RAGE inhibition blunts insulin-induced oncogenic signals in breast cancer.

The receptor for advanced glycation end products (RAGE) is implicated in diabetes and obesity complications, as well as in breast cancer (BC). Herein, we evaluated whether RAGE contributes to the oncogenic actions of Insulin, which plays a key role in BC progression particularly in obese and diabetic patients. Analysis of the publicly available METABRIC study, which collects gene expression and clinical data from a large cohort (n = 1904) of BC patients, revealed that RAGE and the Insulin Receptor (IR) are co-expressed and associated with negative prognostic parameters. In MCF-7, ZR75 and 4T1 BC cells, as well as in patient-derived Cancer-Associated Fibroblasts, the pharmacological inhibition of RAGE as well as its genetic depletion interfered with Insulin-induced activation of the oncogenic pathway IR/IRS1/AKT/CD1. Mechanistically, IR and RAGE directly interacted upon Insulin stimulation, as shown by in situ proximity ligation assays and coimmunoprecipitation studies. Of note, RAGE inhibition halted the activation of both IR and insulin like growth factor 1 receptor (IGF-1R), as demonstrated in MCF-7 cells KO for the IR and the IGF-1R gene via CRISPR-cas9 technology. An unbiased label-free proteomic analysis uncovered proteins and predicted pathways affected by RAGE inhibition in Insulin-stimulated BC cells. Biologically, RAGE inhibition reduced cell proliferation, migration, and patient-derived mammosphere formation triggered by Insulin. In vivo, the pharmacological inhibition of RAGE halted Insulin-induced tumor growth, without affecting blood glucose homeostasis. Together, our findings suggest that targeting RAGE may represent an appealing opportunity to blunt Insulin-induced oncogenic signaling in BC.

Effects of two diet techniques and delivery mode on weight loss, metabolic profile and food intake of obese adolescents: a fixed diet plan and a calorie-counting diet.

The aim of this study is to compare the weight loss of obese adolescents on two different low-calorie diets: fixed diet plan and calorie-counting diet. This is a randomized clinical study with 66 obese adolescents (body mass index Z score (ZBMI)>+3, 13.7±0.7 years, 60.6% male) with anthropometric, food intake, physical activity, laboratory, body composition and stage of pubertal development data evaluated. There was a reduction in the ZBMI in both groups (P<0.0001), without significant difference between them (P=0.87). There was a significant reduction in insulin, and homeostasis model assessment insulin resistance (HOMA-IR), with no difference between groups. A reduction in total energy intake of the groups was found, with an increase in the proportion of protein and reduction in carbohydrates. In this cohort of severely obese adolescents, fixed diet plan and calorie-counting diet led to a similar reduction of ZBMI, metabolic markers and total energy intake.

A comparison of the efficacy of organic and mixed-organic polymers with polyaluminium chloride in chemically assisted primary sedimentation (CAPS).

CAPS is the acronym for chemically assisted primary sedimentation, which consists of adding chemicals to raw urban wastewater to increase the efficacy of coagulation, flocculation and sedimentation. The principal benefits of CAPS are: upgrading of urban wastewater treatment plants; increasing efficacy of primary sedimentation; and the major production of energy from the anaerobic digestion of primary sludge. Metal coagulants are usually used because they are both effective and cheap, but they can cause damage to the biological processes of anaerobic digestion. Generally, biodegradable compounds do not have these drawbacks, but they are comparatively more expensive. Both metal coagulants and biodegradable compounds have preferential and penalizing properties in terms of CAPS application. The problem can be solved by means of a multi-criteria analysis. For this purpose, a series of tests was performed in order to compare the efficacy of several organic and mixed-organic polymers with that of polyaluminium chloride (PACl) under specific conditions. The multi-criteria analysis was carried out coupling the simple additive weighting method with the paired comparison technique as a tool to evaluate the criteria priorities. Five criteria with the following priorities were used: chemical oxygen demand (COD) removal > turbidity, SV60 > coagulant dose, and coagulant cost. The PACl was the best alternative in 70% of the cases. The CAPS process using PACl made it possible to obtain an average COD removal of 68% compared with 38% obtained, on average, with natural sedimentation and 61% obtained, on average, with the best PACl alternatives (cationic polyacrylamide, natural cationic polymer, dicyandiamide resin).

A simple method to equalize the workload when operating several small wastewater treatment plants: a case study.

The aim of the present study is to define a simple (and easy to use) method to equalize the workload of personnel operating several small wastewater treatment plants (SWWTPs). The approach is illustrated through a case study which is the result of collaboration between researchers and a water and wastewater management company operating in Southern Italy. The topic is important since personnel have a significant impact on the operating costs of SWWTPs, and the approach outlined results in the minimum number of staff being required to assure the management of the service. Four kinds of work units are considered: plant managers, assistant plant managers, laboratory technicians and executives. In order to develop a practical, feasible and easy to use method, the workload was evaluated considering only the population equivalent (PE) and the number of plants managed. The core of the method is the evaluation of the percentage of time that the personnel units devote to the operation of SWWTPs of the municipality considered. The proposed procedure offers a useful tool to equalize the workload, both in terms of PE and the number of plants managed, the procedure being easily modifiable to introduce other evaluation criteria. By using familiar concepts such as PE and number of plants managed, the approach of the method can easily be understood by management. It can also be readily adapted to other similar situations.

Groundwater recharge in an endoreic basin with reclaimed municipal wastewater.

The aim of this paper was to evaluate the groundwater pollution in an endoreic basin in southern Italy. The aquifer circulation was carried out on two different levels: a shallow groundwater, with a water table of about 10 m, and a deep groundwater in a karst aquifer, with a water table of 140-190 m. Reclaimed municipal wastewater and superficial water collected in the catchment area were both drained in a swallow hole linked with the deep groundwater. The agricultural practice conducted in the endoreic basin produced an excess of nitrate in the soil. Nitrate was subsequently washed out and displaced in the groundwater. With regard to the EU Drinking Water Directive (98/83/EC), the research activity conducted during 2003 showed the absence of pollution in the deep groundwater used for drinking water supply. The shallow groundwater, instead, was strongly influenced by agricultural and pasture activities, with detectable levels of nitrates and bacteria. In order to reduce the load of pollution entering the swallow hole and then in the deep groundwater, the realisation of a constructed wetland plant was proposed to improve the quality of reclaimed wastewater, as well as to pursue the wastewater reuse in agriculture.

Prenatal and postnatal findings of acrania.

INTRODUCTION: Acrania is a rare congenital anomaly in which the flat bones of the cranial vault are partial or complete absent with complete but abnormal development of the cerebral hemispheres. CASE REPORT: We report two cases, diagnosed prenatally by ultrasound and followed by medical pregnancy termination, one with isolated acrania and one with associated cleft lip and palate and spina bifida. DISCUSSION: The acrania pathogenesis is unknown and differential diagnosis should be made with other conditions like anencephaly and acalvaria. Genetic counselling is not easy because there is no evidence for a specific genetic origin, but the extreme rarity and sporadic nature suggests a low recurrence risk.

Prenatal diagnosis of congenital diaphragmatic hernia in a McFadden Kalousek type 2 triploid fetus.

Triploidy is characterized by an extra haploid set. We report a rare case of a prenatally diagnosed digynic type (McFadden/Kalousek type 2) triploid fetus with congenital diaphragmatic hernia (CDH). Prenatal ultrasonographic examination allows reliable detection of CDH and it is therefore of great importance for proper parental counselling, as well as performing fetal karyotype for associated chromosomal anomalies.

[Skin metastasis from renal carcinoma: epidemiologic and clinical features]. / Metastasi cutanee da carcinoma renale: aspetti epidemiologici e clinici.

The Authors present an additional case of a 70-years old male patient with skin metastasis from renal cell carcinoma, 15 months after nephrectomy. The patient died ten months later for advanced neoplastic disease. Skin metastasis affect 3-6% of patients with renal cell carcinoma. The surgical treatment is mandatory for single nodule. Chemotherapy is often ineffectual. The prognosis is poor, with a 0-8% 5-years survival for patients with multiple nodules.

Electro-oculography in pigment dispersion syndrome.

Clinical data and the common embryological origin of the pigment epithelium of the iris and retinal pigment epithelium suggest that the latter may be involved in the pigment dispersion syndrome (PDS). In our study functional changes of the retinal pigment epithelium were evaluated by electro-oculography in 10 PDS patients, 10 patients with chronic open-angle glaucoma (COAG) and 10 normal subjects. The mean dark-trough latency and Arden ratio for the PDS group were significantly lower than those of the COAG and control groups. Analysis of the Arden ratios based on the Kruskall-Wallis test revealed that 30% of the PDS values were subnormal with respect to the control group mean -1 SD. These results indicate primary involvement of the retinal pigment epithelium in PDS.

[Appendiceal mucoceles. A case report]. / Il mucocele appendicolare. Analisi di un caso.

A case of benign mucocele presenting as a subocclusive syndrome is reported; the surgical treatment consisted in a right colectomy. Appendiceal mucoceles are rare lesions of the appendix, characterized by a gross enlargement of the appendix from accumulation of mucoid substance within the lumen. It is a rare condition, encountered in only 0.1-0.4% of all appendectomies with a female predominance and an average age at the time of diagnosis over 50 years. Following careful review of the literature, the difficulties in differential diagnosis are underlined, especially between benign and malignant forms and the possibility of a pre-operative diagnosis is examined. Abdominal ultrasound and CT scan of the abdomen or colonoscopy may suggest the diagnosis. However, often the diagnosis is an incidental event. The pathogenesis, histologic aspect of the lesion and the different surgical strategies are discussed. A frozen section examination should be performed in all patients, while the abdomen is open, because it may be impossible to predict the underlying pathology merely by inspecting the serosal surface of a dilated appendix. If a simple retentional cyst is uncovered, without atypia, appendectomy should be curative; if cystadenoma or cystadenocarcinoma of the appendix are found, more extended resection was necessary. However if the operation is done as an emergency, a frozen section examination is not always available: in those patients a more extended resection is suggested.

Hypertension in a group of centenarians.

The aim of the present study was to evaluate the frequency of hypertension in a group of extremely old persons: 73 centenarians resident in Calabria were studied, aged 100-110 years, (mean age 102.95 +/- 2.0), 54 women (73.9%) and 19 men (26.1%), in the period January 1st and April 30th, 1994. All the centenarians were visited at their homes. Hypertensive centenarians were 14 (19.1%), women 12 (85.7%) and men 2 (14.3%). Even if blood pressure progressively increases with age, we have not found it in centenarians. The lesser frequency of hypertension in centenarians certainly depends on genetic and constitutional factors, but also their way of life and alimentary habits (decreased sodium intake, increase of calcium and potassium intake), as we have proven in a recent work, could be determinant. Moreover, the presence of hypertensive centenarians might mean that hypertension does not prevent to reach an age by far above the average, if it is the only risk factor.

Multidimensional evaluation in a group of centenarians.

A group of 40 Calabrian centenarians (12 males, 28 females) was studied from June 1 to December 31, 1993. Their age was ranging from 100 to 107 years (mean age: 102.3 +/- 1.8 years). Clinical evaluation was performed by recording anamnestic data, an objective examination, and by assessing cognitive performance, emotional and nutritional status, as well as functional activity. The following tests were applied: mini mental state examination (MMSE), geriatric depression scale (CDS), activity of daily living (ADL), and instrumental activity of daily living (IADL). The nutritional status was estimated by collecting answers to a questionnaire regarding nutritional habits and by measuring body weight, height and calculating body mass index (BMI). In assessment of the socio-economic conditions, the following parameters were considered: years of education, type of job, type of residence ownership, heating system used, toilet set available and the regular income. Our multidimensional evaluation has revealed, in agreement with other findings, that the Calabrian centenarians seem to be a selected population with an amazingly high activity, who overcame severe age-dependent diseases, probably because of their genetic pattern and of the environmental factors which may favor longevity.

[Male breast cancer. Histochemical and flow cytometry analysis of 6 cases]. / Carcinoma mammario maschile. Analisi immunoistochimica e citofluorimetrica di 6 casi.

The authors studied immunohistochemical and flow cytometric parameters of 6 male breast carcinomas. All patients were ER positive and frequency of aneuplody was 66%. Diploid tumors showed low proliferative activity and S-phase fraction < 3%.

Doppler velocimetry of the fetal middle cerebral artery in patients with preterm labor and intact membranes.

A prospective cohort study was conducted to determine whether preterm labor is associated with changes in the impedance to blood flow of the fetal middle cerebral artery. Doppler velocimetry studies were performed in 194 consecutive patients with preterm labor and intact membranes. Pulsatility indices of the middle cerebral artery and umbilical artery were determined on admission. Results were expressed as ratio of the observed pulsatility index to mean value for gestational age expressed as delta MCA PI and delta UA PI, respectively. The prevalence of preterm delivery (< 37 weeks) and delivery within 24 hours of admission was 55.2% (107/194) and 15.5% (30/194), respectively. Patients with an examination-to-delivery interval within 24 hours had significantly lower mean delta MCA PI than that of patients delivered at > or = 37 weeks and > or = 4 weeks after the examination (P < 0.01). Fetuses with a delta MCA PI at or below 0.88 had a relative risk of 2 (95% confidence interval, 1.32 to 2.9) who were delivered within 24 hours compared to controls (sensitivity 76.7%, specificity 62.8%, positive predictive value 26.7%, negative predictive value 93.5%). Stepwise logistic regression analysis indicated that the relationship between delta MCA PI and examination-to-delivery interval remained statistically significant after correcting for cervical dilatation (P < 0.001). Our data indicate that preterm parturition is associated with a decrease in the impedance to flow in the fetal cerebral circulation.

The value of amniotic fluid interleukin-6, white blood cell count, and gram stain in the diagnosis of microbial invasion of the amniotic cavity in patients at term.

PROBLEM: Subclinical microbial invasion of the amniotic cavity occurs in 18.8% of women with term labor and intact membranes and in 34% of patients with term PROM and is a risk factor for the development of puerperal infection related morbidity. Although amniotic fluid white blood cell count, interleukin-6 determination, and Gram stain examination have been used for the diagnosis of intrauterine infection in patients with preterm labor and preterm premature rupture of membranes, no information is available about the accuracy and specific cut-off values for these tests in patients at term. The purpose of this study was to compare the performance of the amniotic fluid Gram stain examination, white blood cell count, and interleukin-6 determination in the identification of microbial invasion of the amniotic cavity in patients at term with and without PROM. METHOD: Amniotic fluid was retrieved from 148 patients with term gestations (90 patients with spontaneous labor and intact membranes and 58 patients with PROM). Samples were cultured for bacteria and Mycoplasma species. Amniotic fluid Gram stain, white blood cell count, and interleukin-6 determinations (ELISA, sensitivity: 43 pg/ml) were performed in all samples. Microbial invasion of the amniotic cavity was defined as a positive amniotic fluid culture for microorganisms. Analysis was conducted using Mann-Whitney U test, Fisher's exact test, receiver operating characteristic curves and logistic regression. RESULTS: Patients with spontaneous labor and intact membranes: The prevalence of microbial invasion of amniotic cavity in this group was 15.6% (14/90). The most sensitive test for the detection of microbial invasion of the amniotic cavity was amniotic fluid interleukin-6 determination (sensitivity for: interleukin-6 > or = 5.7 ng/ml = 86%, white blood cell count > or = 20 cells/mm3 = 64%, Gram stain = 28%). The most specific test was the Gram stain of the amniotic fluid (specificity for: Gram stain = 84%, interleukin-6 = 79% and white blood cell count = 63%). Multiple logistic regression demonstrated that amniotic fluid interleukin-6 concentration was the only covariate that retained statistical significance when intrauterine infection was used as outcome variable. Patients with PROM: The prevalence of a positive amniotic fluid culture in this group was 39.7% (23/58). Logistic regression demonstrated that only interleukin-6 retained a significant relationship with the results of amniotic culture when all variables were entered simultaneously into a model to predict amniotic fluid culture results. The most sensitive tests for the detection of intrauterine infection were interleukin-6 determination and white blood cell count (sensitivity for interleukin-6 > or = 3.4 ng/ml and white blood cell count > or = 20 cells/mm3 = 69.6% for both). The most specific test was Gram stain (97.1%). CONCLUSIONS: Amniotic fluid interleukin-6 determination is the best rapid test for the detection of microbial invasion of the amniotic cavity in patients at term with and without PROM. When this test is not available, amniotic fluid Gram stain and white blood cell count represent valid diagnostic tools to assess the microbial state of amniotic cavity.

The natural interleukin-1 receptor antagonist in the fetal, maternal, and amniotic fluid compartments: the effect of gestational age, fetal gender, and intrauterine infection.

OBJECTIVES: The interleukin-1 receptor antagonist is a newly discovered cytokine that blocks the biologic effects of interleukin-1 in vitro and in vivo. This cytokine is a physiologic component of amniotic fluid and is considered to be of critical importance in the homeostasis of the cytokine network. This study was undertaken to systematically examine the bioavailability of interleukin-1 receptor antagonist in the maternal, fetal, and amniotic fluid compartments during term and preterm parturition in women with and without microbial invasion of the amniotic cavity. STUDY DESIGN: The patient population consisted of (1) pregnant women in the midtrimester (n = 42), (2) patients who underwent cordocentesis for diagnostic purposes (n = 39), (3) patients with preterm labor (n = 126), (4) women with term gestation (n = 102), and (5) healthy nonpregnant women (n = 8). Amniotic fluid was cultured for aerobic and anaerobic bacteria, as well as Mycoplasma sp. Interleukin-1 receptor antagonist concentrations were determined by enzyme-linked immunoassay in maternal and fetal plasma, amniotic fluid, and neonatal urine. Microbial invasion of the amniotic cavity was defined as the presence of a positive amniotic fluid culture for microorganisms. RESULTS: (1) Interleukin-1 receptor antagonist was normally present in fetal plasma samples obtained by cordocentesis, and its concentration increased with advancing gestational age (n = 39; r = 0.61, p < 0.001). (2) Patients at term not in labor had higher amniotic fluid interleukin-1 receptor antagonist concentrations than patients in the midtrimester (median 40.1 ng/ml, range 5.7 to 213.1 vs median 16.2 ng/ml, range 3.2 to 62.2, respectively, p < 0.001). (3) Amniotic fluid and cord plasma interleukin-1 receptor antagonist concentrations were significantly higher in patients with preterm labor and microbial invasion of the amniotic cavity than in those without microbial invasion of the amniotic cavity (amniotic fluid: median 219.9 ng/ml, range 35.4 to 504 vs median 80.6 ng/ml, range 24.3 to 399, respectively, p < 0.001; umbilical cord plasma: median 4.8 ng/ml, range 0.3 to 167.0 vs median 1.0 ng/ml, range 0 to 276.0, respectively, p < 0.05). In contrast, these differences were not found in patients with term labor either with or without microbial invasion of the amniotic cavity. (4) In both term and preterm patients the amniotic fluid and neonatal urine concentrations of interleukin-1 receptor antagonist were significantly higher in female fetuses than in male fetuses (amniotic fluid, preterm: median 191.9 ng/ml, range 51.6 to 504.0 vs median 61.1 ng/ml, range 11.5 to 284.9, respectively, p < 0.001; amniotic fluid, term: median 58.7 ng/ml, range 25.5 to 264.0 vs median 33.9 ng/ml, range 3.4 to 132.4, respectively, p < 0.001; neonatal urine: median 317 ng/ml, range 59.0 to 440.8 vs median 12.2 ng/ml, range 2.5 to 61.6, respectively, p < 0.005). CONCLUSIONS: (1) Interleukin-1 receptor antagonist is physiologically present in the fetal, maternal, and amniotic fluid compartments; (2) microbial invasion of the amniotic cavity in the preterm gestation is associated with a significant increase in the concentrations of this cytokine in the fetal and amniotic fluid compartments but not in maternal plasma; (3) fetal urine is a source of amniotic fluid interleukin-1 receptor antagonist; (4) fetal plasma interleukin-1 receptor antagonist concentrations increase with gestational age; (5) there is a significant effect of fetal gender in amniotic fluid and neonatal urine concentrations of interleukin-1 receptor antagonist.

Ultrasonographic examination of the uterine cervix is better than cervical digital examination as a predictor of the likelihood of premature delivery in patients with preterm labor and intact membranes.

OBJECTIVE: The purpose of this study was to compare the diagnostic performance of ultrasonographic and digital examination of the cervix in the prediction of preterm delivery in patients presenting with preterm labor and intact membranes. STUDY DESIGN: Endovaginal ultrasonography was performed in 59 patients admitted with preterm labor (20 to 35 weeks) and cervical dilatation of < 3 cm. Cervical parameters evaluated included endocervical length, the presence of funneling, funnel length, funnel width, cervical index ([Funnel length + 1]/Endocervical length), and cervical dilatation and effacement as determined by digital examination. Outcome variables were the occurrence of preterm delivery (< 36 weeks) and the admission-to-delivery interval. RESULTS: The prevalence of preterm delivery was 37.3% (22/59). Receiver-operator characteristic curve and logistic regression analyses indicated a significant relationship between the occurrence of preterm delivery and ultrasonographic cervical parameters (p < 0.005 for each) but not with the results of digital examination of the cervix. Survival analysis demonstrated a shorter admission-to-delivery interval for patients with an abnormal cervical index or endocervical length (p < 0.005 for each). CONCLUSIONS: Endovaginal ultrasonographic examination of the uterine cervix is more accurate than digital examination of the cervix in the assessment of the risk for preterm delivery in patients with preterm labor and intact membranes.

Macrophage inflammatory protein-1 alpha in term and preterm parturition: effect of microbial invasion of the amniotic cavity.

PROBLEM: This study was conducted to determine whether: (1) gestational age, parturition, and microbial invasion of the amniotic cavity (MIAC) are associated with changes in amniotic fluid concentrations of immunoreactive macrophage inflammatory protein-1 alpha; (2) amniotic fluid concentrations of macrophage inflammatory protein-1 alpha are correlated with the white blood cell count and the concentrations of interleukin-8 in amniotic fluid. METHOD: Amniotic fluid was retrieved by amniocentesis from 126 patients; 54 women with preterm labor and intact membranes (no MIAC-delivery at term, N = 21; no MIAC-preterm delivery, N = 16; MIAC-preterm delivery, N = 17); 62 patients at term (no labor, N = 19; labor-no MIAC, N = 20; labor-MIAC, N = 23); and 10 patients in the midtrimester of pregnancy. Amniotic fluid was cultured for aerobic, anaerobic and Mycoplasma species. Determinations of amniotic fluid macrophage inflammatory protein-1 alpha and interleukin-8 were performed with immunoassays validated for amniotic fluid (sensitivity: 14.2 pg/ml and 0.3 ng/ml, respectively). Kruskal-Wallis analysis of variance (ANOVA) for censored data, Mann-Whitney U test and Spearman's rank correlation were performed for analysis. RESULTS: 1) Amniotic fluid macrophage inflammatory protein-1 alpha was present in only 31.0% (9/29) of patients not in labor (midtrimester and term). 2) Patients with preterm labor and MIAC had higher amniotic fluid concentrations of macrophage inflammatory protein-1 alpha than those without MIAC (no MIAC-delivery at term: median 0.0 pg/ml, range 0.0-221.2; no MIAC-preterm delivery: median 37.4 pg/ml, range 0.0-494.6; MIAC-preterm delivery: median 7171.0 pg/ml, range 402.5-37994.0; P < 0.00001). 3) Among patients at term, MIAC was associated with higher concentrations of amniotic fluid macrophage inflammatory protein-1 alpha than patients without MIAC (no labor: median 0.0 pg/ml, range 0.0-25.6; labor-no MIAC: median 16.7 pg/ml, range 0.0-161.6; labor-MIAC: median 103.8 pg/ml, range 0.0-4349.0, P < 0.001). 4) Among patients in preterm labor, a strong correlation was found between amniotic fluid concentrations of macrophage inflammatory protein-1 alpha and interleukin-8 (r = 0.9, P < 0.00001) and between amniotic fluid macrophage inflammatory protein-1 alpha concentrations and amniotic fluid white blood cell count (r = 0.6, P < 0.0001). CONCLUSIONS: (1) Macrophage inflammatory protein-1 alpha is undetectable in most amniotic fluid samples from patients in the midtrimester of pregnancy and at term not in labor. (2) Microbial invasion of the amniotic cavity is associated with increased concentrations of immunoreactive amniotic fluid macrophage inflammatory protein-1 alpha in both term and preterm gestations. (3) Amniotic fluid macrophage inflammatory protein-1 alpha concentrations significantly correlate with interleukin-8 levels and white blood cell count in amniotic fluid. Our data strongly suggest a role for macrophage inflammatory protein-1 alpha in the mechanisms responsible for the recruitment of leukocytes into the amniotic cavity during the course of intrauterine infection.