Vaccinia virus dissemination requires p21-activated kinase 1.

The orthopoxvirus vaccinia virus (VACV) interacts with both actin and microtubule cytoskeletons in order to generate and spread progeny virions. Here, we present evidence demonstrating the involvement of PAK1 (p21-activated kinase 1) in the dissemination of VACV. Although PAK1 activation has previously been associated with optimal VACV entry via macropinocytosis, its absence does not affect the production of intracellular mature virions (IMVs) and extracellular enveloped virions (EEVs). Our data demonstrate that low-multiplicity infection of PAK1(-/-) MEFs leads to a reduction in plaque size followed by decreased production of both IMVs and EEVs, strongly suggesting that virus spread was impaired in the absence of PAK1. Confocal and scanning electron microscopy showed a substantial reduction in the amount of VACV-induced actin tails in PAK1(-/-) MEFs, but no significant alteration in the total amount of cell-associated enveloped virions (CEVs). Furthermore, the decreased VACV dissemination in PAK1(-/-) cells was correlated with the absence of phosphorylated ARPC1 (Thr21), a downstream target of PAK1 and a key regulatory subunit of the ARP2/3 complex, which is necessary for the formation of actin tails and viral spread. We conclude that PAK1, besides its role in virus entry, also plays a relevant role in VACV dissemination.

Whistle comparison of four delphinid species in Southeastern Brazil.

The present study evaluates variations in frequency and duration parameters of whistles of four dolphin species (Sotalia guianensis, Steno bredanensis, Stenella frontalis, and Tursiops truncatus), recorded in the Rio de Janeiro State Coast, Southeastern Brazil. A total of 487 whistles were analyzed. Acoustic parameters of the whistles were classified to species by discriminant function analysis. Overall classification score was 72.5%, with the highest classification score obtained for whistles of S. bredanensis and the lowest obtained for S. frontalis. Most differences were among S. bredanensis and S. guianensis, species that did not have their repertoires compared in other studies.

Activation of the PI3K/Akt pathway early during vaccinia and cowpox virus infections is required for both host survival and viral replication.

Viral manipulation of the transduction pathways associated with key cellular functions such as actin remodeling, microtubule stabilization, and survival may favor a productive viral infection. Here we show that consistent with the vaccinia virus (VACV) and cowpox virus (CPXV) requirement for cytoskeleton alterations early during the infection cycle, PBK/Akt was phosphorylated at S473 [Akt(S473-P)], a modification associated with the mammalian target of rapamycin complex 2 (mTORC2), which was paralleled by phosphorylation at T308 [Akt(T308-P)] by PI3K/PDK1, which is required for host survival. Notably, while VACV stimulated Akt(S473-P/T308-P) at early (1 h postinfection [p.i.]) and late (24 h p.i.) times during the infective cycle, CPXV stimulated Akt at early times only. Pharmacological and genetic inhibition of PI3K (LY294002) or Akt (Akt-X and a dominant-negative form of Akt-K179M) resulted in a significant decline in virus yield (from 80% to >/=90%). This decline was secondary to the inhibition of late viral gene expression, which in turn led to an arrest of virion morphogenesis at the immature-virion stage of the viral growth cycle. Furthermore, the cleavage of both caspase-3 and poly(ADP-ribose) polymerase and terminal deoxynucleotidyl transferase-mediated deoxyuridine nick end labeling assays confirmed that permissive, spontaneously immortalized cells such as A31 cells and mouse embryonic fibroblasts (MEFs) underwent apoptosis upon orthopoxvirus infection plus LY294002 treatment. Thus, in A31 cells and MEFs, early viral receptor-mediated signals transmitted via the PI3K/Akt pathway are required and precede the expression of viral antiapoptotic genes. Additionally, the inhibition of these signals resulted in the apoptosis of the infected cells and a significant decline in viral titers.

Whistles emitted by Atlantic spotted dolphins (Stenella frontalis) in Southeastern Brazil.

The whistles of Atlantic spotted dolphins have been studied in a few localities of the North Atlantic Ocean and those studies revealed that the species emits whistles within a broad frequency range, with a high number of inflection points and presence of harmonics. In the South Atlantic Ocean, there is no information about the sounds produced by Atlantic spotted dolphins. A total of 1092 whistles emitted by free-ranging Atlantic spotted dolphins in Southeastern Brazilian coastal waters were analyzed. Whistles recorded in this study had a broad frequency range from 1.15 to 23.44 kHz. Whistles without harmonics were frequently emitted (N=701; 64.2%) and those signals with zero up to two inflection points corresponded to 94% of all whistles. Some differences in whistle characteristics (inflection points and duration) were found in relation to areas in North Atlantic Ocean and whistles were shorter and with a smaller number of inflection points in Brazil. Whistles produced by Atlantic spotted dolphins varied between the two behavioral states in which dolphins were engaged. Whistles were more frequently emitted when dolphins presented behaviors that included fast movement at surface, prey pursuit, aerial behavior, and physical contact. In these situations, whistles were on average longer and had higher frequency parameters than those emitted when animals were engaged in slowly and moderate traveling. The findings presented herewith reveal that dolphins modified whistle structures within behavioral states.

Underwater noise in an impacted environment can affect Guiana dolphin communication.

This study focused on whistles produced by Guiana dolphin under different noise conditions in Guanabara Bay, southeastern Brazil. Recording sessions were performed with a fully calibrated recording system. Whistles and underwater noise levels registered during two behavioral states were compared separately between two areas. Noise levels differed between the two areas across all frequencies. Whistle duration differed between areas and was negatively correlated with noise levels. Whistling rate was positively correlated with noise levels, showing that whistling rate was higher in noisier conditions. Results demonstrated that underwater noise influenced Guiana dolphin acoustic behavior.