Integrity of prokaryotic mRNA isolated from complex samples for in vivo bacterial transcriptome analysis.

Even though several in vitro studies have focused on bacterial biology, the extent of such knowledge is not complete when considering an actual infection. As culture-independent microbiology methods such as high-throughput sequencing became available, important aspects of host-bacterium interactions will be elucidated. Based on microbiological relevance, we considered Bacteroides fragilis in a murine experimental infection as a model system to evaluate the in vivo bacterial transcriptome in host exudates. A disproportionate number of reads belonging to the host genome were retrieved in the first round of pyrosequencing, even after depletion of ribosomal RNA; the average number of reads related to the eukaryotic genome was 71.924-67.7%, whereas prokaryotic reads represented 34.338-32.3% in host exudates. Thus, different treatments were used to improve the prokaryotic RNA yield: i) centrifugation; ii) ultrasonic treatment; and iii) ultrasonic treatment followed by centrifugation. The latter treatment was found to be the most efficient in generating bacterial yields, as it resulted in a higher number of Bacteroides cells. However, the RNA extracted after this treatment was not of sufficient quality to be used in cDNA synthesis. Our results suggest that the methodology routinely used for RNA extraction in transcriptional analysis is not appropriate for in vivo studies in complex samples. Furthermore, the most efficient treatment for generating good bacterial cell yields was not suitable to retrieve high-quality RNA. Therefore, as an alternative methodological approach to enable in vivo studies on host-bacterium interactions, we advise increasing the sequencing depth despite the high costs.

Indoor and outdoor biomonitoring using lichens at urban and rural primary schools.

Monitoring particulate matter (PM) and its chemical constituents in classrooms is a subject of special concern within the scientific community in order to control and minimize child exposure. Regulatory sampling methods have presented several limitations in their application to larger number of classrooms due to operational and financial constraints. Consequently, passive sampling methodologies using filters were developed for indoor sampling. However, such methodologies could not provide parallel information for outdoors, which is important to identify pollution sources and assess outdoor contribution to the indoors. Therefore, biomonitoring with transplanted lichens, a technique usually applied for outdoor studies, was used both indoor and outdoor of classrooms. Three main objectives were proposed, to (i) characterize simultaneously indoor and outdoor of classrooms regarding inorganic air pollutants, (ii) investigate spatial patterns of lichen conductivity, and (iii) assess pollution sources that contribute to a poor indoor air quality in schools. Lichens Flavoparmelia caperata were transplanted to indoor and outdoor of classrooms for 59 d. After exposure, electric conductivity of lichens leachate was measured to evaluate lichen vitality and cell damage. Outdoors lichen conductivity was higher near the main highways, and indoors there was great variability in levels, which indicates different emissions sources and different ventilation patterns. Chemical content of lichens was assessed by instrumental neutron activation analysis (INAA), and As, Br, Ca, Ce, Co, Cr, Cs, Eu, Fe, Hf, K, La, Na, Rb, Sb, Sc, Sm, Sr, Ta, Th, Yb, and Zn were determined. Element accumulation, crustal enrichment factors, and spatial variability of elements were analyzed and contaminants from anthropogenic sources, such as traffic (As, Sb, and Zn) and indoor chalk (Ca) found. Classrooms with potential indoor air quality problems were identified by presenting higher accumulations of inorganic pollutants in exposed biomonitors.

Transcriptome meta-analysis of peripheral lymphomononuclear cells indicates that gestational diabetes is closer to type 1 diabetes than to type 2 diabetes mellitus.

We performed a meta-analysis of the transcription profiles of type 1, type 2 and gestational diabetes to evaluate similarities and dissimilarities among these diabetes types. cRNA samples obtained from peripheral blood lymphomononuclear cells (PBMC) of 56 diabetes mellitus patients (type 1 = 19; type 2 = 20; gestational = 17) were hybridized to the same whole human genome oligomicroarray platform, encompassing 44,000 transcripts. The GeneSpring software was used to perform analysis and hierarchical clustering, and the DAVID database was used for gene ontology. The gene expression profiles showed more similarity between gestational and type 1 diabetes rather than between type 2 and gestational diabetes, a finding that was not influenced by patient gender and age. The meta-analysis of the three types of diabetes disclosed 3,747 differentially and significantly expressed genes. A total of 486 genes were characteristic of gestational diabetes, 202 genes of type 1, and 651 genes of type 2 diabetes. 19 known genes were shared by type 1, type 2 and gestational diabetes, highlighting EGF, FAM46C, HBEGF, ID1, SH3BGRL2, VEPH1, and TMEM158 genes. The meta-analysis of PBMC transcription profiles characterized each type of diabetes revealing that gestational and type 1 diabetes were transcriptionally related.

Winter ventilation rates at primary schools: comparison between Portugal and Finland.

This study focused on examination of ventilation rates in classrooms with two different types of ventilation systems: natural and mechanical. Carbon dioxide (CO2) measurements were conducted in primary schools of Portugal characterized by natural ventilation and compared to Finland where mechanical ventilation is the norm. The winter period was selected since this season exerts a great influence in naturally ventilated classrooms, where opening of windows and doors occurs due to outdoor atmospheric conditions. The ventilation rates were calculated by monitoring CO2 concentrations generated by the occupants (used as a tracer gas) and application of the buildup phase method. A comparison between both countries' results was conducted with respect to ventilation rates and how these levels corresponded to national regulatory standards. Finnish primary schools (n = 2) registered a mean ventilation rate of 13.3 L/s per person, which is higher than the recommended ventilation standards. However, the Finnish classroom that presented the lowest ventilation rate (7.2 L/s per person) displayed short-term CO2 levels above 1200 ppm, which is the threshold limit value (TLV) recommended by national guidelines. The Portuguese classrooms (n = 2) showed low ventilation rates with mean values of 2.4 L/s per person, which is markedly lower than the minimum recommended value of 7 L/s per person as defined by ASHRAE and 20% less than the REHVA minimum of 3 L/s per person. Carbon dioxide levels of 1000 ppm, close to the TLV of 1200 ppm, were also reached in both Portuguese classrooms studied. The situation in Portugal indicates a potentially serious indoor air quality problem and strengthens the need for intervention to improve ventilation rates in naturally ventilated classrooms.

Impact of Sahara dust transport on Cape Verde atmospheric element particles.

The objectives of this study were to (1) conduct an elemental characterization of airborne particles sampled in Cape Verde and (2) assess the influence of Sahara desert on local suspended particles. Particulate matter (PM(10)) was collected in Praia city (14°94'N; 23°49'W) with a low-volume sampler in order to characterize its chemical composition by k0-INAA. The filter samples were first weighed and subsequently irradiated at the Portuguese Research Reactor. Results showed that PM(10) concentrations in Cape Verde markedly exceeded the health-based air quality standards defined by the European Union (EU), World Health Organization (WHO), and U.S. Environmental Protection Agency (EPA), in part due to the influence of Sahara dust transport. The PM(10) composition was characterized essentially by high concentrations of elements originating from the soil (K, Sm, Co, Fe, Sc, Rb, Cr, Ce, and Ba) and sea (Na), and low concentrations of anthropogenic elements (As, Zn, and Sb). In addition, the high concentrations of PM measured in Cape Verde suggest that health of the population may be less affected compared with other sites where PM(10) concentrations are lower but more enriched with toxic elements.

Impact of maritime air mass trajectories on the Western European coast urban aerosol.

Lisbon is the largest urban area in the Western European coast. Due to this geographical position the Atlantic Ocean serves as an important source of particles and plays an important role in many atmospheric processes. The main objectives of this study were to (1) perform a chemical characterization of particulate matter (PM2.5) sampled in Lisbon, (2) identify the main sources of particles, (3) determine PM contribution to this urban area, and (4) assess the impact of maritime air mass trajectories on concentration and composition of respirable PM sampled in Lisbon. During 2007, PM2.5 was collected on a daily basis in the center of Lisbon with a Partisol sampler. The exposed Teflon filters were measured by gravimetry and cut into two parts: one for analysis by instrumental neutron activation analysis (INAA) and the other by ion chromatography (IC). Principal component analysis (PCA) and multilinear regression analysis (MLRA) were used to identify possible sources of PM2.5 and determine mass contribution. Five main groups of sources were identified: secondary aerosols, traffic, calcium, soil, and sea. Four-day backtracking trajectories ending in Lisbon at the starting sampling time were calculated using the HYSPLIT model. Results showed that maritime transport scenarios were frequent. These episodes were characterized by a significant decrease of anthropogenic aerosol concentrations and exerted a significant role on air quality in this urban area.

Disruption of Type-I IFN pathway ameliorates preservation damage in mouse orthotopic liver transplantation via HO-1 dependent mechanism.

Ischemia/reperfusion injury (IRI) remains unresolved problem in clinical organ transplantation. We analyzed the role of Type-I interferon (IFN) pathway in a clinically relevant murine model of extended hepatic cold preservation followed by orthotopic liver transplantation (OLT). Livers from Type-I IFN receptor (IFNAR) knockout (KO) or wild-type (WT) mice (C57/BL6) were harvested, preserved at 4°C in UW solution for 20 h and transplanted to groups of syngeneic IFNAR KO or WT recipients. Liver graft but not recipient IFNAR deficiency was required to consistently ameliorate IRI in OLTs. Indeed, disruption of Type-I IFN signaling decreased serum alanine aminotransferase (sALT) levels (p < 0.001), diminished Suzuki's score of histological OLT damage (p < 0.01) and improved 14-day survival (from 42%[5/12] in WT to 92%[11/12] in IFNAR KO; p < 0.05). Unlike in WT group, IFNAR deficiency attenuated OLT expression of TNF-α, IL-1ß, IL-6, MCP-1, CXCL-10, ICAM-1; diminished infiltration by macrophages/PMNs; and enhanced expression of antioxidant HO-1/Nrf2. The frequency of TUNEL+ apoptotic cells and caspase-3 activity/expression selectively decreased in IFNAR KO group. Small interfering (si)RNA-directed targeting of HO-1 restored cardinal features of liver IRI in otherwise resistant IFNAR-deficient OLTs. Thus, intact Type-I IFN signaling is required for hepatic IRI, whereas HO-1 is needed for cytoprotection against innate immunity-dominated organ preservation damage in IFNAR-deficient liver transplants.

Seasonal evaluation of outdoor/indoor air quality in primary schools in Lisbon.

The aim of this study was to evaluate the indoor (I) and outdoor (O) levels of NO2, speciated volatile organic compounds (VOCs) and carbonyls at fourteen primary schools in Lisbon (Portugal) during spring, autumn and winter. Three of these schools were also selected to be monitored for comfort parameters, such as temperature and relative humidity, carbon dioxide (CO2), carbon monoxide (CO), total VOCs, and both bacterial and fungal colony-forming units per cubic metre. The concentration of CO2 and bioaerosols greatly exceeded the acceptable maximum values of 1800 mg m⁻³ and 500 CFU m⁻³, respectively, in all seasons. Most of the assessed VOCs and carbonyls occurred at I/O ratios above unity in all seasons, thus showing the importance of indoor sources and building conditions in indoor air quality. However, it has been observed that higher indoor VOC concentrations occurred more often in the colder months, while carbonyl concentrations were higher in the warm months. In general, the I/O NO2 ratios ranged between 0.35 and 1, never exceeding the unity. Some actions are suggested to improve the indoor air quality in Lisbon primary schools.

Arsenic, antimony, and other trace element contamination in a mine tailings affected area and uptake by tolerant plant species.

The study was conducted to characterize mineralogical and elemental composition of mine tailings in order to evaluate the environmental hazards, and identify the metal accumulation potential of native plant species from São Domingos mine, one of the long-term activity mines of the Iberian Pyrite Belt dating back to pre-Roman times. The mine tailings including soils and different plant species from São Domingos were analyzed for determination of tailings characteristics and chemical element contents in tailings and plants. The large amounts of mining wastes are causing significant adverse environment impacts due to acid mine drainage production and mobilization of potentially toxic metals and metalloids in residential areas, agricultural fields, downstreams, and rivers. The typical mineralogical composition is as follows: quartz, micas, K-feldspar, olivine-group minerals, magnetite, goethite, hematite, jarosite, and sulfides. The mine tailings were highly contaminated by As, Ag, Cr, Hg, Sn, Sb, Fe, and Zn; and among them, As and Sb, main contaminants, attained the highest concentrations except Fe. Arsenic has exhibited very good correlations with Au, Fe, Sb, Se, and W; and Sb with As, Au, Fe, Se, Sn, and W in tailings. Among the all plant species, the higher concentrations of all the metals were noted in Erica andevalensis, Erica australis, Echium plantagium, and Lavandula luisierra. Considering the tolerant behavior and abundant growth, the plant species Erica australis, Erica andevalensis, Lavandula luisierra, Daphne gnidium, Rumex induratus, Ulex eriocladus, Juncus, and Genista hirsutus are of major importance for the rehabilitation and recovery of degraded São Domingos mining area.

Indoor air quality in elementary schools of Lisbon in spring.

Analysis of indoor air quality (IAQ) in schools usually reveals higher levels of pollutants than in outdoor environments. The aims of this study are to measure indoor and outdoor concentrations of NO(2), speciated volatile organic compounds (VOCs) and carbonyls at 14 elementary schools in Lisbon, Portugal. The investigation was carried out in May-June 2009. Three of the schools were selected to also measure comfort parameters, such as temperature and relative humidity, carbon dioxide (CO(2)), carbon monoxide (CO), total VOCs, and bacterial and fungal colony-forming units per cubic metre. Indoor concentrations of CO(2) in the three main schools indicated inadequate classroom air exchange rates. The indoor/outdoor (I/O) NO(2) ratio ranged between 0.36 and 0.95. At the three main schools, the total bacterial and fungal colony-forming units (CFU) in both indoor and outdoor air were above the advised maximum value of 500 CFU/m(3) defined by Portuguese legislation. The aromatic compounds benzene, toluene, ethylbenzene and xylenes, followed by ethers, alcohols and terpenes, were usually the most abundant classes of VOCs. In general, the indoor total VOC concentrations were markedly higher than those observed outdoors. At all locations, indoor aldehyde levels were higher than those observed outdoors, particularly for formaldehyde. The inadequate ventilation observed likely favours accumulation of pollutants with additional indoor sources.

Adiponectin predicts the antioxidant capacity and size of high-density lipoprotein (HDL) in individuals with diabetes mellitus.

AIMS: The relationship between adiponectin and type 2 diabetes mellitus (T2DM) is established; however the evidence on its role in high-density lipoprotein (HDL) functionality is still scant. The aim of this study was to assess the association of adiponectin with HDL functionality especially on the antioxidant capacity and HDL subfractions in individuals with T2DM. METHODS: This case-control study enrolled 356 individuals who were divided into two groups: diabetics [T2DM (n = 188)] and non-diabetic [nT2DM (n = 168)]. The association of adiponectin level on HDL functionality parameters was done in function of the cut-off point for adiponectin [percentile p < 75 = 12.9 µg/mL versus p ≥ 75 = 12.9 µg/mL] and multiple adjustments applied in the logistic regression models. RESULTS: Body mass index (BMI), waist circumference (WC) and body fat mass (FM) were higher in T2DM. The larger HDL particles (HDLLARGE) were lower in T2DM group in comparison with nT2DM (28.20% versus 30.40%; p = 0.016). Individuals with T2DM and simultaneous highest adiponectin (p ≥ 75) had 2.25 OR (95% CI = 1.03-4.91) and 5.14 OR (95% CI = 2.37-11.15) to present higher HDL-C and HDLLARGE concentrations. After adjustment for multiple confounders, high level of adiponectin was independently related with improvement of the HDL antioxidant capacity (OR = 2.78; 95% CI = 1.16-6.67). CONCLUSIONS: High adiponectin level associates with a lesser negative impact of T2DM on HDL functionality by increase in APO AI, particles size, and cholesterol content. On the same token, higher adiponectin was associated with greater odds to have high antioxidant capacity.

Effect of particulate matter, atmospheric gases, temperature, and humidity on respiratory and circulatory diseases' trends in Lisbon, Portugal.

This study addresses the significant effects of both well-known contaminants (particles, gases) and less-studied variables (temperature, humidity) on serious, if relatively common, respiratory and circulatory diseases. The area of study is Lisbon, Portugal, and time series of health outcome (daily admissions in 12 hospitals) and environmental data (daily averages of air temperature, relative humidity, PM(10), SO(2), NO, NO(2), CO, and O(3)) have been gathered for 1999-2004 to ascertain (1) whether concentrations of air pollutants and levels of temperature and humidity do interfere on human health, as gauged by hospital admissions due to respiratory and circulatory ailments; and (2) whether there is an effect of population age in such admissions. In general terms, statistically significant (p < 0.001) correlations were found between hospital admissions and temperature, humidity, PM(10), and all gaseous pollutants except CO and NO. Age appears to influence respiratory conditions in association with temperature, whereas, for circulatory conditions, such an influence likely involves temperature as well as the gaseous pollutants NO(2) and SO(2).

The role of non-invasive dynamic tests in the diagnosis of Cushing's syndrome.

OBJECTIVE: To evaluate the role of non-invasive dynamic tests in the diagnosis and differential diagnosis of Cushing's syndrome (CS). METHODS: We studied laboratory features of 74 patients with endogenous CS, subdivided as follows: 46 (62.1%) with Cushing's disease (CD), 21 (28.3%) with an adrenal tumor, and 7 (9.5%) with ectopic ACTH syndrome (EAS). RESULTS: In 100% of cases of CS we found serum cortisol levels greater than 1.8 microg/dl after low-dose dexamethasone suppression tests (LDDST), as well as elevation of midnight serum or salivary cortisol. However, urinary free cortisol was normal in 11.5% of patients. ACTH levels were suppressed in patients with adrenal tumors, normal or high in CD and invariably increased in EAS. After the 8-mg overnight dexamethasone suppression test (HDDST), serum cortisol suppression >50% was observed in 79.5% of cases of CD and in 28.6% of subjects with EAS, whereas cortisol suppression >80% was only found in CD. After stimulation with CRH or desmopressin an ACTH rise > or =35% occurred in 86.5% of individuals with CD and 14.3% of those with EAS, whereas an ACTH rise > or =50 achieved 100% specificity. Moreover, the combination of serum cortisol suppression >50% after HDDST and an ACTH increase > or =35% after the administration of CRH or desmopressin only occurred in CD. CONCLUSION: Our findings demonstrate that LDDST had 100% sensitivity for the diagnosis of CS and that HDDST and stimulation tests with CRH or desmopressin may be very useful for confirmation of CS etiology when analyzed together or when more stringent cut-offs are used.

Diagnosis and management of hyperprolactinemia: results of a Brazilian multicenter study with 1234 patients.

OBJECTIVE: The aim of the study was to evaluate clinical and laboratorial features of 1234 patients with different etiologies of hyperprolactinemia, as well as the response of 388 patients with prolactinomas to dopamine agonists. DESIGN, SETTING, AND PATIENTS: A total of 1234 hyperprolactinemic patients from 10 Brazilian endocrine centers were enrolled in this retrospective study. MAIN OUTCOME MEASURE: PRL measurement, thyroid function tests, and screening for macroprolactin were conducted. RESULTS: Patients were subdivided as follows: 56.2% had prolactinomas, 14.5% drug-induced hyperprolactinemia, 9.3% macroprolactinemia, 6.6% non-functioning pituitary adenomas, 6.3% primary hypothyroidism, 3.6% idiopathic hyperprolactinemia, and 3.2% acromegaly. Clinical manifestations were similar irrespective of the etiology of the hyperprolactinemia. The highest PRL levels were observed in patients with prolactinomas but there was a great overlap in PRL values between all groups. However, PRL>500 ng/ml allowed a clear distinction between prolactinomas and the other etiologies. Cabergoline (CAB) was more effective than bromocriptine (BCR) in normalizing PRL levels (81.9% vs 67.1%, p<0.0001) and in inducing significant tumor shrinkage and complete disappearance of tumor mass. Drug resistance was observed in 10% of patients treated with CAB and in 18.4% of those that used BCR (p=0.0006). Side-effects and intolerance were also more common in BCR treated patients. CONCLUSION: Prolactinomas, drug induced hyperprolactinemia, and macroprolactinemia were the 3 most common causes of hyperprolactinemia. Although PRL levels could not reliably define the etiology of hyperprolactinemia, PRL values >500 ng/ml were exclusively seen in patients with prolactinomas. CAB was significantly more effective than BCR in terms of prolactin normalization, tumor shrinkage, and tolerability.

Correlating element atmospheric deposition and cancer mortality in Portugal: data handling and preliminary results.

This study, framed within geographical epidemiology, presents preliminary findings concerning the association between the concentrations of chemical elements obtained through atmospheric biomonitoring with lichens and cancer mortality in the Portuguese population. Exploratory analyses were performed to identify potential confounders for the relationships between chemical elements and neoplasm mortality and to assess the extent of their interference. The results of this study highlight some methodological and conceptual difficulties inherent to observational and geographical studies, in the specific context of the Portuguese population, and the challenge posed by the large numbers of pollutants considered.

Bioaccumulation behaviour of transplants of the lichen Flavoparmelia caperata in relation to total deposition at a polluted location in Portugal.

This experiment compares the short and long time element accumulation behaviour of transplants of Flavoparmelia caperata lichen thalli and total deposition in an atmospheric polluted area. It was found that lichens exposed for a short time behaved differently from lichens in cumulative exposition suggesting the presence of acclimatization behaviour. The lichen transplant elemental content does not unequivocally represent the average or cumulative environmental availability of the exposure period. Reflection characteristics depend on the element and the lichen physiological conditions. Good correlations between lichen elemental contents and total deposition were obtained when a physiological lichen parameter was introduced in a mathematical model, suggesting that metabolically mediated accumulation is important.

Elemental levels in tree-bark and epiphytic-lichen transplants at a mixed environment in mainland Portugal, and comparisons with an in situ lichen.

Samples of Platanus hybrida Brot. bark and Flavoparmelia caperata (L.) Hale thalli, from a clean area in northern Portugal (Baião), were transplanted into an exposure location at the south-western Atlantic coast, impacted by urban-industrial emissions (Sines), for a 10-month long experiment. Bark pieces were confined into nylon bags (2-mm mesh), and lichen thalli kept with their bark substrate (Pinus pinaster (Ait.) Sol.). Every two months, a double set of transplants (one for bark, one for lichens) was brought back into the laboratory, together with native samples of Evernia prunastri (L.) Ach. Following suitable cleansing and preparation procedures, field samples were put through INAA for elemental assessment. The results indicate that, regardless of signal magnitude, (1) concentrations in bark and lichen transplants are significantly correlated with atmospheric deposition for an appreciable number of elements; (2) there are a number of significant correlations between transplanted and native samples, and again between the latter and the deposition; and (3) the elements with biological patterns that follow the deposition in either transplanted or native samples (Co, V) are the very ones whose bioaccumulation seems to benefit from an alternation of wet-dry periods, which fits the precipitation record of the test site during the exposure term.

Impaired cytokine production by peripheral blood mononuclear cells in type 1 diabetic patients.

AIMS: The objective of the present investigation was to study the production of IL-1, IL-6, IL-10, IFNgamma and TNFalpha in cultures of peripheral blood mononuclear cells (PBMC) taken from type 1 diabetic patients with inadequate metabolic control. METHODS: Seventeen type 1 diabetic patients and a gender- and age-matched group of 17 healthy individuals were studied. PBMC cultures were stimulated with phytohemagglutinin (PHA; 20 microg/ml) and lipopolysaccharide (LPS; 10 microg/ml), and enzyme immunoassay (Elisa) was used to measure IL-1, IL-6, IL-10, IFNgamma and TNFalpha in the cell-culture supernatants. RESULTS: IFNgamma levels in PHA-stimulated cultures were lower in the type 1 diabetics than in the non-diabetic controls (P<0.0001) while, in contrast, IL-10 levels were increased in the PHA-stimulated culture supernatants of the diabetics compared with the controls (P<0.0001). In addition, supernatant levels of the cytokines IL-1, IL-6 and TNFalpha released in the presence of LPS in the cell cultures from the diabetic patients were significantly lower than in the non-diabetic subjects (P<0.0001, P<0.0001 and P<0.03, respectively). CONCLUSIONS: The impaired production of IL-1, IL-6, TNFalpha and IFNgamma, and the increased production of IL-10, in PBMC cultures from type 1 diabetics with inadequate metabolic control compared with healthy subjects may be an indication of a deficiency in mononuclear cell activation and, consequently, a deficient immune cellular adaptive response that, in turn, may be the cause of the increased incidence of infections in people with type 1 diabetes.

Clinical and laboratory features greatly overlap in patients with macroprolactinemia or monomeric hyperprolactinemia.

AIM: The aim of this study was to evaluate the clinical and laboratory features of 64 patients with macroprolactinemia and to compare them to those of individuals with monomeric hyperprolactinemia. METHODS: The study included 64 patients (54 women and 10 men) with macroprolactinemia and 96 patients (70 women and 26 men) with monomeric hyperprolactinemia (32 with prolactinomas). RESULTS: Symptoms related to prolactin (PRL) excess were found in about 44% of individuals from the macroprolactinemia group and in 88.5% of patients with monomeric hyperprolactinemia (P<0.0001). However, the frequency of menstrual disturbances (oligomenorrhea or amenorrhea), galactorrhea and erectile dysfunction did not differ in both groups. In contrast, the association of galactorrhea and menstrual disturbances was significantly more prevalent in women with monomeric hyperprolactinemia. Although mean PRL levels were higher in patients with monomeric hyperprolactinemia (565.9+/-2726.4 vs 113.3+/-94.5 ng/mL, P<0.001), there was a great overlap between both groups. Among macroprolactinemic patients, pituitary magnetic resonance imaging revealed an image suggestive of a microadenoma in 7 (10.9%) and a macroadenoma in 1 (1.6%). Normalization of PRL levels during therapy with dopamine agonists was significantly more frequent in patients with monomeric hyperprolactinemia than in subjects with macroprolactinemia (78.6% vs 32%, P=0.0006). CONCLUSION: Our data show that symptoms related to PRL excess are frequently found in subjects with macroprolactinemia. Moreover, no clinical or laboratory features could reliably differentiate macroprolactinemic patients from those with monomeric hyperprolactinemia. Therefore, the screening for macroprolactin should not be restricted to asymptomatic patients.

Effect of metabolic control on interferon-gamma and interleukin-10 production by peripheral blood mononuclear cells from type 1 and type 2 diabetic patients.

The objective of the present study was to evaluate the production of cytokines, interferon-gamma (INF-gamma) and interleukin-10 (IL-10), in cultures of peripheral blood mononuclear cells (PBMC) from type 1 and type 2 diabetic patients and to correlate it with inadequate and adequate metabolic control. We studied 11 type 1 and 13 type 2 diabetic patients and 21 healthy individuals divided into two groups (N = 11 and 10) paired by sex and age with type 1 and type 2 diabetic patients. The PBMC cultures were stimulated with concanavalin-A to measure INF-gamma and IL-10 supernatant concentration by ELISA. For patients with inadequate metabolic control, the cultures were performed on the first day of hospitalization and again after intensive treatment to achieve adequate control. INF-gamma levels in the supernatants of type 1 diabetic patient cultures were higher compared to type 2 diabetic patients with adequate metabolic control (P < 0.001). Additionally, INF-gamma and IL-10 tended to increase the liberation of PBMC from type 1 and 2 diabetic patients with adequate metabolic control (P = 0.009 and 0.09, respectively). The increased levels of INF-gamma and IL-10 released from PBMC of type 1 and 2 diabetic patients with adequate metabolic control suggest that diabetic control improves the capacity of activation and maintenance of the immune response, reducing the susceptibility to infections.