Accurate calibration of a molecular beam time-of-flight mass spectrometer for on-line analysis of high molecular weight species.

RATIONALE: Time-of-Flight (TOF) Mass Spectrometry is a powerful analytical technique, provided that an accurate calibration by standard molecules in the same m/z range of the analytes is performed. Calibration in a very large m/z range is a difficult task, particularly in studies focusing on the detection of high molecular weight clusters of different molecules or high molecular weight species. METHODS: External calibration is the most common procedure used for TOF mass spectrometric analysis in the gas phase and, generally, the only available standards are made up of mixtures of noble gases, covering a small mass range for calibration, up to m/z 136 (higher mass isotope of xenon). In this work, an accurate calibration of a Molecular Beam Time-of Flight Mass Spectrometer (MB-TOFMS) is presented, based on the use of water clusters up to m/z 3000. RESULTS: The advantages of calibrating a MB-TOFMS with water clusters for the detection of analytes with masses above those of the traditional calibrants such as noble gases were quantitatively shown by statistical calculations. A comparison of the water cluster and noble gases calibration procedures in attributing the masses to a test mixture extending up to m/z 800 is also reported. In the case of the analysis of combustion products, another important feature of water cluster calibration was shown, that is the possibility of using them as "internal standard" directly formed from the combustion water, under suitable experimental conditions. CONCLUSIONS: The water clusters calibration of a MB-TOFMS gives rise to a ten-fold reduction in error compared to the traditional calibration with noble gases. The consequent improvement in mass accuracy in the calibration of a MB-TOFMS has important implications in various fields where detection of high molecular mass species is required. In combustion products analysis, it is also possible to obtain a new calibration spectrum before the acquisition of each spectrum, only modifying some operative conditions. Copyright © 2016 John Wiley & Sons, Ltd.

Multiphoton ionization of large water clusters.

Water clusters are multimers of water molecules held together by hydrogen bonds. In the present work, multiphoton ionization in the UV range coupled with time of flight mass spectrometry has been applied to water clusters with up to 160 molecules in order to obtain information on the electronic states of clusters of different sizes up to dimensions that can approximate the bulk phase. The dependence of ion intensities of water clusters and their metastable fragments produced by laser ionization at 355 nm on laser power density indicates a (3+1)-photon resonance-enhanced multiphoton ionization process. It also explains the large increase of ionization efficiency at 355 nm compared to that at 266 nm. Indeed, it was found, by applying both nanosecond and picosecond laser ionization with the two different UV wavelengths, that no water cluster sequences after n = 9 could be observed at 266 nm, whereas water clusters up to m/z 2000 Th in reflectron mode and m/z 3000 Th in linear mode were detected at 355 nm. The agreement between our findings on clusters of water, especially true in the range with n > 10, and reported data for liquid water supports the hypothesis that clusters above a critical dimension can approximate the liquid phase. It should thus be possible to study clusters just above 10 water molecules, for getting information on the bulk phase structure.

Double laparotomy wound recurrence of endometrial carcinoma.

INTRODUCTION: Abdominal scar recurrence of endometrial carcinoma after abdominal total hysterectomy is very rare. We report a case of a 65-year-old woman who had two recurrences in the abdominal incisional scar after total hysterectomy. CASE REPORT: A 65-year-old woman underwent total hysterectomy with bilateral salpingo-oophorectomy and pelvic lymphadenectomy because of well-differentiated endometrial adenocarcinoma (Stage IIB). Thus, the patient was treated by external beam radiotherapy. She developed two recurrences in the abdominal incisional scar two and three years after total hysterectomy, respectively. Surgery plus chemotherapy and surgery plus hormonal therapy were used for treatment of the first and second scar recurrence, respectively. CONCLUSIONS: It is a very intriguing and controversial biologic question how neoplastic cells can implant and grow in an abdominal scar without other concomitant metastases. We report a review of the literature and the possible mechanism of recurrences in laparotomy wounds.

Female adnexal tumor of probable wolffian origin: clinicopathological, immunohistochemical and cytofluorimetric analyses of a 22-year-old virgin. case report.

The term female adnexal tumor of probable Wolffian origin "FATWO" designs this tumor wich arises by the rare persisting remnants of the mesonephric duct (Wolffian duct). About 40 cases have been reported in literature. Few cases of recurrence have been reported, FATWO usually shows no signs of hormonal activity. We report a case of the youngest patient affected by FATWO in October 2002. At laparotomy the left adnexa were deformed by a well-capsulated mass, totally removed and sent to the pathologist with a specimen of peritoneal fluid and of the omentum. The histological examination showed a prevalent tubular structure with focal retiform area, without intraluminal mucines. Immunohistochemical findings of the case reported are similar to those described by other authors, except for inhibin which has not been detected by us. The cytofluorimetry showed the low presence of aneuploid cells, with a very low prolifing component (< 1%).

Vascular endothelial growth factor (VEGF): can we use it as prognostic factor in endometrial cancer?

AIM: The aim of the study was to investigate if VEGF levels reflect the severity of endometrial cancer and the clinic relationship between microvasal density (MVD) and concentration of VEGF in tumor. METHODS: The study was conducted on 22 patients affected by endometrial cancer who were submitted to total abdominal radical hysterectomy plus bilateral salpingo-ophorectomy. VEGF (pg/mL) and MVD values were measured on histologic specimens of endometrial cancer obtained during the surgical treatment. The means and standard deviations of estimated values were calculated and a statistical comparison was effected by student t test for not coupled data. Pearson correlation test was used to analyze the eventual correlation among VEGF and MVD values in overall patients. RESULTS: We have documented that VEGF expression and MVD change according to FIGO stage, lympho-vascular infiltration and lymph node involvement. Pearson correlation test shows a good linear positive correlation in overall patients between VEGF and MVD values. CONCLUSIONS: Results obtained show a possible use of VEGF as prognostic factor in endometrial cancer. Confirmation of these data may permit both to identify high-risk patients, who must be treated with a more aggressive treatment, and to use an angiogenic therapy in endometrial cancer.

Insights on Clusters Formation Mechanism by Time of Flight Mass Spectrometry. 2. The Case of Acetone-Water Clusters.

This paper is the second of a series dealing with clusters formation mechanism. In part 1, water clusters with the addition of an electrophilic molecule such as ethanol were studied by Time Of Flight Mass Spectrometry (TOFMS). Mass distributions of molecular clusters of ethanol, water and ethanol-water mixed clusters, were obtained by means of two different ionization methods: Electron Ionization (EI) and picosecond laser Photo-Ionization (PI) at a wavelength of 355 nm. In part 2, the same experimental approach was employed to obtain mass spectra of clusters generated by acetone-water binary mixtures with a different composition. Strong dependence of the mass spectra of clusters with EI and PI on the acetone-water mixing ratio was observed. It was shown that the spectral pattern changes gradually and water-rich cluster signals become fainter while acetone-rich cluster signals become more intensive with increasing acetone concentrations from 0.3% to 40%. Owing to the hydrogen bond acceptor character of acetone, its self-association is discouraged with respect to ethanol. The autocorrelation function (AF) was used to analyze the variation of the water clusters composition with the increase of the acetone concentration in terms of fundamental periodicities. However, although acetone and ethanol present a very different hydrogen-bonding ability, similarly to ethanol-water system, in acetone-water system the formation of water-rich clusters and subsequent metastable fragmentation are the dominant process that determine the clusters distribution, irrespective of the ionization process, while the ionization process significantly affects the acetone-rich clusters distribution. Graphical Abstract ᅟ.

Comparative evaluation of standard criteria and CA-125 in ovarian cancers treated with platinum or paclitaxel.

PURPOSE: To assess CA-125 in defining tumor response in patients treated with paclitaxel. PATIENTS AND METHODS: We analyzed 150 women treated for epithelial ovarian carcinoma with platinum or paclitaxel. We compared the patients treated with two agents, using a precise definition of CA-125 response, determined by 50% and 75% reductions, like other authors have published. RESULTS: CA-125 criteria gave response rates very similar to the standard response rates, both for patients treated with platinum (75% vs 63%) and also for those treated with paclitaxel (40% vs 39%). Rates of false-positive prediction of response by CA-125 were also similar for patients treated with these two agents. CONCLUSION: Precise 50% or 75% CA-125 response criteria are as sensitive as standard criteria for assessing activity of therapy for the ovarian cancers treated with platinum or paclitaxel. We propose that they may be useful in defining response in lieu of or in addition to standard response criteria in clinical trials involving epithelial ovarian cancer.

Unrecognized diabetes among hospitalized patients.

OBJECTIVE: To evaluate the hospital care rendered to hyperglycemic individuals who did not have a diagnosis of diabetes before admission. RESEARCH DESIGN AND METHODS: A total of 1,034 consecutively hospitalized adult patients at a 750-bed inner-city teaching hospital were evaluated. Patients with one or more plasma glucose values > 200 mg/dl were identified by the laboratory data system on a daily basis. Patients without a diagnosis of diabetes at the time of admission were evaluated to determine if and how physicians addressed the hyperglycemia, whether a new diagnosis of diabetes was made during admission, and whether follow-up was planned to address the hyperglycemia. RESULTS: After excluding patients who were admitted for a primary diagnosis of diabetes, 37.5% of all hyperglycemic medical patients and 33% of hyperglycemic surgical patients were without a diagnosis of diabetes at the time of admission. These patients had a mean peak glucose of 299 mg/dl, and 66% had two or more elevated values during their hospitalization. Fifty-four percent received insulin therapy, and 59% received bedside glucose monitoring, yet 66% of daily patient progress notes failed to comment on the presence of hyperglycemia or diabetes. Diabetes was documented in only three patients (7.3%) as a possible diagnosis in the daily progress notes. CONCLUSIONS: Despite marked hyperglycemia, most medical records made no reference to the possibility of unrecognized diabetes. Given the average delay of a decade between the onset and diagnosis of type 2 diabetes, further evaluation of hyperglycemic hospitalized patients may present an important opportunity for earlier detection and the initiation of therapy.

Effect of physician specialty on outcomes in diabetic ketoacidosis.

OBJECTIVE: More than 100,000 people are hospitalized annually in the U.S. with diabetic ketoacidosis (DKA). Outcome differences have not been examined for these patients based on whether their primary care provider is a generalist or a diabetes specialist. The objective of this study was to investigate hospital charges and hospital length of stay (LOS) for patients with DKA according to the specialty of their primary care provider. RESEARCH DESIGN AND METHODS: We investigated all patients with a primary diagnosis of DKA during a 3.5-year period (n = 260) in a large urban teaching hospital. Hospital charges and LOS were studied regarding the specialty of the primary care provider. Demographic factors, severity of illness, laboratory data, and readmission rates were compared. RESULTS: Patients cared for by generalists and endocrinologists had a similar case mix and severity of DKA. The age-adjusted mean LOS for patients of generalists was 4.9 days (95% CI 4.5-5.4), and the mean LOS for patients of endocrinologists was 3.3 days (2.6-4.2) (P < 0.0043). Mean hospital charges differed (P < 0.0001) with an age- and sex-adjusted mean for patients of endocrinologists of $5,463 ($4,179-7,141) and a mean for patients of generalists of $10,109 ($9,151-11,166). The additional charges incurred by generalists were due in part to patients undergoing more procedures. No differences in diabetes-related complications occurred during admission, but the endocrinologist-treated group had a lower readmission rate for DKA during the study period than the generalist-treated group (2 vs. 6%, respectively) (P = 0.03). CONCLUSIONS: Endocrinologists provide more cost-effective care than generalists do when serving as primary care providers for patients hospitalized with DKA.

Factitious Cushing syndrome.

There have been few reports of factitious Cushing syndrome. To characterize the clinical and laboratory features leading to this unusual diagnosis, we describe 6 patients (5 women, 1 man), ages 31-44, identified retrospectively among 860 patients evaluated for hypercortisolism at the National Institutes of Health Clinical Center. All six patients had multiple surgeries unrelated to Cushing syndrome and a history of depression or anxiety. Four patients had close contact with the medical profession, three a history of drug abuse, and three had undergone previous treatment for Cushing syndrome. The physical features of Cushing syndrome were variable and not helpful in the differential diagnosis with endogenous Cushing syndrome. Four patients had striking variability in urine-free cortisol (UFC) and 17-hydroxysteroid (17-OHCS) values from low to high. Adrenal computed tomography, performed in two patients, showed small adrenal glands (n = 1) or a left-sided mass (n = 1), and adrenal magnetic resonance imaging, performed in one patient, showed atrophic glands. Pituitary magnetic resonance imaging, carried out in four patients, was either normal (n = 1) or exhibited questionable signs of microadenoma (n = 3). Determination of synthetic glucocorticoids by high pressure liquid chromatography (HPLC) was positive in the four patients in whom it was performed. Factitious Cushing syndrome is a difficult diagnosis. To conserve time and resources, high pressure liquid chromatography analysis of urine steroids, the most definitive test for the factitious disorder, should be performed whenever there is clinical suspicion of glucocorticoid abuse.

Maturational changes in sympathetic and sensory innervation of the rat uterus: effects of neonatal capsaicin treatment.

The plasticity of the sympathetic and sensory innervation of the rat uterus was examined, before and after puberty, in controls and in animals where primary sensory nerves had been destroyed by neonatal capsaicin treatment. Immunohistochemical and histochemical methods were used in association with nerve density measurements and biochemical assays. The main findings were as follows: (1) Puberty was associated with a marked increase in the weight of the uterine horn, uterine cervix and parametrial tissue. This was unaffected by capsaicin treatment. (2) The sympathetic innervation of the uterine horn and parametrial tissue was reduced following puberty as revealed by a decrease in the density of noradrenaline-containing nerves and a marked decrease in the tissue concentration of noradrenaline. Sympathetic nerves supplying the uterine cervix and the blood vessels of the uterus appeared to be unaffected by puberty. (3) In contrast, the sensory supply of the uterus by substance P and calcitonin gene-related peptide-containing nerves increased in parallel with uterine growth during puberty resulting in no change in nerve density and only a slight reduction in peptide concentration. (4) Neonatal capsaicin treatment caused a long-lasting depletion of substance P- and calcitonin gene-related peptide-containing nerves. In the uterine horn and parametrial tissue, capsaicin-resistant calcitonin gene-related peptide, but not substance P, still increased with tissue weight during puberty, indeed, in the uterine horn, the relative increase was greater than in controls. (5) Sensory denervation resulted in an increase in the non-vascular sympathetic supply of the uterus, although there was a regional variation in the time course of the response. Perivascular sympathetic nerves were unaffected by capsaicin treatment. The pattern of change in non-vascular noradrenaline-containing nerves associated with puberty was similar in nature to controls. Thus, there is considerable plasticity in the innervation of the uterus both during puberty and following sensory denervation. A complex pattern of change occurs with differential responses in vascular and nonvascular nerves and in different regions of the uterus. Such differences may be due in part to the different origins of individual nerve populations and/or to their relative sensitivities to sex hormones.

Plasticity of autonomic nerves: differential effects of long-term guanethidine sympathectomy on the sensory innervation of the rat uterus during maturation.

The sensory nerves, containing substance P and calcitonin gene-related peptide, and noradrenaline-containing sympathetic nerves of the rat uterus were analyzed following long-term sympathectomy with guanethidine in prepubertal (four weeks), young adult (eight weeks) and fully adult animals (18 weeks). Immunohistochemical and histochemical methods were used in association with nerve density measurements and biochemical assays. The main findings were as follows: (1) long-term guanethidine treatment completely abolished the noradrenergic innervation of the uterine horn and parametrial tissue and markedly reduced the tissue levels of noradrenaline in both regions at the three ages analysed; (2) in the uterine horn guanethidine treatment had no effect on the tissue levels of either calcitonin gene-related peptide or substance P or on the density of calcitonin gene-related peptide-containing nerves, at any of the three ages studied; (3) in the parametrial tissue increased levels of calcitonin gene-related peptide were observed at 8 and 18 weeks of age, together with a significant increase in the density of calcitonin gene-related peptide-containing nerves. Substance P levels showed a transient increase in this tissue at eight weeks. In conclusion, long-term sympathectomy with guanethidine resulted in an increase in calcitonin gene-related peptide and substance P in sensory nerves in the parametrial tissue, but not in the uterine horn. The changes in the parametrial tissue only occurred after puberty. It is suggested that sensory nerves in the uterine horn may be less responsive to sympathetic denervation since loss of sympathetic nerves occurs as part of a normal physiological process during pregnancy in this region.

Lactiferous fistula.

Lactiferous fistula, or Zuska's disease, is a rare recurrent condition characterized by draining abscesses about the nipple on one or both breasts. Because little is known about the disease, it is often misdiagnosed and inappropriately treated. We reviewed the medical records of 51 women diagnosed as having lactiferous fistula at The Cleveland Clinic Foundation between 1961 and 1991. The clinical features were tabulated. An experienced breast pathologist reviewed the specimens of all the patients, and surgical techniques were compared. Patients ranged in age from 14 to 66 years, with a mean age of 40 years. Thirty-six patients had a swelling or mass at the areola, 51 had a draining fistula from the subareolar tissue, 40 had a chronic thick, pasty discharge from the nipple and 35 reported pain with the discharge. Fourteen patients had unsuccessful operations elsewhere, including four women who had subcutaneous mastectomies before coming to us. The average duration of symptoms was 3.2 years and the average follow-up period postoperatively was 51 months. On histologic examination, we found that, in all instances, keratinizing squamous epithelium had replaced the lining of one or more lactiferous ducts for a variable distance into the subareolar tissue. Core excision of the fistula and all of the retroareolar fibroglandular tissue and the ductal tissue within the nipple proved to be the definitive therapy in 47 of the 48 patients who had follow-up evaluation.

Time length of negativization of hCG serum values after either surgical or medical treatment of ectopic pregnancy.

OBJECTIVE: The aim of this study was to compare the time length until the human chorionic gonadotropin titer became negative after medical or surgical treatment of ectopic pregnancy. EXPERIMENTAL DESIGN: Prospective, randomized study. PARTECIPANTS AND INTERVENTIONS: We compared time to resolution in 30 cases of tubal pregnancies successfully treated, 15 underwent laparoscopic linear salpingostomy (group 1) and 15 medical treatment with single-ose methotrexate (group 2). The patients of both groups had no meaningful differences of age, gestational age and human chorionic gonadotropin mean values. MEASURES: During the follow-up human chorionic gonadotropin serum values were obtained every two days until the titer was negative. RESULTS: The time to resolution was 33.6 +/- 6.6 days in group 1, 31.5 +/- 7.8 in group 2 with no statistically meaningful differences. CONCLUSION: The data, comparable with results obtained by laparoscopic treatment, suggest that human chorionic gonadotropin value becomes negative independently of type of treatment and residual trophoblast.

Bone density in women with endometriosis.

OBJECTIVE: To evaluate the impact of endometriosis on bone metabolism. MATERIALS AND METHODS: We compared bone mineral density and biochemical markers (plasma osteocalcin, bone alkaline phosphatase, fasting urinary hydroxyproline, urinary excretion of cross-linked N-telopeptide of type I collagen) of bone turnover in forty-nine perimenopausal women undergoing laparotomy because of benign gynecologic pathology: in twenty-four of them (group A) endometriosis was diagnosed, the remaining twenty-five represented the control group (group B). Statistical analysis was performed by means of Student "t"-test; significance was set at p < 0.05. RESULTS: Bone density of the lumbar spine (0.898 +/- 0.325 vs 0.940 +/- 0.350) and bone markers failed to show statistically significant differences between the two groups. No significant correlation was observed between any bone density measurement and severity of endometriosis. CONCLUSION: Endometriosis does not seem to induce even in advanced stages, a reduction of bone density.

Effects of hormone replacement therapy on glucose metabolism.

OBJECTIVE: To evaluate the relationship between hormone replacement therapy and glucose metabolism, and the possible role played by the administration route. DESIGN: Prospective randomized study. MATERIALS AND METHODS: Eighty-four patients in either surgical or spontaneous menopause were randomly allocated into four groups: 15 patients in surgical menopause were treated with estrogens alone administered transdermically (17 beta-estradiol, 50 micrograms/day); 15 patients in surgical menopause were treated with oral conjugated equine estrogens (0.625 mg/day); 18 patients in spontaneous menopause were treated with transdermic estrogens plus oral progestagen (17 beta-estradiol 50 micrograms/day, acetate medroxyprogestogen 10 mg/day for 12 days per month); 16 patients in spontaneous menopause were treated with oral conjugated equine estrogens (0.625 mg/day) plus oral progestagen (10 mg/day for 12 days per month). Twenty patients not given any medication represented the control group. Before starting the trial and after six months of therapy, glucose and insulin levels before and after an oral glucose loading test were evaluated. Data analysis was performed by means of Student's "t"-test, ANOVA was used to compare mean levels between the groups. Significance was set at p < 0.005. RESULTS: Dosages made after 6 months of therapy showed comparable basal glucose levels in all the studied group, while basal insuline levels in patients taking estrogen alone were lower (76.88 +/- 23.66 vs 95.91 +/- 24.57 in group 1, 80 +/- 13.34 vs 96.91 +/- 18.97 in group 2) than pretreatment values. No significant difference in glucose levels was found after glucose load in the four groups. CONCLUSION. Women treated with estrogen alone have a tendency to a lower insulin response, which could indicate a greater insulin sensitivity. This effect seems to be more evident in transdermic administration than with oral administration. The addition of progestagen seems to wane the increase in insulin sensitivity induced by estrogens.

Differentiation of smooth muscle in the genital tract of the female mouse and its temporal relation with the development of the Wolffian nerve.

The development of the smooth muscle in the genital tract of the female mouse was studied by light and electron microscopy before and after birth. These studies showed that: a) between 13 days of fetal development and 2 days after birth the cells surrounding the Mullerian duct were undifferentiated and showed a fibroblast-like appearance; b) between 3 and 10 days after birth the cells acquired several characteristics of smooth muscle but they did not seem fully mature; c) between 30 and 180 days after birth the cells acquired a mature appearance; and d) the Wolffian nerve reached the Mullerian duct surrounding tissue before the start of smooth muscle differentiation.

Long-term silastic catheters and chest pain.

There are multiple reports in the literature of vascular erosion in the innominate vein or superior vena cava from the use of temporary central venous catheters. Catheter malposition is likely to precede the development of superior vena cava perforations, a catastrophic complication of central venous catheters. Catheter malposition after initial adequate placement is a very unusual long-term complication and delayed recognition of this complication may have disastrous consequences. Should the catheter change position so the tip is angled toward the sidewall, the repetitive movement of the catheter tip that occurs with respiratory excursion and the cardiac cycle may lead to endothelial injury and eventual erosion of the vein. These problems are thought to be alleviated in the patient receiving long-term intravenous therapy by using a soft Silastic catheter, which may not cause as much damage to the endothelium of the vein. We report three patients with left-sided long-term indwelling Silastic catheters that had changed position over time who presented with chest pain upon infusion of their total parenteral nutrition solutions. In each case, chest x-ray revealed that the tip of the catheter had migrated and was directed against the sidewall of the superior vena cava. In each case, catheter removal and replacement with a new catheter into the right side (subclavian and jugular systems) led to prompt relief of the patient's symptoms.

[HPV infection. Clinical features and treatment]. / Infezione da HPV. Aspetti clinici e trattamento.

The aspecific and exiguous symptoms and the lacking information are among the reasons of the diffusion of the vulvo-vaginal papillomatosis. We carried out the present study between 1995 and 1999 in the outpatient clinic of cervico-vaginal pathology of the Second University of Naples. 680 patients (aged between 18 and 56 years) underwent vulvoscopic and colposcopic examination. The did not show any relevant symptoms specific for HPV infection. Among the viral strains, HPV-16 and HPV 18 are able to induce a cervical cancer. To eliminate the pathology the primary prevention is necessary: it consist of both an adequate information about the micro-condilomatosis, the role of the activator agent, the modality of the infection, and the annual screening examinations such as pap-test and colposcopy. As first line treatment during secondary prevention, we utilize Roferon A, and perform diathermocoagulation according with the local diffusion and the degree of the disease (mild, moderate, severe). At the end of the therapy with Roferon A we observed that the infective focus was eliminated in about 60% of the cases and, only for moderate and severe micro-condilomatosis a diathermocoagulation was necessary.

[Role of intestinal resection in primary cytoreduction of ovarian cancer]. / Il ruolo della resezione intestinale nella citoriduzione primaria del cancro dell'ovaio.

BACKGROUND: The aim of this study is to define the role of the intestinal removal for the therapy of ovarian cancer in advanced stadium. METHODS: We have examined 247 females with epithelial ovarian cancer in advanced stadium, that had intestinal removal. RESULTS: The survival in the females that had a very good intestinal removal is greatest than in the females that hadn't a very good intestinal removal.