Effects of leptin on in vitro maturation, fertilization and embryonic cleavage after ICSI and early developmental expression of leptin (Ob) and leptin receptor (ObR) proteins in the horse

Background  

The identification of the adipocyte-derived obesity gene product, leptin (Ob), and subsequently its association with reproduction in rodents and humans led to speculations that leptin may be involved in the regulation of oocyte and preimplantation embryo development. In mice and pigs, in vitro leptin addition significantly increased meiotic resumption and promoted preimplantation embryo development in a dose-dependent manner. This study was conducted to determine whether leptin supplementation during in vitro maturation (IVM) to horse oocytes could have effects on their developmental capacity after fertilization by IntraCytoplasmic Sperm Injection (ICSI).     

Cytokine profile of autologous conditioned serum for treatment of osteoarthritis, <Emphasis Type="Italic">in vitro</Emphasis>effects on cartilage metabolism and intra-articular levels after injection

Introduction  

Intraarticular administration of autologous conditioned serum (ACS) recently demonstrated some clinical effectiveness in treatment of osteoarthritis (OA). The current study aims to evaluate the in vitro effects of ACS on cartilage proteoglycan (PG) metabolism, its composition and the effects on synovial fluid (SF) cytokine levels following intraarticular ACS administration.     

Nitrogen fluxes from treefrogs to tank epiphytic bromeliads: an isotopic and physiological approach

Diverse invertebrate and vertebrate species live in association with plants of the large Neotropical family Bromeliaceae. Although previous studies have assumed that debris of associated organisms improves plant nutrition, so far little evidence supports this assumption. In this study we used isotopic (¹⁵N) and physiological methods to investigate if the treefrog Scinax hayii, which uses the tank epiphytic bromeliad Vriesea bituminosa as a diurnal shelter, contributes to host plant nutrition. In the field, bromeliads with frogs had higher stable N isotopic composition (δ¹⁵N) values than those without frogs. Similar results were obtained from a controlled greenhouse experiment. Linear mixing models showed that frog feces and dead termites used to simulate insects that eventually fall inside the bromeliad tank contributed, respectively, 27.7% (±0.07 SE) and 49.6% (±0.50 SE) of the total N of V. bituminosa. Net photosynthetic rate was higher in plants that received feces and termites than in controls; however, this effect was only detected in the rainy, but not in the dry season. These results demonstrate for the first time that vertebrates contribute to bromeliad nutrition, and that this benefit is seasonally restricted. Since amphibian–bromeliad associations occur in diverse habitats in South and Central America, this mechanism for deriving nutrients may be important in bromeliad systems throughout the Neotropics.     

Chromosome Banding and 18S rDNA in situ Hybridization Analysis of Seven Species of the Family Achiridae (Teleostei: Pleuronectiformes)

Achiridae is an important family of the order Pleuronectiformes widely distributed in North, Central, and South America with freshwater and marine species. In the present study cytogenetic analyses comprising conventional and molecular techniques were carried out in seven species of this family. The following diploid numbers (2n) and fundamental numbers (FN) were obtained: Achirus declivis 2n = 34, FN = 52; Achirus lineatus 2n = 40, FN = 66; Catathyridium jenynsi 2n = 40 and FN = 50; Gymnachirus nudus 2n = 36 and FN = 50; Hypoclinemus mentalis 2n = 38 and FN = 54; Trinectes paulistanus 2n = 42 and FN = 52; and Trinectes sp. 2n = 38 and FN = 54. All species presented a single nucleolar organizer region (NOR) bearing chromosome pair and C-band positive segments mainly distributed at the pericentromeric position. The wide variation observed in chromosome number and FN suggests the occurrence of larger chromosome rearrangements in the family Achiridae if compared with other families of the same order.     

Comparative Analysis of a Mitochondrial DNA Control Region Fragment Amplified from Three Adriatic Flatfish Species and Molecular Phylogenesis of Pleuronectiformes

The 5′-end of the mitochondrial control region of three Pleuronectiformes from the Adriatic Sea, Platichthys flesus italicus (Adriatic flounder), Solea vulgaris (common sole), and Solea kleini (Klein's sole), was sequenced and compared with that of six other flatfish species from the families Pleuronectidae and Bothidae. The sequence structures of all flatfishes appear very similar and consist of alternate short segments with low, medium, and high rates of nucleotide substitution. Four conserved 19-bp repeats occur at the beginning of the European and Adriatic flounder sequences. The common occurrence of tandem arrays in fish control regions could be related to a stable secondary structure. Molecular phylogenetic relationships among Pleuronectiformes agree well with previous morphologic data at all taxonomic levels. Molecular analyses could therefore contribute to resolving phylogenetic and taxonomic debates within the Pleuronectiformes. Received December 1, 1997; accepted June 30, 1998.     

Feedback control of the limbs position during voluntary rhythmic oscillation

The mechanisms that control the limbs position during rhythmic voluntary oscillations were investigated in ten subjects, who were asked to synchronise the lower peak of their hand or foot rhythmic oscillations to a metronome beat. The efficacy of the “position control” was estimated by measuring the degree of synchronisation between the metronome signal and the requested limb position and how it was affected by changing both the oscillation frequency (between 0.4 and 3.0 Hz) and the limbs inertial properties. With the limbs unloaded, the lower peak of both the hand and foot oscillations lagged the metronome beat of a slight amount that remained constant over the whole frequency range (mean phase delay −13.2° for the hand and −4.7° for the foot). The constancy was obtained by phase-advancing, at each frequency increment, the electromyogram (EMG) activation with respect of the clock beat of the amount necessary to compensate for the simultaneous increase of the lag between the EMG and the movement, produced by the limb mechanical impedance. After loading of either limb, the increase of the oscillation frequency induced larger EMG-movement delays and the anticipatory compensation became insufficient, so that the movement progressively phase-lagged the clock beat. The above results have been accurately simulated by a neural network connected to a pendulum model that shared the same mechanical properties of the moving limb. The network compares a central command (the intended position) to the actual position of the effector and acts as a closed-loop proportional, integrative and derivative controller. It is proposed that the synchronisation of rhythmic oscillations of either the hand or the foot is sustained by a feed-back control that conforms the position of each limb to that encoded in the central voluntary command.     

Glutamate-cysteine ligase attenuates TNF-induced mitochondrial injury and apoptosis

Glutathione (GSH) is important in free radical scavenging, maintaining cellular redox status, and regulating cell survival in response to a wide variety of toxicants. The rate-limiting enzyme in GSH synthesis is glutamate-cysteine ligase (GCL), which is composed of catalytic (GCLC) and modifier (GCLM) subunits. To determine whether increased GSH biosynthetic capacity enhances cellular resistance to tumor necrosis factor-alpha- (TNF-alpha-) induced apoptotic cell death, we have established several mouse liver hepatoma (Hepa-1) cell lines overexpressing GCLC and/or GCLM. Cells overexpressing GCLC alone exhibit modest increases in GCL activity, while cells overexpressing both subunits have large increases in GCL activity. Importantly, cells overexpressing both GCL subunits exhibit increased resistance to TNF-induced apoptosis as judged by a loss of redox potential; mitochondrial membrane potential; translocation of cytochrome c to the cytoplasm; and activation of caspase-3, caspase-8, and caspase-9. Analysis of the effects of TNF on these parameters indicates that maintaining mitochondrial integrity mediates this protective effect in GCL-overexpressing cells.     

Proteomic evaluation of sheep serum proteins

Background

Mammary tumours frequently develop in female domestic cats being highly malignant in a large percentage of cases. Chemokines regulate many physiological and pathological processes including organogenesis, chemotaxis of inflammatory cells, as well as tumour progression and metastasization. In particular, the chemokine/receptor pair SDF-1/CXCR4 has been involved in the regulation of metastatic potential of neoplastic cells, including breast cancer. The aim of this study was the immunohistochemical defininition of the expression profile of CXCR4 in primary and metastatic feline mammary carcinomas and the evaluation of the role of SDF-1 in feline mammary tumour cell proliferation.

Results

A total of 45 mammary surgical samples, including 33 primary tumours (31 carcinomas and 2 adenomas), 6 metastases, and 4 normal mammary tissues were anlyzed. Tumor samples were collected from a total number of 26 animals, as in some cases concurrent occurrence of neoplasm in more than one mammary gland was observed. Tissues were processed for standard histological examination, and all lesions were classified according to the World Health Organization criteria. CXCR4 expression in neoplastic cells was evaluated by immunohistochemistry. The level of CXCR4 immunoreactivity was semi-quantitatively estimated as CXCR4 score evaluating both the number of positive cells and the intensity of staining. Six primary, fibroblast-free primary cultures were obtained from fresh feline mammary carcinomas and characterized by immunofluorescence for CXCR4 and malignant mammary cell marker expression. SDF-1-dependent in vitro proliferative effects were also assayed. CXCR4 expression was observed in 29 out of 31 malignant tissues with a higher CXCR4 score observed in 4 out of 6 metastatic lesions than in the respective primary tumours. In 2 benign lesions analyzed, only the single basaloid adenoma showed a mild positive immunostaining against CXCR4. Normal tissue did not show CXCR4 immunoreactivity. CXCR4 score was statistically significantly associated with the histological features of the samples, showing an increase accordingly with the degree of neoplastic transformation (from normal tissue to metastatic lesions). Finally, in the primary cultures obtained from 6 primary feline mammary carcinomas CXCR4 expression was detected in all cells and its activation by SDF-1 in vitro treatment caused a significant increase in the proliferation rate in 5 out of 6 tumours.

Conclusions

These results indicate that malignant feline mammary tumours commonly express CXCR4, with a higher level in malignant tumours, and, in most of the cases analysed, metastatic cells display stronger immunoreactivity for CXCR4 than the corresponding primary tumours. Moreover, CXCR4 activation in primary cultures of feline mammary carcinomas causes increase in the proliferative rate. Thus, SDF-1/CXCR4 system seems to play a tumorigenic in feline mammary gland malignancy and in vitro cultures from these tumour samples may represent an experimental model to investigate the biological and pharmacological role of this chemokinergic axis.     

Morphological convergence and molecular divergence: the taxonomic status of Sceloporus serrifer (squamata,phrynosomatidae) subspecies

Martínez‐Méndez, N., Mejía, O., Rocha‐Gómez, A. & Méndez‐De La Cruz, F. R. (2012) Morphological convergence and molecular divergence: the taxonomic status of Sceloporus serrifer (squamata, phrynosomatidae) subspecies. —Zoologica Scripta, 41, 97–108. The systematics of the subspecies included in Sceloporus serrifer has undergone several taxonomic and nomenclatural rearrangements. This species previously comprised two recognized subspecies, Sceloporus serrifer serrifer and Sceloporus serrifer prezygus, which inhabit contrasting habitats. In this study, we re‐evaluate the taxonomic status of both subspecies using molecular and morphological evidence. Sceloporus serrifer serrifer was recovered as a paraphyletic group, whereas S. s. prezygus was recovered as a polyphyletic group. The molecular results indicate the existence of two independent lineages that diverged at least 1.29 Myr, with the populations currently recognized as S. s. prezygus from the Cuchumatanes Mountains representing a secondary invasion of S. s. serrifer from the lowlands. The lack of meristic differences among the highland populations of different species suggests convergence in similar environments achieved by two different lineages.     

Plant plasma membrane‐resident receptors: Surveillance for infections and coordination for growth and development

As sessile organisms, plants are exposed to pathogen invasions and environmental fluctuations. To overcome the challenges of their surroundings, plants acquire the potential to sense endogenous and exogenous cues, resulting in their adaptability. Hence, plants have evolved a large collection of plasma membrane-resident receptors, including RECEPTOR-LIKE KINASEs(RLKs) and RECEPTOR-LIKE PROTEINs(RLPs) to perceive those signals and regulate plant growth,development, and immunity. The ability of RLKs and RLPs to recognize distinct ligands relies on diverse categories of extracellular domains evolved. Co-regulatory receptors are often required to associate with RLKs and RLPs to facilitate cellular signal transduction. RECEPTOR-LIKE CYTOPLASMIC KINASEs(RLCKs) also associate with the complex, bifurcating the signal to key signaling hubs, such as MITOGEN-ACTIVATED PROTEIN KINASE(MAPK) cascades, to regulate diverse biological processes. Here, we discuss recent knowledge advances in understanding the roles of RLKs and RLPs in plant growth, development, and immunity, and their connection with co-regulatory receptors, leading to activation of diverse intracellular signaling pathways.