Drug-induced death of the asexual blood stages of Plasmodium falciparum occurs without typical signs of apoptosis

There is clear evidence that most antimalarial drugs, while acting through different mechanisms, are associated with parasite growth/development inhibition and eventual parasite death. However, the exact mode of parasite death remains unclear. In the present study, we investigated the ability of various drugs, including two antimalarial drugs (chloroquine and atovaquone), a topoisemerase II inhibitor (etoposide) and a nitric oxide donor (S-nitro-N-acetyl-D, L-penicillamine), to induce apoptosis in a laboratory strain of Plasmodium falciparum. Results obtained from flow cytometric analysis showed a significant reduction in the percent of parasitemia and parasite growth in all drug-treated parasite cultures, including those treated with etoposide and S-nitro-N-acetyl-D, L-penicillamine. For further investigation, we used various biochemical approaches including the terminal dUTP nick-end labeling assay, determination of mitochondrial membrane integrity and DNA degradation/fragmentation, to analyze the changes occurring during parasite-drug interactions and eventual death. We observed that loss of membrane potential was induced in parasite cultures treated with atovaquone, while S-nitro-N-acetyl-D, L-penicillamine induced abnormal parasite forms, "crisis forms", and minor DNA degradation. However, these features were not observed in the parasite cultures treated with chloroquine nor were other features of apoptosis-like death associated with any of the drugs used in this study. The death resulting from the various drug treatments is atypical of apotosis. More studies will be needed to define the precise mode of death exhibited by P. falciparum.     

Absence of a paternally inherited FOXP2 gene in developmental verbal dyspraxia

Mutations in FOXP2 cause developmental verbal dyspraxia (DVD), but only a few cases have been described. We characterize 13 patients with DVD--5 with hemizygous paternal deletions spanning the FOXP2 gene, 1 with a translocation interrupting FOXP2, and the remaining 7 with maternal uniparental disomy of chromosome 7 (UPD7), who were also given a diagnosis of Silver-Russell Syndrome (SRS). Of these individuals with DVD, all 12 for whom parental DNA was available showed absence of a paternal copy of FOXP2. Five other individuals with deletions of paternally inherited FOXP2 but with incomplete clinical information or phenotypes too complex to properly assess are also described. Four of the patients with DVD also meet criteria for autism spectrum disorder. Individuals with paternal UPD7 or with partial maternal UPD7 or deletion starting downstream of FOXP2 do not have DVD. Using quantitative real-time polymerase chain reaction, we show the maternally inherited FOXP2 to be comparatively underexpressed. Our results indicate that absence of paternal FOXP2 is the cause of DVD in patients with SRS with maternal UPD7. The data also point to a role for differential parent-of-origin expression of FOXP2 in human speech development.     

The BPI-like/PLUNC family proteins in cattle

Members of the protein family having similarity to BPI (bactericidal/permeability increasing protein) (the BPI-like proteins), also known as the PLUNC (palate, lung and nasal epithelium clone) family, have been found in a range of mammals; however, those in species other than human or mouse have been relatively little characterized. Analysis of the BPI-like proteins in cattle presents unique opportunities to investigate the function of these proteins, as well as address their evolution and contribution to the distinct physiology of ruminants. The present review summarizes the current understanding of the nature of the BPI-like locus in cattle, including the duplications giving rise to the multiple BSP30 (bovine salivary protein 30?kDa) genes from an ancestral gene in common with the single PSP (parotid secretory protein) gene found in monogastric species. Current knowledge of the expression of the BPI-like proteins in cattle is also presented, including their pattern of expression among tissues, which illustrate their independent regulation at sites of high pathogen exposure, and the abundance of the BSP30 proteins in saliva and salivary tissues. Finally, investigations of the function of the BSP30 proteins are presented, including their antimicrobial, lipopolysaccharide-binding and bacterial aggregation activities. These results are discussed in relation to hypotheses regarding the physiological role of the BPI-like proteins in cattle, including the role they may play in host defence and the unique aspects of digestion in ruminants.     

Auswirkungen von abgestufter pflanzenschutzmittel‐intensität und unkrautbesatz auf laufkäferzönosen (coleoptera; carabidae) während einer fruchtfolgerotation unter den spezifischen Bedingungen des mitteldeutschen trockengebietes*

Die Untersuchungen fanden von 1991 bis 1995 auf einem 43 ha großen Schlag in Obhausen (Querfurter Platte) statt. Zielstellung der Untersuchungen war es, Auswirkungen abgestufter Pflanzenschutzintensitäten auf Laufkäfer zu erfassen. Dazu wurden auf dem Feld 6 Parzellen von 72 × 200 m Größe angelegt, von denen zwei als pflanzenschutzmittelfreie Kontrollflächen dienten. Auf zwei weiteren Teilstücken erfolgten intensive chemische Pflanzenschutzmaßnahmen. In den anderen beiden Varianten wurden im Sinne eines integrierten Pflanzenschutzes unter Beachtung von Schadschwellenwerten (Unkräuter, pilzliche und tierische Schaderreger) gezielte Pflanzenschutzmaßnahmen praktiziert. Zur Erfassung der Laufkäfer dienten Bodenfallen. Die Fruchtfolge begann 1991 mit Wintergerste, die aber wegen Trockenheit umgebrochen werden mußte. Die Intensitätsabstufungen des Pflanzenschutzmitteleinsatzes erfolgten ab 1992 im Mais und 1993 sowie 1994 in Winterweizen. Letztes Fruchtfolgeglied war 1995 Winterroggen, bei dem alle Parzellen wieder konventionell bewirtschaftet wurden. Im Mais kam es in den unbehandelten Kontrollen zu einer starken Verunkrautung. In diesen Parzellen erreichte die Fangsumme der Laufkäfer den 2,5 fachen Wert gegenüber den anderen Prüfgliedern. In den darauffolgenden Jahren, in den Winterweizenbeständen, lagen die höchsten Fangsummen wiederum in den Kontrollen vor, doch fielen die Differenzen zu den anderen Prüfgliedern nicht so groß aus. Die wenigsten Laufkäfer wurden in den Intensiv‐Varianten erfaßt. Die Bewirtschaftung nach dem Schadschwellenprinzip erbrachte wesentlich höhere Fangsummen. Während der Intensitätsabstufung besiedelten die Arten Calosoma auropunc‐tatum, Harpalus affinis, Pseudoophonus rufipes, Harpalus distinguendus, Poecilus punctulatus, Harpalus tardus sowie Harpalus zabroides überwiegend die Kontrollparzellen. Bei den abschließenden Untersuchungen im Winterroggen 1995 traf dies nur noch für Harpalus affinis zu. Ob für das Überwiegen dieser Arten in den Kontrollen der starke Unkrautbesatz im Jahre 1992 bzw. nachfolgend der Vorrat an Diasporen oder der völlige Verzicht auf Pflanzenschutzmittel ausschlaggebend war, konnte nicht eindeutig geklärt werden. Entgegen allen Erwartungen lag im Winterroggen die höchste Fangsumme der Laufkäfer in den ehemals intensiv behandelten Parzellen vor.

Ein Verzicht auf Pflanzenschutzmittel geht oft mit nicht mehr tolerierbaren Ertragsverlusten einher, wie z.B. im Mais 1992. Bei einem Einsatz von Pflanzenschutzmitteln nach Schadschwellenwerten entsprechend den Prinzipien eines integrierten Pflanzenschutzes sind langfristig keine negativen Effekte auf den Naturhaushalt zu erwarten.     

Ground Beetle (Coleoptera,Carabidae) Communities of the Exhausted Open-Cast Mining Area K™nigsaue and of Agricultural Areas in the District Aschersleben-staβfurt (Saxony-Anhalt)

In the years 1995 to 1997, following recultivation activities, investigations refering to the colonisation by ground beetles were lead through at four sites in the open-cast mining area Königsaue by means of pitfall traps. Altogether 74 species could be proved, 19% of these species are considered as endangered. The occurrence of the tiger beetle Cicindela arenaria viennensis is remarkable for nature conservation reasons, because this species is threaten by extinction in Germany. The most frequent species in the exhausted open-cast mining area were Calathus ambiguus and Bembidion femoratum . On a site nearly without any vegetation Amara fulva belonged to the dominant species. Species- and dominance-identities between the ground beetle communities of the four sites were low. Also the species turnover between the years of investgation was relatively high. A Benjes-hedge, which was planted in 1996, was investigated additionally. Pitfall trap catches within stands of various crops (winter wheat, summer wheat, maize, peas, sugar beet) nearby the open-cast mining area were carried out in 1995 to 1997 as well. The fields were treated conventionally on tschernosem soils and peaty soils. 67 species of ground beetles were registrated in the fields. The results with regard to species composition and dominance structure resembled those which had been obtained in former investigations lead through near Halle (Saale) and Obhausen (Querfurter Platte). On the peaty soils Dyschirius globosus was very numerous, whereas it was not found on the other fields. An extremly high activity density of ground beetles with more than 20 individuals per day and pitfall trap could be observed in a sugar beet field near Schadeleben. Additional investigations on field edges at another site were realized in 1996. As a whole in the cause of this research 101 species of ground beetles could be proved in the district Aschersleben-Staáfurt, among these many endangered species.     

Identification and differentiation of Cryptosporidium species by capillary electrophoresis single-strand conformation polymorphism

Cryptosporidium species generally lack distinguishing morphological traits, and consequently, molecular methods are commonly used for parasite identification. Various methods for Cryptosporidium identification have been proposed, each with their advantages and disadvantages. In this study, we show that capillary electrophoresis coupled with single-strand conformation polymorphism (CE-SSCP) is a rapid, simple and cost-effective method for the identification of Cryptosporidium species and genotypes. Species could be readily differentiated based on the SSCP mobility of amplified 18S rRNA gene molecules. Clones that differed by single-nucleotide polymorphisms could be distinguished on CE-SSCP mobility. Profiles of species known to have heterogenic copies of 18S rRNA gene contained multiple peaks. Cloning and sequencing of Cryptosporidium parvum, Cryptosporidium hominis, Cryptosporidium fayeri and Cryptosporidium possum genotype 18S rRNA gene amplicons confirmed that these multiple peaks represented type A and type B 18S rRNA gene copies. CE-SSCP provides a reliable and sensitive analysis for epidemiological studies, environmental detection and diversity screening.     

The ketogenic diet preserves skeletal muscle with aging in mice

The causes of the decline in skeletal muscle mass and function with age, known as sarcopenia, are poorly understood. Nutrition (calorie restriction) interventions impact many cellular processes and increase lifespan and preserve muscle mass and function with age. As we previously observed an increase in life span and muscle function in aging mice on a ketogenic diet (KD), we aimed to investigate the effect of a KD on the maintenance of skeletal muscle mass with age and the potential molecular mechanisms of this action. Twelve‐month‐old mice were assigned to an isocaloric control or KD until 16 or 26 months of age, at which time skeletal muscle was collected for evaluating mass, morphology, and biochemical properties. Skeletal muscle mass was significantly greater at 26 months in the gastrocnemius of mice on the KD. This result in KD mice was associated with a shift in fiber type from type IIb to IIa fibers and a range of molecular parameters including increased markers of NMJ remodeling, mitochondrial biogenesis, oxidative metabolism, and antioxidant capacity, while decreasing endoplasmic reticulum (ER) stress, protein synthesis, and proteasome activity. Overall, this study shows the effectiveness of a long‐term KD in mitigating sarcopenia. The diet preferentially preserved oxidative muscle fibers and improved mitochondrial and antioxidant capacity. These adaptations may result in a healthier cellular environment, decreasing oxidative and ER stress resulting in less protein turnover. These shifts allow mice to better maintain muscle mass and function with age.