Comparing the characteristics of recurrent versus primary odontogenic keratocysts - A single center retrospective cohort study.

The high recurrence rate of odontogenic keratocysts (OKC) entails a large number of follow-up interventions after primary surgery. This study aimed to compare recurrent with primary OKC in regard to recurrence rate, treatment modality, radiographic and clinical findings. A single center retrospective cohort study with surgically treated OKC between 2012 and 2021 was conducted. The primary predictor was recurrence type of the lesion: primary (P-OKC), first recurrence (R1-OKC) and second recurrence (R2-OKC). The primary outcome variables were recurrence and time to recurrence. 68 surgeries were identified. Recurrence was not significantly associated with recurrence type (p = 0.906) but with the method of surgery (p < 0.001). Marsupialized R1-OKC recurred earlier than marsupialized P-OKC. Diameter was significantly associated with recurrence type (p = 0.002). R1-OKC had a smaller median diameter than P-OKC and R2-OKC were smaller than R1-and P-OKC. P-OKC were associated with teeth in 79% of surgeries, R1-OKC in 61% and R2-OKC in 27% (p = 0.007). Postoperative numbness was present after 40% of surgeries and decreased to 15% at follow-up, independently of recurrence type. Time to recurrence may be shorter for recurrent marsupialized OKC. Recurrent OKC are smaller and less often associated with teeth. Postoperative numbness after OKC surgery has a good prognosis.

Transfer of membrane(s) matter(s)-non-genetic inheritance of (metabolic) phenotypes?

Glycosylphosphatidylinositol-anchored proteins (GPI-APs) are anchored at the outer phospholipid layer of eukaryotic plasma membranes exclusively by a glycolipid. GPI-APs are not only released into extracellular compartments by lipolytic cleavage. In addition, certain GPI-APs with the glycosylphosphatidylinositol anchor including their fatty acids remaining coupled to the carboxy-terminus of their protein components are also detectable in body fluids, in response to certain stimuli, such as oxidative stress, radicals or high-fat diet. As a consequence, the fatty acid moieties of GPI-APs must be shielded from access of the aqueous environment by incorporation into membranes of extracellular vesicles or into micelle-like complexes together with (lyso)phospholipids and cholesterol. The GPI-APs released from somatic cells and tissues are transferred via those complexes or EVs to somatic as well as pluripotent stem cells with metabolic consequences, such as upregulation of glycogen and lipid synthesis. From these and additional findings, the following hypotheses are developed: i) Transfer of GPI-APs via EVs or micelle-like complexes leads to the induction of new phenotypes in the daughter cells or zygotes, which are presumably not restricted to metabolism. ii) The membrane topographies transferred by the concerted action of GPI-APs and interacting components are replicated by self-organization and self-templation and remain accessible to structural changes by environmental factors. iii) Transfer from mother cells and gametes to their daughter cells and zygotes, respectively, is not restricted to DNA and genes, but also encompasses non-genetic matter, such as GPI-APs and specific membrane constituents. iv) The intergenerational transfer of membrane matter between mammalian organisms is understood as an epigenetic mechanism for phenotypic plasticity, which does not rely on modifications of DNA and histones, but is regarded as molecular mechanism for the inheritance of acquired traits, such as complex metabolic diseases. v) The missing interest in research of non-genetic matter of inheritance, which may be interpreted in the sense of Darwin's "Gemmules" or Galton's "Stirps", should be addressed in future investigations of the philosophy of science and sociology of media.

Effect of textile colour on vector mosquito host selection: a simulated field study in Mali, West Africa.

BACKGROUND: The effect of clothing colour on the biting rates of different vector mosquito species is not well understood. Studies under tropical field conditions are lacking. This study aimed to determine the influence of clothing colours on mosquito biting rates in rural and suburban settings in West Africa. METHODS: We performed a simulated field study in a suburban and a rural site in Mali using Mosquito-Magnet traps utilizing CO2 and other attractants, which were covered with black, white, and black/white striped textile sheets covers. These targets operated continuously for 10 consecutive days with bright nights (around full moon) and 10 consecutive days with dark nights (around new moon). Trapped mosquitoes were collected and catch rates counted hourly. Mosquitoes were morphologically identified to the species complex level (Anopheles gambiae s.l. and Culex pipiens s.l.) or species level (Aedes aegypti). A subset of Anopheles specimens were further identified by molecular methods. RESULTS: Under bright-night conditions, An. gambiae s.l. was significantly more attracted to black targets than to white and striped targets; during dark nights, no target preference was noted. During bright nights, Cx. pipiens s.l. was significantly more attracted to black and striped targets than to white targets; a similar trend was noted during dark nights (not significant). For day-active Ae. aegypti, striped targets were more attractive than the other targets and black were more attractive than white targets. CONCLUSIONS: The study firstly demonstrated that under field conditions in Mali, West Africa, mosquito catch rates were influenced by different clothing colours, depending on mosquito species and light conditions. Overall, light colours were least attractive to host-seeking mosquitoes. Using white or other light-coloured clothing can potentially reduce bite exposure and risk of disease transmission in endemic tropical regions.

[Conspicuous Conditions of the Tongue: Normal Variants vs. Pathological Findings].

INTRODUCTION: With its sensitivity, taste buds and complex anatomical structure of various muscles, the tongue is a central organ for speaking, tasting and food intake, especially oral food transport, chewing and swallowing. Changes in the tongue 's condition are frequent and often lead to uncertainty among patients and eventually to a visit to the family doctor, to the ear, nose and throat specialist, dentist or maxillofacial surgeon. The question whether the condition of the tongue is a lesion requiring treatment or just a variant can quite often prove a major challenge. The differential diagnoses are wide-ranging from harmless changes to alarming signs of disease. The time and duration of occurrence, the accompanying symptoms such as a burning sensation or taste disorders as well as risk factors such as nicotine and alcohol consumption are important anamnestic elements. Possible causes can be malnutrition, systemic diseases, inflammatory processes or malignancies. Accordingly, a blood test and a smear or a biopsy may be necessary as the first diagnostic step. The aim of this review is to explain the different types and causes of tongue problems and to explain in which cases further clarifications are necessary.

Notes on Central Asian Furcula with description of a new species.

Genitalia of the female holotype of the Central Asian species Furcula terminata (Wiltshire, 1958) (TL: northeastern Afghanistan) were studied for the first time. They showed a clear similarity with the genitalia of Furcula gorbunovi Schintlmeister, 1998 (TL: Gissar Range, Tajikistan) and Furcula mimonovi Schintlmeister, 1998 (TL: Karakalpakstan, Uzbekistan). Males of F. terminata from Afghanistan were not found in the collections. However, the specimens from neighboring Tajikistan and topotypes of F. gorbunovi and F. mimonovi were studied and all of them have pointed apex of the costal process of valva specific for the bifida-group, not the furcula-group as suggested previously. Considering the morphological similarity and overlapping distribution areas of the three studied taxa, two new synonyms are established for F. terminata: F. gorbunovi syn. n. and Furcula mimonovi syn. n. The population of Tian Shan and Alay Range in Kyrgyzstan and Uzbekistan, previously regarded as F. terminata, are described here as a new species based on differences in the genitalia of both sexes.

Five new species of the genus Meganaclia Aurivillius, 1892 (Lepidoptera: Erebidae: Arctiinae: Syntomini).

The Afrotropical genus Meganaclia Aurivillius, 1892 is revised, five new species M. smithi sp. n., M. grehani sp. n., M. josephi sp. n., M. johannae sp. n. and M. gaerberfesti sp. n. are described with a key for identification of species based on male genitalia structures. Habitus and genitalia of all taxa are illustrated and a distribution map is provided.

New records of Lasiocampidae (Lepidoptera) from Zanzibar Island with taxonomic notes and description of one new species of Odontopacha Aurivillius, 1909.

Seven genera and seven species of Lasiocampidae are newly recorded from the Zanzibar Island (Unguja): Bombycopsis C. & R. Felder, 1874 with Bombycopsis nigrovittata Aurivillius, 1927; Pallastica Zolotuhin & Gurkovich, 2009 with an unidentified species; Dollmania Tams, 1930 with an unidentified species; Mallocampa Aurivillius, 1902 with Mallocampa leighi Aurivillius, 1922; Eucraera Tams, 1930 with Eucraera witti Prozorov, 2016; Philotherma Möschler, 1887 with Philotherma montibia Strand, 1912; and Odontopacha Aurivillius, 1909 with Odontopacha fenestrata Aurivillius, 1909. The species are followed with taxonomic notes updating the status and distribution of the taxa. Bombycopsis nigrovittata is shown to have the maximum p-distance of 0.3% in cytochrome c oxidase I from Bombycopsis pallida Joannou & Krüger, 2009. Two specimens of Pallastica sp. from Zanzibar are different in wing coloration but identical genetically, both are 0.8-1.2% far from sequenced specimens collected in southern Malawi and eastern Zimbabwe and altogether 3.0-3.8% far from the Zambian and Malawian populations considered to be Pallastica pallens (Bethune-Baker, 1908). The barcoding revealed two distinct lineages of Dollmania in Tanzania with a p-distance of 3.5-3.7% between them, neither can be attributed to either Dollmania marwitzi (Strand, 1913) or Dollmania reussi (Strand, 1913) until the primary types or fresh topotypes are sequenced. The species Ph. montibia is taken out from the synonymy to Philotherma rosa (Druce, 1887) and is stated to be a bona species because of the difference in wing pattern and p-distance of 5.7-5.9%. A new species of the genus Odontopacha - Odontopacha dargei sp. n. - is described from southern Kenya and northern Tanzania where it occurs sympatrically with O. fenestrata. It differs from O. fenestrata by the paler coloration with the spotted external fascia on both wings and a p-distance of 3.04-3.65%. Lectotypes for D. marwitzi and Ph. montibia are established. Mallocampa leighi is recorded from Tanzania for the first time. Females of Chrysopsyche lutulenta Tams, 1923 earlier recorded from Zanzibar Island are figured and the species is recorded from DRC for the first time.

Three new Chinese species of Palpifer Hampson from the collection of Franz Daniel (Lepidoptera: Hepialidae).

New Chinese Palpifer species are described from Yunnan and Fujian provinces. The male of Palpifer nielseni sp. n. is described from specimens housed at the Witt Museum Weiden and the Zoologisches Forschungsmuseum Alexander Koenig, while a male of P. chui sp. n. and a male and female of Palpifer climoi sp. n., are described from specimens in the latter collection only. Specimens were originally part of the Franz Daniel collection, collected in 1934-1935 from elevations of 2,300 and 3,000 m. The new species are diagnosed primarily by differences in the male genitalia. The female genitalia of P. climoi sp. n. represent the second published description for Palpifer. Four unique features of the forewing supporting monophyly of Palpifer are discussed.

(Patho)Physiology of Glycosylphosphatidylinositol-Anchored Proteins II: Intercellular Transfer of Matter (Inheritance?) That Matters.

Glycosylphosphatidylinositol (GPI)-anchored proteins (APs) are anchored at the outer leaflet of the plasma membrane (PM) bilayer by covalent linkage to a typical glycolipid and expressed in all eukaryotic organisms so far studied. Lipolytic release from PMs into extracellular compartments and intercellular transfer are regarded as the main (patho)physiological roles exerted by GPI-APs. The intercellular transfer of GPI-APs relies on the complete GPI anchor and is mediated by extracellular vesicles such as microvesicles and exosomes and lipid-free homo- or heteromeric aggregates, and lipoprotein-like particles such as prostasomes and surfactant-like particles, or lipid-containing micelle-like complexes. In mammalian organisms, non-vesicular transfer is controlled by the distance between donor and acceptor cells/tissues; intrinsic conditions such as age, metabolic state, and stress; extrinsic factors such as GPI-binding proteins; hormones such as insulin; and drugs such as anti-diabetic sulfonylureas. It proceeds either "directly" upon close neighborhood or contact of donor and acceptor cells or "indirectly" as a consequence of the induced lipolytic release of GPI-APs from PMs. Those displace from the serum GPI-binding proteins GPI-APs, which have retained the complete anchor, and become assembled in aggregates or micelle-like complexes. Importantly, intercellular transfer of GPI-APs has been shown to induce specific phenotypes such as stimulation of lipid and glycogen synthesis, in cultured human adipocytes, blood cells, and induced pluripotent stem cells. As a consequence, intercellular transfer of GPI-APs should be regarded as non-genetic inheritance of (acquired) features between somatic cells which is based on the biogenesis and transmission of matter such as GPI-APs and "membrane landscapes", rather than the replication and transmission of information such as DNA. Its operation in mammalian organisms remains to be clarified.

(Patho)Physiology of Glycosylphosphatidylinositol-Anchored Proteins I: Localization at Plasma Membranes and Extracellular Compartments.

Glycosylphosphatidylinositol (GPI)-anchored proteins (APs) are anchored at the outer leaflet of plasma membranes (PMs) of all eukaryotic organisms studied so far by covalent linkage to a highly conserved glycolipid rather than a transmembrane domain. Since their first description, experimental data have been accumulating for the capability of GPI-APs to be released from PMs into the surrounding milieu. It became evident that this release results in distinct arrangements of GPI-APs which are compatible with the aqueous milieu upon loss of their GPI anchor by (proteolytic or lipolytic) cleavage or in the course of shielding of the full-length GPI anchor by incorporation into extracellular vesicles, lipoprotein-like particles and (lyso)phospholipid- and cholesterol-harboring micelle-like complexes or by association with GPI-binding proteins or/and other full-length GPI-APs. In mammalian organisms, the (patho)physiological roles of the released GPI-APs in the extracellular environment, such as blood and tissue cells, depend on the molecular mechanisms of their release as well as the cell types and tissues involved, and are controlled by their removal from circulation. This is accomplished by endocytic uptake by liver cells and/or degradation by GPI-specific phospholipase D in order to bypass potential unwanted effects of the released GPI-APs or their transfer from the releasing donor to acceptor cells (which will be reviewed in a forthcoming manuscript).

Two-screw osteosynthesis of the mandibular condylar head with different screw materials: a finite element analysis.

This study compared the biomechanical behavior of titanium, magnesium, and polylactic acid screws for two-screw osteosynthesis of mandibular condylar head fractures using finite element analysis. Von Mises stress distribution, fracture displacement, and fragment deformation were evaluated. Titanium screws performed the best in terms of carrying the highest load, resulting in the least fracture displacement and fragment deformation. Magnesium screws showed intermediate results, while PLA screws were found to be unsuitable with stress values exceeding their tensile strength. These findings suggest that magnesium alloys could be considered a suitable alternative to titanium screws in mandibular condylar head osteosynthesis.

Transfer of Proteins from Cultured Human Adipose to Blood Cells and Induction of Anabolic Phenotype Are Controlled by Serum, Insulin and Sulfonylurea Drugs.

Glycosylphosphatidylinositol-anchored proteins (GPI-APs) are anchored at the outer leaflet of eukaryotic plasma membranes (PMs) only by carboxy-terminal covalently coupled GPI. GPI-APs are known to be released from the surface of donor cells in response to insulin and antidiabetic sulfonylureas (SUs) by lipolytic cleavage of the GPI or upon metabolic derangement as full-length GPI-APs with the complete GPI attached. Full-length GPI-APs become removed from extracellular compartments by binding to serum proteins, such as GPI-specific phospholipase D (GPLD1), or insertion into the PMs of acceptor cells. Here, the interplay between the lipolytic release and intercellular transfer of GPI-APs and its potential functional impact was studied using transwell co-culture with human adipocytes as insulin-/SU-responsive donor cells and GPI-deficient erythroleukemia as acceptor cells (ELCs). Measurement of the transfer as the expression of full-length GPI-APs at the ELC PMs by their microfluidic chip-based sensing with GPI-binding α-toxin and GPI-APs antibodies and of the ELC anabolic state as glycogen synthesis upon incubation with insulin, SUs and serum yielded the following results: (i) Loss of GPI-APs from the PM upon termination of their transfer and decline of glycogen synthesis in ELCs, as well as prolongation of the PM expression of transferred GPI-APs upon inhibition of their endocytosis and upregulated glycogen synthesis follow similar time courses. (ii) Insulin and SUs inhibit both GPI-AP transfer and glycogen synthesis upregulation in a concentration-dependent fashion, with the efficacies of the SUs increasing with their blood glucose-lowering activity. (iii) Serum from rats eliminates insulin- and SU-inhibition of both GPI-APs' transfer and glycogen synthesis in a volume-dependent fashion, with the potency increasing with their metabolic derangement. (iv) In rat serum, full-length GPI-APs bind to proteins, among them (inhibited) GPLD1, with the efficacy increasing with the metabolic derangement. (v) GPI-APs are displaced from serum proteins by synthetic phosphoinositolglycans and then transferred to ELCs with accompanying stimulation of glycogen synthesis, each with efficacies increasing with their structural similarity to the GPI glycan core. Thus, both insulin and SUs either block or foster transfer when serum proteins are depleted of or loaded with full-length GPI-APs, respectively, i.e., in the normal or metabolically deranged state. The transfer of the anabolic state from somatic to blood cells over long distance and its "indirect" complex control by insulin, SUs and serum proteins support the (patho)physiological relevance of the intercellular transfer of GPI-APs.

The West Africa ICEMR Partnerships for Guiding Policy to Improve the Malaria Prevention and Control.

The Mali National Malaria Control Program (NMCP) recently established a phased set of goals for eliminating malaria in Mali by 2030. Over the past decade, the scale-up of NMCP-led malaria control interventions has led to considerable progress, as evidenced by multiple malariometric indicators. The West Africa International Center of Excellence in Malaria Research (WA-ICEMR) is a multidisciplinary research program that works closely with the NMCP and its partners to address critical research needs for malaria control. This coordinated effort includes assessing the effectiveness of control interventions based on key malaria research topics, including immune status, parasite genetic diversity, insecticide and drug resistance, diagnostic accuracy, malaria vector populations and biting behaviors, and vectorial capacity. Several signature accomplishments of the WA-ICEMR include identifying changing malaria age demographic profiles, testing innovative approaches to improve control strategies, and providing regular reporting on drug and insecticide resistance status. The NMCP and WA-ICEMR partnership between the WA-ICEMR and the NMCP offers a comprehensive research platform that informs the design and implementation of malaria prevention and control research programs. These efforts build local expertise and capacity for the next generation of malaria researchers and guide local policy, which is crucial in sustaining efforts toward eliminating malaria in West Africa.

A Decade of Progress Accelerating Malaria Control in Mali: Evidence from the West Africa International Center of Excellence for Malaria Research.

This article highlights over a decade of signature achievements by the West Africa International Centers for Excellence in Malaria Research (WA-ICEMR) and its partners toward guiding malaria prevention and control strategies. Since 2010, the WA-ICEMR has performed longitudinal studies to monitor and assess malaria control interventions with respect to space-time patterns, vector transmission indicators, and drug resistance markers. These activities were facilitated and supported by the Mali National Malaria Control Program. Research activities included large-scale active and passive surveillance and expanded coverage of universal long-lasting insecticide-treated bed nets and seasonal malaria chemoprevention (SMC). The findings revealed substantial declines in malaria occurrence after the scale-up of control interventions in WA-ICEMR study sites. WA-ICEMR studies showed that SMC using sulfadoxine-pyrimethamine plus amodiaquine was highly effective in preventing malaria among children under 5 years of age. An alternative SMC regimen (dihydroartemisinin plus piperaquine) was shown to be potentially more effective and provided advantages for acceptability and compliance over the standard SMC regimen. Other findings discussed in this article include higher observed multiplicity of infection rates for malaria in historically high-endemic areas, continued antimalarial drug sensitivity to Plasmodium falciparum, high outdoor malaria transmission rates, and increased insecticide resistance over the past decade. The progress achieved by the WA-ICEMR and its partners highlights the critical need for maintaining current malaria control interventions while developing novel strategies to disrupt malaria transmission. Enhanced evaluation of these strategies through research partnerships is particularly needed in the wake of reported artemisinin resistance in Southeast Asia and East Africa.

Biological Role of the Intercellular Transfer of Glycosylphosphatidylinositol-Anchored Proteins: Stimulation of Lipid and Glycogen Synthesis.

Glycosylphosphatidylinositol-anchored proteins (GPI-APs), which are anchored at the outer leaflet of plasma membranes (PM) only by a carboxy-terminal GPI glycolipid, are known to fulfill multiple enzymic and receptor functions at the cell surface. Previous studies revealed that full-length GPI-APs with the complete GPI anchor attached can be released from and inserted into PMs in vitro. Moreover, full-length GPI-APs were recovered from serum, dependent on the age and metabolic state of rats and humans. Here, the possibility of intercellular control of metabolism by the intercellular transfer of GPI-APs was studied. Mutant K562 erythroleukemia (EL) cells, mannosamine-treated human adipocytes and methyl-ß-cyclodextrin-treated rat adipocytes as acceptor cells for GPI-APs, based on their impaired PM expression of GPI-APs, were incubated with full-length GPI-APs, prepared from rat adipocytes and embedded in micelle-like complexes, or with EL cells and human adipocytes with normal expression of GPI-APs as donor cells in transwell co-cultures. Increases in the amounts of full-length GPI-APs at the PM of acceptor cells as a measure of their transfer was assayed by chip-based sensing. Both experimental setups supported both the transfer and upregulation of glycogen (EL cells) and lipid (adipocytes) synthesis. These were all diminished by serum, serum GPI-specific phospholipase D, albumin, active bacterial PI-specific phospholipase C or depletion of total GPI-APs from the culture medium. Serum inhibition of both transfer and glycogen/lipid synthesis was counteracted by synthetic phosphoinositolglycans (PIGs), which closely resemble the structure of the GPI glycan core and caused dissociation of GPI-APs from serum proteins. Finally, large, heavily lipid-loaded donor and small, slightly lipid-loaded acceptor adipocytes were most effective in stimulating transfer and lipid synthesis. In conclusion, full-length GPI-APs can be transferred between adipocytes or between blood cells as well as between these cell types. Transfer and the resulting stimulation of lipid and glycogen synthesis, respectively, are downregulated by serum proteins and upregulated by PIGs. These findings argue for the (patho)physiological relevance of the intercellular transfer of GPI-APs in general and its role in the paracrine vs. endocrine (dys)regulation of metabolism, in particular. Moreover, they raise the possibility of the use of full-length GPI-APs as therapeutics for metabolic diseases.

Two-versus three-screw osteosynthesis of the mandibular condylar head: A finite element analysis.

Titanium screws are commonly used for osteosynthesis of mandibular condylar head fractures. Evidence suggests that the insertion of three screws may result in better fracture stability. Two screws only, on the other hand, could reduce adverse effects, mainly bone resorption. This study aimed to investigate the biomechanical differences in mandibular condylar head osteosynthesis with two versus three titanium screws using finite element analysis. A finite element model of the mandible with a right type P condylar head fracture fixed with two or three titanium screws was analyzed in ANSYS Mechanical. The geometry of the model assembly was constructed in ANSYS Spaceclaim. Biomechanical load boundary conditions were obtained from a validated musculoskeletal model in AnyBody Modeling System™. The preprocessing of the finite element model and mapping of the obtained boundary conditions was done in docq VIT. Fracture displacement, fragment deformation, von Mises stress distribution, and reaction forces within the screws were evaluated in ANSYS for three different loading scenarios. Finite element analysis showed similar results when comparing osteosynthesis with two versus three titanium screws for all three loading scenarios. Contralateral molar loading resulted in the highest stress on both the fracture and the screws with the maximum von Mises stress being found at the condylar neck. Stress concentration within the screws was found in the fracture gap and was higher in the lateral fragment. In all scenarios, maximum von Mises stress values were smaller when forces were distributed among three screws. However, stability was also adequate when two screws were used. Mandibular condylar head osteosynthesis with two titanium screws appears to provide sufficient fracture stability. Further clinical studies are needed to clarify the implications of these results.

Revision of the Lemonia taraxaci complex, with a description of a new species from Italy and clarification of the status of Lemonia strigata (Lepidoptera: Brahmaeidae: Lemoniinae).

The species Lemonia taraxaci (Denis & Schiffermüller, 1755) is divided into three species based on distribution areas and cytochrome c oxidase subunit 1 (COI) distances above 2%: Central European Lemonia taraxaci (type locality: Austria, Vienna area), Eastern European to South Siberian Lemonia sibirica Wnukowsky, 1934 stat. rev., bona sp. (type locality: Russia, Novosibirsk Oblast, Ozero-Karachi and Russia, Altai Krai, Kornilovo), and Italian Lemonia italiana sp. n. (type locality: Italy, Calabria centr., Sila grd., Casali del Manco, San Nicola Silano (CS)). Authorship of Lemonia strigata Antoshin & Zolotuhin, 2011 is reconsidered. The close relation between the typical L. strigata and taraxaci-like adults from the Balkans is proven genetically. According to this, L. strigata is suggested to be a junior synonym of taraxaci-looking Lemonia taraxaci var. montana Buresch, 1915 (type locality: Bulgaria, Rhodope Mts, Musala), although further DNA investigation is needed to prove this hypothesis.

Chip-Based Sensing of the Intercellular Transfer of Cell Surface Proteins: Regulation by the Metabolic State.

Glycosylphosphatidylinositol (GPI)-anchored proteins (GPI-APs) are anchored at the surface of mammalian blood and tissue cells through a carboxy-terminal GPI glycolipid. Eventually, they are released into incubation medium in vitro and blood in vivo and subsequently inserted into neighboring cells, potentially leading to inappropriate surface expression or lysis. To obtain first insight into the potential (patho)physiological relevance of intercellular GPI-AP transfer and its biochemical characterization, a cell-free chip- and microfluidic channel-based sensing system was introduced. For this, rat or human adipocyte or erythrocyte plasma membranes (PM) were covalently captured by the TiO2 chip surface operating as the acceptor PM. To measure transfer between PM, donor erythrocyte or adipocyte PM were injected into the channels of a flow chamber, incubated, and washed out, and the type and amount of proteins which had been transferred to acceptor PM evaluated with specific antibodies. Antibody binding was detected as phase shift of horizontal surface acoustic waves propagating over the chip surface. Time- and temperature-dependent transfer, which did not rely on fusion of donor and acceptor PM, was detected for GPI-APs, but not typical transmembrane proteins. Transfer of GPI-APs was found to be prevented by α-toxin, which binds to the glycan core of GPI anchors, and serum proteins in concentration-dependent fashion. Blockade of transfer, which was restored by synthetic phosphoinositolglycans mimicking the glycan core of GPI anchors, led to accumulation in the chip channels of full-length GPI-APs in association with phospholipids and cholesterol in non-membrane structures. Strikingly, efficacy of transfer between adipocytes and erythrocytes was determined by the metabolic state (genotype and feeding state) of the rats, which were used as source for the PM and sera, with upregulation in obese and diabetic rats and counterbalance by serum proteins. The novel chip-based sensing system for GPI-AP transfer may be useful for the prediction and stratification of metabolic diseases as well as elucidation of the putative role of intercellular transfer of cell surface proteins, such as GPI-APs, in (patho)physiological mechanisms.

Interaction of Full-Length Glycosylphosphatidylinositol-Anchored Proteins with Serum Proteins and Their Translocation to Cells In Vitro Depend on the (Pre-)Diabetic State in Rats and Humans.

Glycosylphosphatidylinositol (GPI)-anchored proteins (GPI-APs), which are anchored at the surface of mammalian cultured and tissue cells through a carboxy-terminal GPI glycolipid, are susceptible to release into incubation medium and (rat and human) blood, respectively, in response to metabolic stress and ageing. Those GPI-APs with the complete GPI still attached form micelle-like complexes together with (lyso)phospholipids and cholesterol and are prone to degradation by serum GPI-specific phospholipase D (GPLD1), as well as translocation to the surface of acceptor cells in vitro. In this study, the interaction of GPI-APs with GPLD1 or other serum proteins derived from metabolically deranged rat and humans and their translocation were measured by microfluidic chip- and surface acoustic wave-based sensing of micelle-like complexes reconstituted with model GPI-APs. The effect of GPI-AP translocation on the integrity of the acceptor cell surface was studied as lactate dehydrogenase release. For both rats and humans, the dependence of serum GPLD1 activity on the hyperglycemic/hyperinsulinemic state was found to be primarily based on upregulation of the interaction of GPLD1 with micelle-like GPI-AP complexes, rather than on its amount. In addition to GPLD1, other serum proteins were found to interact with the GPI phosphoinositolglycan of full-length GPI-APs. Upon incubation of rat adipocytes with full-length GPI-APs, their translocation from the micelle-like complexes (and also with lower efficacy from reconstituted high-density lipoproteins and liposomes) to acceptor cells was observed, accompanied by upregulation of their lysis. Both GPI-AP translocation and adipocyte lysis became reduced in the presence of serum proteins, including (inhibited) GPLD1. The reduction was higher with serum from hyperglycemic/hyperinsulinemic rats and diabetic humans compared to healthy ones. These findings suggest that the deleterious effects of full-length GPI-APs following spontaneous release into the circulation of metabolically deranged rats and humans are counterbalanced by upregulated interaction of their GPI anchor with GPLD1 and other serum proteins. Thereby, translocation of GPI-APs to blood and tissue cells and their lysis are prevented. The identification of GPI-APs and serum proteins interacting within micelle-like complexes may facilitate the prediction and stratification of diseases that are associated with impaired cell-surface anchorage of GPI-APs, such as obesity and diabetes.

Testing configurations of attractive toxic sugar bait (ATSB) stations in Mali, West Africa, for improving the control of malaria parasite transmission by vector mosquitoes and minimizing their effect on non-target insects.

BACKGROUND: Application methods of |Attractive Toxic Sugar Baits (ATSB) need to be improved for wide-scale use, and effects on non-target organisms (NTOs) must be assessed. The goals of this study were to determine, at the village level, the effect of different configurations of bait stations to (1) achieve < 25% Anopheles mosquito vector daily feeding rate for both males and females and (2) minimize the effect on non-target organisms. METHODS: Dye was added to Attractive Sugar Bait Stations (without toxin) to mark mosquitoes feeding on the baits, and CDC UV light traps were used to monitor for marked mosquitoes. An array of different traps were used to catch dye marked NTOs, indicating feeding on the ASB. Stations were hung on homes (1, 2, or 3 per home to optimize density) at different heights (1.0 m or 1.8 m above the ground). Eight villages were chosen as for the experiments. RESULTS: The use of one ASB station per house did not mark enough mosquitoes. Use of two and three stations per house gave feeding rates above the 25% goal. There was no statistical difference in the percentage of marked mosquitoes between two and three stations, however, the catches using two and three bait stations were both significantly higher than using one. There was no difference in An. gambiae s.l. feeding when stations were hung at 1.0 and 1.8 m. At 1.8 m stations sustained less accidental damage. ASB stations 1.8 m above ground were fed on by three of seven monitored insect orders. The monitored orders were: Hymenoptera, Lepidoptera, Coleoptera, Diptera, Hemiptera, Neuroptera and Orthoptera. Using one or two stations significantly reduced percentage of bait-fed NTOs compared to three stations which had the highest feeding rates. Percentages were as follows: 6.84 ± 2.03% Brachycera followed by wasps (Hymenoptera: Vespidae) 5.32 ± 2.27%, and Rhopalocera 2.22 ± 1.79%. Hanging the optimal number of stations per house for catching mosquitoes (two) at 1.8 m above ground, limited the groups of non-targets to Brachycera, Chironomidae, Noctuoidea, Rhopalocera, parasitic wasps and wasps (Hymenoptera). Feeding at 1.8 m only occurred when stations were damaged. CONCLUSIONS: The goal of marking quarter of the total Anopheles population per day was obtained using 2 bait stations at 1.8 m height above the ground. This configuration also had minimal effects on non-target insects.