A robust and versatile mass spectrometry platform for comprehensive assessment of the thiol redox metabolome.

Several diseases are associated with perturbations in redox signaling and aberrant hydrogen sulfide metabolism, and numerous analytical methods exist for the measurement of the sulfur-containing species affected. However, uncertainty remains about their concentrations and speciation in cells/biofluids, perhaps in part due to differences in sample processing and detection principles. Using ultrahigh-performance liquid chromatography in combination with electrospray-ionization tandem mass spectrometry we here outline a specific and sensitive platform for the simultaneous measurement of 12 analytes, including total and free thiols, their disulfides and sulfide in complex biological matrices such as blood, saliva and urine. Total assay run time is < 10 min, enabling high-throughput analysis. Enhanced sensitivity and avoidance of artifactual thiol oxidation is achieved by taking advantage of the rapid reaction of sulfhydryl groups with N-ethylmaleimide. We optimized the analytical procedure for detection and separation conditions, linearity and precision including three stable isotope labelled standards. Its versatility for future more comprehensive coverage of the thiol redox metabolome was demonstrated by implementing additional analytes such as methanethiol, N-acetylcysteine, and coenzyme A. Apparent plasma sulfide concentrations were found to vary substantially with sample pretreatment and nature of the alkylating agent. In addition to protein binding in the form of mixed disulfides (S-thiolation) a significant fraction of aminothiols and sulfide appears to be also non-covalently associated with proteins. Methodological accuracy was tested by comparing the plasma redox status of 10 healthy human volunteers to a well-established protocol optimized for reduced/oxidized glutathione. In a proof-of-principle study a deeper analysis of the thiol redox metabolome including free reduced/oxidized as well as bound thiols and sulfide was performed. Additional determination of acid-labile sulfide/thiols was demonstrated in human blood cells, urine and saliva. Using this simplified mass spectrometry-based workflow the thiol redox metabolome can be determined in samples from clinical and translational studies, providing a novel prognostic/diagnostic platform for patient stratification, drug monitoring, and identification of new therapeutic approaches in redox diseases.

[Pseudomyxoma peritonei with appendiceal and ovarian mucinous cystadenoma. A case report. Correlations with synchronous or metachronous colorectal carcinomas]. / Pseudomixoma peritonei con cistoadenoma mucinoso appendicolare ed ovarico. Contributo clinico. Rapporti con i carcinomi colo-rettali sincroni o metacroni.

Mucocele is a rare pathology which is difficult to diagnose prior to surgery. The clinical symptoms are aspecific. Diagnosis is generally made at the time of surgery. Explorative laparoscopy is not advised owing to the possibility of encouraging metastatic diffusion. Surgery associated with ex tempore freezer tests must be carried out with the greatest caution to avoid rupture of organs and the spread of mucin. Appendectomy and removal of the adnexa is recommended in benign forms, with or without omentectomy. Omentectomy is necessary together with hystero-adnexectomy and right hemicolectomy in malignant forms, as well as lymph node cleaning and removal of all mucin in pseudomyxoma peritonei.

[Low ileorectal anastomosis in the surgery of ulcerative rectocolitis]. / L'ileorettoanastomosi bassa nella chirurgia della rettocolite ulcerosa.

The authors report their experience of 23 patients with ulcerative colitis treated with surgery from 1991 to 1994. At admission 8 patients had a high-grade illness, 10 patients had a middle-grade illness, 5 patients had a toxic megacolon. All the patients were treated with systemic medical management (mesalazine + methylprednisolone) and topical medical management with rectal steroid (methylprednisolone) and rectal mesalazine by enema, and with nutritional support (Total Parenteral Nutrition). The 5 patients with toxic megacolon, after 48 hours of unsuccessful medical management, underwent surgery with deferred urgency; the other 18 patients underwent surgery after one-four months. The authors prefer the ileorectal anastomosis (IRA), since the rectal lesions are more susceptible to topical therapy. A single-stage IRA was performed in 17 patients. A sigmoid resection has been employed in the first stage in the 18th patient with a local development of the disease in the sigma; a following relapse of the lesions required a total colectomy with a low ileorectal anastomosis. A first-stage subtotal colectomy with ileostomy and Hartmann closure of the rectum with low ileorectal anastomosis at a later date was performed in the 5 patients with toxic megacolon. One of these, 18 years old, died after a heart failure. After surgery, as soon as the patients start moving their bowels all of these had again a systemic therapy with mesalazine for a short period and after they had a topical therapy with mesalazine + methylprednisolone by enema, for a long period. The average postoperative period of admission was 16.3 days. The functional results have been encouraging with an average of 1.6 bowel movements daily and an average of 0.4 nocturnal bowel movements. All the patients had a normal anal sphincter function with an acceptable stool frequency. In male patients there no urinary or sexual defects. In one case of these, there was a low grade of dysplasia, revealed by endoscopic biopsy. The authors conclude that today surgical treatment of ulcerative colitis is not well established.

[Heredity and colorectal cancer]. / Familiarità e cancro colorettale.

The authors underline the role of inheritance in colorectal cancer and reporting the genealogical study of two families with high incidence of colorectal cancers personally observed. They thought that the genetic endowment (oncogenes, suppressor genes, microsatellite instability, protein-kinase C) was implicated directly in the genesis of disease.

[The utility of isolating the recurrent nerve and parathyroids in the prevention of complications in thyroid surgery. The role of the superior laryngeal nerve and C cells]. / Utilità dell'isolamento del nervo ricorrente e delle paratiroidi nella prevenzione delle complicanze in chirurgia tiroidea. Il ruolo del laringeo superiore e delle cellule C.

The authors emphasize systematic isolation of the recurrent nerve and of the parathyroids, to prevent complications in surgery of the thyroid. Moreover they try to underline the significance of the superior laryngeal nerve and the role of parafollicular cells (today unknown) for a rational and modern therapy of the non-neoplastic diseases of the thyroid.

Protective effect of zinc acexamate on experimental gastric ulcers: a histochemical study.

The antiulcerogenic effect of zinc acexamate on gastric ulcers induced by reserpine and changes in the morphology of gastric mucosa were studied in rats by histochemical methods. Histochemistry revealed that zinc acexamate preserved reserpine-depleted neutral and acid glycoproteins. ATPase reaction remained strong in nearly normal periglandular capillaries. The reaction intensity of SDH and NADH2-tetrazolium reductase, and the number and size of the DH-positive parietal cells were decreased, illustrating the decline of energy metabolism involved in acid secretion. The decreased height and weaker staining of the pyroninophile chief cell layer corresponded to the lower amount of RNA, an indirect indicator of pepsinogen synthesis. The significant correlation indices "r" between the severity of gastric lesions and histochemical parameters of the defensive (glycoproteins and microvascular ATPase) and aggressive factors (parietal cell DH and chief cell RNA) confirmed the pathogenic effect of reserpine and the protection provided by zinc acexamate. These findings confirm the multifactorial mechanism of action described for zinc acexamate in several previous works.

[Surgical chemoprophylaxis with ceftriaxone]. / Chemioprofilassi chirurgica con ceftriaxone.

UNLABELLED: Ceftriaxone is a cephalosporin of third generation. It is characterized by a broad spectrum, a long half-life and a good capacity of diffusion in tissue. We have studied 67 patients. Age ratio was 44 years. 25 patients did not receive short term prophylaxis; 42 patients did (2 gr iv of ceftriaxone one hour before the operation). In clean surgery, only patients immunodepressed or malnourished received chemoprophylaxis. Patients who received 2 gr iv of ceftriaxone one hour before incision, received antibiotic therapy only on the appearance of septic complications. RESULTS: two patients submitted to chemoprophylaxis (4.8%) showed complications (cystitis and bronchial pneumonia). In the control group antibiotic therapy was undertaken only if septic complication appeared. 19 patients (76%) did not showed any complications. 6 patients (24%) showed surgical wound infection, acute pharyngo-tracheitis, cystitis. Finally, wound infections were limited exclusively to the control group. The ratio of respiratory infections was not statistically significant in the two groups; cystitis resulted more frequent in the control group than in the prophylaxis group. Prophylaxis with 2 gr single dose of ceftriaxone intravenously is effective in reducing the influence of infective complications in clean/contaminated surgery and clean risk surgery.

[Staplers in surgery: state of the art]. / Le suturatrici meccaniche in chirurgia: stato dell'arte.

After a review of the technical development and application of staplers from their introduction to the present day, the indications to the use of this instrument in all gastroenterological areas from the oesophagus to the rectum as well as in chest, gynaecological and urological surgery specified. The advantages offered by staplers in the creation of intestinal anastomoses are undeniable, but these instruments must be considered alternatives to traditional surgery, the techniques of which should be well known to all surgeons. In certain areas (gastric, ileal and colonic resections etc.), the now widespread return to single thread manual stitches sheds a new light on staplers and reflects the fact that an anastomosis can be performed just as quickly by hand but will be softer and less rigid than a stapled one. It is only in oesophageal and lower rectal surgery that staplers offer a significant advantage, solving problems where hand suturing cannot.

Ultrasonographic diagnosis of obstructive jaundice.

The diagnostic accuracy of ultrasonography in differentiating between medical and surgical jaundice (99%), and in revealing the site (80%) and nature of the obstruction (79.8%) was assessed in a study on 128 jaundiced patients. The validity of this diagnostic procedure is underlined and its use is recommended in patients with obstructive jaundice.

Effects of zinc-aspartate and zinc-glycinate in healthy rats and on reserpine-induced gastric lesions.

Experimental studies performed on 227 rats showed that Zn-aspartate and Zn-glycinate administered ip lowered the incidence, number, and severity of the reserpine-induced gastric lesions ensuring significant protection indices. Histochemical methods revealed increased amount of mucosal glycoproteins. The activity of dehydrogenases involved in energy metabolism that modulates acid secretion in the parietal cells was depressed. RNA content in the chief cells, as premises of pepsinogen synthesis, was decreased. ATPase reaction in the periglandular capillaries was uniform and stronger, showing an improvement of gastric mucosal microcirculation. Since these histochemical changes were also noted in healthy rats receiving Zn salts, it might be suggested that they are not the mere expression of an anti-ulcer protective effect of zinc, but rather reflect its mechanism of action, relating to the complex metabolic events induced by the trace element. Our results are in agreement with those previously reported concerning the noxious influence of Zn depletion, the accelerated healing of peptic ulcer patients after Zn treatment, and the protective effect of Zn against ulcerogenesis in several experimental models involving different pathomechanisms.

Short-term systems to assess thiophosphoryl chloride standards.

The rat acute oral LD50 value of PSCl3 is 750 mg per kg body weight. A rat 90-day oral feeding study at dose levels of 7.5 mg (P1) and 37.5 mg (P5) per kg resulted in increased body weight gain. Other responses inconsistently varied between dose levels. At 7.5 mg kg-1, a trend towards enhancement of relative weight of liver, spleen, kidney and adrenals was noted; at 37.5 mg kg-1, decreasing relative spleen and adrenal weights were observed. The most of the hematological and biochemical changes in blood serum cannot be believed to be transitory adaptive changes, but valid toxic effects. On the other hand, there are some indicators that suggest a possible trophic effect of this compound. However, the reversible dystrophic changes in liver, as well as the histochemical and histoenzymical results, require us to take into account that inactive chronic hepatitis could be reactivated by further toxic insult. A teratology study demonstrated increased number, body weight and length of fetuses, without apparent pathological events (tumors, external malformations). A mutagenicity test performed on rat femoral bone marrow after 30-days feeding of 37.5 mg kg-1 did not indicate any significant change in the incidence of chromosomal aberrations, compared with the control group. The recommended maximum allowable concentration in the working zone for thiophosphoryl chloride in air was set at 3 mg m-3.

Effect of silymarin on experimental liver lesions.

The effect of Silymarin (Legalon) upon liver lesions was investigated using four experimental models: In acute galactosamine-hepatitis, Silymarin administration achieved protection of the liver structure (electron-microscopy included), liver cell glycogen, RNA and enzymatic activity, Galactosamine-depressed gluconeogenesis in the isolated perfused rat liver was significantly preserved by Silymarin treatment. In lead and cadmium poisoning the structural damage and histochemical and histoenzymatic changes were partly but significantly prevented. The complex noxious effects of Imuran overdoses were favourably influenced by Silymarin, without diminishing the cytostatic-immunosuppressive action of Imuran.

[The action of silymarin on Galactosamine-induced hepatitis in the rat (author's transl)]. / Die Wirkung von Silymarin auf die experimentelle Galaktosamin-Hepatitis bei der Ratte

The hepatoprotective action of Silymarin was studied in 65 male Wistar rats, prior to and following D-galactosamine intoxication. There was a marked reduction in the histological and ultrastructural changes in the nucleolus, nuclear membrane, mitochondria, granular and agranular endoplasmic reticulum and lysosomes of the liver cell and also in the Kupffer stellate cells. The reduction in glycogen and RNA loss was determined biochemically. The activities of many enzymes were kept constant (oxidoreductases, NADH2 diaphorase, G-6-phosphatase, Mg++ and K+/Na+-dependent ATPases, acid phosphatases).

Protecting action of aspartate on the hepatic changes induced by D-galactosamine.

The protective action of aspartic acid on isolated and perfused rat liver was studied. In case of D-galactosamine intoxication the GOT, GPT and SDH activity and the lactate and pyruvate concentration in the perfusion medium were less augmented and the glycogen level in hepatic tissue was less diminished in animals treated with aspartic acid, as compared to controls. The histochemical applied (PAS reaction for glycogen, nucleic acids, NADH2-diaphorase, glucose-6-phosphatase and membrane-ATP-ase), also stated a protecting effect in the treated animals. The protective action of aspartate is hypothetically considered to be exerted by its capacity to reestablish the cellular deficit of pyridine nucleotides and thus to improve the synthesis of nucleic acids, glycoprotein and glycolipids or/and by its participation in various metabolic pathways.