Challenging the Wigglesworthia, Sodalis, Wolbachia symbiosis dogma in tsetse flies: Spiroplasma is present in both laboratory and natural populations.

Profiling of wild and laboratory tsetse populations using 16S rRNA gene amplicon sequencing allowed us to examine whether the "Wigglesworthia-Sodalis-Wolbachia dogma" operates across species and populations. The most abundant taxa, in wild and laboratory populations, were Wigglesworthia (the primary endosymbiont), Sodalis and Wolbachia as previously characterized. The species richness of the microbiota was greater in wild than laboratory populations. Spiroplasma was identified as a new symbiont exclusively in Glossina fuscipes fuscipes and G. tachinoides, members of the palpalis sub-group, and the infection prevalence in several laboratory and natural populations was surveyed. Multi locus sequencing typing (MLST) analysis identified two strains of tsetse-associated Spiroplasma, present in G. f. fuscipes and G. tachinoides. Spiroplasma density in G. f. fuscipes larva guts was significantly higher than in guts from teneral and 15-day old male and female adults. In gonads of teneral and 15-day old insects, Spiroplasma density was higher in testes than ovaries, and was significantly higher density in live versus prematurely deceased females indicating a potentially mutualistic association. Higher Spiroplasma density in testes than in ovaries was also detected by fluorescent in situ hybridization in G. f. fuscipes.

Maggot therapy in treatment of a complex hand injury complicated by mycotic infection.

Complex injuries of the hand remain a therapeutic challenge for surgeons. We present the case of a male who suffered a devastating injury of the hand caused by a conveyor belt. The patient developed a progressive Absidia corymbifera infection of the affected soft tissues. Initial treatments with serial surgical debridement and topical and intravenous itraconazole were unsuccessful in eliminating the infection. We decided to use maggot debridement therapy in a new special design to debride all necrotic, devitalized tissue and preserve only healthy tissue and functioning structures. This maneuverer followed by negative pressure therapy allowed progressive healing. In such complex hand injuries, maggot debridement combined with negative pressure therapy could be considered to achieve effective and considerable results, although future functional morbidity may occur (Fig. 4, Ref. 18).

Growth and survival of blowfly Lucilia sericata larvae under simulated wound conditions: implications for maggot debridement therapy.

Maggot debridement therapy has become a well-established method of wound debridement. Despite its success, little information is available about the optimum duration of the treatment cycle and larval growth in wounds. This study examines the development of Lucilia sericata (Diptera: Calliphoridae) larvae under two containment conditions (bagged and free range) under simulated wound conditions and assesses the impact of transport and further storage of larvae on their survival and growth. There was no significant difference in size between bagged and free-range larvae over the 72-h experimental period. Larvae grew fastest 8-24 h after inoculation and completed their growth at 40-48 h. Mortality rates were similar (0.12-0.23% per hour) in both containment conditions and did not differ significantly (P = 0.3212). Survival of free-range larvae was on average 16% lower than survival of bagged larvae. Refrigeration of larvae upon simulated delivery for > 1 day reduced their survival to < 50% and caused a reduction in growth of up to 30% at 12 h, but not at 48 h, of incubation. Therefore, it is recommended that free-range larvae are left in the wound for a maximum of 40-48 h, and bagged larvae for 48-72 h. Larvae should be used within 24 h of delivery to avoid high mortality caused by prolonged refrigeration.

Midgut lysozymes of Lucilia sericata - new antimicrobials involved in maggot debridement therapy.

Larvae of Lucilia sericata are used for maggot debridement therapy (MDT) because of their ability to remove necrotic tissue and eradicate bacterial pathogens of infected wounds. So far, very few antibacterial factors have been fully characterized (eg lucifensin). Using a molecular approach, some other putative antimicrobial compounds, including three novel lysozymes, have been previously identified and predicted to be involved in MDT. Nevertheless, data on lysozymes tissue origin and their functions have never been elucidated. Therefore, the aim of this study was to investigate the expression of three lysozymes in L. sericata and confirm their antibacterial effects within MDT. Moreover, we characterized the eradication process of bacteria within the digestive system of maggots and determined the role of lysozymes in this process. We found that three lysozymes are expressed in specific sections of the L. sericata midgut. Recombinant lysozymes displayed comparable antibacterial activity against Micrococcus luteus. Furthermore, the majority of Gram-positive bacteria were destroyed in vivo within the particular section of the L. sericata midgut where lysozymes are produced. Larval ingestion and subsequent eradication of wound pathogens during their passage through the intestine of maggots are due to, at least in part, antibacterial action of three midgut lysozymes.

Position paper on PRM and persons with long term disabilities.

In the current population we observe a rise of chronic health problems often with multiple characteristics. This results in a growing number of people who are experiencing long-term disabilities or difficulties in functioning because of disability. These conditions require a complex response over an extended period of time, that involves coordinated inputs from a wide range of health professionals. This paper argues the central role and benefit of rehabilitation and describes the rehabilitation as an integral component in the management of people with chronic disabilities. It also presents the most important related definitions: long-term care, rehabilitation for chronic disease and disability, the aim of physical and rehabilitation medicine (PRM). An interdisciplinary team is ideal for an effective implementation of rehabilitation for chronic disease and disability. However, the article mainly focuses on defining the role and contribution of the PRM physician in the rehabilitation of persons with long-term disabilities. The article includes: descriptions of his/her key role and competencies, particularly with regard to medical and functional status and prognosis, of the ability to comprehensively define the rehabilitation needs of the patient/person with respect to ICD-WHO classification domains, of the cooperation with other medical specialists and health professionals, of determining the rehabilitation potential, of developing the rehabilitation plan tailored to specific needs, as well as of the contribution of PRM physician in the follow-up care pathways.

Effect of spa therapy after intervertebral disc surgery in the cervical spine.

OBJECTIVE: The aim of this paper is to determine whether standard spa treatment significantly affects disability, functional status, and quality of life of patients after surgery of intervertebral discs in the cervical spine. MATERIAL AND METHODS: The study was performed in patients who underwent an artificial intervertebral disc replacement in the cervical spine.The study involved a group of 88 patients with mean age of 49.49 (range 32 to 74, standard deviation 9.12) while the experimental group consisted of 40 patients who underwent spa treatment with mean age of 49.35 (range 32 to 72, standard deviation 7.69). The control group was not treated by spa treatment, and consisted of 48 patients with mean age of 49.6 (range 36 to 74, standard deviation 8.73). These patients underwent standard medical rehabilitation at rehabilitation departments.Dysfunction of the cervical spine was scaled according to The Neck Disability Index (NDI). Functional status of patients was evaluated by Pulses Profile, and quality of life was evaluated by test of the quality of life according to Spitzer. RESULTS: The statistical evaluation parameter for NDI, Pulses and Spitzer found significant difference at the significance level of 0.01, suggesting that experimental group of patients who received spa treatment during follow-up examination had significantly better results in comparison with the control group. CONCLUSION: As a result of this study we have concluded that after implantation of the artificial intervertebral disc replacement in the cervical spine it is appropriate to recommend spa treatment for its positive effect on disability, functional status, and quality of life (Tab. 3, Fig. 3, Ref. 24).

The influence of housefly Musca domestica embryo age on viability in water and at low temperatures.

The sensitivity of housefly Musca domestica L. (Diptera: Muscidae) embryos to storage at low temperatures (5 and 10 °C on moist sponges in Petri dishes) and in water at 26 °C was investigated to develop suitable protocols for the storage and transport of housefly eggs. The youngest embryos (aged 0-3 h) were the most sensitive to storage at 5 °C, with 45% survival after storage for 24 h. Storage of embryos aged 3-12 h at 5 °C for 24 h had no negative effect; longer storage resulted in significantly decreased larval survival (30-34% after 48-72 h, compared with 61% in the control group) and reduced hatching rates (83% after 72 h storage). No negative effects were observed when embryos aged 0-9 h were stored at 10 °C for 24 h, but this temperature did not completely inhibit development and eggs began to hatch if stored for longer than 24 h. All age groups of embryos showed high mortality after storage in water at 26 °C for 24 h, with the youngest embryos being least resistant to submersion.

Improvement of survival of the house fly (Musca domestica L.) larvae under mass-rearing conditions.

Two new approaches were examined, aimed at increasing survival of the house fly (Musca domestica L.) larvae under mass-rearing conditions of a biodegradation facility: modification of the larval substrate and dispersal of the eggs during inoculation. The two types of pig manure used in this study (manure with sawdust and manure without sawdust) differed in terms of larval survival and nutritional value for the house fly larvae. Larval survival in manure without sawdust in the control treatment was low (46.8 ± 2.1%) and its nutritional value for the larvae were high. Addition of 5.7% of previously biodegraded manure did not significantly affect larval survival (52.3 ± 1.9%), but larval development was faster and the pupae were significantly smaller (14.28 ± 0.4 mg) compared to the control (16.29 ± 0.5 mg). Using alternative substrate for incubation of eggs and first-instar larvae significantly increased larval survival (63.3 ± 3.3%) and decreased the mean weight of produced pupae (14.39 ± 0.71 mg). Overall, the weight of recovered biomass in the alternative substrate treatment increased by 14.3 kg ton-1 of manure compared to the control. Larval survival in manure with sawdust was generally higher than 70%, but its nutritional value for the larvae was lower than in manure without sawdust. Dispersal of eggs over the surface of manure with sawdust significantly affected larval survival and mean weight of pupae. Larval survival was significantly lower (59.2 ± 4.0%) and pupae were significantly heavier (18.45 ± 0.8 mg) when eggs were applied to a small area on the manure surface (spot treatment), as compared to diagonal, Z-line and multiple zig-zag dispersal (72.5 ± 2.4 to 74.6 ± 3.0% and 14.76 ± 0.6 to 15.97 ± 0.6 mg, respectively). No significant differences were observed in larval survival or mean weight of pupae when comparing the diagonal, Z-line and multiple zig-zag dispersal patterns. Implementation of the techniques which improve larval survival and increase the weight of produced fly biomass may decrease demand for production of house fly eggs and, therefore, reduce the maintenance costs of adult colony, as well as increase the revenue earned by selling the products.

[Functionality of pelvic floor in patients after vaginal and abdominal hysterectomy, valuation by EMG-Biofeedback according to Perfect Scheme]. / Hodnotenie funkcného stavu panvového dna metódou EMG-Biofeedback podl'a Perfect schémy u pacientiek po abdominálnej a vaginálnej hysterektómii.

OBJECTIVE: Our research deals with the efficiency of rehabilitation in patients after vaginal and abdominal hysterectomy. The activation of pelvic floor muscles has been evaluated and comparison of individual parameters according has been carried out. Also the occurrence of incontinence has been assessed. Subjects consisted of 66 women, out of which there were 21 women after vaginal hysterectomy and 45 women after abdominal hysterectomy. TYPE OF STUDY: Prospective experimental study. METHODS USE: EMG-biofeedback has been used for the evaluation of the functionality of pelvic floor according to Perfect Scheme. For the quantification of leaked urine one hour PW test has been used. REHABILITATION METHODS: We applied the individual exercise on strengthening and relaxation of pelvic floor muscles with the aid of repeated, selective voluntary contractions or relaxation of pelvic floor muscles conducted two times a week, with the duration time of 15 minutes. Group exercise on strengthening and relaxation of pelvic floor muscles conducted two times a week, with the duration time of 30 minutes. Biofeedback (EMG-Module) as a therapeutic method was applied. RESULTS: In patients after vaginal hysterectomy there was a higher occurrence of incontinence than in women after abdominal hysterectomy. In both groups a significant increase in pelvic floor muscle strength has been achieved after the therapy. In both of these groups there was no statistically significant relaxation of m. rectus abdominis after the therapy. The patients after abdominal hysterectomy had more statistically significant relaxed abdomen muscles before treatment than patients after vaginal hysterectomy. It means, that performed exercise in the right manner, with reduction of intra abdominal pressure. We suppose that this phenomenon may be related to the higher percentage of occurrence of incontinence in patients after vaginal hysterectomy.

Effects of horsefly (Tabanidae) salivary gland extracts on isolated perfused rat heart.

The speed with which horseflies (Diptera: Tabanidae) obtain a bloodmeal suggests they have potent vasodilators. We used isolated perfused rat heart to examine the vasoactivity of salivary gland extracts (SGEs) of three horsefly species, Hybomitra bimaculata Macquart, Tabanus bromius Linnaeus and Tabanus glaucopis Meigen. Administration of horsefly SGEs to the heart produced biphasic coronary responses: a decrease and subsequent increase in coronary flow (CF), characterized by initial vasoconstriction followed by prolonged vasodilation of coronary vessels. However, although SGEs of H. bimaculata induced a significant decrease in left ventricular pressure (LVP), the effect on changes in CF was not significant except at the highest dose tested. The ability to reduce LVP without significantly lowering CF, or affecting heart rate and rhythm, represents a unique set of properties that have considerable therapeutic potential if they can be reproduced by a single molecule.

Maggot debridement therapy.

AIM: To establish a maggot debridement therapy unit and assess efficacy of this method in chronic conservative non-treatable leg ulcers in patient in whom conventional therapy failed. MATERIAL AND METHODS: From August 2004 to December 2005 we treated with Maggot debridement therapy (MDT) 10 patients, 6 men and 4 women, with 13 leg ulcers treated first with conventional and then with maggot therapy. Average age of these patients was 55 years, 11 wounds were of venous and 2 of arterial origin. 7 patients had diabetes mellitus. RESULTS: 1 patient with arterial ulcers underwent supracondylar amputation, 2 patients were re-admitted for recurrent ulcers, one extremely obese patient with cardiovascular co-morbidity did not heal, but all other ulcers have been healed, cleared or minimised during 4-8 week MDT. We observed massive growth of granulation tissue and microbiological cleaning of these wounds. No adverse effects were observed. CONCLUSION: Maggot therapy was more effective and efficient in debriding non-healing leg ulcers than a conservative treatment (Fig. 3, Ref. 6).

Vasodilatory activity in horsefly and deerfly salivary glands.

Salivary gland extract (SGE) of four horsefly species (Hybomitra bimaculata Macquart, Hybomitra ciureai Séguy, Tabanus bromius L., Tabanus glaucopis Meigen) and one deerfly species (Chrysops relictus Meigen) (Diptera: Tabanidae) were shown to contain vasodilatory activity. Aliquots equivalent to 1, 5 and 10 pairs of salivary glands (SG) relaxed rat femoral artery (with intact endothelium) pre-constricted with phenylephrine. Vasodilatory activity was dose-dependent. SGE of one horsefly species (Haematopota pluvialis L.) did not induce relaxation. The kinetics of vasodilation induced by SGE of four horsefly species differed from the deerfly. These results indicate that tabanid species may produce more than one type of vasodilator to aid blood feeding.

Conservation of ecdysis-triggering hormone signalling in insects.

Pre-ecdysis- and ecdysis-triggering hormones (PETH and ETH) from endocrine Inka cells initiate ecdysis in moths and Drosophila through direct actions on the central nervous system (CNS). Using immunohistochemistry, we found Inka cells in representatives of all major insect orders. In most insects, Inka cells are numerous, small and scattered throughout the tracheal system. Only some higher holometabolous insects exhibit 8-9 pairs of large Inka cells attached to tracheae in each prothoracic and abdominal segment. The number and morphology of Inka cells can be very variable even in the same individuals or related insects, but all produce peptide hormones that are completely released at each ecdysis. Injection of tracheal extracts prepared from representatives of several insect orders induces pre-ecdysis and ecdysis behaviours in pharate larvae of Bombyx, indicating functional similarity of these peptides. We isolated several PETH-immunoreactive peptides from tracheal extracts of the cockroach Nauphoeta cinerea and the bug Pyrrhocoris apterus and identified the gene encoding two putative ETHs in the mosquito Anopheles gambiae. Inka cells also are stained with antisera to myomodulin, FMRFamide and other peptides sharing RXamide carboxyl termini. However, our enzyme immunoassays show that these antisera cross-react with PETH and ETH. Our results suggest that Inka cells of different insects produce only peptide hormones closely related to PETH and ETH, which are essential endocrine factors required for activation of the ecdysis behavioural sequence.

Identification of anticoagulant activities in salivary gland extracts of four horsefly species (Diptera, tabanidae).

Anticoagulant activities against the extrinsic and intrinsic coagulation pathways were identified in salivary gland extracts (SGE) prepared from four tabanids (Hybomitra muehlfeldi, Tabanus autumnalis, Haematopota pluvialis, Heptatoma pellucens). All extracts prolonged human plasma clotting time in a dose-dependent manner and inhibited thrombin activity in the chromogenic substrate assay. Horsefly SGE did not inhibit factor Xa. Partial purification of SGE proteins using reversed-phase high-performance liquid chromatography revealed species-specific differences in the elution profiles and range of fractions with anticoagulant activities.

Effect of Methyl Farnesoate on Late Larval Development and Metamorphosis in the Prawn Macrobrachium rosenbergii (Decapoda, Palaemonidae): A Juvenoid-like Effect?

Methyl farnesoate (MF), the unepoxidated form of insect juvenile hormone III, was detected in larvae of the freshwater prawn Macrobrachium rosenbergii, which metamorphose to post-larvae following 11 larval stages. The possible role of MF as a morphogen was studied by administering the compound to M. rosenbergii larvae via an Artemia vector. Higher MF levels caused earlier retardation of late larval growth, and the highest dose retarded larval development. Furthermore, MF significantly affected the patterns of metamorphosis and the appearance of intermediate individuals exhibiting both larval and post-larval morphology and behavior. Three intermediate types were defined, two of which were found only at the MF-treated groups and one that was exclusive to the higher dose treatments. The relative abundance of intermediate specimens increased from 2% in the control to 32% in the high MF concentration, which suggests that MF has a juvenoid-like effect in this decapod crustacean.

[Angiotensin converting enzyme inhibitors in ischemic heart disease]. / Inhibítory angiotenzín konvertujúceho enzýmu pri ischemickej chorobe srdca.

Angiotensin formed in the renin-angiotensin system is involved in the genesis and development of atherosclerosis and coronary heart disease. It has a negative impact on the process of ischaemization/reperfusion. The renin-angiotensin system is activated during a new myocardial infarction and has an impact on the process of remodelling of the left ventricle after myocardial infarction which causes its dysfunction and heart failure. ACE inhibitors are one of the important means which influence the formation of angiotensin II and thus prevent its action on heart and vessels. They prevent the development of atherosclerosis, reduce the extent of necroses during myocardial infarction and reduce the extent of left ventricular dysfunction. They diminish also stunning of the heart muscle during ischaemia/reperfusion and significantly prolong the patients life after infarction. They do not influence the development of restenosis after percutaneous transluminal angioplasty. ACE inhibitors have a positive effect on heart failure, they reduce the rate of reinfarctions and the mortality rate.

Dispersion population models discrete in time and continuous in space.

We analyze a discrete-time model of populations that grow and disperse in separate phases. The growth phase is a nonlinear process that allows for the effects of local crowding. The dispersion phase is a linear process that distributes the population throughout its spatial habitat. Our study quantifies the issues of survival and extinction, the existence and stability of nontrivial steady states, and the comparison of various dispersion strategies. Our results show that all of these issues are tied to the global nature of various model parameters. The extreme strategies of staying-in-place and going-everywhere-uniformly are compared numerically to diffusion strategies in various contexts. We approach the mathematical analysis of our model from a functional analysis and an operator theory point of view. We use recent results from the theory of positive operators in Banach lattices.