Ehlers-Danlos syndrome (cutaneous asthenia) in a Campbell's dwarf hamster (Phodopus campbelli).

A case of cutaneous asthenia in a Campbell's dwarf hamster is described. The animal was found to have hyperextensible skin, glaucoma and lens dislocation. Histopathological examination revealed an irregular, haphazard arrangement of collagen fibres in the dermis. The animal underwent surgical reduction of the skin folds which provided only temporary relief.

Un cas d'asthénie cutanée chez un hamster nain de Campbell est décrit. L'animal présente une peau hyperextensible, un glaucome et une luxation du cristallin. L'examen histopathologique révèle une disposition irrégulière et désordonnée des fibres de collagène dans le derme. L'animal a subi une réduction chirurgicale des plis cutanés qui n'a apporté qu'un soulagement temporaire.

Descreve­se um caso de astenia cutânea em um hamster anão de Campbell. O animal apresentou hiperextensibilidade cutânea, glaucoma e deslocamento de lentes. Ao exame histopatológico, observou­se um arranjo irregular e aleatório das fibras colágenas na derme. O animal foi submetido à redução cirúrgica das dobras cutâneas que resultou em alívio apenas temporário.

Se describe un caso de astenia cutánea en un hámster enano Campbell. Se examinó al animal obsevando piel hiperextensible, glaucoma y luxación del cristalino. El examen histopatológico reveló una orientación aleatoria e irregular de las fibras de colágeno en la dermis. Se utilizó una técnica de reducción quirúrgica de los pliegues de la piel que solo indujo una mejora temporal.

Prevalence of Encephalitozoon cuniculi Infection in Guinea Pigs (Cavia porcellus) in Poland with Different Clinical Disorders-A Pilot Study.

Encephalitozoonosis is a disease caused by E. cuniculi. It is diagnosed primarily in rabbits but is less frequently so in other animal species. E. cuniculi is classified among Microsporidia-fungi frequently found in the environment, that are resistant to numerous external factors. Apart from rabbits, rodents form the next group of animals most exposed to infection with these pathogens. The objective of the study was to analyze the prevalence of E. cuniculi infection in guinea pigs with different clinical disorders. The study included 67 animals with E. cuniculi infection confirmed via real-time PCR. The infected animals most frequently exhibited nervous and urinary system symptoms, as well as issues with vision organs, while several animals were also recorded as having problems with the respiratory system and thyroid gland dysfunction. The study shows that encephalitozoonosis constitutes a significant problem in rodents kept as domestic animals, which in turn may be a source of infection for humans.

Biocompatible curdlan-based biomaterials loaded with gentamicin and Zn-doped nano-hydroxyapatite as promising dressing materials for the treatment of infected wounds and prevention of surgical site infections.

A topical application of antibiotic-loaded wound dressings is recommended only for chronically infected wounds with poor vascularization. Thus, more often dressing materials loaded with antibacterial metal ions are produced. In turn, gentamicin sponges are commonly used to prevent surgical site infections. The aim of this study was to produce curdlan-based biomaterials enriched with gentamicin and zinc (Zn)-doped nano-hydroxyapatite to prevent wound and surgical site infections. Developed biomaterials were subjected to basic microstructural characterization, cytotoxicity test against human skin fibroblasts (BJ cell line), and comprehensive microbiological experiments using Staphylococcus aureus and Pseudomonas aeruginosa strains. To evaluate the in vivo healing capacity of the developed biomaterials, severely infected chronic wound in a veterinary patient was treated with the use of gentamicin-loaded dressing. Fabricated biomaterials were characterized by a highly porous microstructure with high plasma absorption capacity (approx. 7 mL/g for Zn-loaded biomaterial and 13 mL/g for gentamicin-enriched dressing) and optimal water vapor transmission rate (approx. 1700 g/m2/day). Due to the presence of bioceramics, material containing Zn showed slightly higher compressive strength (0.37 MPa) and Young's modulus (3.33 MPa) values compared to gentamicin-loaded biomaterial (0.12 MPa and 1.29 MPa, respectively). Gentamicin-enriched biomaterial showed burst release of the drug within the first 5 h, while, the zinc-loaded biomaterial exhibited a constant gradual release of the zinc ions. Conducted assays showed that developed biomaterials were non-toxic against human skin fibroblasts (cell viability in the range of 71-95 %) and revealed strong bactericidal activity (99.9 % reduction in the number of viable bacterial CFUs in direct contact test) against S. aureus. In the case of P. aeruginosa, only gentamicin-loaded biomaterial exhibited bactericidal effect. Additionally, biomaterials had the ability to uptake, lock in, and kill bacteria within their gel structure, enabling the cleansing of the wound bed at every dressing change. Finally, the treatment of severely infected wound in veterinary patient confirmed the effectiveness of gentamicin-loaded biomaterial. Biomaterial enriched with gentamicin possesses great potential to be used as a dressing material or sponge for the treatment of chronically infected wounds and surgical site infections. In turn, the zinc-loaded biomaterial may be used as a wound dressing to reduce and prevent microbial contamination.

Granulocytic anaplasmosis in captive ring-tailed lemur (Lemur catta) in Poland.

BACKGROUND: Anaplasma are obligate intracellular bacteria and aetiological agents of tick-borne diseases of both veterinary and medical interest. The genus Anaplasma comprises six species: Anaplasma marginale, Anaplasma centrale, Anaplasma ovis, Anaplasma phagocytophilum, Anaplasma bovis and Anaplasma platys. They can infect humans, carnivores, ruminants, rodents, insectivores, birds and reptiles. The aim of this study was to present the first clinical case of granulocytic anaplasmosis in a captive ring-tailed lemur in Poland. CASE PRESENTATION: A 4-year-old female lemur presented anorexia, epistaxis and tick infestation. The microscopic examination of a blood smear revealed morulae in neutrophils. Polymerase chain reaction test and sequencing of obtained PCR product confirmed infection by the GU183908 Anaplasma phagocytophilum strain. Therapeutic protocol included doxycycline (2.5 mg/kg p.o., b.i.d.) for 3 weeks and the lemur recovered within 24 h. CONCLUSIONS: This is the first report on granulocytic anaplasmosis in a ring-tailed lemur in Europe, indicating that A. phagocytophilum infection must also be considered in differential diagnosis in this animal species, especially in individuals with thrombocytopenia associated with Ixodes ricinus parasitism.

Effect of Bathing in a 0.1% Aqueous Solution of Ethacridine Lactate on Selected Physiological Parameters of Cornu Aspersum Müller Edible Snails.

INTRODUCTION: The prohibition of antibiotic use in edible snails obligates breeders to treat bacterial infections by different means, of which a common one is a bath in Gram-positive- and partially Gram-negative-bactericidal ethacridine lactate solution. The aim of the study was to determine the effect of bathing Cornu aspersum Müller snails in a 0.1% aqueous solution of ethacridine lactate on selected physiological parameters of haemolymph. MATERIAL AND METHODS: The study included 80 snails, divided into two equal groups (study and control). The study group was subjected to bathing in ethacridine lactate and the control group to bathing in tap water. Both groups were treated daily for seven days. The number of haemocytes in the haemolymph, the activity of alanine (ALT) and aspartate (AST) aminotransferases, and the concentration of urea were determined. RESULTS: In the study group, after exposure to ethacridine lactate solution an increase in ALT activity, changes in the De Ritis ratio, an increase in the amount of haemocytes, and a decrease in body weight were found. No such changes were detected in the control group snails or in animals after the first bath. CONCLUSION: Multiple applications of a 0.1% ethacridine lactate bath may adversely affect Cornu aspersum Müller snails.

The diverse roles of the eIF4A family: you are the company you keep.

The eIF4A (eukaryotic initiation factor 4A) proteins belong to the extensive DEAD-box RNA helicase family, the members of which are involved in many aspects of RNA metabolism by virtue of their RNA-binding capacity and ATPase activity. Three eIF4A proteins have been characterized in vertebrates: eIF4A1 and eIF4A2 are cytoplasmic, whereas eIF4A3 is nuclear-localized. Although highly similar, they have been shown to possess rather diverse roles in the mRNA lifecycle. Their specific and diverse functions are often regulated and dictated by interacting partner proteins. The key differences between eIF4A family members are discussed in the present review.

Xenopus laevis zygote arrest 2 (zar2) encodes a zinc finger RNA-binding protein that binds to the translational control sequence in the maternal Wee1 mRNA and regulates translation.

Zygote arrest (Zar) proteins are crucial for early embryonic development, but their molecular mechanism of action is unknown. The Translational Control Sequence (TCS) in the 3' untranslated region (UTR) of the maternal mRNA, Wee1, mediates translational repression in immature Xenopus oocytes and translational activation in mature oocytes, but the protein that binds to the TCS and mediates translational control is not known. Here we show that Xenopus laevis Zar2 (encoded by zar2) binds to the TCS in maternal Wee1 mRNA and represses translation in immature oocytes. Using yeast 3 hybrid assays and electrophoretic mobility shift assays, Zar2 was shown to bind specifically to the TCS in the Wee1 3'UTR. RNA binding required the presence of Zn(2+) and conserved cysteines in the C-terminal domain, suggesting that Zar2 contains a zinc finger. Consistent with regulating maternal mRNAs, Zar2 was present throughout oogenesis, and endogenous Zar2 co-immunoprecipitated endogenous Wee1 mRNA from immature oocytes, demonstrating the physiological significance of the protein-RNA interaction. Interestingly, Zar2 levels decreased during oocyte maturation. Dual luciferase reporter tethered assays showed that Zar2 repressed translation in immature oocytes. Translational repression was relieved during oocyte maturation and this coincided with degradation of Zar2 during maturation. This is the first report of a molecular function of zygote arrest proteins. These data show that Zar2 contains a zinc finger and is a trans-acting factor for the TCS in maternal mRNAs in immature Xenopus oocytes.

Translational control assessed using the tethered function assay in Xenopus oocytes.

The tethered function assay is a method designed to address the role of an RNA-binding protein upon the metabolism of a reporter RNA. The basis of this assay is to artificially tether a test protein to a reporter mRNA by employing an unrelated bacteriophage MS2 or lambda N RNA-protein interaction, and to assess the effects of the test protein on the reporter RNA. In this chapter, we first discuss the principles and validity of the tethered function approach, drawing on appropriate examples from several cell types and of many proteins that regulate RNA in a variety of processes, including RNA processing (splicing, polyadenylation/deadenylation, decay), localisation and protein synthesis. Secondly, we will focus on the use of this approach to monitor translational activation and repression in Xenopus oocytes, giving a detailed protocol, and discussing possible optimizations we have explored.

[Assessment of three-level selective perinatal care based on the analysis of early perinatal death rates and cesarean sections in Poland in 2008]. / Ocena funkcjonowania trójstopniowej selektywnej opieki perinatalnej na podstawie analizy umieralnosci okoloporodowej wczesnej i ciec cesarskich w Polsce w 2008 roku.

AIM: The aim of the following work was to assess three-level selective perinatal care in Polish voivodeships in 2008 on the basis of the following parameters: birth rates as well as perinatal death rates, divided into three classes of neonatal weights, in hospitals on each of the three levels. The goal of selective perinatal care is, among other things, to diagnose threats to the mother and/or fetus and direct women with high-risk pregnancies to higher level obstetrics and neonatology clinics and units. The structure of a regional three-level perinatal care, as well as the rules and procedures governing the process of redirecting patients to different levels of perinatal care have been defined in great detail. MATERIAL AND METHODS: Perinatal death rates analysis has been carried out on the basis of data received from Voivodeship Public Health Centers in sixteen voivodeships in Poland in 2008. The main document constituted MZ-29 form section X, modified by the authors and subdivided into levels of perinatal care. All data contained in the form have been verified: the numbers concerning birth and death rates as well as perinatal deaths and birth weight subgroups from given voivodeship hospitals. Statistic analysis was limited to the presentation of result tables and graphs within voivodeships. RESULTS: Birth rates and perinatal death rates revealed that in the course of ten years the level of perinatal care, introduced gradually in Poland between the years 1997-1999, resulted in its improvement. Perinatal death rates decreased in the course of ten years from 9.5% in 1999 to 6.45% in 2008, i.e. by 0.3% annually. On the first level, the rate of neonates with very low birth weight, 500-999g, decreased by 5.5% and was 21.1% in 2008 and 36.6% in 1999, whereas on the third level, the birth rate in the same group (500-999g) increased by 12.7% and was 47.7% in 2008 and 35.5% in 1999. There is a growing and alarming tendency to perform cesarean sections. The increase amounted up to 1.2% annually (18.2% in 19999 and 30.5% in 2008), with vast differences among hospitals and voivodeships. In 2008 there were 28.4% of cesarean sections in level one-hospitals, 29.3% in level two-hospitals and 40.6% in level three-hospitals. CONCLUSIONS: The results of an overall decrease in perinatal deaths rate and an increase in birth rates in the group of neonates with very low birth weight on the third level are not satisfactory. Reintroduction of the program and strategy from the years 1995 and 1997 will enable us to improve the situation. Particularly this should be the case on the basic level perinatal care. In the context of three-level selective strategy the reintroduction of periodical analysis of perinatal care results is essential.

Abundant and dynamically expressed miRNAs, piRNAs, and other small RNAs in the vertebrate Xenopus tropicalis.

Small regulatory RNAs have recently emerged as key regulators of eukaryotic gene expression. Here we used high-throughput sequencing to determine small RNA populations in the germline and soma of the African clawed frog Xenopus tropicalis. We identified a number of miRNAs that were expressed in the female germline. miRNA expression profiling revealed that miR-202-5p is an oocyte-enriched miRNA. We identified two novel miRNAs that were expressed in the soma. In addition, we sequenced large numbers of Piwi-associated RNAs (piRNAs) and other endogenous small RNAs, likely representing endogenous siRNAs (endo-siRNAs). Of these, only piRNAs were restricted to the germline, suggesting that endo-siRNAs are an abundant class of small RNAs in the vertebrate soma. In the germline, both endogenous small RNAs and piRNAs mapped to many high copy number loci. Furthermore, endogenous small RNAs mapped to the same specific subsets of repetitive elements in both the soma and the germline, suggesting that these RNAs might act to silence repetitive elements in both compartments. Data presented here suggest a conserved role for miRNAs in the vertebrate germline. Furthermore, this study provides a basis for the functional analysis of small regulatory RNAs in an important vertebrate model system.

[Pathogenesis and risk of HIV infection in women--review of recent literature]. / Patogeneza i ryzyko zakazenia HIV u kobiet--przeglad aktualnego pismiennictwa.

In Poland, as well as worldwide, HIV prevalence is increasing among women and their children. Women acquire the infection mainly through heterosexual contacts, while children get it from their mothers. Identification of the factors leading to the infection is essential in order to work out effective strategies for prevention of HIV transmission among women and children. In Polish literature there are no detailed clinical guidelines how to prevent mother-to-child HIV infection--a fact which is not without meaning as it put Poland in an unfavourable position when comparing to other European countries with a higher number of infected children. In this review authors present recent data concerning women susceptibility to HIV infection, virus pathogenesis, the role of co-infections with other pathogens, as well as the role of sex hormones on HIV infection risk. Authors hope that the information will help to establish Polish standards on prevention of HIV infection in women and their children, and that they will soon be put into practice.

Two Piwi proteins, Xiwi and Xili, are expressed in the Xenopus female germline.

The Argonaute superfamily is a large family of RNA-binding proteins involved in gene regulation mediated by small noncoding RNA and characterized by the presence of PAZ and PIWI domains. The family consists of two branches, the Ago and the Piwi clade. Piwi proteins bind to 21-30-nucleotide-long Piwi-interacting RNAs (piRNAs), which map primarily to transposons and repeated sequence elements. Piwi/piRNAs are important regulators of gametogenesis and have been proposed to play roles in transposon silencing, DNA methylation, transcriptional silencing, and/or post-transcriptional control of translation and RNA stability. Most reports to date have concentrated on the Piwi family members in the male germline. We have identified four Piwi proteins in Xenopus and demonstrate that two, namely, Xiwi1b and Xili, are expressed in the oocyte and early embryo. Xiwi1 and Xili are predominantly found in small, separate complexes, and we do not detect significant interaction of Piwi proteins with the cap-binding complex. Putative nuclear localization and export signals were identified in Xiwi1 and Xili, supporting our observation that Xiwi1, but not Xili, is a nucleo-cytoplasmic protein. Furthermore, by immunoprecipitation of small RNAs, we establish Xiwi1 as a bona fide Piwi protein. These results suggest that the Piwi/piRNA pathway is active in translationally repressed oocytes. This is a significant finding as the Xenopus model provides an excellent tool to study post-transcriptional mechanisms.

Function and regulation of the mammalian Musashi mRNA translational regulator.

The evolutionarily conserved RNA-binding protein, Musashi, regulates neural stem cell self-renewal. Musashi expression is also indicative of stem cell populations in breast and intestinal tissues and is linked to cell overproliferation in cancers of these tissues. Musashi has been primarily implicated as a repressor of target mRNAs in stem cell populations. However, little is known about the mechanism by which Musashi exerts mRNA translational control or how Musashi function is regulated. Recent findings in oocytes of the frog, Xenopus, indicate an unexpected role for Musashi as an activator of a number of maternal mRNAs during meiotic cell cycle progression. Given the importance of Musashi function in stem cell biology and the implications of aberrant Musashi expression in cancer, it is critical that we understand the molecular processes that regulate Musashi function.

Hairpin structure within the 3'UTR of DNA polymerase beta mRNA acts as a post-transcriptional regulatory element and interacts with Hax-1.

Aberrant expression of DNA polymerase beta, a key enzyme involved in base excision repair, leads to genetic instability and carcinogenesis. Pol beta expression has been previously shown to be regulated at the level of transcription, but there is also evidence of post-transcriptional regulation, since rat transcripts undergo alternative polyadenylation, and the resulting 3'UTR contain at least one regulatory element. Data presented here indicate that RNA of the short 3'UTR folds to form a strong secondary structure (hairpin). Its regulatory role was established utilizing a luciferase-based reporter system. Further studies led to the identification of a protein factor, which binds to this element-the anti-apoptotic, cytoskeleton-related protein Hax-1. The results of in vitro binding analysis indicate that the formation of the RNA-protein complex is significantly impaired by disruption of the hairpin motif. We demonstrate that Hax-1 binds to Pol beta mRNA exclusively in the form of a dimer. Biochemical analysis revealed the presence of Hax-1 in mitochondria, but also in the nuclear matrix, which, along with its transcript-binding properties, suggests that Hax-1 plays a role in post-transcriptional regulation of expression of Pol beta.

Musashi regulates the temporal order of mRNA translation during Xenopus oocyte maturation.

A strict temporal order of maternal mRNA translation is essential for meiotic cell cycle progression in oocytes of the frog Xenopus laevis. The molecular mechanisms controlling the ordered pattern of mRNA translational activation have not been elucidated. We report a novel role for the neural stem cell regulatory protein, Musashi, in controlling the translational activation of the mRNA encoding the Mos proto-oncogene during meiotic cell cycle progression. We demonstrate that Musashi interacts specifically with the polyadenylation response element in the 3' untranslated region of the Mos mRNA and that this interaction is necessary for early Mos mRNA translational activation. A dominant inhibitory form of Musashi blocks maternal mRNA cytoplasmic polyadenylation and meiotic cell cycle progression. Our data suggest that Musashi is a target of the initiating progesterone signaling pathway and reveal that late cytoplasmic polyadenylation element-directed mRNA translation requires early, Musashi-dependent mRNA translation. These findings indicate that Musashi function is necessary to establish the temporal order of maternal mRNA translation during Xenopus meiotic cell cycle progression.

Identification and expression analysis of alternative splice variants of the rat Hax-1 gene.

Hax-1 protein, which has been studied in mice and humans, shows a potent anti-apoptotic activity and is involved in regulation of cell motility. Cloning of the rat Hax-1 cDNA has revealed seven alternative transcripts, which differ mostly in their 5' region. Alternative splicing concerns exon 1, skipped in 5 transcripts, intron 1 which is partially retained in these transcripts, exon 2, which can be partially skipped, and intron 2, retained in one variant. The existence of different splicing variants was confirmed by exon-junction-specific RT-PCR and RNase protection assay. Analysis of expression indicates that overall Hax-1 mRNA level is relatively low in most tissues and very high in testes, and that the expression pattern of the variants is similar in different tissues. Presence of different transcripts implies the existence of several protein isoforms, with three putative start codons. The existence of at least three protein isoforms was confirmed by Western blot. Interestingly, high mRNA level in testes does not translate into high protein level, suggesting the existence of tissue-specific translational regulation or regulated protein degradation.

Low expression of DNA polymerase beta in human testicular germ cell tumours--impact on foetal gonocytic origin theory.

Different models of pathogenesis of adult testicular germ cell tumours (TGCTs) are presented. Analysis of telomeric length and DNA polymerase beta expression suggests that seminoma and nonseminoma, two main histological types of TGCTs, derive independently from transformed foetal primordial cells.