Large-Scale Spatial Distribution of Virioplankton in the Adriatic Sea: Testing the Trophic State Control Hypothesis

Little is known concerning environmental factors that may control the distribution of virioplankton on large spatial scales. In previous studies workers reported high viral levels in eutrophic systems and suggested that the trophic state is a possible driving force controlling the spatial distribution of viruses. In order to test this hypothesis, we determined the distribution of viral abundance and bacterial abundance and the virus-to-bacterium ratio in a wide area covering the entire Adriatic basin (Mediterranean Sea). To gather additional information on factors controlling viral distribution on a large scale, functional microbial parameters (exoenzymatic activities, bacterial production and turnover) were related to trophic gradients. At large spatial scales, viral distribution was independent of autotrophic biomass and all other environmental parameters. We concluded that in contrast to what was previously hypothesized, changing trophic conditions do not directly affect virioplankton distribution. Since virus distribution was coupled with bacterial turnover times, our results suggest that viral abundance depends on bacterial activity and on host cell abundance.     

Decreased hypertrophic differentiation accompanies enhanced matrix formation in co-cultures of outer meniscus cells with bone marrow mesenchymal stromal cells

Introduction

The main objective of this study was to determine whether meniscus cells from the outer (MCO) and inner (MCI) regions of the meniscus interact similarly to or differently with mesenchymal stromal stem cells (MSCs). Previous study had shown that co-culture of meniscus cells with bone marrow-derived MSCs result in enhanced matrix formation relative to mono-cultures of meniscus cells and MSCs. However, the study did not examine if cells from the different regions of the meniscus interacted similarly to or differently with MSCs.

Methods

Human menisci were harvested from four patients undergoing total knee replacements. Tissue from the outer and inner regions represented pieces taken from one third and two thirds of the radial distance of the meniscus, respectively. Meniscus cells were released from the menisci after collagenase treatment. Bone marrow MSCs were obtained from the iliac crest of two patients after plastic adherence and in vitro culture until passage 2. Primary meniscus cells from the outer (MCO) or inner (MCI) regions of the meniscus were co-cultured with MSCs in three-dimensional (3D) pellet cultures at 1:3 ratio, respectively, for 3 weeks in the presence of serum-free chondrogenic medium containing TGF-β1. Mono-cultures of MCO, MCI and MSCs served as experimental control groups. The tissue formed after 3 weeks was assessed biochemically, histochemically and by quantitative RT-PCR.

Results

Co-culture of inner (MCI) or outer (MCO) meniscus cells with MSCs resulted in neo-tissue with increased (up to 2.2-fold) proteoglycan (GAG) matrix content relative to tissues formed from mono-cultures of MSCs, MCI and MCO. Co-cultures of MCI or MCO with MSCs produced the same amount of matrix in the tissue formed. However, the expression level of aggrecan was highest in mono-cultures of MSCs but similar in the other four groups. The DNA content of the tissues from co-cultured cells was not statistically different from tissues formed from mono-cultures of MSCs, MCI and MCO. The expression of collagen I (COL1A2) mRNA increased in co-cultured cells relative to mono-cultures of MCO and MCI but not compared to MSC mono-cultures. Collagen II (COL2A1) mRNA expression increased significantly in co-cultures of both MCO and MCI with MSCs compared to their own controls (mono-cultures of MCO and MCI respectively) but only the co-cultures of MCO:MSCs were significantly increased compared to MSC control mono-cultures. Increased collagen II protein expression was visible by collagen II immuno-histochemistry. The mRNA expression level of Sox9 was similar in all pellet cultures. The expression of collagen × (COL10A1) mRNA was 2-fold higher in co-cultures of MCI:MSCs relative to co-cultures of MCO:MSCs. Additionally, other hypertrophic genes, MMP-13 and Indian Hedgehog (IHh), were highly expressed by 4-fold and 18-fold, respectively, in co-cultures of MCI:MSCs relative to co-cultures of MCO:MSCs.

Conclusions

Co-culture of primary MCI or MCO with MSCs resulted in enhanced matrix formation. MCI and MCO increased matrix formation similarly after co-culture with MSCs. However, MCO was more potent than MCI in suppressing hypertrophic differentiation of MSCs. These findings suggest that meniscus cells from the outer-vascular regions of the meniscus can be supplemented with MSCs in order to engineer functional grafts to reconstruct inner-avascular meniscus.     

Notes about the uses of plants by one of the last healers in the Basilicata Region (South Italy)

Human societies utilize mollusks for myriad material and spiritual ends. An example of their use in a religious context is found in Brazil's African-derived belief systems. Candomblé, an Afro-Brazilian religion introduced during the 18^th-19^th centuries by enslaved Yoruba, includes various magical and liturgical uses of mollusks. This work inventoried the species utilized by adherents and to analyzed their symbolic and magical context. Data were obtained from Candomblé temples in two cities in the northeast of Brazil-Caruaru, in the state of Pernambuco, and Campina Grande, in the state of Paraíba. Questionnaires administered to eleven adepts revealed that at least nineteen mollusk species are being used. Shells from Monetaria moneta, M. annulus and Erosaria caputserpentis were cited by all of the interviewees. Three uses stood out: divination (jogo de búzios); utilization as ritual objects; and employment as sacrificial offerings (Igbin or Boi-de-Oxalá). The jogo de búzios (shell toss), employed in West Africa, Brazil and Cuba, is of fundamental importance to the cult, representing the means by which the faithful enter in contact with the divinities (Orixás) and consult people's futures (Odu). The utilization of mollusks in Candomblé is strongly influenced by ancient Yoruba myths (Itãs) which, having survived enslavement and generations of captive labor, continue to guide the lives of Brazil's African Diaspora.     

Birds of a Feather: Neanderthal Exploitation of Raptors and Corvids

The hypothesis that Neanderthals exploited birds for the use of their feathers or claws as personal ornaments in symbolic behaviour is revolutionary as it assigns unprecedented cognitive abilities to these hominins. This inference, however, is based on modest faunal samples and thus may not represent a regular or systematic behaviour. Here we address this issue by looking for evidence of such behaviour across a large temporal and geographical framework. Our analyses try to answer four main questions: 1) does a Neanderthal to raptor-corvid connection exist at a large scale, thus avoiding associations that might be regarded as local in space or time?; 2) did Middle (associated with Neanderthals) and Upper Palaeolithic (associated with modern humans) sites contain a greater range of these species than Late Pleistocene paleontological sites?; 3) is there a taphonomic association between Neanderthals and corvids-raptors at Middle Palaeolithic sites on Gibraltar, specifically Gorham''s, Vanguard and Ibex Caves? and; 4) was the extraction of wing feathers a local phenomenon exclusive to the Neanderthals at these sites or was it a geographically wider phenomenon?. We compiled a database of 1699 Pleistocene Palearctic sites based on fossil bird sites. We also compiled a taphonomical database from the Middle Palaeolithic assemblages of Gibraltar. We establish a clear, previously unknown and widespread, association between Neanderthals, raptors and corvids. We show that the association involved the direct intervention of Neanderthals on the bones of these birds, which we interpret as evidence of extraction of large flight feathers. The large number of bones, the variety of species processed and the different temporal periods when the behaviour is observed, indicate that this was a systematic, geographically and temporally broad, activity that the Neanderthals undertook. Our results, providing clear evidence that Neanderthal cognitive capacities were comparable to those of Modern Humans, constitute a major advance in the study of human evolution.     

Captive breeding and reintroduction of the lesser kestrel Falco naumanni: a genetic analysis using microsatellites

We used microsatellites to assess ongoing captive breeding and reintroduction programs of the lesser kestrel. The extent of genetic variation within the captive populations analysed did not differ significantly from that reported in wild populations. Thus, the application of widely recommended management practices, such as the registration of crosses between individuals in proper stud books and the introduction of new individuals into the genetic pools, has proven satisfactory to maintain high levels of genetic variation. The high rates of hatching failure occasionally documented in captivity can therefore not be attributed to depressed genetic variation. Even though genetic diversity in reintroduced populations did not differ significantly when compared to wild populations either, average observed heterozygosities and inbreeding coefficients were significantly lower and higher, respectively, when compared to the captive demes where released birds came. Monitoring of reproductive parameters during single-pairing breeding and paternity inference within colonial facilities revealed large variations in breeding success between reproductive adults. The relative number of breeding pairs that contributed to a large part of captive-born birds (50–56%) was similar in both cases (28.6 and 26.9%, respectively). Thus, the chances for polygyny (rarely in this study), extra-pair paternity (not found in this study) and/or social stimulation of breeding parameters do not seem to greatly affect the genetically effective population size. Independently of breeding strategies, the release of unrelated fledglings into the same area and the promotion of immigration should be mandatory to counteract founder effects and avoid inbreeding in reintroduced populations of lesser kestrels.     

Effects of geolocator attachments on breeding parameters of Lesser Kestrels

ABSTRACT Light level geolocators, also known as GLS loggers, are electronic devices intended for tracking the location of wide‐ranging animals using ambient light to estimate latitude and longitude. Miniaturized geolocators that can be used on relatively small migratory birds have recently become available, but little is known about the potential harmful effects of geolocators on birds. We examined the possible effects of 1.5‐g geolocators (dimensions: 21 × 6.5 × 9 mm) on the breeding success and survival of migratory Lesser Kestrels (Falco naumanni). During the 2007 breeding season, kestrels were fitted with geolocators using two attachment methods (Teflon wing harnesses and darvic bands), and geolocators were removed in 2008 after the birds returned to the breeding grounds. We found no differences in the breeding success of control and tagged pairs during the 2007 breeding season, but tagged pairs had greater fledgling mortality in the following breeding season. Furthermore, nestlings of tagged individuals had higher triglyceride and uric acid concentrations in their blood than control nestlings during the breeding season following tagging. As for return rates, 75% of tagged birds came back to the colony after the nonbreeding period, a proportion similar to that reported in previous studies. Although back‐mounts are slightly heavier and require more skill to attach, we recommend their use on small migratory raptors because most leg‐mounted geolocators in this study were damaged or rendered useless by dirt obscuring the light sensor.     

DJ-1 Modulates α-Synuclein Aggregation State in a Cellular Model of Oxidative Stress: Relevance for Parkinson's Disease and Involvement of HSP70

Background

Parkinson''s disease (PD) is a neurodegenerative pathology whose molecular etiopathogenesis is not known. Novel contributions have come from familial forms of PD caused by alterations in genes with apparently unrelated physiological functions. The gene coding for alpha-synuclein (α-syn) (PARK1) has been investigated as α-syn is located in Lewy bodies (LB), intraneuronal inclusions in the substantia nigra (SN) of PD patients. A-syn has neuroprotective chaperone-like and antioxidant functions and is involved in dopamine storage and release. DJ-1 (PARK7), another family-PD-linked gene causing an autosomal recessive form of the pathology, shows antioxidant and chaperone-like activities too.

Methodology/Principal Findings

The present study addressed the question whether α-syn and DJ-1 interact functionally, with a view to finding some mechanism linking DJ-1 inactivation and α-syn aggregation and toxicity. We developed an in vitro model of α-syn toxicity in the human neuroblastoma cell line SK-N-BE, influencing DJ-1 and α-syn intracellular concentrations by exogenous addition of the fusion proteins TAT-α-syn and TAT-DJ-1; DJ-1 was inactivated by the siRNA method. On a micromolar scale TAT-α-syn aggregated and triggered neurotoxicity, while on the nanomolar scale it was neuroprotective against oxidative stress (induced by H₂O₂ or 6-hydroxydopamine). TAT-DJ-1 increased the expression of HSP70, while DJ-1 silencing made SK-N-BE cells more susceptible to oxidative challenge, rendering TAT-α-syn neurotoxic at nanomolar scale, with the appearance of TAT-α-syn aggregates.

Conclusion/Significance

DJ-1 inactivation may thus promote α-syn aggregation and the related toxicity, and in this model HSP70 is involved in the antioxidant response and in the regulation of α-syn fibril formation.