ABSTRACT Using genetic data, we confirm for the first time the presence of the Tanzanian helmeted terrapin Pelomedusa kobe in Akagera National Park, Rwanda, extending their known distribution. Our study provides the first natural history observations, population and morphological data for this species from a wild population. As species in the genus Pelomedusa are morphologically difficult to distinguish, we gain the first insights into intraspecific and interspecific patterns of size and shape variation of plastron morphology between two morphologically‐similar species living in Eastern Africa, P. kobe and P. subrufa , using geometric morphometrics. Species affiliation, rather than sex, is the dominant factor influencing size and shape variation. P. kobe exhibits female‐based sexual size dimorphism (SSD), while P. subrufa shows a tendency toward male‐based SSD. Furthermore, sexual shape dimorphism (SShD) was observed in P. kobe but not in P. subrufa . Our results provide the first integrative data (genetic, morphological and natural history) on P. kobe , however further information on their distribution in the park is needed for effective conservation of this data‐deficient species. Our findings lay the groundwork for future ecological studies on P. kobe , potentially revealing unique adaptations and conservation needs previously overlooked due to its cryptic nature.

This article aims to reconstruct the invasion of the ear-shaped pond snail, Radix auricularia (Linnaeus, 1758), to Lake Baikal, East Siberia. This species is widely distributed in the Palaearctic and Northern America, and since the early 20th century has formed abundant and sustained populations in Lake Baikal. The data provided on the morphological and genetic variability of R. auricularia help to better describe and delineate the species. With an integrative approach involving morphological and molecular data, we improved the knowledge of the intraspecific variability of R. auricularia in the most important characteristics used for its determination. Molecular sequences of nuclear spacer fragment ITS-2 and mitochondrial gene fragment cyt–b were obtained from 32 individuals of Radix (including seven outgroup Radix species) collected from various parts of Lake Baikal and adjacent waterbodies and compared with sequences of 32 individuals of R. auricularia from different regions of the Palaearctic as well as with individuals determinated as R. intercisa from Lake Baikal, R. iturupica from the Kurile Islands, R. ussuriensis from the Khabarovsk region, R. narzykulovi from Tajikistan, and R. schubinae from the Amur region. Molecular genetic analyses revealed that all specimens collected from Lake Baikal belong to R. auricularia. There are no genetically distinct groups of snails that would correspond to two morphospecies previously recorded in Lake Baikal (e.g., R. auricularia s. str. and R. intercisa). Variability of the characteristics that are commonly used for species identification (shell morphology, mantle pigmentation, shape and position of the bursa copulatrix, length and position of the bursa duct, length ratio of preputium to penial sheath) were found in individuals analysed with molecular genetics to be broader than recognised in the current literature. Some shells of R. auricularia collected from Lake Baikal resemble shells of another lymnaeid species, R. balthica, and without molecular assessment can be confused with the latter. Geometric morphometric analysis of more than 250 shells revealed no observed hiatus between Baikalian and non-Baikalian R. auricularia. The probable stages and pathways of R. auricularia invasion to Lake Baikal’s ecosystem are outlined and discussed. Factors such as global climate warming and human activity stimulated and facilitated the ongoing dispersal of ear pond snails within Lake Baikal.

There is no clear empirical evidence to support the general assumption that genetic diversity favours successful invasions. Many invading species disperse and establish successfully despite low genetic diversity, a phenomenon known as the genetic paradox of biological invasion. Model systems that allow comparison of genetic patterns between exotic and native source populations are still scarce. This is particularly true for amphibians. Here we compare genetic patterns of the widely introduced Johnstone’s Whistling Frog, Eleutherodactylus johnstonei, with its successful alien congener E. antillensis and the single island endemic E. portoricensis. Genetic diversity and population differentiation in native and introduced populations of the three taxa were inferred from mitochondrial D-loop sequences (235 bp). Our results reveal that exotic populations of the two alien taxa, E. johnstonei and E. antillensis, are not only genetically impoverished due to founder effects, but that, moreover, their native range source-populations exhibit low genetic diversity and inter-population differentiation in the first place. Populations of the endemic E. portoricensis, on the other hand, are genetically more diverse and show marked inter-population differentiation. These observed genetic patterns are consistent with geological processes and invasion histories. We argue that the establishment success of the alien taxa in our model system is better explained by ecological factors and anthropogenic drivers than by genetic diversity. As these factors provide more parsimonious explanations, they should be given priority in management decisions. However, molecular studies with higher resolution are needed to fully test possible genetic and epigenetic components that could promote the invasion process.

Larval dispersal and limited knowledge of physical boundaries challenge our understanding of the processes that drive genetic divergence and potential speciation in the marine environment. Divergence, both within and between populations of marine taxa, is not uncommon, but spatial and temporal stability of observed genetic structure is not well known. Previously, we detected large genetic differences among populations of the cardinalfish species Ostorhinchus doederleini inhabiting adjacent coral reefs. Here, we determined the spatial and temporal persistence of these genetic structures over the course of ten consecutive generations. Using microsatellite markers, we detected large changes (genetic population distance, D est , ranged from 0.04 to 0.46) in the genetic structure in some years, but some reefs maintained the same populations for nearly all sampling years. As this species’ life span does not exceed 1 yr, persistence of distinct reef populations suggests natal homing. Mitochondrial identity based on two mtDNA markers corroborates the nuclear genetic evidence for genetic differences large enough to constitute different clades and even cryptic species in O. doederleini , which, based on gross morphology, was thought to be a single taxon. Habitat specialization was observed in one clade that exclusively inhabited reef lagoons, while all clades could be observed on reef slopes. We suggest that local habitat recognition combined with local population recognition and selection against hybrids can form barriers that maintain a cryptic species complex.

The evolution of cooperative behaviour requires mechanisms to avoid investing in conspecifics that are not increasing an individual's direct or indirect fitness. This suggests that selection should favour the capability of recognizing kin.One mechanism to discriminate between kin and nonkin is based on phenotype matching, when an individual learns a template of itself or of its kin and can later use this template to recognize even unfamiliar kin. In this study, we found that in zebrafish olfactory kin recognition depended on an imprinting process that required the two-step learning process of olfactory as well as visual cues of kin. Larvae that were exposed to either visual or olfactory cues or to both cues of nonkin did not show imprinting. This capability of imprinting on kin but not on nonkin cues can be explained by genetic predisposition or self-referencing. Through this combined imprinting process larvae can avoid false imprinting on unrelated individuals. (c) 2013 The Association for the Study of Animal Behaviour. Published by Elsevier Ltd. All rights reserved.

Olfactory imprinting on environmental, population- and kin-specific cues is a specific form of life-long memory promoting homing of salmon to their natal rivers and the return of coral reef fish to natal sites. Despite its ecological significance, natural chemicals for olfactory imprinting have not been identified yet. Here, we show that MHC peptides function as chemical signals for olfactory imprinting in zebrafish. We found that MHC peptides consisting of nine amino acids elicit olfactory imprinting and subsequent kin recognition depending on the MHC genotype of the fish. In vivo calcium imaging shows that some olfactory bulb neurons are highly sensitive to MHC peptides with a detection threshold at 1 pM or lower, indicating that MHC peptides are potent olfactory stimuli. Responses to MHC peptides overlapped spatially with responses to kin odour but not food odour, consistent with the hypothesis that MHC peptides are natural signals for olfactory imprinting.

The systematic position of the rare Indian turtle Geoemyda silvatica Henderson is examined by a phylogenetic analysis of mtDNA sequence data (cytochrome-b gene) of most species of Geoernydidae. Geoemyda silvatica represents a basal and isolated taxon within Geoemydidae, definitely not a close relative of any species of Geoemyda or Heosenlys, the genera in which G. silvatica has been placed in the past. Therefore, the new genus Vijayachelys is proposed for G silvatica. Cranial morphology and some other osteological characters of Vijayachelys silvatica are described and illustrated. Differential diagnoses are given for the type species of Melanochelys and the respective type species of the superficially similar genera Geoemyda, Heosemys, and Leueocephalon. According to Bayesian analysis of mtDNA data, Melanochelys tryuga could be distantly related to V. silvatica, whereas the morphological similarity of the other species probably is the result of a similar mode of life. The discovery of the phylogenetically isolated position of V silvatica highlights the importance of the Western Ghats as a biodiversity hotspot rich in higher-level endemics. (c) 2006 Gesellschaft fur Biologische Systematik. Published by Elsevier GmbH. All rights reserved.

Sequence data of the mitochondrial cytochrome b gene of stripe-necked terrapins (Mauremys leprosa) have been compared from localities north and south of the Atlas Mts. (Ceuta; Morocco) and from Donana National Park (Spain). A low maximum sequence divergence (approximately 1%) corresponds to two major clades; one is represented by localities to the north of the Atlas Mts. and in Donana National Park and the other by localities to the south of the Atlas Mts. Differentiation between populations north and south of the Atlas Mts. is much more pronounced than that found between samples from each side of the Strait of Gibraltar. These findings suggest that the Strait of Gibraltar is, in contrast to the Atlas Mts., not a significant barrier to gene flow in stripe-necked terrapins. The major clades could reflect taxonomic segregation between populations north and south of the Atlas Mts. Sequences from Marrakech (corresponding to M. l. marokkensis), Ceuta, and the Donana National Park (M. l. leprosa) are only weakly differentiated. South of the Atlas Mts. we found no consistent differences between samples from catchment basins of the Oued Drâa (M. l. vanmeerhaghei) and the Oued Noun (M. l. saharica). Our findings imply that taxonomic differentiation within M. leprosa is currently overestimated.

South American leptodactylid frogs of the genus Phrynopus occur in cloud‐forest, páramo, subpáramo and puna habitats (1000–4400 m elevation) from Colombia to Bolivia. Currently, there are 34 described species; however, many additional species new to science have been reported from Colombia, Peru, and Bolivia. The phylogeny of the species‐diverse Phrynopus is unknown and the position of the genus within Leptodactylidae is poorly understood. We present the results of a phylogenetic study based on 12S and 16S mitochondrial rDNA. Fifteen species of Phrynopus from Bolivia to Ecuador are included, along with several other genera of Leptodactylidae and representatives of other frog families. Our results indicate that Phrynopus is phylogenetically nested within Eleutherodactylus, whereas Phyllonastes is phylogenetically nested within Phrynopus. Based on the recovered phylogeny, we transfer Phrynopus simonsii to Eleutherodactylus, and show that Phrynopus carpish needs to be removed from Phrynopus. Eleven of the 13 Phrynopus species occurring in Central Peru share the absence of an external and internal ear.

DNA alterations at chromosome 5q22 occur frequently in different types of tumors including urological cancers. Previously, we narrowed a common target region between loci D5S659 and D5S2055 at chromosome 5q22.3 by microsatellite allelotyping. After constructing a BAC contig and shot-gun sequencing we identified a putative exon by the NIX software package. By PCR cloning using the putative exon (5qex5) specific primers and primers directed to the vector (λZAPII) sequence of human brain and kidney cDNA libraries, we obtained a full-length cDNA of 4074 bp of the new gene RBCC728/TRIM36 (GenBank accession no.AJ272269) with an ORF coding for a protein of 728 amino acids. The TRIM36, which is a new member of the tripartite motif (TRIM) gene family, shows a RING finger C3HC4 structure, two B-box, a coiled-coil, a fibronectin type III and a C-terminal domain of unknown function (SPRY). The TRIM36 has a weak homology to MID1. Immunohistochemistry of recombinant and native TRIM36 displays a cytoplasmic, slightly filamentous staining pattern in COS-7 cells. The TRIM36 is expressed in adult testis, brain, prostate, kidney, heart and lung. A variable level of TRIM36 expression was detected by Q-RT-PCR in conventional RCC, while the gene was consequently upregulated in PCs. We did not find mutation in the open reading frame of the TRIM36 in cancer cells. The overexpression of the TRIM36 in the vast majority of prostate cancer suggest that this gene might be involved in the prostate tumorigenesis.

We established a mice tumor model to investigate the effects of continuous cancer gene therapy, including antigen-presenting cell (APC) engineering and local stimulation of the immune system. B16 melanoma or Lewis lung carcinoma cells were injected intradermally on the back of C57/BL6 mice. The overlaying dermis or the tumor was shot with a gene gun (particle-mediated gene transfer) starting 8 days after tumor implantation in the case of the melanoma (Lewis lung carcinoma start day 7), continuing every fourth day thereafter until death. Control groups were mice without any therapy (A) or gene therapy with the empty plasmid (B). Therapy groups (Melanoma) received the genes as follows: group C — day 8, IL-12; day 12, IL-2…; group D — day 8, IFN- γ/B7.1; day 12, IFN-γ/B7.1…; group E — day 8, IFN-γ/B7.1; day 12, IL-12, day 16, IL-2…. Melanoma: Mean survival time was enhanced in all therapy groups significantly, whereby the greatest survival time was found in group C. Tumor growth was reduced in all therapy groups similarly (C and D significant). Lewis Lung: Only mice of group C had an enhanced survival and reduced tumor growth (both significant). An antimetastatic effect was seen in all therapy groups.

Objective. We investigated the effects of continuous cancer gene therapy, including APC engineering and local stimulation of the immune system in a mice melanoma model. Materials and methods. B16 melanoma cells were injected into C57/Bl6 mice intradermally. The overlying dermis or the tumor were shot with different plasmids using a gene gun every 4th day starting 8 days after tumor implantation. Control groups were mice without any therapy or gene therapy as described above with the empty plasmid. Therapy was: group I, IL-12 and IL-2; group II, IFN-γ/B7.1; group III, IFN-γ/B7.1, IL-12, and IL-2. Results. The median survival time of all therapy groups was significantly enhanced. The longest median survival was in the IL-12/IL-2 group. Tumor growth was reduced in all therapy groups. Control groups suffered a higher rate of metastasis and had fewer inflammatory cells at the tumor site. Conclusions. Continuous therapy with the gene gun is easy to handle and shows good results. Therapy with genes for IL-12 and IL-2 was superior to that with additional IFN-γ/B7.1 or IFN-γ/B7.1 alone. APC engineering is clearly less sufficient in the case of the B16 melanoma.

We describe a patient who underwent thoracic radiation therapy for biopsy-proven pulmonary spindle cell sarcoma in the left lower lobe, 15 months after birth. At the age of 37 she developed shoulder pain, fatigue, and progressive exertion dyspnoea. Chest X-ray revealed a pulmonary mass in the left lower lobe due to a cytology-proven malignant tumour.The patient underwent left pneumonectomy. Histology revealed a myosarcoma of the lung, similar to the previous sarcoma. Furthermore, the patient was diagnosed to have Turner syndrome mosaic and chromosomal analysis revealed a translocation t(1;13) in 3/50 metaphases. However a germline mutation of the p53 tumour suppressor gene was excluded. After 2 years of follow-up the patient is stable and there are no signs of recurrence of the tumour.We conclude a re-occurrence of this very rare malignant disorder of the lung after a 36-year interval in a patient with Turner syndrome mosaic. Following initial curative radiation therapy, with a remission over 36 years, lung resection was now successfully performed.

Pulmonale Blastome sind eine seltene, heterogene Gruppe maligner pulmonaler Mischtumoren. Unter diesen stellt das gut differenzierte fetale Adenokarzinom der Lunge eine Sonderform ohne sarkomatöse Entartung dar, dessen Diagnostik an eine subtile pathohistologische Untersuchung gebunden ist. In der vorgestellten Kasuistik werden erstmalig ein bilaterales Vorliegen dieses Tumors referiert und therapeutische Strategien dieser Tumorentität diskutiert.

Viral IL-10 (vIL-10), the IL-10 homologue of Epstein-Barr virus, has so far been described as a cytokine that solely inhibits T cell function. Here we show in vivo and in vitro that after longterm secretion vIL-10 has a stimulatory effect on T cells. For this purpose we employed transfectants derived from a mastocytoma cell line (P815 cells, H-2(d)) constitutively secreting vIL-10 (P815-vIL-10) or expressing the co-stimulatory molecule B7-1 (P815-B7). After in vitro stimulation of splenocytes from syngeneic DBA/2 mice for 7 days in the presence of P815-vIL-10 cells we could detect a marked reduction of proliferation as well as cytotoxicity against P815 target cells. However, this inhibitory effect was reversed when stimulation with P815-vIL-10 cells was extended to 14 days. In vivo P815-vIL-10 cells were rejected whereas P815 cells transfected with a control plasmic were tumorigenic after injection into syngeneic DBA/2 mice. Furthermore, this stimulatory effect of constitutive vIL-10 secretion could be exploited to irradicate already established P815 tumors which were smaller than 5 x 5 mm. In contrast, paracrine vIL-10 secretion for a limited time period of 8-9 days was associated with inhibitory effects in vivo: P815-B7 cells, which are normally eliminated in DBA/2 mice, could grow if exposed to temporarily secreted vIL-10 . These time-dependent immunomodulatory effects have to be considered in potential therapeutic applications of vIL-10.

Herpes simplex virus type 1 (HSV1), a large DNA-containing virus, is endemic in all human populations investigated. After infection of mucocutaneous surfaces, HSV1 establishes a latent infection in nerve cells. Recently, it was demonstrated that HSV1 can also infect activated T lymphocytes. However, the consequences of T cell infection for viral pathogenesis and immunity are unknown. We have observed that in contrast to the situation in human fibroblasts, in human T cell lines antigen presentation by major histocompatibility complex class I molecules is not blocked after HSV1 infection. Moreover, HSV1 infection of T cells results in rapid elimination of antiviral T cells by fratricide. To dissect the underlying molecular events, we used a transgenic mouse model of HSV1 infection to demonstrate that CD95 (Apo-1, Fas)-triggered apoptosis is essential for HSV1-induced fratricide, whereas tumor necrosis factor (TNF) also contributes to this phenomenon but to a lesser extent. By contrast, neither TRAIL (TNF-related apoptosis-inducing ligand) nor perforin were involved. Finally, we defined two mechanisms associated with HSV1-associated fratricide of antiviral T cells: (a) T cell receptor-mediated upregulation of CD95 ligand and (b) a viral "competence-to-die" signal that renders activated T lymphocytes susceptible to CD95 signaling. We propose that induction of fratricide is an important immune evasion mechanism of HSV1, helping the virus to persist in the host organism throughout its lifetime.

The Epstein-Barr virus (EBV) encodes an open reading frame with significant homology to the cellular IL-10 gene. This viral IL-10 (vIL-10) might enable EBV to evade antiviral T cells. We employed transfectants of a murine tumor cell line (P815) to investigate whether vIL-10 interferes with the first (antigenic) or second (co-stimulatory) signal of T cell activation. Untransfected P815 cells caused tumors in syngeneic DBA/2 mice after s.c. inoculation. In contrast, transfectants that provided either a strong antigenic stimulus (P815-Kb cells) or a strong co-stimulatory signal (P815-B7 cells) were rejected. Injection of double-transfected P815 cells expressing Kb and secreting high levels of vIL-10 (P815-Kb-vIL-10) did not result in tumor growth. We then investigated whether vIL-10 could paralyse co-stimulation by B7 under the same conditions. Therefore P815-B7 cells were mixed with vIL-10-secreting P815-Kb cells and co-injected into DBA/2 animals. Most of these mice developed a tumor. Explanted tumor cells expressed the B7 molecule but not the Kb antigen. These observations in vivo were mirrored by experiments in vitro: vIL-10 could induce T cell tolerance towards P815-B7 cells but not P815-Kb cells. Taken together our results suggest that vIL-10 acts directly on T cells to inhibit co-stimulatory signals mediated via B7 receptors such as CD28 or CTLA-4.