Post-translational targeting of Rab35 by the effector IcsB of Shigella determines intracellular bacterial niche formation.

Escape from the bacterial-containing vacuole (BCV) is a key step of Shigella host cell invasion. Rab GTPases subverted to in situ-formed macropinosomes in the vicinity of the BCV have been shown to promote its rupture. The involvement of the BCV itself has remained unclear.

Microtubules provide force to promote membrane uncoating in vacuolar escape for a cyto-invasive bacterial pathogen.

Intracellular bacterial pathogens gain entry to mammalian cells inside a vacuole derived from the host membrane. Some of them escape the bacteria-containing vacuole (BCV) and colonize the cytosol. Bacteria replicating within BCVs coopt the microtubule network to position it within infected cells, whereas the role of microtubules for cyto-invasive pathogens remains obscure.

Shigella generates distinct IAM subpopulations during epithelial cell invasion to promote efficient intracellular niche formation.

The facultative intracellular pathogen Shigella flexneri invades non-phagocytic epithelial gut cells. Through a syringe-like apparatus called type 3 secretion system, it injects effector proteins into the host cell triggering actin rearrangements leading to its uptake within a tight vacuole, termed the bacterial-containing vacuole (BCV). Simultaneously, Shigella induces the formation of large vesicles around the entry site, which we refer to as infection-associated macropinosomes (IAMs).

An automated microscopy workflow to study Shigella-neutrophil interactions and antibiotic efficacy in vivo.

Shigella are Gram-negative bacterial pathogens responsible for bacillary dysentery (also called shigellosis). The absence of a licensed vaccine and widespread emergence of antibiotic resistance has led the World Health Organisation (WHO) to highlight Shigella as a priority pathogen requiring urgent attention. Several infection models have been useful to explore the Shigella infection process; yet, we still lack information regarding events taking place in vivo.

Explainable artificial intelligence for assault sentence prediction in New Zealand.

The judiciary has historically been conservative in its use of Artificial Intelligence, but recent advances in machine learning have prompted scholars to reconsider such use in tasks like sentence prediction. This paper investigates by experimentation the potential use of explainable artificial intelligence for predicting imprisonment sentences in assault cases in New Zealand's courts. We propose a proof-of-concept explainable model and verify in practice that it is fit for purpose, with predicted sentences accurate to within one year.

Genetic Programming for Evolving a Front of Interpretable Models for Data Visualization.

Data visualization is a key tool in data mining for understanding big datasets. Many visualization methods have been proposed, including the well-regarded state-of-the-art method t-distributed stochastic neighbor embedding. However, the most powerful visualization methods have a significant limitation: the manner in which they create their visualization from the original features of the dataset is completely opaque.

Genetic Programming for Evolving Similarity Functions for Clustering: Representations and Analysis.

Clustering is a difficult and widely studied data mining task, with many varieties of clustering algorithms proposed in the literature. Nearly all algorithms use a similarity measure such as a distance metric (e.g.

Transmission-reducing immunity is inversely related to age in Plasmodium falciparum gametocyte carriers.

Immunity to the sexual stages of Plasmodium falciparum is induced during natural infections and can significantly reduce the transmission of parasites to mosquitoes (transmission reducing activity; TRA) but little is known about how these responses develop with increasing age/exposure to malaria. Routinely TRA is measured in the standard membrane feeding assay (SMFA). Sera were collected from a total of 199 gametocyte carriers (median age 4 years, quartiles 2 and 9 years) near Ifakara, Tanzania; 128 samples were tested in the SMFA and generated TRA data classified as a reduction of > 50% and > 90% of transmission.

Mechanisms that reduce transmission of Plasmodium falciparum malaria in semiimmune and nonimmune persons.

Transmission of Plasmodium falciparum can be reduced by immune factors present in the mosquito blood meal. Specific antibodies and white blood cells (WBCs) can interact with the sexual stages of the parasite inside the mosquito midgut. The relative contribution of serum factors and WBCs on transmission reduction in gametocyte carriers from an endemic area in Cameroon and in travelers with a first malaria experience was studied.

Leukocytes in a Plasmodium falciparum-infected blood meal reduce transmission of malaria to Anopheles mosquitoes.

Mosquitoes are infected with Plasmodium falciparum by taking a blood meal from a gametocyte carrier. Since a mosquito takes a volume of 1 to 2 microl, a blood meal may contain 1 x 10(4) to 3 x 10(4) leukocytes (WBC). The majority of WBC are composed of neutrophils which may phagocytose and kill developing gametes inside the mosquito midgut.

Infectivity of malarial parasites to mosquitoes: 'the interdependent roles of parasite, vector and host'.

It has been possible to infect mosquitoes routinely with cultured gametocytes of Plasmodium falciparum since 1980. This has enabled the development of a reliable bio-assay for potential transmission-blocking vaccines and research on the role of specific antibodies from the host on the parasitic stages in the mosquito midgut. After some development and fine-tuning of the assay, it became apparent that the immune responses of the human host, as well as factors from the parasite and the mosquito, determined the final outcome of the mosquito infection.

Measurement by membrane feeding of reduction in Plasmodium falciparum transmission induced by endemic sera.

The standard laboratory test for reduction in malaria transmission is based on the measurement of oocyst numbers in mosquitoes fed on blood meals containing test and control sera. Interpretation of the results, however, is often hampered by the large variation in numbers of infected mosquitoes and oocysts. The objective of this study was to compare 3 measures for the assessment of transmission reduction (so-called R values) and to define the experimental criteria that allow interpretation of the results.

Transmission-blocking antibodies against multiple, non-variant target epitopes of the Plasmodium falciparum gamete surface antigen Pfs230 are all complement-fixing.

We have studied the properties of 16 newly derived monoclonal antibodies (MoAbs) against Pfs230, a gamete surface protein of Plasmodium falciparum and a target of transmission-blocking antibodies. All 16 MoAbs recognized Pfs230 by immunoprecipitation from non-ionic detergent extracts of the protein radio-labelled with 125Iodine. The MoAbs also recognized this protein on Western blots under non-reducing conditions but none of them recognized the protein under reducing conditions.

Transmission blocking antibody of the Plasmodium falciparum zygote/ookinete surface protein Pfs25 also influences sporozoite development.

The Plasmodium falciparum zygote/ookinete surface protein, Pfs25, persists in the oocyst wall throughout its development. Anti-25 kD transmission blocking antibody, given to infected Anopheles stephensi or A. gambiae mosquitoes in an additional bloodmeal, 3-6 days after being fed gametocyte infected blood, penetrated the oocyst and reacted with the 25 kD protein within it.

A scanning electron microscopic study of the sporogonic development of Plasmodium falciparum in Anopheles stephensi.

The full development of Plasmodium falciparum in Anopheles stephensi mosquitoes was studied by scanning electron microscopy. Ookinetic development was described from in vitro cultures. Growing oocysts beneath the basal lamina of the midgut wall mechanically stretch this lamina until it is torn and displaced by day 7.

Feeding behaviour and sporozoite ejection by infected Anopheles stephensi.

Anopheles stephensi mosquitoes infected with Plasmodium falciparum sporozoites were allowed to feed individually through fresh whole thickness mouse skin. More sporozoites were ejected into the skin in clusters than into the blood. Deposition of sporozoites in the blood was an infrequent occurrence and always coincided with ejection of these stages into the skin--perhaps a spill-over effect.

Large-scale production of Plasmodium vivax sporozoites.

Mass-scale production of Plasmodium vivax sporozoites in Anopheles stephensi was achieved using the chimpanzee (Pan troglodytes) as a source of infective blood. Membrane feeding was as successful as feeding mosquitoes directly on the animal so long as the time between drawing the blood and feeding was restricted to 45 min. Longer delays such as 2-3 h resulted in loss of infectivity in terms of oocyst production.

Infectivity of cultured Plasmodium falciparum gametocytes to mosquitoes.

Various factors that may influence routine and high levels of mosquito infection with cultured Plasmodium falciparum gametocytes are considered in this paper. One of the most important is the choice of an appropriate isolate, with facilities for cryopreservation and a good technique for initiation of cultures. The use of automated culture systems with strict adherence to detail and routine has eliminated much of the variability.

Plasmodium falciparum ookinetes migrate intercellularly through Anopheles stephensi midgut epithelium.

The migration of Plasmodium falciparum and P. berghei ookinetes through the midgut epithelium in Anopheles stephensi was studied by transmission electron microscopy. With ruthenium red (RR) staining, the results of previous studies were confirmed: P.

Sporozoite load of mosquitoes infected with Plasmodium falciparum.

In the laboratory, mosquitoes given a second blood meal 5-11 d after an infective one have more sporozoites in their salivary glands than do those given a single infective blood meal only. The presence of specific anti-sporozoite antibody in the second blood meal does not reduce the number of sporozoites in salivary glands. On the contrary, the presence of the raised immunoglobulin levels--even non-specific ones--may result in higher gland infections.

DNA synthesis in gametocytes of Plasmodium falciparum.

The DNA content of Plasmodium falciparum gametocytes during intra-erythrocytic development and during gametogenesis was established by cytophotometric methods. Intraerythrocytic micro- and macrogametocytes (Stage I-Stage VB) contain about twice the amount of DNA of haploid sporozoites and ringstages, indicating that DNA is synthesized during transformation of ringforms into Stage I gametocytes. Microgametocytes, after activation at pH 8, rapidly duplicate their genome several times, while the DNA content of macrogametocytes remains constant during gametogenesis.

Transmission blockade of Plasmodium falciparum: its variability with gametocyte numbers and concentration of antibody.

The transmission of Plasmodium falciparum through Anopheles stephensi was measured in the presence of anti-gamete (anti-45/48 kDa) and anti-zygote/ookinete (anti-25 kDa) antibodies. With lowering numbers of infectious gametocytes in the presence of the same concentration of antibodies, different results were obtained with the two types of antibodies. Transmission blocking appeared to weaken when using anti-45/48 kDa antibody, whilst, with the anti-25 kDa antibody, transmission inhibition was markedly strengthened with lower parasite numbers.