Effects of an intrauterine copper device on serum copper, endometrial histology, and ovarian, hepatic, and renal functions in the Japanese monkey (Macaca fuscata fuscata).

A copper intrauterine device (Cu-IUD) was inserted transabdominally into the uterine cavity of eight Japanese monkeys (Macaca fuscata fuscata) for 4 to 6 months, and effects on various organ functions were examined. Results showed no significant effects on the menstrual cycle length, serum levels of LH, estradiol-17 beta, progesterone or clinical biochemical data such as serum copper, glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, lactic dehydrogenase, and blood urea nitrogen. Histology revealed edema and infiltration of eosinophilic leukocytes in the endometrium treated with a Cu-IUD.     

Thiosulfate promotes hair growth in mouse model

The present study describes the hair growth-promoting effects of sodium thiosulfate (STS), a widely used compound, in mice. STS accelerated hair growth in the “telogen model”, suggesting that it stimulates telogen hair follicles to reenter the anagen phase of hair growth. In the same model, STS potentiated hair growth in an additive manner with minoxidil (MXD), a drug used for the treatment of androgenic alopecia. Furthermore, in the “anagen model”, STS promoted hair growth, probably by promoting hair follicle proliferation. Since STS elevated the skin surface temperature, its hair growth-promoting activity may be partly due to vasorelaxation, similar to MXD. In addition, STS is known to generate a gaseous mediator, H₂S, which has vasorelaxation and anti-inflammatory/anti-oxidative stress activities. Therefore, STS and/or provisionally its metabolite, H₂S, may aid the hair growth process. Collectively, these results suggest that salts of thiosulfate may represent a novel and beneficial remedy for hair loss.     

The properties and interrelationships of various force-time parameters during maximal repeated rhythmic grip

The purpose of this study was to examine the properties and interrelationships of various force-time parameters including the inflection point for the rate of decline in force during a maximal repeated rhythmic grip. Fifteen healthy males (age M=21.5, SD=2.1 yr, height M=172.4, SD=5.7 cm, body mass M=68.2, SD=9.2 kg) participated in this study. Subjects performed a maximal repeated rhythmic grip with maximal effort with a target frequency of 30 grip.min(-1) for 6 min. The force value decreased linearly and markedly until about 70% of maximal strength for about 55 s after the onset of a maximal repeated rhythmic grip, and then decreased moderately. Because all parameters showed fair or good correlations between 3 min and 6 min, they are considered to be able to sufficiently evaluate muscle endurance for 3 min instead of 6 min. However, there were significant differences between 3 min and 6 min in the integrated area, the final force, the rate of the decrement constant (k) fitting the force decreasing data to y=ae(-kx)+b and the force of maximal difference between the force and a straight line from peak force to the final force. Their parameters may vary generally by the length of a steady state, namely, a measurement time. The final force value before finishing and the rate of the decrement constant (k) reflect the latter phase during a maximal repeated rhythmic grip. Although many parameters show relatively high mutual relationships, the rate constant (k) shows relatively low correlations with other parameters. We inferred that decreasing the time until 80% of maximal strength and the amount of the decrement force for the first 1 min reflect a linear decrease in the initial phase.     

Two Rab7 isotypes, EhRab7A and EhRab7B, play distinct roles in biogenesis of lysosomes and phagosomes in the enteric protozoan parasite Entamoeba histolytica

Rab7 small GTPase plays a crucial role in the regulation of trafficking to late endosomes, lysosomes and phagosomes. While most eukaryotes encode a single Rab7, the parasitic protist Entamoeba histolytica possesses nine Rab7. In this study, to understand the significance of the presence of multiple Rab7 isotypes, a role of two representative Rab7 isotypes, EhRab7A and EhRab7B, was investigated. EhRab7B was exclusively localized to acidic vacuoles containing lysosomal proteins, e.g. amoebapore-A and cysteine protease. This lysosome localization of EhRab7B was in good contrast to EhRab7A, localized to a non-acidic compartment in steady state, and only partially colocalized with lysosomal proteins. Overexpression of EhRab7B resulted in augmentation of late endosome/lysosome acidification, similar to the EhRab7A overexpression. Expression of EhRab7B-GTP mutant caused dominant-negative phenotypes including decrease in late endosome/lysosome acidification and missecretion of lysosomal proteins, while EhRab7A-GTP enhanced acidification but did not affect either intracellular or secreted cysteine protease activity. Expression of either EhRab7B or EhRab7B-GTP mutant caused defect in phagocytosis, concomitant with the disturbed formation and disassembly of prephagosomal vacuoles, the compartment previously shown to be linked to efficient ingestion. Altogether, these data indicate that the two Rab7 isotypes play distinct but co-ordinated roles in lysosome and phagosome biogenesis.     

Small GTPase Rab21 Mediates Fibronectin Induced Actin Reorganization in Entamoeba histolytica: Implications in Pathogen Invasion

The protozoan parasite Entamoeba histolytica causes a wide spectrum of intestinal infections. In severe cases, the trophozoites can breach the mucosal barrier, invade the intestinal epithelium and travel via the portal circulation to the liver, where they cause hepatic abscesses, which can prove fatal if left untreated. The host Extra Cellular Matrix (ECM) plays a crucial role in amoebic invasion by triggering an array of cellular responses in the parasite, including induction of actin rich adhesion structures. Similar actin rich protrusive structures, known as ‘invadosomes’, promote chemotactic migration of the metastatic cancer cells and non-transformed cells by remodeling the ECM. Recent studies showed a central role for Rab GTPases, the master regulators of vesicular trafficking, in biogenesis of invadosomes. Here, we showed that fibronectin, a major host ECM component induced actin remodeling in the parasite in a Rab21 dependent manner. The focalized actin structures formed were reminiscent of the mammalian invadosomes. By using various approaches, such as immunofluorescence confocal microscopy and scanning electron microscopy, along with in vitro invasion assay and matrix degradation assay, we show that the fibronectin induced formation of amoebic actin dots depend on the nucleotide status of the GTPase. The ECM components, fibronectin and collagen type I, displayed differential control over the formation of actin dots, with fibronectin positively and collagen type I negatively modulating it. The cell surface adhesion molecule Gal/GalNAc complex was also found to impose additional regulation on this process, which might have implication in collagen type I mediated suppression of actin dots.     

EVIDENCE FOR TUBULAR MATING STRUCTURES INDUCED IN EACH MATING TYPE OF HETEROTHALLIC GONIUM PECTORALE (VOLVOCALES,CHLOROPHYTA)1

Gametes were induced separately in cultures of each mating type of the heterothallic, isogamous colonial volvocalean Gonium pectorale O. F. Müll. to examine the tubular mating structure (TMS) of both mating types plus and minus (plus and minus), referred to as “bilateral mating papillae.” Addition of dibutyryl cyclic adenosine monophosphate (DcAMP or db‐cAMP) and 3‐isobutyl‐1‐methylxanthine (IBMX) to approximately 3‐week‐old cultures of each mating type induced immediate release of naked gametes from the cell walls. Both plus and minus gametes formed a TMS in the anterior region of the protoplasts. Accumulation of actin was visualized by antibody staining in the TMS of both mating types as occurs in the TMS (fertilization tubule) of the plus gametes of the unicellular volvocalean Chlamydomonas reinhardtii P. A. Dang. Induction of naked gametes with a TMS in each mating type will be useful for future cell biological and evolutionary studies of the isogametes of colonial volvocalean algae.     

Role of conserved active site tryptophan-101 in functional activity and stability of phosphoserine aminotransferase from an enteric human parasite

Site-directed mutagenesis study was performed to elucidate the role of conserved tryptophan-101 present at the active site of phosphoserine aminotransferase from an enteric human parasite Entamoeba histolytica. Fluorescence resonance energy transfer and molecular dynamic simulation show that the indole ring of Trp101 stacks with the cofactor PLP. Loss of enzymatic activity and PLP polarization values suggest that Trp101 plays a major role in maintaining a defined PLP microenvironment essentially required for optimal enzymatic activity. Studies on W101F, W101H and W101A mutants show that only the indole ring of the conserved Trp101 forms most favorable stacking interaction with the pyridine ring of the cofactor PLP. Protein stability was compromised on substitution of Trp101 with Phe/His/Ala amino acids. A difference in conformational free energy of 1.65?kcal?mol(-1) was observed between WT-protein and W101A mutant.