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RNA viruses in insects are targets of an RNA interference (RNAi)-based antiviral immune response, in which viral replication intermediates or viral dsRNA genomes are processed by Dicer-2 (Dcr-2) into viral small interfering RNAs (vsiRNAs). Whether dsDNA virus infections are controlled by the RNAi pathway remains to be determined. Here, we analyzed the role of RNAi in DNA virus infection using Drosophila melanogaster infected with Invertebrate iridescent virus 6 (IIV-6) as a model. We show that Dcr-2 and Argonaute-2 mutant flies are more sensitive to virus infection, suggesting that vsiRNAs contribute to the control of DNA virus infection. Indeed, small RNA sequencing of IIV-6-infected WT and RNAi mutant flies identified abundant vsiRNAs that were produced in a Dcr-2-dependent manner. We observed a highly uneven distribution with strong clustering of vsiRNAs to small defined regions (hotspots) and modest coverage at other regions (coldspots). vsiRNAs mapped in similar proportions to both strands of the viral genome, suggesting that long dsRNA derived from convergent overlapping transcripts serves as a substrate for Dcr-2. In agreement, strand-specific RT-PCR and Northern blot analyses indicated that antisense transcripts are produced during infection. Moreover, we show that vsiRNAs are functional in silencing reporter constructs carrying fragments of the IIV-6 genome. Together, our data indicate that RNAi provides antiviral defense against dsDNA viruses in animals. Thus, RNAi is the predominant antiviral defense mechanism in insects that provides protection against all major classes of viruses.

We study a blood testing procedure for detecting viruses like HIV, HBV and HCV. In this procedure, blood samples go through two screening steps. The first test is ELISA (antibody Enzyme Linked Immuno-Sorbent Assay). The portions of blood which are found not contaminated in this first phase are tested in groups through PCR (Polymerase Chain Reaction). The ELISA test is less sensitive than the PCR test and the PCR tests are considerably more expensive. We model the two test phases of blood samples as services in two queues in series; service in the second queue is in batches, as PCR tests are done in groups. The fact that blood can only be used for transfusions until a certain expiration date leads, in the tandem queue, to the feature of customer impatience. Since the first queue basically is an infinite server queue, we mainly focus on the second queue, which in its most general form is an S-server M=G[k;K]=S + G queue, with batches of sizes which are bounded by k and K. Our objective is to maximize the expected profit of the system, which is composed of the amount earned for items which pass the test (and before their patience runs out), minus costs. This is done by an appropriate choice of the decision variables, namely, the batch sizes and the number of servers at the second service station. As will be seen, even the simplest version of the batch queue, the M=M[k;K]=1 + M queue, already gives rise to serious analytical complications for any batch size larger than 1. These complications are discussed in detail. In view of the fact that we aim to solve realistic optimization problems for blood screening procedures, these analytical complications force us to take recourse to either a numerical approach or approximations. We present a numerical solution for the queue length distribution in theM=M[k;K]=S+M queue and then formulate and solve several optimization problems. The power-series algorithm, which is a numerical-analytic method, is also discussed.

The mitochondrial DNAs (mtDNAs) from 116 Oriental and Caucasian blood samples were analyzed for their Hinc II restriction endonuclease cleavage patterns using Southern analysis and 32P human mtDNA probes. Seven distinct patterns were found, all of which could be interrelated by single nucleotide changes. The predominant pattern (mtHinc II-2) was found in 97% of the Caucasians and 73% of the Orientals. This mtDNA morph had one more Hinc II site than did the second most common morph (mtHinc II-1), which was found only in 20% of the Orientals. Three additional patterns were in a single Oriental sample, a fourth in a single Caucasian sample, and a fifth in one member of each population. The polymorphic site that differentiated mtHinc II-1 and mtHinc II-2 was cloned and sequenced. A single nucleotide change was found that created an Hinc II site and changed the amino acid sequence of the URF5 gene. Comparison of these sequences with those of other primates [15] revealed that the Asian mtHinc II-1 and mtHinc II-4 mtDNAs were identical in this region with those of chimpanzees and orangutans. These results suggest that the Asian mtHinc II-1 mtDNA may have been ancestral to other human mtDNAs.

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