Canine spermatozoa—What do we know about their morphology and physiology? An overview

Spermatozoa are unique cells because of their morphological and physiological characteristics. They are produced during the process called spermatogenesis. Spermatogenesis consists of three phases: spermatocytogenesis, spermiogenesis and spermiation, during which spermatozoa undergo several changes. Spermatogenesis takes place within the seminiferous tubules containing two types of cells—the germ cells and the Sertoli cells—that alongside the Leydig cells, which play an important role when it comes to normal fertility. Everything is regulated by the hypothalamic–pituitary–gonadal axis and specific hormones due to multi-hormonal feedback systems. Spermatozoa possess morphological and physiological features, which are sometimes completely different from what is observed in various somatic cells. What is more, canine spermatozoa have specific characteristics making them special compared to the spermatozoa of other mammalian species. The metabolic energy production, which is crucial for the appropriate functioning of spermatozoa, can be fuelled by different metabolic pathways utilizing different chemical substrates. Inseparable from the oxidative phosphorylation process is the production of reactive oxygen species, which are both essential and toxic to spermatozoa. Furthermore, epididymis is a very important structure, responsible for the transport and maturation of spermatozoa, which are then stored in the last segment of epididymis—the epididymal cauda. Moreover, the retrieval of spermatozoa from the epididymides is crucial for the development of assisted reproduction techniques and sperm cryopreservation methods. The information gained from the research on domestic dogs might be transferred to their wild relatives, especially those species categorized as endangered.     

Structural observations during androgenic microspore culture of the 4c1 genotype of Zea mays L.

Summary The capacity of the maize genotype 4c1 to regenerate microcalli and embryos from cultured microspores has been examined by comparing various cold pretreatments and culture media, using microspores and pollen at different stages of development. Viability of cultured cells was tested with FDA and their development was traced with light and fluorescence microscopy using DAPI as a nuclear dye.It was found that a pre-incubation of dissected flowers floating in a liquid nutrient medium at 8°C during 10–14 days was most successful for the induction of cell division. Among the developmental stages tested only the microspores appeared to regenerate. Subculture at 25°C in the same liquid medium, supplemented with 0.1 mg/l TIBA, gave highest rates of microspore division, i.e. up to 70% at 4 to 6 days of culture.All pathways described earlier for maize androgenic embryogenesis were observed within the 4c1 genotype. Symmetric divisions occurred in cultured microspores but most frequently asymmetric divisions lead to the formation of microcalli within 12 days of culture. In at least 60% of all dividing microspores cells were derived from the generative nucleus. Microcalli further developed either into loose or compact calli. Compact calli formed embryo-like structures.Abbreviations DAPI 4,6-diamidino-2-phenylindole - Dicamba 3,6-dichloro-2-methoxy benzoic acid - 2,4D 2,4 dichlorophenoxyacetic acid - FDA fluorescein diacetate - PAA phenylacetic acid - TIBA 2,3,5-triiodobenzoic acid - YP medium Yu-Pei basal salt medium     

Effects of green tea polyphenol (-)-epigallocatechin-3-gallate on newly developed high-fat/Western-style diet-induced obesity and metabolic syndrome in mice

The aim of this study was to investigate the effects of (-)-epigallocatechin-3-gallate (EGCG) on newly developed high-fat/Western-style diet-induced obesity and symptoms of metabolic syndrome. Male C57BL/6J mice were fed a high fat/Western-style (HFW; 60% energy as fat and lower levels of calcium, vitamin D(3), folic acid, choline bitartrate, and fiber) or HFW with EGCG (HFWE; HFW with 0.32% EGCG) diet for 17 wks. As a comparison, two other groups of mice fed a low-fat diet (LF; 10% energy as fat) and high-fat diet (HF; 60% energy as fat) were also included. The HFW group developed more body weight gain and severe symptoms of metabolic syndrome than the HF group. The EGCG treatment significantly reduced body weight gain associated with increased fecal lipids and decreased blood glucose and alanine aminotransferase (ALT) levels compared to those of the HFW group. Fatty liver incidence, liver damage, and liver triglyceride levels were also decreased by the EGCG treatment. Moreover, the EGCG treatment attenuated insulin resistance and levels of plasma cholesterol, monocyte chemoattractant protein-1 (MCP-1), C-reactive protein (CRP), interlukin-6 (IL-6), and granulocyte colony-stimulating factor (G-CSF). Our results demonstrate that the HFW diet produces more severe symptoms of metabolic syndrome than the HF diet and that the EGCG treatment can alleviate these symptoms and body fat accumulation. The beneficial effects of EGCG are associated with decreased lipid absorption and reduced levels of inflammatory cytokines.     

Can mineral and organic fertilization help sequestrate carbon dioxide in cropland?

The soil organic matter content represents a huge reservoir of plant nutrients and an effective safeguard against pollution; beside it can sequestrate atmospheric CO₂. Since 1966 up to now in the Southeast Po valley (Italy), the soil organic C (SOC) and total N (TN) dynamics in the 0–0.40 m soil layer under a maize–wheat rainfed rotation are studied as influenced by organic and mineral N fertilizations. Every year in the same plots cattle manure, cattle slurry, and crop residues (i.e. wheat straw and maize stalk) are ploughed under to 0.40 m depth at a same dry matter rate (6.0 and 7.5 t DM ha⁻¹ year⁻¹ after wheat and maize, respectively) and are compared to an unamended control. Each plot is splitted to receive four rates of mineral fertilizer (0–100–200–300 kg N ha⁻¹). In the whole experiment, in 2000 SOC concentration was lower than in 1966 (6.77 and 7.72 g kg⁻¹, respectively), likely for the deeper tillage that diluted SOC and favoured mineralization in deeper soil layer. From 1972 to 2000 SOC stock did not change in the control and N fertilized plots, while it increased at mean rates of 0.16, 0.18, and 0.26 t ha⁻¹ year⁻¹ with the incorporation of residues, slurry and manure, corresponding to sequestration efficiencies of 3.7, 3.8 and 8.1% of added C with the various materials. TN followed the same SOC dynamic, demonstrating how it depends on the soil organic matter. Manure thus confirmed its efficacy in increasing both SOC content and soil fertility on the long-term. In developed countries, however, this material has become scarcely available; slurry management is expensive and implies high environmental risks. Moreover, in a C balance at a farm (or regional) scale, the CO₂ lost during manure and slurry stocking should be considered. For these reasons, the incorporation of cereal residues, even if only a little of their C content was found capable of soil accumulation, appears the best way to obtain a significant CO₂ sequestration in developed countries. Our long-term experiment clearly shows how difficult it is to modify SOC content. Moreover, because climate and soil type can greatly influence SOC dynamic, to increase CO₂ sequestration in cropland, it is important to optimize the fertilization within an agricultural management that includes all the agronomic practices (e.g. tillage, water management, cover crops, etc.) favouring the organic matter build up in the soil.     

PPR‐B‐31: a new maintainer allele in the male‐fertility restorer gene of radish (Raphanus sativus) Ogura cytoplasm

The PPR‐B gene is responsible for male‐fertility restoration of the Ogura‐type male‐sterile radish plants, and it is located in the complex Rfo locus in the vicinity of similar PPR‐A gene and PPR‐C pseudogene. The aim of this study was to identify PPR‐B alleles and understand the structure of the Rfo locus in radish breeding lines. Five lines of radish with normal male‐fertile cytoplasm were tested. The entire PPR‐B gene was amplified, sequenced and allelic PPR‐B sequences were identified. The results indicated that the maintainer lines 7, 15 and 21 contained a non‐restoring form of PPR‐B protein. A unique PPR‐B was found in lines 24/15 and 31 that are restorer and maintainer lines, respectively. The substitutions might be responsible for the loss of a restoring function of the PPR‐B‐31 allele. Amplification of the PPR‐A/PPR‐B and PPR‐B/PPR‐C intergenic regions allowed to identify rearrangements within Rfo locus. Obtained results confirm the wide allelic variation within the Rfo locus, as well as high genetic complexity of the fertility restoration mechanism in radish.     

Artificially induced polyploidization in Humulus lupulus L. and its effect on morphological and chemical traits

Chemically induced polyploids were obtained by the colchicine treatment of shoot tips of Humulus lupulus L. ‘Sybilla’. Flow cytometry revealed that most of the treatments resulted in the production of tetraploids. The highest number of tetraploids was obtained when explants were immersed in 0.05% colchicine for 48 h. A field experiment was conducted to compare diploid and tetraploid plants and assess the effect of genome polyploidization on the morphological and chemical characteristics. Tetraploids showed significant differences in relation to diploids. They had thinner and shorter shoots. The influence of chromosome doubling was also reflected in the length, width and area of leaves. The length of female flowers in the tetraploids was significantly shorter than that observed in diploids. Tetraploids produced a diverse number of lupuline glands that were almost twice as large as those observed in diploids. The most distinct effect of genome polyploidization was a significant increase in the weight of cones and spindles. Contents of major chemical constituents of hop cones was little affected by ploidy level. Total essential oils were significantly lower than those in diploids. However there was a significant increase in the proportion of humulene, caryophyllene and farnesene, oils desired by the brewing industry.     

A novel extracellular peroxidase and nucleases from a milky sap of Chelidonium majus

Using affinity chromatography, SDS-PAGE, peroxidase activity assay and mass spectrometry data, a new extracellular peroxidase (CMP) from Chelidonium majus milky sap was isolated and characterized. The protein has a molecular weight of about 40 kDa and belongs to secretory class III plant peroxidases. The peroxidase activity is also accompanied by DN-ase activities. A novel CMP combined with other proteins is probably involved in development and differentiation of the plant and defence against different pathogens. It suggests that the biological activity of C. majus whole plants and extracts may depend not only on its alkaloidal content but also on the presence of biologically active proteins.     

Iridoid, phenylethanoid and flavonoid glycosides from Sideritis trojana

From the MeOH extract of Sideritis trojana, a new iridoid glycoside, 10-O-(E)-feruloylmelittoside (1) was obtained in addition to four known iridoid glycosides [melittoside (2), 10-O-(E)-p-coumaroylmelittoside (3), stachysosides E (4) and G (5)]. Moreover, five phenylethanoid glycosides [verbascoside (6), isoacteoside (7), lamalboside (8), leonoside A (9), isolavandulifolioside (10), three flavone glycosides (isoscutellarein 7-O-[6'-O-acetyl-β-allopyranosyl-(1→2)]-β-glucopyranoside (11), 4'-O-methyisoscutellarein 7-O-[6'-O-acetyl-β-allopyranosyl-(1→2)]-β-glucopyranoside (12), 3'-hydroxy-4'-O-methyisoscutellarein 7-O-[6'-O-acetyl-β-allopyranosyl-(1→2)]-β-glucopyranoside (13) and a benzylalcohol derivative (di-O-methylcrenatin) were obtained and identified. The structures were elucidated on the basis of NMR and HRMS data. All compounds were tested for their antioxidant activity by in vitro TEAC assay and some of them exhibited moderate activity (0.97-1.44 mM) when compared with the reference compound (quercetin 1.86 mM). Glycosides 6-13, the most active compounds in the TEAC assay, were also tested by flow cytometry to evaluate their ability to affect the levels of reactive oxygen species (ROS) in human prostate cancer cells (PC3).