Determining acrosomal status of the cynomolgus monkey (Macaca fascicularis) sperm by fluorescence microscopy

The acrosome of Macaca fascicularis sperm cannot be distinguished by conventional light microscopy, so determining whether sperm are acrosome-intact or-reacted is difficult. We describe methods for labeling the acrosomal region of sperm with two different probes: fluoresceinated Pisum sativum agglutinin and anti-sperm antiserum. Acrosome-intact sperm are much more heavily labeled in the acrosomal region than are acrosome-reacted sperm, providing a simple means of differentiating the two types of sperm. The two probes detect similar numbers of acrosome-reacted sperm following treatment with the divalent cation ionophore, A23187.     

An Attachment for Nondestructive, Fast Quenching of Samples in Rapid-Mixing Experiments

The present paper describes a quenching-and-washing chamber (QWC) to be used with a rapid-mixing apparatus (RMA) for the study of processes in the millisecond time scale. The QWC enables fast, nondestructive quenching by cooling and dilution of reactants in particulate systems that can be trapped on a filter. The reaction mixture (e.g., at 25 degrees C) is injected from the RMA into the QWC where it is immediately mixed with a stream of ice-cold solution flowing at a rate of 15-40 ml s-1. Quenching requires that the process studied is slowed considerably by cooling to 0-2 degrees C and/or by removal of reactants by dilution. The equipment was characterized through a study of the tight binding (occlusion) of 86Rb+ to purified, membrane-bound Na+/K+-ATPase. Millipore filters of 0.22-0.80 microm pore size trapped close to 100% of the enzyme protein. Enzyme with occluded 86Rb+ was formed in the RMA under conditions where the rate constant for release of Rb+ at 25 degrees C is up to 25 s-1 and then injected into the QWC. The high off-rate constant is due to the presence of 2.5 mM ATP, which accelerates release of Rb+. The recovery of occluded 86Rb+ on the filter was at least 90%, indicating that both cooling of the reactants and dilution of ATP are fast enough to stop the reaction. The quenching time was 3-4 ms.     

Biological performance of Diclidophlebia smithi (Hemiptera: Psyllidae), a potential biocontrol agent for the invasive weed Miconia calvescens

Diclidophlebia smithi Burckhardt, Morais and Picanço (Hemiptera: Psyllidae) is a promising biological control agent of Miconia calvescens DC. (Melastomataceae), a neotropical invasive weed in forest ecosystems in French Polynesia and Hawaii and a threat in Australia, where it was also introduced. A study on the reproductive performance of D. smithi under laboratory conditions through life expectancy and fertility tables is presented. Results indicated that this psyllid has a high reproductive capacity (R ₀>1 and r _m >0) and a short life cycle (46–47 days) and can have up to nine generations per year. The critical period of its life cycle is during the nymphal stage which is clearly inadequate for field introductions. The best age for introducing D. smithi against M. calvescens is the 4th day of the adult stage. D. smithi is easily mass-reared and has a short life cycle and a high reproductive capacity, which are desirable characteristics for a biological control agent.     

The pollen of Caesalpinia pyramidalis Tul. is toxic to honeybees (Apis mellifera)

Caesalpinia pyramidalis Tul. (syn. Caesalpinia gardneriana Benth.) is an endemic tree from northeastern Brazil whose flowers are visited by a number of bee species, including the honeybee (Apis mellifera). The present study aimed to determine whether the pollen of C. pyramidalis is toxic to honeybees. Pollen samples were collected, dried, and added to food at doses of 0 (control), 2.5, 5.0, and 10 %. Each dose of food was fed to 60 honeybees, which were observed daily until the last one died. Statistical comparison of the survival curves was performed using the log-rank test. The mean survival times of the bees fed with C. pyramidalis pollen were 6.40 ± 0.28 days for the 2.5 % group, 5.70 ± 0.18 days for the 5.0 % group, 5.72 ± 0.23 days for the 10.0 % group, and 13.7 ± 0.61 days for the control group. All the pollen concentrations of C. pyramidalis significantly (P < 0.0001) reduced the survival of honeybees in comparison with control group. In conclusion, the results of our study show that the pollen produced by C. pyramidalis is toxic to A. mellifera under laboratory conditions.     

Ovarian morphology and folliculogenesis and ovulation process in the flat-faced fruit-eating bat Artibeus planirostris and the Argentine brown bat Eptesicus furinalis: A comparative analysis

The Neotropical bat species Artibeus planirostris and Eptesicus furinalis present a different morphology of the female reproductive organs: the first presents a simplex uterus, while the second presents a bicornuate uterus, but there is no information about their ovaries. Our aim was to compare the general ovary morphology and the folliculogenesis process in these species to increase the knowledge about the reproductive diversity of tropical bats. We observed a morphological distinction between the ovaries of both species: A. planirostris presents the primordial follicles located in a cranial portion of the ovary and the interstitial gland cells are not distinctive, while in E. furinalis, the primordial follicles are located throughout the cortex, and there is an abundance of interstitial gland cells. Both species present binovular or triovular follicles. Artibeus planirostris is a monovular species, with a preferential ovulation in the left side. Some females of E. furinalis exhibited two corpora lutea in the same ovary, and others presented a corpus luteum in both right and left ovaries at the same time; thus, E. furinalis is a polyovular species. Our results express the variation between two Neotropical species, reflecting the great variation in the reproductive aspects in Chiroptera.     

Association of TYK2 polymorphisms with autoimmune diseases: A comprehensive and updated systematic review with meta-analysis

Autoimmune diseases are characterized by the loss of self-tolerance, leading to immune-mediated tissue destruction and chronic inflammation. Tyrosine kinase 2 (TYK2) protein plays a key role in immunity and apoptosis pathways. Studies have reported associations between single nucleotide polymorphisms (SNPs) in the TYK2 gene and autoimmune diseases; however, results are still inconclusive. Thus, we conducted a systematic review followed by meta-analysis. A literature search was performed to find studies that investigated associations between TYK2 SNPs and autoimmune diseases (multiple sclerosis, systemic lupus erythematosus, Crohn’s disease, ulcerative colitis, psoriasis, rheumatoid arthritis, type 1 diabetes, and inflammatory bowel disease). Pooled odds ratios (OR) with 95 % CI were calculated using random (REM) or fixed (FEM) effects models in the Stata 11.0 Software. Thirty-four articles were eligible for inclusion in the meta-analyses, comprising 9 different SNPs: rs280496, rs280500, rs280523, rs280519, rs2304256, rs12720270, rs12720356, rs34536443, and rs35018800. Meta-analysis results showed the minor alleles of rs2304256, rs12720270, rs12720356, rs34536443, and rs35018800 SNPs were associated with protection against autoimmune diseases. Moreover, the A allele of the rs280519 SNP was associated with risk for systemic lupus erythematosus. Our meta-analyses demonstrated that the rs2304256, rs12720270, rs12720356, rs34536443, rs35018800, and rs280519 SNPs in the TYK2 gene are associated with different autoimmune diseases.