Effects of supplemental flaxseed on the ovarian and uterine functions of adult cycling mice.

Dietary fatty acids (FA) have an effect on animal reproduction. The purpose of the study was to find out how supplemental flaxseed can modulate the FA metabolism and how FA can influence the release of ovarian hormones and functions of the uteri through the modulation of their specific receptors. Cycling mice were fed a basal diet (control) and basal diet fortified with 10% flaxseed for 6 weeks to examine its influence on the structure and function of the ovaries and uteri, and serum levels of FA. Unlike controls (30%), 100% of mice fed flaxseed exposed oestrus at the end of the supplemental period. Serum FA were analysed using gas chromatography. The ovaries and uteri underwent histological and immunohistochemical analyses, respectively. The ovarian fragments were cultured with or without follicle-stimulating hormone and culture media were analysed for progesterone (P4) and oestradiol-17ß (E2) using immunoassays. Dietary flaxseed increased the serum FA concentration, sizes of the ovaries and primary follicles, the release of P4 and E2, the thickness of endometrium and myometrium, and altered the expression of oestrogen and progesterone receptors in all uterine compartments. Dietary flaxseed can promote ovarian steroidogenesis and uterine activity in the mouse.

Carbon Monoxide (CO) as a Retinal Regulator of Heme Oxygenases -1, and -2 (HO's) Expression.

Carbon monoxide (CO) has been proposed as a chemical light signal and neural system modulator via heme oxygenases -1 and -2 (HO-1 and HO-2). Many papers have proven the CO-HO circuit to be important for such physiological pathways as the molecular biological clock and the GnRH axis, but also in such pathological occurrences as ischemic injuries, or inflammation as a regenerative and neuroprotective factor. In this in vivo experiment, we used three groups of pigs: control-housed in natural conditions without any procedures; without CO-adapted and kept in constant darkness, infused with blank plasma; and with CO-adapted and kept in constant darkness infused with CO-enriched plasma. After the experiments, each animal was slaughtered and its eyes were collected for further analysis. Quantitative PCR and Western blot analysis were performed to show statistical differences in the expressions between the experimental groups. Our data revealed that exogenous CO is regulator of mRNA transcription for HO-1 and HO-2 and PCNA. Moreover, the mRNA abundance of analyzed factors in the experimental group after CO elevation revealed a restored gene-expression level similar to the control group, which we had observed in the group's restored protein level after CO elevation. In conclusion, exogenous CO regulates HO's and PCNA gene expression on transcriptional and translational levels in a similar way as a light cue.

Clinical outcomes of potential high responders after individualized FSH dosing based on anti-Müllerian hormone and body weight.

RESEARCH QUESTION: How does the efficacy and safety of individualized follitropin delta dosing compare with conventional dosing for ovarian stimulation in potential high responders? DESIGN: Retrospective analysis of 153 potential high responders identified on the basis of baseline serum anti-Müllerian hormone (AMH) levels above 35 pmol/l, who were originally randomized to an individualized fixed dose of follitropin delta based on AMH and body weight (n = 78) or to a daily starting dose of 150 IU follitropin alfa (n = 75). RESULTS: At the end of stimulation, patients treated with individualized follitropin delta or conventional follitropin alfa had 12.1 ± 7.0 and 18.3 ± 7.0 (P < 0.001) follicles measuring 12 mm or wider, and 27.3% and 62.7% had serum progesterone levels higher than 3.18 nmol/l (P < 0.001), respectively. Overall number of oocytes in these two respective arms was 9.3 ± 6.7 and 17.9 ± 8.7 (P < 0.001), and the ongoing pregnancy rate per started cycle after fresh blastocyst transfer was 28.2% and 24.0%. The risk of ovarian hyperstimulation syndrome (OHSS) for all cases was three times higher in the conventional follitropin alfa arm at 16.0% versus 5.1% with individualized follitropin delta treatment (P = 0.025) and 26.7% versus 7.7% (P = 0.001) for early moderate or severe OHSS, preventive interventions for early OHSS, or both. CONCLUSIONS: Treatment with individualized follitropin delta provides an improved efficacy-safety balance in women with high ovarian reserve, as it normalizes the ovarian response and decreases the risk of OHSS without compromising the chance of pregnancy.

Dietary flaxseed's protective effects on body tissues of mice after oral exposure to xylene.

Xylene is a common pollutant in the environment that enters the body of animals and humans in various ways, but most often through the respiratory tract and adversely affects their overall health. However, xylene effects after oral exposure have not been sufficiently studied. This study aimed to investigate the effects of xylene exposure on the mouse organism and to identify possible beneficial effects of flaxseed on such exposure. Eighty mice were divided into four groups: control group C (basal diet + no xylene exposure), group X (oral exposure by 400 mg/kg/day xylene), group F (10% flaxseed supplementation of basal diet), and group XF (10% dietary flaxseed + oral exposure by xylene). Experimental trial took 14 days. Clinical examination, spectroscopic analysis of tissue aminotransferases, total lactate dehydrogenase (TLDH), and acetylcholinesterase (AchE) activities, electrophoretic analysis of LDH isoenzymes, western blot and immunohistochemical analysis of apoptosis as well as routine histology of the kidneys and jejunum, and transmission electron microscopy of the liver were performed. Marked restlessness in group X and high weight losses in mice of all groups were recorded during the experiment. Xylene promoted apoptosis (caspase-3 expression) without causing marked structural changes in the liver and jejunum, although renal cortex structure was affected adversely. In the brain, liver, and kidney of mice, xylene increased levels of liver transaminases, LDH, and decreased AchE activities, reflecting cell membrane damage. Flaxseed feeding improved animal behaviour, leakage of enzymes and prevented selected tissue toxic damage induced by xylene by protecting cell membrane integrity and fluidity and by suppressing apoptosis. These results point at the protective effect of flaxseed consumption on mice.

Assisted reproductive medicine in Poland, 2013-2016: Polish Society of Reproductive Medicine and Embryology (PTMRiE) and Fertility and Sterility Special Interest Group of the Polish Society of Gynaecologists and Obstetricians (SPiN PTGiP) report.

OBJECTIVES: The purpose of this publication is to present data on the results and complications associated with infertility treatment using assisted reproductive technology (ART) and intrauterine insemination (IUI) in Poland between 2013 and 2016. MATERIAL AND METHODS: The report was prepared by the Polish Society of Reproductive Medicine and Embryology (PTMRiE) and the Fertility and Sterility Special Interest Group of the Polish Society of Gynaecologists and Obstetrics (SPiN PTGiP) as a part of the European IVF Monitoring program (EIM) for the European Society of Human Reproduction and Embryology (ESHRE). Reporting was voluntary and the data was not subject to external control. The report presents the availability and structure of infertility treatment services, the number of procedures, their effectiveness and complications. RESULTS: Between 2013 and 2016, a total of 106,718 treatment cycles using ART [64,413 classical in vitro fertilization and in vitro fertilization with intracytoplasmic sperm injection (IVF + ICSI), 36,041 frozen embryo replacements (FER)] and 51,405 IUI were recorded. The clinical pregnancy rates per embryo transfer in IVF, ICSI and FER were 38.3%, 38.1% and 32.4%, respectively. The effectiveness of IUI with husband/partner's semen (IUI-H) was 11.1% and with donor semen (IUI-D) 16.7%. Multiple delivery rates were 11.3% and 6.2% in IVF + ICSI and IUI, respectively. The most common complication was the ovarian hyperstimulation syndrome (OHSS) (0.34%). CONCLUSIONS: PTMRiE and SPiN PTGiP report is the only national study documenting Polish reproductive medicine. The results of infertility treatment effectiveness in Poland are comparable with the European data, complications are less frequent than in other countries. The low percentage of multiple pregnancies, and so perinatal complications, is especially valuable. However, due to the lack of a central database and register, the possibility of external control and monitoring of pregnancies and births is limited. Thus, a fully reliable assessment of the treatment quality in our country is not possible.

Altered circadian dynamics of Per2 after cystathionine-ß-synthase and/or cystathionine-γ-lyase pharmacological inhibition in serum-shocked NIH-3T3 cells.

Circadian clock genes are found in almost every cell that has a nucleus; they regulate the rhythmic nature of all processes that are cyclical. Among the genes controlled by the circadian clock, there are numerous factors that regulate key processes in the functioning of the cell. Disturbances in the functioning of the circadian clock are associated with numerous disorders. A recent study has shown the key role of H2S in regulating circadian rhythm. In this study, we investigated the in vitro effect of pharmacological inhibition of cystathionine-ß-synthase (CBS) and/or cystathionine-γ-lyase (CSE) on the circadian dynamics of Per2 expression in serum-shocked NIH-3T3 cells. Alternatively, Cbs and Cse were knocked down by transfection with siRNA. The 48-h treatment of serum-shocked NIH-3T3 cells with 1 mM dl-propargylglycine (PAG), a specific CSE inhibitor, significantly decreased the amplitude and baseline expression of Per2. During exposure to an effective CBS and CSE inhibitor (aminooxyacetic acid [AOAA]), the amplitude of oscillation and baseline expression of Per2 significantly increased. Incubation of NIH-3T3 cells with both inhibitors also significantly increased the amplitude and baseline expression of Per2 messenger RNA (mRNA). siCbs or siCse knockdowan significantly reduced the baseline and amplitude of oscillation of Per2. In conclusion, we showed that CBS/CSE/H2S pathway participates in the regulation of the circadian clock system. PAG and AOAA, change the general expression and dynamics of Per2 genes, but the increase of amplitude and overall Per2 mRNA level due to exposure to AOAA is probably caused by factors other than CBS and CSE activity.

Changes in γH2AX and H4K16ac levels are involved in the biochemical response to a competitive soccer match in adolescent players.

The aim of this study was to examine novel putative markers of the response to the competitive soccer match in adolescent players, such as changes in global levels of γH2AX and H4K16ac in the chromatin of peripheral mononuclear blood cells (PMBCs) and a Fourier-transform infrared spectroscopy (FTIR)-based biochemical fingerprint of serum. These characteristics were examined with reference to the physiological and metabolic aspects of this response. Immediately post-match we noticed: (1) a systemic inflammatory response, manifesting as peaks in leukocyte count and changes in concentrations of IL-6, TNFα, and cortisol; (2) a peak in plasma lactate; (3) onset of oxidative stress, manifesting as a decline in GSH/GSSG; (4) onset of muscle injury, reflected in an increase in CK activity. Twenty-four hours post-match the decrease in GSH/GSSG was accompanied by accumulation of MDA and 8-OHdG, macromolecule oxidation end-products, and an increase in CK activity. No changes in SOD1 or GPX1 levels were found. Repeated measures correlation revealed several associations between the investigated biomarkers. The FTIR analysis revealed that the match had the greatest impact on serum lipid profile immediately post-game. In turn, increases in γH2AX and H4K16ac levels at 24 h post-match indicated activation of a DNA repair pathway.

Treating honey bees with an extremely low frequency electromagnetic field and pesticides: Impact on the rate of disappearance of azoxystrobin and λ-cyhalothrin and the structure of some functional groups of the probabilistic molecules.

The purpose of these laboratory tests was to assess the impact of 50 Hz EMF (electromagnetic field) on the disappearance of azoxystrobin (active ingredient (AI) of Amistar 250 SC) and λ-cyhalothrin (AI of Karate Zeon 050 CS) in the body of honey bees (Apis mellifera) and the structure of some functional groups of the probabilistic molecules in their organisms. Amistar 250 SC (an azoxystrobin-based fungicide; ABF) and Karate Zeon 050 CS (a λ-cyhalothrin-based insecticide; CBI) are plant protection products (PPPs) applied to bee-pollinated-crops. Chromatographic methods were used to assess the rate of AI disappearance. EMF affected the rate of disappearance of azoxystrobin and λ-cyhalothrin in bees within 6 h of intoxication. When these substances were used separately their disappearance in the presence of EMF slowed from 12.6% to 10.5% h-1 and from 9.2% to 4.8% h-1, respectively, and accelerated when used in a mixture, from 14.1% to 14.7% h-1 and from 9.3% to 11.5% h-1 respectively. Fourier Transform Infrared (FTIR) spectroscopy was used to analyze changes in the functional groups of the probabilistic molecules of the tested bees. To obtain the information about the spectra variations we used the Principal Component Analysis. It has been shown, that EMF statistically significantly interferes with amide I and II, symmetric PO32- group from DNA, RNA and phospholipids vibrations. It also increased the number of changes of functional groups of the probabilistic molecules caused by ABF, but at the same time limited the changes in the functional groups studied in bees treated with CBI and a mixture containing both of them. In addition, exposure to EMF in bees treated with fungicide or insecticide, separately, and with both preparations caused differences (p < 0.05) in the secondary structure of proteins compared to controls. The obtained results indicate that EMF may affect the rate of metabolism and the detoxification process of pesticides in bees, depending on the AI of PPPs, applied individually or together. However, further detailed research is required to explain the mechanism of EMF as a detoxification modulator.

Melatonin concentration in peripheral blood and melatonin receptors (MT1 and MT2) in the testis and epididymis of male roe deer during active spermatogenesis.

Melatonin regulates male reproductive function in seasonal and non-seasonal breeder mammals. The presence of melatonin membrane receptors (MT1 and MT2) in the testis and epididymis has been demonstrated in several species. Wild roe deer are a short-day breeding species characterised by a short rutting season lasting from mid-July to mid-August. The aim of this study was to determine the concentration of melatonin in the peripheral blood and the presence of MT1 and MT2 receptors in the testis and epididymis in male roe deer during the pre-rut (May), rut (July/August) and post-rut (September) periods. The melatonin concentration was higher in May (522.50 ± 54.20 pg/mL) compared to July/August (258.50 ± 36.82 pg/mL; P < 0.05). During September, the melatonin concentration was higher (393.50 ± 36.77 pg/mL) than in July/August (P < 0.05) but lower than in May (P < 0.05). Immunohistochemical analysis showed the presence of MT1 and MT2 receptors in Leydig cells, Sertoli cells and germ cells in the testis, in addition to the epithelial cells of the epididymis caput, corpus and cauda. MT1 and MT2 receptor expression in the testis and epididymis, assessed by Western blot, was higher in May and July/August (when spermatogenic and steroidogenic activity restarts and reaches its peak, respectively) compared to September (when spermatogenic and steroidogenic activity decreases). This could indicate a stimulatory effect of melatonin on testicular (i.e., steroidogenesis and spermatogenesis) and epididymal (i.e., spermatozoa maturation) function in male roe deer through the MT1 and MT2 receptors. Our results form the basis for further studies into the detailed mechanism of action of melatonin through MT1 and MT2 receptors for optimal reproductive activity in male roe deer and other mammals.

In vitro study of effects of ELF-EMF on testicular tissues of roe deer (Capreolus capreolus) - FTIR and FT-Raman spectroscopic investigation.

Electromagnetic fields (EMF) are classified as an environmental factor affecting living organisms. The aim of the study was to evaluate the effect of EMF at different frequencies (50 and 120 Hz), durations of treatment (2 and 4 h) and with the magnetic induction of 8 m T on testicular tissues of roe deer (Capreolus capreolus) in vitro by comparison with the control samples. Fourier Transform Raman Spectroscopy (FT-Raman) and Fourier Transform Infrared Spectroscopy (FTIR) were utilized in this study to identify the chemical changes in the testicular tissues. The FTIR and FT-Raman spectroscopy methods were used to evaluate differences in spectra of the treated tissues compared to the control group. The results from the analysis of the spectra indicated there were characteristic differences in the testicular tissue compared with the control samples. There was identification of peaks attributed to different biochemical components. Comparing the spectra for different frequencies and treatment times, there was a greater intensity of peaks originating from most of the functional groups in the tissues evaluated. With the FTIR spectra, there were five of 15 peaks, while with the FT-Raman spectra, there were six of ten peaks that were shifted. For FTIR and FT-Raman analyzed spectral ranges, results from the PCA analysis indicate there was no similarity between control groups (2 and 4 h) and samples treated with EMF at a frequency of 120 Hz for 2 and 4 h. In conclusion, therefore, EMF is an environmental factor affecting the testis of roe deer.

Correction to: Xenopus LAP2ß protein knockdown affects location of Lamin B and nucleoporins and has effect on assembly of cell nucleus and cell viability.

The original publication of this paper contains a mistake.

Altered dynamics in the circadian oscillation of clock genes in serum-shocked NIH-3T3 cells by the treatment of GYY4137 or AOAA.

BACKGROUND AND PURPOSE: Several members of the core clock mechanism are equipped with a Per-Arnt-Sim (PAS) domain through which they can bind haem [Fe(II)]. Haem is a ligand for the orphan receptors REV-ERBα/ß (NR1D1/2), which regulate circadian rhythm and metabolism. The ability to bind haem sensitises these clock components to the action of small molecule gases, including NO, CO and H2S. Studies conducted with European hamsters revealed that during winter sleep, key clock genes stop oscillating. At the same time, H2S, when administered at subtoxic concentrations, can induce a hypometabolic state in the cell. We suppose that core clock components, including the nuclear receptors REV-ERBs, neuronal PAS domain protein 2 (nPAS2) and PER2, can be H2S targets. The general objective of this study was to investigate the effect of the H2S system on the expression profile of the core clock genes in cells in vitro. EXPERIMENTAL APPROACH: We analysed the expression of Per1, Per2, Per3, Bmal1, Cry1, Cry2, Nr1d1, Nfil-3 and Dbp messenger RNA (mRNA) in serum-shocked NIH-3T3 cells treated with a slow-releasing H2S donor (GYY4137) or the cystathionine beta-synthase (CBS) inhibitor (AOAA) cultured under constant darkness and collected during 3 days in 3 h interval. KEY RESULTS AND CONCLUSIONS AND IMPLICATIONS: We found that pharmacological CBS inhibition increased the general expression and dynamics of several clock genes. On the other hand, increased H2S decreased Per2 expression. These data suggest that CBS can affect circadian clock and effect on clock-controlled transcription output.

From epithelial remodelling to carcinogenesis.

The novel cancer theory named 'the tissue organization field theory' (TOFT) suggests that carcinogenesis is a process analogous to embryonic development, whereby organs are formed through interactions among different cell types. The suggested 'morphological remodelling' of the epithelium under hypoxia in gut breathing fish (GBF) has many common features with carcinogenesis. It appears that research into the relationship among epidermal growth factor receptor (EGFR), hypoxia inducible factor (HIF) as well as hypoxia and normoxia states in GBF fishes can be crucial in learning about the steering mechanisms of squamous epithelium proliferation, leading to a better understanding of carcinogenesis.

Chemoreceptors as a key to understanding carcinogenesis process.

The tissue organization field theory (TOFT) presented completely new, different from the previous one, perspective of research on neoplasm processes. It implicates that secretory neuroepithelial-like cells (NECs), putative chemoreceptors are probably responsible for the control of squamous epithelial cells proliferation in the digestive tract during hypoxia in gut breathing fish (GBF). On the other hand, chemoreceptors dysfunction can lead to uncontrolled proliferation and risk of cancer development in mammals, including humans. The studies on NECs like cells (signal capturing and transduction) may be crucial for understanding the processes of controlling the proliferation of squamous epithelial cells in the digestive tract of GBF fish during hypoxia states. This knowledge can contribute to the explanation of cancer processes.

The seasonal changes of the heme oxygenase in the retina pig.

The eye is a very important organ in the human body which is affected by various external factors. One of these factors is the sunlight which can cause the visual impairment and as well as the increase in the oxidative stress. The heme oxygenase I (HO-1) plays a very important role in the fight against the oxidative stress. The HO enzyme catalyses the degradation of the heme to the ferrous iron, the biliverdin and the carbon monoxide (CO). The HO-2 is the isoform HO-1 and is mainly constitutively expressed. We have studied the changes in the HO-1 and the HO-2 in the retina on the level of the RNA and the protein in the summer and in the winter season (the biggest difference is in the length of the day light). The retina of the eye was obtained from the breeding pigs in concern (Sus scrofa f. domestica) posthumously. The expression of the HO-1 genes in the retina cells is higher in the winter and the amount of protein decreases. However, the HO enzyme concentration definitely increases in the summer, when the production of the free radicals (the oxidative stress) related to the exposition to the sunlight is greater. The obtained results suggest that various factors have the influence on the protein synthesis. One of the factors, can be the miRNA which blocks the synthesis of the HO. Another factors, influencing the HO are the biological clock, the sunlight and the UV radiation associated with it.

Emerin Is Required for Proper Nucleus Reassembly after Mitosis: Implications for New Pathogenetic Mechanisms for Laminopathies Detected in EDMD1 Patients.

Emerin is an essential LEM (LAP2, Emerin, MAN1) domain protein in metazoans and an integral membrane protein associated with inner and outer nuclear membranes. Mutations in the human EMD gene coding for emerin result in the rare genetic disorder: Emery⁻Dreifuss muscular dystrophy type 1 (EDMD1). This disease belongs to a broader group called laminopathies-a heterogeneous group of rare genetic disorders affecting tissues of mesodermal origin. EDMD1 phenotype is characterized by progressive muscle wasting, contractures of the elbow and Achilles tendons, and cardiac conduction defects. Emerin is involved in many cellular and intranuclear processes through interactions with several partners: lamins; barrier-to-autointegration factor (BAF), ß-catenin, actin, and tubulin. Our study demonstrates the presence of the emerin fraction which associates with mitotic spindle microtubules and centrosomes during mitosis and colocalizes during early mitosis with lamin A/C, BAF, and membranes at the mitotic spindle. Transfection studies with cells expressing EGFP-emerin protein demonstrate that the emerin fusion protein fraction also localizes to centrosomes and mitotic spindle microtubules during mitosis. Transient expression of emerin deletion mutants revealed that the resulting phenotypes vary and are mutant dependent. The most frequent phenotypes include aberrant nuclear shape, tubulin network mislocalization, aberrant mitosis, and mislocalization of centrosomes. Emerin deletion mutants demonstrated different chromatin binding capacities in an in vitro nuclear assembly assay and chromatin-binding properties correlated with the strength of phenotypic alteration in transfected cells. Aberrant tubulin staining and microtubule network phenotype appearance depended on the presence of the tubulin binding region in the expressed deletion mutants. We believe that the association with tubulin might help to "deliver" emerin and associated membranes to decondensing chromatin. Preliminary analyses of cells from Polish patients with EDMD1 revealed that for several mutations thought to be null for emerin protein, a truncated emerin protein was present. We infer that the EDMD1 phenotype may be strengthened by the toxicity of truncated emerin expressed in patients with certain nonsense mutations in EMD.

Differential expression of epidermal growth factor receptor (EGFR) in stomach and diverticulum of Otocinclus affinis (Steindachner, 1877) as a potential element of the epithelium remodeling mechanism.

It is suggested that due to the several stomach modifications, Otocinculus affinis (dwarf sucking catfish) possess the ability to breathe air during hypoxia, however, the exact mechanism remains unknown. The aim of this study was detailed analysis of the expression of EGFR in the stomach and diverticulum of the O. affinis at the mRNA and protein levels together with the immunohistochemical localization of EGFR in these organs. The intensity of band fluorescence corresponding to the EGFR gene expression level is significantly higher in the stomach than in the diverticulum. Further, quantitative analysis of EGFR protein abundance also revealed its higher synthesis in the stomach than in the diverticulum and the immunohistochemistry method confirmed these results. As regional localization of respiratory function in gut air-breathing fishes seems to be connected with "morphological remodeling" of the epithelium of their gut, the present research demonstrated the potential efficiency of the O. affinis stomach as a respiratory organ. Having the potential possibility to create an air-blood barrier in the gastrointestinal tract allowing gas diffusion and respiration in hypoxic states seems to be very beneficial for these fish. It seems that facultative gut air breathing fish species are a relevant vertebrate model for high throughput screening, vascular biology and evolution.

Supplemental flaxseed modulates ovarian functions of weanling gilts via the action of selected fatty acids.

The aim of this study was to examine the influence of dietary flaxseed on the endocrine and ovarian functions of weanling gilts challenged with E. coli and Coronavirus infections treated with dietary probiotic cheeses and to understand the possible mechanisms of its effects on ovarian function. Probiotics were used as a natural substitution for antibiotics and 10% dietary flaxseed is an effective prebiotic which supports the action of probiotics and has other beneficial effects on the organism. Probiotics with or without flaxseed were fed to weanling gilts starting 10 days before and lasting up until 14 days after weaning. The ovaries were measured and histologically analysed. The blood samples for the levels of steroid hormones and insulin-like growth factor I (IGF-I) were assessed using immunoassays and the levels of fatty acids were assessed using gas chromatography. All samples were collected on the day of weaning and 14 days after weaning. On the day of weaning, increased levels of linoleic acid and IGF-I was associated with higher body weight. The steroid hormones were not affected by the diet. The conversion of alpha-linolenic acid (ALA) to timodonic (EPA) and cervonic (DHA) acids were lower compared to controls, and together with high levels of myristic, palmitic and palmitoleic acids was associated with the higher proliferation and lower apoptosis in the primordial, primary and secondary follicles; although the inhibition of the cell cycle was observed in relation to the low level of eicosadienoic acid. The high levels of ALA, EPA and DHA and the low levels of myristic, palmitic and palmitoleic acids may have been the effect of flaxseed feeding 14 days post-weaning and may have had a reverse effect on the proliferation and apoptosis of ovarian follicles. These data suggest that flaxseed may suppress the follicle development in weanlings via the stimulation of apoptosis and the inhibition of proliferation via the modulation of the metabolism of selected fatty acids.

Confirmation of the immunoreactivity of monoclonal anti-human C-terminal EGFR antibodies in bronze Corydoras Corydoras aeneus (Callichthyidae Teleostei) by Western Blot method.

Bronze corydoras (Corydoras aeneus) uses the distal part of the intestine as accessory respiratory organ. Our previous study showed the presence of epidermal growth factor receptor (EGFR) cytoplasmic domain in the digestive tract of the bronze corydoras. In this study, using Western Blot method, we validated the results presented in the previous research. In detail, results of Western Blot analysis on digestive and respiratory part of bronze corydoras intestine homogenates confirmed the immunoreactivity of anti-cytoplasmic domain (C-terminal) human EGFR antibodies with protein band of approximately 180kDa (EGFR molecular weight). This indicates a high homology of EGFR domain between these species and the possibility of such antibody use in bronze corydoras. Statistically significantly higher EGFR expression was observed in the respiratory part of intestine when compared to the digestive part. This implies higher proliferation activity and angiogenesis of epithelium in this part of intestine, creating conditions for air respiration. Therefore, Corydoras aeneus may be considered as a model organism for the molecular studies of the mechanisms of epithelial proliferation initiation and inhibition depending on hypoxia and normoxia.

Squamous epithelium formation in the respiratory intestine of the bronze Corydoras Corydoras aeneus (Callichthyidae Teleostei).

Accessory respiratory organs in fish exhibit great diversity but share the presence of numerous capillaries covered by a simple squamous epithelium. The adoption of the intestine for respiratory function needs certain special modifications. In this study, we explored immunohistochemical and metabolic fingerprint features that could underlay this adaptation in bronze corydoras Corydoras aeneus. Immunohistochemical localization of the cytoplasmic domain of epidermal growth factor receptor (EGFR) in the respiratory part of intestine demonstrated a strong positive immunoreaction in epithelial cells and connective tissue. Fourier Transfer Infrared (FTIR) spectroscopy coupled with chemometrics discriminated between anterior and posterior region of intestine in terms of secondary structure of proteins and the abundance of p-cresol and other phenolics. The latter were reduced in the posterior part of intestine, indicating the cessation of digestive function in this region. It has been suggested that aquatic hypoxia via endocrine cells (hypoxia-sensitive) activate EGFR, which induce proliferation of squamous epithelial cells, thereby enabling gas diffusion in the posterior part of intestine. It seems that hypoxia and normoxia are opposed conditions adjusting the production of squamous epithelial cells in this intestine. The physiological role of EGFR in the respiratory intestine of bronze corydoras is of interest not only from an evolutionary aspect but also in terms of a potential model for observations process proliferation squamous epithelial cells. Future investigations on the molecular responses to different water oxygen levels in air-breathing bronze corydoras fish are required to clarify the mechanism responsible for squamous cell proliferation.