Squamous cell carcinoma and basal cell carcinoma of the lips: 25 years of experience in a northeast Brazilian population.

BACKGROUND: The lips are the transition zone between the facial skin and the oral mucosa and are the site of alterations related to a broad spectrum of etiologies. Squamous cell carcinoma (SCC) and basal cell carcinoma (BCC) are the most prevalent neoplasms affecting lips. This study evaluated the demographic and clinicopathological features of the SCC and BCC in the lip. MATERIAL AND METHODS: A retrospective cross-sectional descriptive study (1994-2019) was carried out. Demographic and clinicopathologic data were collected from a hospital's dermatological service and an oncologic hospital. The data were submitted to descriptive analysis and Pearson's chi-square and Fisher's exact tests (p ≤ 0.05). RESULTS: 417 medical records were analyzed, of which 323 corresponded to SCC (77.5%) and 94 to BCC (22.5%). SCC showed more frequency in males (58.8%) and BCC in females (54.3%). The lower lip was significantly affected in male patients (p < 0.0001) and by both neoplasms (70.6% and 56.4%, respectively; p = 0.014). SCC and BCC were mainly treated with surgery (88.3% and 93.2%, respectively). Surgical margin was frequently negative in SCC and BCC (87%; 72.3%, respectively), and no recurrence was observed in 79.9% of SCC and 69.1% of BCC cases. CONCLUSIONS: SCC was more frequent in male patients, while BCC showed more frequency in female patients. Both neoplasms mainly affect the lower lip. Understanding the epidemiological profile of these lesions in the lip, as well as their etiology and clinical features, is fundamental for appropriate clinical conduct and the creation and/or amplification of preventive measures.

Identification and characterisation of de novo germline structural variants in two commercial pig lines using trio-based whole genome sequencing.

BACKGROUND: De novo mutations arising in the germline are a source of genetic variation and their discovery broadens our understanding of genetic disorders and evolutionary patterns. Although the number of de novo single nucleotide variants (dnSNVs) has been studied in a number of species, relatively little is known about the occurrence of de novo structural variants (dnSVs). In this study, we investigated 37 deeply sequenced pig trios from two commercial lines to identify dnSVs present in the offspring. The identified dnSVs were characterised by identifying their parent of origin, their functional annotations and characterizing sequence homology at the breakpoints. RESULTS: We identified four swine germline dnSVs, all located in intronic regions of protein-coding genes. Our conservative, first estimate of the swine germline dnSV rate is 0.108 (95% CI 0.038-0.255) per generation (one dnSV per nine offspring), detected using short-read sequencing. Two detected dnSVs are clusters of mutations. Mutation cluster 1 contains a de novo duplication, a dnSNV and a de novo deletion. Mutation cluster 2 contains a de novo deletion and three de novo duplications, of which one is inverted. Mutation cluster 2 is 25 kb in size, whereas mutation cluster 1 (197 bp) and the other two individual dnSVs (64 and 573 bp) are smaller. Only mutation cluster 2 could be phased and is located on the paternal haplotype. Mutation cluster 2 originates from both micro-homology as well as non-homology mutation mechanisms, where mutation cluster 1 and the other two dnSVs are caused by mutation mechanisms lacking sequence homology. The 64 bp deletion and mutation cluster 1 were validated through PCR. Lastly, the 64 bp deletion and the 573 bp duplication were validated in sequenced offspring of probands with three generations of sequence data. CONCLUSIONS: Our estimate of 0.108 dnSVs per generation in the swine germline is conservative, due to our small sample size and restricted possibilities of dnSV detection from short-read sequencing. The current study highlights the complexity of dnSVs and shows the potential of breeding programs for pigs and livestock species in general, to provide a suitable population structure for identification and characterisation of dnSVs.

The stromal immunoexpression of CLIC4 may be related to the difference in the biological behavior between oral squamous cell carcinoma and oral verrucous carcinoma.

BACKGROUND: Oral squamous cell carcinoma (OSCC) has high morbidity and mortality rates while oral verrucous carcinoma (OVC), an uncommon variant of OSCC, exhibits a distinct biological behavior. CLIC4 protein plays a role in the cell cycle and apoptosis regulation and participates in the myofibroblasts transdifferentiation process, which are the main cells of the tumor stroma. This study analyzed the immunoexpression of CLIC4 and α-SMA in 20 OSCC cases and 15 OVC cases. MATERIAL AND METHODS: A semiquantitative analysis of CLIC4 and α-SMA immunoexpression was performed in the parenchyma and stroma. Nuclear and cytoplasmic reactivity was analyzed separately for the CLIC4 immunostaining. The data were submitted to Pearson's chi-square and Spearman's correlation tests (p ≤ 0.05). RESULTS: In the CLIC4 analysis, there was a significant difference in the immunoexpression of this protein between OSCC and OVC stroma (p < 0.001). It was observed a higher expression of α-SMA in the OSCC stroma. There was a positive and significant correlation between CLIC4 and α-SMA immunoexpression in the OVC stroma (r = 0,612; p = 0,015). CONCLUSIONS: The decrease or absence of nuclear CLIC4 immunoexpression in the neoplastic epithelial cells and the increase of its expression in the stroma may influence the difference in biological behavior between OSCC and OVC.

Peripheral ossifying fibroma: A 20-year retrospective study with focus on clinical and morphological features.

BACKGROUND: Peripheral Ossifying Fibroma (POF) is a reactive hyperplastic lesion that exclusively occurs in the gingiva and is characterized by the deposition of dystrophic calcification, cementum-like tissue, and immature and mature bone within the connective tissue. The objective of the present study was to perform a retrospective analysis of clinicopathologic features of POF. MATERIAL AND METHODS: Clinical and histopathological data were obtained from biopsy records and histopathological reports from a Brazilian reference service in Oral Pathology (1999 - 2020). Morphological analysis was performed to evaluate features related to the mesenchymal component, inflammatory infiltrate, ulceration, and mineralized tissue. RESULTS: A total of 270 POFs were diagnosed during the study period. A higher frequency was observed in females (71.9%) between the third (22.9%) and fourth (23.3%) decades of life. The anterior upper gingiva (29.1%) was the most affected region. Mature (86.7%) and immature (52.6%) bone tissue were the most frequent. There was a significant association between immature bone deposition and lesions with size ≤ 1.7 cm (p = 0.041); immature bone and cement-like tissue deposition with an evolution time ≤ 16 months (p < 0.001); deposition of immature bone and mesenchymal hypercellularization (p < 0.001); deposition of dystrophic calcification and the presence of ulceration (p < 0.001). CONCLUSIONS: The clinical characteristics corroborate the findings in the literature. The heterogeneous distribution and quantity of mineralized tissues found in the analyzed cases support the theory that the different mineralized tissues constitute a spectrum of clinical maturation of POF.

Angina bullosa haemorrhagica: A 14-year multi-institutional retrospective study from Brazil and literature review.

BACKGROUND: Angina bullosa haemorrhagica (ABH) is characterized by the recurrent appearance of blood blisters on the oral mucosa, mainly in adults' soft palate. In general, the blisters rupture spontaneously, lacking the necessity for biopsy. We report the clinical features of 23 ABH cases, emphasizing the clinical behavior and the management of these conditions. MATERIAL AND METHODS: A retrospective descriptive cross-sectional study was performed. A total of 12,727 clinical records of oral and maxillofacial lesions from four dental services in Brazil were analyzed. Clinical data were collected from the clinical records and evaluated. RESULTS: The series comprised 12 males (52.2%) and 11 females (47.8%), with a mean age of 56.8 ± 14.6 years (ranging: 24-82 years) and a 1.1:1 male-to-female ratio. Most of the lesions affected the soft palate (n = 15, 65.2%). Clinically, the lesions presented mainly as an asymptomatic (n = 17, 73.9%) blood-filled blister that ruptured after a few minutes or hours, leaving an erosion. The masticatory trauma was the most frequent triggering event. No patient had coagulation disorders. A biopsy was performed in only four cases (17.4%). Treatment was symptomatic with a favorable outcome. CONCLUSIONS: ABH is still poorly documented in the literature, and its etiology remains uncertain. ABH mainly affects the soft palate of elderly adults and has a favorable evolution in a few days. The therapeutic approach is often focused only on the relief of symptoms. However, it can share some clinical features with more serious diseases. Therefore, clinicians must recognize these lesions to avoid misdiagnosis.

Features of aerobic granular sludge formation treating fluctuating industrial saline wastewater at pilot scale.

A pilot-scale sequencing batch reactor, with a working volume of 3 m3, was installed in a fish cannery to develop aerobic granular sludge treating the produced effluents. Depending on the nitrogen (N) and organic matter (COD) concentration, the effluents were named in this study as medium-low-strength (Stage I) and high-strength (Stage II) wastewater. The composition of the wastewater was found to be a crucial factor to select granule-forming organisms. With medium-low-strength wastewater as feeding, the first granules were observed after 30 days, but the extremely high COD/N ratios of the wastewater provoked the overgrowth of filamentous bacteria after 4 months of operation (Stage I). When treating high-strength wastewater, stable aggregates with good settleability appeared, but well-shaped granules were not observed since the granulation process was not completed. The system was able to remove both COD (70-95%) and N (30-90%) treating both types of effluents. Biomass growth was the main N removal pathway. The reactor was found to be robust against factory production stops and, thus, a suitable alternative to treat wastewater from industries with discontinuous operation.

Molossinema wimsatti infection in the brain of Pallas's mastiff bats (Molossus molossus).

The present report describes two cases of infection by Molossinema wimsatti in the brain of Pallas's mastiff bats (Molossus molossus). The first bat was captured and killed by a domestic cat in a suburban area of the municipality of Patos, Paraiba, northeastern Brazil. The second bat was found crawling on the ground in the same area before dying. No gross lesions were found at necropsy. Histology of the central nervous system revealed filarioid nematodes in the brain ventricles and cerebellum. There were adults, subadults and eggs, the latter sometimes containing microfilariae. No inflammatory response was observed in bat 1, while bat 2 presented a mild lymphoplasmacytic meningoencephalitis. Three nematodes were recovered and submitted for parasitological examination. The diagnosis of M. wimsatti infection was based on the histomorphological and parasitological characteristics of the agent and its location in the brain ventricular system of insectivorous bats. The infection likely occurs in other insectivorous bats from South American and Caribbean countries but may be overlooked.

Unpaved road conservation planning at the catchment scale.

In addition to soil losses on hillslopes, unpaved rural roads, especially when poorly designed and maintained, can be a significant contributor to the erosive processes seen at the catchment scale. In areas with deep soils, the solutions primarily focus on channeling excess surface runoff into settling ponds or terraces. However, few studies have addressed runoff control from roads on steep slopes in areas of shallow soil. Modeling hydrological processes at the catchment scale is a useful strategy for choosing the most effective and least costly conservation practices to control surface runoff. This study applies a mathematical model to a monitored catchment in southern Brazil to better understand the effects of conservation practices on unpaved roads and their impact on the hydrological and erosive dynamics of a small rural catchment. We calibrated the LISEM model using data from eight stormwater events and evaluated how three different road conservation scenarios-low (LI), medium (MI), and high intensity (HI)-contributed to sediment yield (SY), surface runoff volume (Qe), and peak flow (Qp) reduction. The LI and MI scenarios involved installation of hydraulic structures to control the road surface runoff (i.e. road ditch graveling, diversion weirs and grass waterways) while the HI scenario added surface runoff control practices (grass strips) to surrounding crop fields, in addition to the practices included in the MI scenario. Based on these scenarios, the results showed a Qe reduction at the catchment outlet from - 3.5% (LI) to - 22.5% (HI). The Qp and SY varied from + 6.0% (LI) to - 292.5% (HI) and from + 20.0% (LI) to - 963.9% (HI), respectively. These results show that the low- and medium-intensity practices were not effective in controlling surface runoff from roads, based on the Qe, Qb, and SY observed at the catchment's outlet. On the other hand, when MI scenarios were complemented with practices to control surface runoff in the cultivated areas, a significant reduction in surface runoff (Qe and Qp) and SY was verified.

Clinicopathologic features of nasopalatine duct cysts: A retrospective study in two Brazilian oral and maxillofacial pathology referral centers.

BACKGROUND: Nasopalatine duct cyst (NDC) is the most common non-odontogenic cyst in the oral cavity. Clinically it is not difficult to suspect these lesions based on clinical and radiographic appearance. However, the histopathological diagnosis may be difficult due to the broad morphological diversity of these lesions. The objective was to analyze the clinicopathological features of NDCs diagnosed in two oral and maxillofacial pathology services in the Brazilian northeast. MATERIAL AND METHODS: A retrospective clinicopathologic study was performed. A total of 18,121 clinical records of oral lesions from two oral and maxillofacial pathology services in Brazil were analyzed (2000-2020). All NDCs cases were revised and demographic, clinical, radiographic, and histopathological data were collected. RESULTS: Among 18,121 diagnoses in the oral pathology services, 45 (0.2%) were NDCs. The series comprises 24 males (53.3%) and 21 females (46.7%), with a mean age of 43.2 years-old. Most lesions were asymptomatic (n = 27, 60%) with an mean size of 2.1 cm. Microscopically, the non-keratinized stratified squamous epithelium was the most common (66.7%). However, in 88.9% of cases, the epithelial lining was varied and composed of two or more types of epithelium. There was no significant association between the type of epithelium and the size of the cysts (p = 0.389). Nerve, blood vessels, hemorrhage, and chronic inflammatory infiltrate were commonly observed. In contrast, there was a low frequency of mucous glands, sebaceous glands, cholesterol clefts, and multinucleated giant cells. CONCLUSIONS: The clinical, radiographic, and microscopic findings observed in this study are similar to those reported in the literature. Due to the morphological diversity of NDC, it is needed to correlate its histopathological features with the clinical and radiographic findings to establish a correct diagnosis.

Postprandial glycemia in healthy subjects: Which probiotic dairy food is more adequate?

The consumption of probiotic-enriched dairy products has been associated with many health benefits, including anti-hyperglycemic activity. The effect on health is dependent on the type of probiotic culture used and the dairy product consumed. This study evaluated the effect of different probiotic-enriched dairy matrices (Minas Frescal cheese, Prato cheese, and whey dairy beverage) containing Lactobacillus casei on in vitro and in vivo anti-hyperglycemic activity. For this purpose, in vitro anti-hyperglycemic activity was determined by the inhibition of α-glucosidase and α-amylase activities, and a human study was performed with healthy individuals (n = 15, consumption of bread as a control; bread + Minas Frescal cheese; bread + Prato cheese; bread + dairy beverage) to assess the effects of different probiotic foods on postprandial glycemia. In vitro data showed that Prato cheese presented the highest lipid (36.9 g/100 g) and protein (26.5 g/100 g) contents as well as the highest α-amylase (60.7%) and α-glucosidase (52.6%) inhibition. The consumption of Prato cheese resulted in a lesser increase in blood glucose level (13 mg/dL) compared with the consumption of bread alone (19 mg/dL), Minas Frescal cheese (20 mg/dL), and whey dairy beverage (30 mg/dL), with glycemic indices similar to that observed for the control. The present results demonstrated a good correlation between in vitro and in vivo data, in which the type of dairy matrix affects the anti-hyperglycemic activity. It is concluded that the consumption of probiotic Prato cheese can contribute to the reduction of postprandial glycemia in healthy individuals.

Dermoid and epidermoid cysts of the oral cavity: a 48-year retrospective study with focus on clinical and morphological features and review of main topics.

BACKGROUND: Dermoid and epidermoid cysts are slow-growing, benign developmental cysts that arise from ectodermal tissue and can occur anywhere in the body. Less than 7% of these cysts involve the head and neck region, with only 1.6% of cases presenting in the oral cavity. To evaluate the clinical and histopathological features of dermoid (DCs) and epidermoid (ECs) cysts stored in the archives of a referred Oral Pathology Service over a 48-year-period, and to review current concepts about these cysts. MATERIAL AND METHODS: All DCs and ECs were reviewed, and clinical data were obtained from the patient records. Fourteen cases of DCs and thirteen cases of ECs were re-evaluated microscopically by 2 oral pathologists. RESULTS: Among 15.387 cases, 14 (0.09%) had a histopathological diagnosis of DCs and 13 (0.08%) of ECs. For DCs, ten (71.4%) patients were women, with the mean age of 37.2 years. All DCs were lined by a stratified squamous epithelium (100%), with gut and respiratory epithelium observed in 1 (7.1%) and 2 (14.3%) cases, respectively. Chronic inflammatory cells, melanin, multinucleated giant cell reaction, and Pacini bodies were also observed. For ECs, eight (61.5%) cases were in women, and the mean age was 38.2 years. All ECs were lined by a stratified squamous epithelium (100%). Chronic inflammatory cells, melanin pigmentation, and adipose tissue were observed in the fibrous capsule. CONCLUSION: Our results suggest that stratified squamous epithelium is the predominant epithelial lining of these cystic lesions. Also, we may find some unusual findings in DCs, such as Pacini bodies.

A retrospective 11-year study on lip lesions attended at an oral diagnostic service.

BACKGROUND: The objective of this study was to describe the upper and lower lip lesion occurrence in an oral diagnostic service. MATERIAL AND METHODS: Retrospective descriptive sectional study was performed. Clinical records were obtained from the archives of an Oral Diagnostic Service referral center between 2006 and 2016. Data such as gender, age, anatomical location, and diagnosis were collected and categorized. The collected data were submitted to a descriptive analysis and Pearson's chi-square test (p ≤ 0.05). RESULTS: A total of 587 patient records of lip lesions were analyzed. Most lesions were diagnosed in female (52.1%) and adults (56.9%) patients in the lower lip (76.2%). Among all lip lesions, the reactive/inflammatory lesions (n = 238; 40.5%) and oral potentially malignant disorders (n = 164; 28%) were the most frequent group lesions. Mucocele (n = 147; 25%), actinic cheilitis (n = 136; 23.1%) and vascular lesions (n = 51; 8.7%) were the most frequent lesion in the sample. Actinic cheilitis was significant in relation to gender (p < 0.001), all three most frequent lesions were significant in concerning to age group and anatomical site. CONCLUSIONS: Mucocele was the most common lower lip lesion in all age groups, followed by actinic cheilitis and vascular lesions, which mainly affected adults and the elderly.

Follicle populations and gene expression profiles of Nelore and Angus heifers with low and high ovarian follicle counts.

The number of antral follicles counted (AFC) by ultrasound is associated with fertility in cattle. Cows with higher follicle count (HFC) have higher performance in reproductive-assisted technologies than cows with lower follicle count (LFC). In this study, we aimed to define the preantral follicle count by histology and to identify differentially expressed genes (DEGs) using a microarray in Nelore and Angus heifers with HFC and LFC. The ovaries of each animal were scanned with an ultrasound device 12 to 24 hr after estrus. The groups were formed based on the average number of total follicles (≥3 mm) counted in each breed consistently ± the standard deviation. For the histological analysis, preantral follicles were counted and classified under a stereo microscope, and follicle density was determined. Microarray analysis was performed on pools of three follicles dissected from the ovaries of 15 Nelore (6 HFC and 9 LFC) and 17 Angus heifers (9 HFC and 8 LFC). Angus heifers have increased total and primordial follicle density. Nelore heifers have increased antral follicle count. Different patterns of gene expression regulate follicle recruitment and development in Angus and Nelore heifers and may be associated with the different follicle densities observed in Angus versus Nelore heifers. Furthermore, HFC heifers presented increased expression of genes associated with cellular development and metabolism.

Inhibition of STEP61 ameliorates deficits in mouse and hiPSC-based schizophrenia models.

The brain-specific tyrosine phosphatase, STEP (STriatal-Enriched protein tyrosine Phosphatase) is an important regulator of synaptic function. STEP normally opposes synaptic strengthening by increasing N-methyl D-aspartate glutamate receptor (NMDAR) internalization through dephosphorylation of GluN2B and inactivation of the kinases extracellular signal-regulated kinase 1/2 and Fyn. Here we show that STEP61 is elevated in the cortex in the Nrg1+/- knockout mouse model of schizophrenia (SZ). Genetic reduction or pharmacological inhibition of STEP prevents the loss of NMDARs from synaptic membranes and reverses behavioral deficits in Nrg1+/- mice. STEP61 protein is also increased in cortical lysates from the central nervous system-specific ErbB2/4 mouse model of SZ, as well as in human induced pluripotent stem cell (hiPSC)-derived forebrain neurons and Ngn2-induced excitatory neurons, from two independent SZ patient cohorts. In these selected SZ models, increased STEP61 protein levels likely reflect reduced ubiquitination and degradation. These convergent findings from mouse and hiPSC SZ models provide evidence for STEP61 dysfunction in SZ.

Can psychosocial work factors influence psychologists' positive mental health?

BACKGROUND: Working in healthcare can entail intense emotional demands that increases susceptibility to occupational risk factors. Psychosocial risk assessment can contribute to promoting awareness of the effects of work on positive mental health. AIMS: To explore and analyse the influence of psychosocial work factors on positive mental health among psychologists. METHODS: A cross-sectional study of 339 psychologists was conducted. Two instruments were used for data collection: the Mental Health Continuum-Short Form (MHC-SF) to assess well-being and the Health and Work Survey (INSAT) to assess psychosocial work factors. RESULTS: This study identified psychosocial work factors that affect psychologists' positive mental health, namely, emotional well-being was affected by 'Need help from colleagues' (ß = -1.091), 'Have no one I can trust' (ß = -1.253) and 'Complex work' (ß = 0.751); psychological well-being was affected by 'Intense work pace' (ß = 1.151), 'Not able to participate in decisions' (ß = -3.695) and 'Complex work' (ß = 1.520); and social well-being was affected by 'Always changing roles and tasks' (ß = -1.810) and 'Not able to participate in decisions' (ß = -2.470). CONCLUSIONS: Psychosocial work factors such as work organization, work relationships and emotional demands influence psychologists' positive mental health. Social support at the workplace and having challenging and autonomous work can promote mental health. It is important to develop better organizational practices to promote mental health and well-being among these professionals.

Low efficacy of atopy patch test in predicting tolerance development in non-IgE-mediated cow's milk allergy.

BACKGROUND: The food atopy patch (APT) test has been used in previous studies to help the diagnosis of non-IgE mediated food allergies (FA). The aim of this study was to evaluate the efficacy of different cow's milk APT preparations to predict oral tolerance in children with previous non-IgE-mediated cow's milk allergy (CMA) diagnosis. METHODS: Thirty-two patients non-IgE-mediated CMA diagnosed by oral food challenge (OFC) were enrolled to perform APT with three different cow's milk preparations (fresh, 2% in saline solution, 2% in petrolatum) and comparing with a new OFC after at least three months of diet exclusion. RESULTS: Only six (18.7%) subjects presented positive OFC to cow's milk. No differences in gender, onset symptoms age, OFC age, Z-score, and exclusion period were found between positive and negative OFC patients. Preparations using fresh milk and powdered milk in petrolatum presented sensitivity equal to zero and specificity 92.3% and 96.1%. The preparation using powdered milk in saline solution showed sensitivity and specificity of 33.3% and 96.1%. Two patients presented typical IgE symptoms after OFC. CONCLUSION: Cow's milk APT presented a low efficacy to predict tolerance in patients with previous non-IgE-mediated CMA and should not be used in clinical routine. The presence of typical IgE reactions after OFC hallmark the necessity of previous IgE-mediated investigation for this patient group.

Epidemiologic study of odontogenic and non-odontogenic cysts in children and adolescents of a Brazilian population.

BACKGROUND: the objective of this study was to describe the frequency of cystic lesions in a Brazilian population of patients histopathologically diagnosed in the first and second decade of life. MATERIAL AND METHODS: Retrospective descriptive cross-sectional study was performed. Biopsy records were obtained from the archives of a Brazilian referral center between 1980 and 2016. RESULTS: A total of 2.114 biopsy records of pediatric patients were analyzed with oral and maxillofacial lesions. Data such as gender, age, anatomical location, and histopathological diagnosis were collected and categorized. Among all oral and maxillofacial lesions (n=2.114), were diagnosed 294 cases of odontogenic cysts (13.9%) and 16 cases of non-odontogenic cysts (0.8%). The most frequent lesions in each group were, respectively: radicular cyst (n=145) and epidermoid cyst (n=4). These lesions were most common in female (n=158), with a mean age of 14 years. For intraosseous lesions, the mandible (n=148) was the most affected anatomic site; moreover, the floor of the mouth (n=6) was most affected by cysts in soft tissues. CONCLUSIONS: Odontogenic cysts were relatively common in population studied, but non-odontogenic cysts were rare in these patients.

Evidence that fibroblast growth factor 10 plays a role in follicle selection in cattle.

There is evidence that regulation of follicle selection in cattle involves locally produced growth factors. In the present study, we investigated the expression of members of the fibroblast growth factor (FGF) 7 family during follicle deviation. The largest and second largest follicles were recovered during the second day of a synchronised follicle wave and the future dominant and future subordinate follicles were identified based on diameter and cytochrome P450, family 19, subfamily A, polypeptide 1 (CYP19A1) mRNA levels in granulosa cells. Theca cells of the future dominant follicle contained less mRNA encoding FGF7 and FGF10 compared with those from the future subordinate follicle 2.5 days after ovulation, before a significant difference between the diameters of the future dominant and future subordinate follicles could be observed, but FGF22 mRNA levels did not change. Levels of mRNA encoding FGF receptors FGFR1B and FGFR2B in theca and granulosa cells, respectively, were lower in the future dominant follicle compared with the future subordinate follicle. Addition of FGF10 to granulosa cells in vitro significantly decreased oestradiol secretion, as well as CYP19A1, FSH receptor (FSHR) and insulin-like growth factor 1 receptor (IGF1R) mRNA abundance, whereas FGF22 had no effect. We conclude that FGF10 and FGFR2B expression is increased in the future subordinate follicle before morphological deviation, which may contribute to follicle selection.

Gene expression profile in heat-shocked Holstein and Nelore oocytes and cumulus cells.

The present study determined the transcriptome profile in Nelore and Holstein oocytes subjected to heat shock during IVM and the mRNA abundance of selected candidate genes in Nelore and Holstein heat-shocked oocytes and cumulus cells (CC). Holstein and Nelore cows were subjected to in vivo follicle aspiration. Cumulus-oocyte complexes were assigned to control (38.5°C, 22h) or heat shock (41°C for 12h, followed by 38.5°C for 10h) treatment during IVM. Denuded oocytes were subjected to bovine microarray analysis. Transcriptome analysis demonstrated 127, nine and six genes were differentially expressed between breed, temperature and the breed×temperature interaction respectively. Selected differentially expressed genes were evaluated by real-time polymerase chain reaction in oocytes and respective CC. The molecular motor kinesin family member 3A (KIF3A) was upregulated in Holstein oocytes, whereas the pro-apoptotic gene death-associated protein (DAP) and the membrane trafficking gene DENN/MADD domain containing 3 (DENND3) were downregulated in Holstein oocytes. Nelore CC showed increased transcript abundance for tight junction claudin 11 (CLDN11), whereas Holstein CC showed increased transcript abundance for antioxidant metallothionein 1E (MT1E) . Moreover, heat shock downregulated antioxidant MT1E mRNA expression in CC. In conclusion, oocyte transcriptome analysis indicated a strong difference between breeds involving organisation and cell death. In CC, both breed and temperature affected mRNA abundance, involving cellular organisation and oxidative stress.