A Preliminary Study on Hepatoprotective, Hypolipidemic and Aortic Morphometric Effects of Omega-3-Rich Fish Oil in Hypercholesterolemic Mice.

This study aims to evaluate the hepatoprotective, hypolipidemic and aortic morphometric effects of fish oil rich in omega-3 in hypercholesterolemic BALB/c mice. This is an experimental model that included 16 male BALB/c mice (Mus musculus) divided into three groups (G1 (standard commercial chow and 0.9% saline solution), G2 (hypercholesterolemic diet and 0.9% saline solution) and G3 (hypercholesterolemic diet and fish oil)) for 8 weeks. There was no significant difference in the treatment with omega-3-rich fish oil in the lipid profile (p > 0.05). In the histological analysis, group G2 detected the presence of hepatitis and liver tissue necrosis, but this was not observed in group G3. As for the morphometry in the light area of the vessel, the G1 group had a higher score (2.62 ± 0.36 mm2) when compared to G2 (2.10 ± 0.16 mm2) and G3 (2.26 ± 0.25 mm2) (p < 0.05). The vessel wall thickness did not differ between the groups (p > 0.05). It is concluded that supplementation with fish oil rich in omega-3 carried out in this study may have a protective effect on liver tissue, but it has not yet improved the lipid and morphometric profile. Despite this research being preliminary, it is a relevant study with future prospects for improving the doses of EPA and DHA in order to better elucidate the benefits of fish oil in models of dyslipidemia.

Old but still good: genetic diversity of ancient pecan genotypes from southern Brazil.

Pecan [Carya illinoinensis (Wangenh.) K. Koch] is a crop fruit native to the USA and Mexico currently cultivated in several countries, including Brazil, Uruguay, Argentina, Chile, Peru, China, South Africa, and Australia. Supported by the increasing consumption and market prices, the interest in the cultivation of this fruit crop is strongly growing around the world. In this study, AFLP and S-SAP markers were employed to characterize the genetic diversity of ancient accessions of pecan from southern Brazil. The evaluated plants were selected and preserved by the farmers and are remnants of the first introduction of seedlings from the U.S.A into southern Brazil aiming at developing research towards establishing commercial orchards. High levels of genetic diversity were estimated, suggesting that these plants have an important genetic background for the establishment of a germplasm collection with a wide genetic basis, for the development of breeding programs for this fruit crop. Cluster analysis of the genetic datasets revealed some correlation between the nuts' morphometric traits and genetic markers. Such correlation should be further exploited. These ancient genotypes must be evaluated for other agronomic traits of interest and included in core collections of pecans.

Medicina Veterinaria Forense, Bienestar y Maltrato Animal: Percepciones y Conocimiento de Estudiantes de Medicina Veterinaria Colombianos y Brasileños.

Los médicos veterinarios juegan un papel fundamental en la detección de casos de maltrato animal y violencia doméstica. Por lo tanto, es esencial que las escuelas de medicina veterinaria brinden una formación adecuada en bienestar animal y medicina veterinaria forense. El objetivo del presente artículo es caracterizar la percepción y el conocimiento de los estudiantes de medicina veterinaria sobre la formación en medicina veterinaria forense, bienestar animal y la asociación entre maltrato animal y violencia humana. Un cuestionario online fue distribuido a los estudiantes de las 227 y 22 facultades de medicina veterinaria en Brasil y Colombia, respectivamente. Pruebas de chi-cuadrado fueron realizadas para comparar las respuestas de las variables categóricas entre los estudiantes de Brasil y Colombia. La mayoría de los estudiantes encuestados indicaron que su escuela ofrecía capacitación en bienestar animal. Sin embargo, solo el 21.8% (47/216) de los estudiantes colombianos y el 43.1% (216/523) de los brasileños mencionaron que su facultad ofrecía formación en medicina veterinaria forense. En ambos países fueron identificadas deficiencias en la capacitación sobre la identificación de traumas no accidentales, denuncia de maltrato animal y el conocimiento sobre la asociación entre maltrato animal y violencia interpersonal. A pesar de esto, más del 90% de los estudiantes relataron ser conscientes de la relación que existe entre estos dos crímenes y la importancia de recibir capacitación obligatoria sobre maltrato animal y medicina veterinaria forense. Adicionalmente, la mayoría de los encuestados reconocieron que el maltrato animal incluye tanto el abuso físico como el mental. Nuestros resultados destacan la necesidad de mejorar la educación en bienestar animal, maltrato animal, violencia humana y medicina veterinaria forense en las facultades de medicina veterinaria de Brasil y Colombia.

Characterizing protein conformers by cross-linking mass spectrometry and pattern recognition.

MOTIVATION: Chemical cross-linking coupled to mass spectrometry (XLMS) emerged as a powerful technique for studying protein structures and large-scale protein-protein interactions. Nonetheless, XLMS lacks software tailored toward dealing with multiple conformers; this scenario can lead to high-quality identifications that are mutually exclusive. This limitation hampers the applicability of XLMS in structural experiments of dynamic protein systems, where less abundant conformers of the target protein are expected in the sample. RESULTS: We present QUIN-XL, a software that uses unsupervised clustering to group cross-link identifications by their quantitative profile across multiple samples. QUIN-XL highlights regions of the protein or system presenting changes in its conformation when comparing different biological conditions. We demonstrate our software's usefulness by revisiting the HSP90 protein, comparing three of its different conformers. QUIN-XL's clusters correlate directly to known protein 3D structures of the conformers and therefore validates our software. AVAILABILITYAND IMPLEMENTATION: QUIN-XL and a user tutorial are freely available at http://patternlabforproteomics.org/quinxl for academic users. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.

Veterinary Forensics, Animal Welfare and Animal Abuse: Perceptions and Knowledge of Brazilian and Colombian Veterinary Students.

Veterinarians have a fundamental role to play in the detection of animal abuse and domestic violence cases. Therefore, it is essential that veterinary colleges provide appropriate training in animal welfare and veterinary forensics. The aim of this article is to characterize the perception and knowledge of veterinary students about training in veterinary forensics, animal welfare and the association between animal abuse and human violence. An online survey was made available to veterinary students at 227 veterinary colleges in Brazil and 22 in Colombia. The Chi-square test of independence was performed to compare responses of Brazilian and Colombian students for categorical survey items. Most of the surveyed students indicated that their college offered animal welfare training. However, only 21.8% (47/216) of the Colombian and 43.1% (216/523) of the Brazilian students mentioned that their veterinary colleges offered veterinary forensics training. Deficits in training in identification of non-accidental traumas, reporting of animal abuse and awareness of the association between interpersonal violence and animal abuse were identified in both countries. Despite this, more than 90% of students were aware of the relationship between these two crimes and in the importance of receiving compulsory training in animal abuse and veterinary forensics. Likewise, most of the respondents recognized that animal abuse includes both physical and mental abuse. The results highlight the need to improve education in animal welfare, animal abuse, human violence and veterinary forensics in veterinary colleges in Brazil and Colombia.

Corrigendum: Revisiting Meiosis in Sugarcane: Chromosomal Irregularities and the Prevalence of Bivalent Configurations.

[This corrects the article DOI: 10.3389/fgene.2018.00213.].

Optical coherence tomography follow-up of patients treated from periodontal disease.

Optical coherence tomography (OCT) is one of the most important imaging modalities for biophotonics applications. In this work, an important step towards the clinical use of OCT in dental practice is reported, by following-up patients treated from periodontal disease (PD). A total of 147 vestibular dental sites from 14 patients diagnosed with PD were evaluated prior and after treatment, using a swept-source OCT and two periodontal probes (Florida probe and North Carolina) for comparison. The evaluation was performed at four stages: day 0, day 30, day 60 and day 90. Exceptionally one patient was evaluated 1-year after treatment. It was possible to visualize in the two-dimensional images the architectural components that compose the periodontal anatomy, and identify the improvements in biofilm and dental calculus upon treatment. In the follow-up after the treatment, it was observed in some cases decrease of the gingival thickness associated with extinction of gingival calculus. In some cases, the improvement of both depth of probing with the traditional probes and the evidence in the images of the region was emphasized. The study evidenced the ability of OCT in the identification of periodontal structures and alterations, being an important noninvasive complement or even alternative for periodontal probes for treatment follow-up. OCT system being used in a clinical environment. Above OCT image (left) prior treatment and (right) 30 days after treatment.

Revisiting Meiosis in Sugarcane: Chromosomal Irregularities and the Prevalence of Bivalent Configurations.

Traditional sugarcane cultivars (Saccharum officinarum) proved highly susceptible to diseases, and this led breeders to progress to interspecific crosses resulting in disease resistance. A backcrossing program to S. officinarum was then required to boost sucrose content. Clonal selection across generations and incorporation of other germplasm into cultivated backgrounds established the (narrow) genetic base of modern cultivars (Saccharum spp.), which have a man-made genome. The genome complexity has inspired several molecular studies that have elucidated aspects of sugarcane genome constitution, architecture, and cytogenetics. However, there is a critical shortage of information on chromosome behavior throughout meiosis in modern cultivars. In this study, we examined the microsporogenesis of a contemporary variety, providing a detailed analysis of the meiotic process and chromosome association at diakinesis, using FISH with centromeric probes. Chromosomal abnormalities were documented by examining high quality preparations of pollen mother cells (700 in total). Approximately 70% of the cells showed abnormalities, such as metaphase chromosomes not lined up at the plate, lagging chromosomes and chromosomal bridges, and tetrad cells with micronuclei. Some dyads with asynchronous behavior were also observed. Due to the hybrid composition of the sugarcane genome, we suggest that bivalent incomplete pairing may occur in the first prophase leading to univalency. The presence of rod bivalents showing the lagging tendency is consistent with a reduction in chiasma frequency. Finally, the presence of chromatin bridges indicates the indirect occurrence of chromosomal inversions, although chromosome fragments were not clearly recognized. Possible reasons for such meiotic abnormalities and the large prevalence of bivalent formation are discussed.

A simple, rapid and green ultrasound assisted and ionic liquid dispersive microextraction procedure for the determination of tin in foods employing ETAAS.

This paper proposes a simple, rapid and green ultrasound assisted and ionic liquid dispersive microextraction procedure using pyrocatechol violet (PV) as complexing reagent and 1-hexyl-3-methylimidazolium bis(trifluoromethylsulfonyl)-imide [C6MIM][Tf2N] as ionic liquid for the detection of tin employing electrothermal atomic absorption spectrometry (ETAAS). The optimization step was performed using a two-level full factorial design involving the following factors: pH of the working media, amount reagents, ionic liquid volume and extraction time and the chemometric response was tin recovery. The procedure allowed the determination of tin with limits of detection and quantification of 3.4 and 11.3 ng L-1, respectively. The relative standard deviation was 4.5% for a tin solution of 0.50 µg L-1. The validation method was confirmed by analysis of rice flour certified reference material. The method was applied for the quantification of tin in several food samples. The concentration range found varied from 0.10 to 1.50 µg g-1.

Original Research: ACE2 activator associated with physical exercise potentiates the reduction of pulmonary fibrosis.

The interstitial lung diseases are poorly understood and there are currently no studies evaluating the association of physical exercise with an ACE2 activator (DIZE) as a possible treatment for this group of diseases. We evaluate the effects of pharmacological treatment with an angiotensin-converting enzyme 2 activator drug, associated with exercise, on the pulmonary lesions induced by bleomycin. From the 96 male Balb/c mice used in the experiment, only 49 received 8 U/kg of bleomycin (BLM, intratracheally). The mice were divided into control (C) and bleomycin (BLM) groups, sedentary and trained (C-SED, C-EXE, BLM-SED, BLM-EXE), control and bleomycin and also sedentary and trained treated with diminazene (C-SED/E, C-EXE/E, BLM-SED/E, BLM-EXE/E). The animals were trained five days/week, 1 h/day with 60% of the maximum load obtained in a functional capacity test, for four weeks. Diminazene groups were treated (1 mg/kg, by gavage) daily until the end of the experiment. The lungs were collected 48 h after the training program, set in buffered formalin and investigated by Gomori's trichrome, immunohistochemistry of collagen type I, TGF-ß1, beta-prolyl-4-hydroxylase, MMP-1 and -2. The BLM-EXE/E group obtained a significant increase in functional capacity, reduced amount of fibrosis and type I collagen, decreased expression of TGF-ß1 and beta-prolyl-4-hydroxylase and an increase of metalloproteinase -1, -2 when compared with the other groups. The present research shows, for the first time, that exercise training associated with the activation of ACE2 potentially reduces pulmonary fibrosis.

Neutrophilic infiltration in lungs of mice with peritonitis in acid or basic medium.

BACKGROUND: Bacterial peritonitis is associated with systemic complications such as pneumonia. OBJECTIVE: To determine in an experimental model of peritonitis whether the pH of peritoneal fluid infection influences the influx of neutrophils into the lung, and whether treatment outcome would be similar in peritonitis with liquid at any pH. MATERIALS AND METHODS: We studied 48 mice with peritonitis induced by cecal ligation and puncture. The animals were distributed randomly into three groups: the first one had an injection into the peritoneal cavity with saline, pH 7.0; the second group was injected with saline, pH 8.0; and the third group with saline, pH 3.0. After 2 hours, half the animals in each group was treated by washing the abdominal cavity with warm saline solution and administration of ceftriaxone every 12 hours, and half of each group was killed by anesthetic overdose, and lung biopsy was done. The animals kept in treatment were killed 24 hours after treatment, and lung biopsy was also performed. The samples were stained with H&E and the number of neutrophils in 20 areas was checked. The mean number of cells in each group was compared between groups and with an untreated one. RESULTS: The group with peritonitis associated with alkaline solution showed a higher population of neutrophils during untreated peritonitis (P = 0.04). The response to treatment by lavage of the peritoneal cavity and antibiotics was more effective in reducing the population of neutrophils in the group with peritonitis at pH 8.0, unlike that observed in animals with peritonitis at pH 3.0 (P = 0.03). CONCLUSION: Peritonitis associated with lower pH solution, despite the lower influx of leukocytes in the first two hours after installation of peritonitis, was not able to reduce the population of these cells in mice's lung in response to standard therapy.

Non-Invasive Periodontal Probing Through Fourier-Domain Optical Coherence Tomography.

BACKGROUND: Periodontitis is a multifactorial and infectious disease that may result in significant debilitation. The aim of this study is to exploit two optical coherence tomography (OCT) systems operating in the Fourier domain at different wavelengths, 930 and 1,325 nm, for structural analysis of periodontal tissue in porcine jaws. METHODS: Five fresh porcine jaws were sectioned and stored in formalin before OCT analysis. Two- and three-dimensional OCT images of the tooth/gingiva interface were performed, and measurements of the gingival structures were obtained. The 930-nm OCT system operates in the spectral domain, whereas the 1,325-nm system is a swept-source model. Stereomicroscope images, the gold standard, were used for direct comparison. RESULTS: Through image analysis, it is possible to identify the free gingiva and the attached gingiva, the calculus deposition over tooth surfaces, and the subgingival calculus that enables the enlargement of the gingival sulcus. In addition, the gingival thickness and the gingival sulcus depth can be non-invasively measured, varying from 0.8 to 4 mm. CONCLUSIONS: Regarding the ability of the two OCT systems to visualize periodontal structures, the system operating at 1,325 nm shows a better performance, owing to a longer central wavelength that allows deeper tissue penetration. The results with the system at 930 nm can also be used, but some features could not be observed due to its lower penetration depth in the tissue.

Development and characterization of nine microsatellite loci for Sisyrinchium micranthum (Iridaceae).

PREMISE OF THE STUDY: Sisyrinchium micranthum is an herb with wide distribution and morphological variation, presenting three known cytotypes (2x, 4x, and 6x), which is naturally occurring from Mexico to Argentina. The development of microsatellite markers for this species is important to provide molecular tools to investigate polyploidy origin, as well as its relationship with phenotypic and genetic variability. • METHODS AND RESULTS: Nine microsatellite markers were isolated using an enriched genomic library and characterized in a diploid population. Eight markers generated polymorphic loci in 27 plants of a population from southern Brazil. The number of alleles per locus ranged from two to nine, and the observed and expected heterozygosities varied from 0.192 to 0.741 and from 0.370 to 0.884, respectively. • CONCLUSIONS: These species-specific DNA markers may be useful for further investigations of the involved polyploidization process and the population genetics of S. micranthum.

Exercise attenuates pulmonary injury in mice with bleomycin-induced pulmonary fibrosis.

Human idiopathic pulmonary fibrosis (IPF) is a disease with unknown etiology and poor prognosis in which patients present a decrease in functional exercise tolerance and quality of life. At present, no treatment which can improve the prognosis of this disease is available. Many biomarkers of pulmonary fibrosis have been studied, and surfactant protein A (SP-A) expression is considered a specific marker of lung disease. This study aimed to investigate the influence of exercise training on exercise endurance capacity and murine-lung lesions induced by bleomycin (BLM). Thirty-four male Balb/c mice were subdivided into four groups: control sedentary (C-SED), bleomycin-treated sedentary (BLM-SED), control exercised (C-EXE) and bleomycin-treated exercised (BLM-EXE). Mice received 6.25 U/kg of BLM or saline via intratracheal instillation. After adaptation in a swimming pool, the animals started training one hour per day, with 60% of maximum load obtained in exercise endurance capacity assessment, five days/week for four weeks. The lungs were collected 48 h after the second endurance capacity assessment, fixed in buffered formalin and embedded in paraffin. Sections were analyzed using histochemical and immunohistochemical reactions for digital morphometry of pulmonary fibrosis, type I collagen, SP-A and type II pneumocytes (PII). The exercise endurance capacity of groups C-EXE (9.20 ± 0.81 min) and BLM-EXE (8.40 ± 0.82 min) increased significantly when compared with groups C-SED (5.84 ± 0.4 min) and BLM-SED (5.67 ± 0.60 min). The amounts of connective tissue, type I collagen, PII and SP-A increased significantly in the BLM-SED group. Exercise training significantly attenuated this response as observed in the BLM-EXE group. The present study shows that exercise training can prevent the decline of exercise endurance capacity and attenuate the progression of IPF.

Susceptibility to Entamoeba histolytica intestinal infection is related to reduction in natural killer T-lymphocytes in C57BL/6 mice.

Entamoeba histolytica is a protozoan that causes amoebiasis. Recent studies demonstrated that natural killer T lymphocytes (NKT) are critical for preventing the development of amoebic liver abscess. In spite of that, there are only a handful of studies in the area. Herein, we explored the role of NKT cells in E. histolytica infection using C57BL/6 wild-type and CD1(-/-) mice. Animals were inoculated with E. histolytica and sacrificed 48 hours later to collect caecum samples that were used for quantitative analyses of lesions, trophozoites, NK1.1(+) T lymphocytes and expression of the mucus protein MUC-2 by immunohistochemistry technique. Quantitative analyses confirmed that the frequency of NK1.1(+) T cells was significantly lower in samples from C57BL/6 CD1(-/-) mice as compared to their wild type (WT) counterparts. The extension of necrotic mucosa was larger and the number of trophozoites higher in Entamoeba (Eh)-infected CD1(-/-) mice when compared with Eh-infected WT mice. In mice from both groups, non-infected (CTRL) and Eh-infected CD1(-/-), there was a reduction in the thickness of the caecal mucosa and in the MUC-2-stained area in comparison with CTRL- and Eh-WT mice. Our results showed that NKT lymphocytes contribute to resistance against Entamoeba histolytica infection and to the control of inflammation in the colitis induced by infection. The presence of a normal epithelial layer containing appropriate levels of mucus had also a protective role against infection.

Toll-like receptor 9 is required for full host resistance to Mycobacterium avium infection but plays no role in induction of Th1 responses.

To investigate the role of Toll-like receptor 9 (TLR9) in innate immunity to Mycobacterium avium, TLR9, TLR2, and MyD88 knockout (KO) mice were infected with this bacterium. Bacterial burdens were higher in the spleens, livers, and lungs of infected TLR9 KO mice than in those of C57BL/6 mice, indicating that TLR9 is required for efficient control of M. avium infection. However, TLR9 KO or TLR2 KO spleen cells displayed normal M. avium-induced tumor necrosis factor alpha (TNF-α) and gamma interferon (IFN-γ) responses. This finding was confirmed by determining the number of splenic CD4(+) T cells producing IFN-γ by flow cytometry. Furthermore, TLR2 and MyD88, but not TLR9, played a major role in interleukin-12 and TNF-α production by M. avium-infected macrophages and dendritic cells (DCs). We also found that major histocompatibility complex class II molecule expression on DCs is regulated by TLR2 and MyD88 signaling but not by TLR9. Finally, lack of TLR9, TLR2, or MyD88 reduced the numbers of macrophages, epithelioid cells, and lymphocytes in M. avium-induced granulomas but only MyD88 deficiency affected the number of liver granulomas. In summary, our data demonstrated that the involvement of TLR9 in the control of M. avium infection is not related to the induction of Th1 responses.

Effects of acute systemic cocaine administration on the cortisol, ACTH and prolactin levels of black tufted-ear marmosets.

The effect of acute systemic cocaine administration on total circulating cortisol, adrenocorticotropin hormone (ACTH) and prolactin levels of nine adult black tufted-ear marmoset monkeys (Callithrix penicillata) was investigated. For each subject, two blood samples were obtained after an i.p. injection of 10 and 20mg/kg of cocaine: one sample was drawn 30-min post-injection, while the second was following a 60-min interval. Blood samples were also obtained 0, 30 and 60 min following a saline injection. The 20 mg/kg dose of cocaine induced a significant decrease in ACTH levels 60 min after being administered. Circulating levels of cortisol, on the other hand, increased significantly 60 min after the 20 mg/kg dose. Prolactin concentrations decreased significantly 30 and 60 min after both doses of cocaine had been administered (10 and 20 mg/kg). Finally, the content of the three hormones analyzed remained constant 0, 30 and 60 min post-saline administrations. Therefore, an acute systemic cocaine challenge alters the basal circulating levels of ACTH, cortisol and prolactin of marmoset monkeys. A distinct response pattern was observed for each hormone analyzed, possibly related to specific negative feedback mechanisms and/or this species glucocorticoid-resistance. The use of this small neotropical primate may thus provide a unique opportunity to investigate cocaine-induced neuroendocrine effects.

The Tec family tyrosine kinases Itk and Rlk regulate the development of conventional CD8+ T cells.

The Tec family tyrosine kinases, Itk and Rlk, are expressed in thymocytes and peripheral T cells and regulate thresholds of T cell receptor signaling. Yet little is known about the specific role of Itk- and Rlk-dependent signals in CD8(+) T cell maturation. We show here that Itk(-/-) and Rlk(-/-)Itk(-/-) mice were nearly devoid of conventional CD8(+) T cells and, instead, contained a large population of CD8(+) T cells that bear striking similarity to lineages of innate lymphocytes. Itk(-/-) and Rlk(-/-)Itk(-/-) CD8(+) thymocytes and T cells were CD44(hi), CD122(+), and NK1.1(+); were able to produce interferon-gamma directly ex vivo; and were dependent on interleukin-15. Itk(-/-) and Rlk(-/-)Itk(-/-) CD8(+) thymocytes expressed abundant transcripts for the T box transcription factor, eomesodermin, correlating with their phenotype and function. These data indicate a critical role for Itk and Rlk in conventional CD8(+) T cell development in the thymus.

Tec kinases Itk and Rlk are required for CD8+ T cell responses to virus infection independent of their role in CD4+ T cell help.

Itk and Rlk are members of the Tec kinase family of nonreceptor protein tyrosine kinases that are expressed in T cells, NK cells, and mast cells. These proteins are involved in the regulation of signaling processes downstream of the TCR in CD4(+) T cells, particularly in the phosphorylation of phospholipase C-gamma1 after TCR activation; furthermore, both Itk and Rlk are important in CD4(+) T cell development, differentiation, function, and homeostasis. However, few studies have addressed the roles of these kinases in CD8(+) T cell signaling and function. Using Itk(-/-) and Itk(-/-)Rlk(-/-) mice, we examined the roles of these Tec family kinases in CD8(+) T cells, both in vitro and in vivo. These studies demonstrate that the loss of Itk and Rlk impairs TCR-dependent signaling, causing defects in phospholipase C-gamma1, p38, and ERK activation as well as defects in calcium flux and cytokine production in vitro and expansion and effector cytokine production by CD8(+) T cells in response to viral infection. These defects cannot be rescued by providing virus-specific CD4(+) T cell help, thereby substantiating the important role of Tec kinases in CD8(+) T cell signaling.

Endoscopic evaluation of the upper gastrointestinal tract is worthwhile in premenopausal women with iron-deficiency anaemia irrespective of menstrual flow.

BACKGROUND: In premenopausal women, iron-deficiency anaemia is common and menstrual flow is often held responsible, but it is not clear whether these women should be submitted to gastrointestinal (GI) evaluation. We aim to prospectively investigate whether premenopausal women with iron-deficiency anaemia benefit from GI evaluation regardless of menstrual flow. METHODS: The study population comprised 59 consecutive premenopausal women with iron-deficiency anaemia. Excluded were women with obvious or suspected causes of anaemia and those < or = 21 years. Heavy menstrual loss was not considered an exclusion criterion. All subjects had: complete blood count, ferritin, non-invasive testing by faecal occult blood (FOB), 13C-urea breath test (13C-UBT), anti-tissue transglutaminase antibodies (tTG) and gastrin levels. Gastroscopy with antral (n = 3), corporal (n = 3) and duodenal (n = 2) biopsies was performed in women with positive 13C-UBT or tTG titre or hypergastrinaemia. RESULTS: Heavy menstrual loss was present in 50.8%. Non-invasive tests were positive in 40/59 (67.8%): 30 had positive 13C-UBT, 12 had hypergastrinaemia, 7 had positive tTG and 3 had positive FOB. Women tested positive were similar to those tested negative as far as concerned age, haemoglobin and ferritin levels and heavy menstrual flow (55% versus 42.1%). All 40 women tested positive underwent gastroscopy with biopsies. Four (10%) had bleeding-associated lesions and 34 (85%) had non-bleeding-associated lesions. As regards upper GI findings, no differences were observed between women with normal and those with heavy menstrual flow. No lower GI tract lesions were detected in the three women with positive FOB. CONCLUSIONS: Our data suggest that premenopausal women with iron-deficiency anaemia benefit from endoscopic evaluation of the upper GI tract irrespective of menstrual flow.