deviations of the mean blood lead concentrations of the population below the "concerned lead levels" (< 10 mg/dl; p = 0.0005). There was no relationship between whole blood ZnPP concentrations and whole blood lead levels. The sensitivity and specificity of ALAD measurements, calculated by using the 2 x 2 contingency table with respect to blood lead concentrations, were of poor predictive diagnostic value.

4,6-O-benzylidene-D-glucopyranose and its sodium salt: new data on their preparation and properties.

An improved method for the preparation of 4,6-O-benzylidene-D-glucopyranose (BG), and new or correlated data on its 1H and 13C NMR spectra, specific rotations, and tautomeric equilibria, and on those of its anomeric sodium salt (BGNa), are reported. Evidence is presented in favour of the hypothesis that crystalline BGNa exists entirely in its beta-anomeric form and that it can be useful in the access to beta-glucosides in reactions with strong electrophiles under strictly heterogeneous conditions.

On the oxidation of cystathionamine and selenocystathionamine by plant amineoxidase.

Cystathionamine and selenocystathionamine, diamines analogous to 1,6-diaminohexane but having the third methylene group of the carbon chain substituted by a S or a Se atom, are asymmetrical thio- (seleno-) ethers. They can give rise by oxidative monodeamination to two different aminoaldehydes. It has been shown that lentil seedlings amineoxidase catalyzes the oxidative deamination of either the one or the other aminogroup of cystathionamine or of selenocystathionamine, giving rise to both possible aminoaldehydes.