THE EFFECT OF COMBINED ACTION OF ANTIBACTERIAL DRUGS WITH LOW-INTENSIVE LASER RADIATION ON CLINICAL STRAINS S. AUREUS AND S. SALIVARIUS IN THE ORAL CAVITY.

The oral mucosa is constantly contaminated by a large number of microorganisms that may cause diseases such as periodontitis and caries. The present paper aims to study the effectiveness of the antimicrobial effect of combined use of antibacterial drugs (AD) and low-intensity laser radiation (LLR) on S. aureus S. salivarius isolated from the oral cavity. The study included 20 individuals with dental caries, 20 individuals with periodontitis and 10 without any signs of dental disease. The material for the microbacterial study was collected from surfaces of the teeth, oral cavity with dental caries and periodontal pockets. The intensity of bacterial isolation was estimated by two factors: the frequency of isolation and percentage of other aerobic microorganisms. The obtained data demonstrated that the use of several antibacterial drugs had a different impact on the strains of S. salivarius and S. aureus, depending on the source of their collection. The collected isolates were used to determine the effect of a 5 minute laser radiation combined with antibacterial drugs. The simultaneous use of antibacterial therapy and laser radiation showed an increase in the therapeutic effect of all investigated antibiotics followed by the inhibition of the growth presentations in S. aureus and S. salivarius. The application of photodynamic therapy, e.g. LLR, combined with antibacterial drugs allowed to achieve a complete inhibition of the microbial growth.

[APPLICATION OF SYNTHETIC OSTEOPLASTIC MATERIAL EASYGRAFT® IN MAXILLA SUBANTRAL AUGMENTATION (SINUS-LIFT)].

The problem of complex dental rehabilitation of a patient after partial or complete teeth loss is an urgent issue for dental practice. The most of patients coming to dentist with to make dentures on dental implants after tooth loss in lateral part of maxilla are often observed with atrophy of the alveolar bone resulting in maxillary sinus (sinus Haymori) enlargement. Creating an additional space for dental implant placement in the maxilla is possible by performing a sinus lift (subantral maxilla augmentation) - artificially thickening the lower and outer wall of the maxillary sinus with usage of osteoplastic materials. A clinical case of lateral (open) sinus-lifting performed by "grind-out" technique with the DASK® kit (Dentium Advanced Sinus Kit®) is described. As an osteoplastic material was used an alloplastic self- hardening in wound material easygraft®.

DYNAMICS OF MORPHOLOGICAL CHANGES OF RATS' ADENOHYPOPHYSIS IN BURN DISEASE.

The article presents data relating to structural changes in rat adenohypophysis in experimental burn disease and its treatment. Experimental studies were conducted for defining peculiarities of structural changes in rats' adenohypophisis without and after skin burn injuries (in 7-14 and 21-30 days) alongside its correction with 0.9% NaCl solution. Application of DNA-cytometry allowed defining subtle mechanisms of pathogenic influence of burn injuries on the organism in general and the cells in particular.

A new in vitro test to evaluate the resistance level against acaricides of the cattle tick, Rhipicephalus (Boophilus) microplus.

In this article we present a new bioassay to assess the resistance status of ticks to acaricides. The Larval Tarsal Test (LTT) is a sensitive, highly time-effective in vitro test. It allows the investigation of a large number of compounds and doses on the cattle tick Rhipicephalus (Boophilus) microplus in a short period of time. The ability of the LTT to assess the lethal concentration at 50% mortality (LC(50)) and resistance ratios (RRs) of a susceptible and a resistant R. microplus strain was compared with the FAO-recommended Larval Packet Test (LPT). Representative compounds of the carbamate, organophosphate (OP), synthetic pyrethroid (SP), formamidine (FOR), macrocyclic lactone and pyrazole classes were used for this comparison. The resistance status against OP, SP and FOR of the resistant R. microplus strain was confirmed in vivo. The LTT resulted in resistance ratios comparable to those obtained with the LPT. However, the lethal concentrations were up to 150-fold lower in the LTT than in the LPT. The advantage of the LTT is to simplify the methodology by avoiding the handling of larvae and using multi-well plates. The LTT is therefore a suitable test for the assessment of the level of resistance of R. microplus and is very promising to evaluate the resistance profile of field strains. Additionally, the LTT is also suitable to test other ixodid species.

Dose confirmation studies for monepantel, an amino-acetonitrile derivative, against fourth stage gastro-intestinal nematode larvae infecting sheep.

Monepantel is the first compound from the recently discovered amino-acetonitrile derivative (AAD) class of anthelmintics to be developed for use in sheep. Nine dose confirmation studies were conducted in Australia, New Zealand and Switzerland to confirm the minimum therapeutic oral dose of monepantel to control fourth stage (L4) gastro-intestinal nematode larvae in sheep (target species were Haemonchus contortus, Teladorsagia (Ostertagia) circumcincta, Teladorsagia trifurcata, Trichostrongylus axei, Trichostrongylus colubriformis, Trichostrongylus vitrinus, Cooperia curticei, Cooperia oncophora, Nematodirusbattus, Nematodirusfilicollis, Nematodirus spathiger, Chabertia ovina and Oesophagostomum venulosum). In each study, sheep infected with a defined selection of the target nematodes were treated with 2.5mg monepantel/kg liveweight. Following euthanasia and worm counting, efficacy was calculated against worm counts from untreated control groups. The results demonstrate high (95<100%) efficacy of monepantel when administered orally to sheep at 2.5mg/kg for most species tested. Efficacy levels against N. spathiger and O. venulosum were variable and failed to meet the required regulatory standard (> or =90%) in some studies. Efficacy was demonstrated against L4 stages of nematodes known to be resistant to either benzimidazole and/or levamisole anthelmintics (macrocyclic lactone resistant isolates were not available for testing). The broad-spectrum activity of monepantel against L4 larvae of common gastro-intestinal nematodes in sheep and its favorable safety profile represents a significant advance in the treatment of parasitic gastro-enteritis in this animal species. No adverse effects related to treatment with monepantel were observed.

Dose determination studies for monepantel, an amino-acetonitrile derivative, against fourth stage gastro-intestinal nematode larvae infecting sheep.

Monepantel is the first compound from the recently discovered amino-acetonitrile derivative (AAD) class of anthelmintics to be developed for use in sheep. Three dose determination studies were conducted in Australia and Switzerland to identify the minimum therapeutic dose of monepantel when formulated for the oral treatment of sheep to control fourth stage (L4) gastro-intestinal nematode larvae. In each study, sheep infected with the target nematodes (selected from Haemonchus contortus, Teladorsagia (Ostertagia) circumcincta, Teladorsagia trifurcata, Trichostrongylus axei, Trichostrongylus colubriformis, Trichostrongylus vitrinus, Cooperia curticei, Cooperia oncophora, Nematodirus battus, Nematodirus filicollis, Nematodirus spathiger, Chabertia ovina and Oesophagostomum venulosum) were treated with either 1.25, 2.5 or 5.0 mg monepantel/kg liveweight. Following euthanasia and worm counting, efficacy was calculated against worm counts from untreated control groups. Monepantel proved highly effective at 2.5 and 5.0 mg/kg, but was only moderately effective against some nematode species (L4 stage) at 1.25 mg/kg. The results also confirmed that monepantel will effectively control L4 stages of nematodes resistant to at least some of the currently available broad-spectrum anthelmintic classes (macrocyclic lactone resistant strains were not included in the studies). It was concluded that 2.5 mg/kg would be a suitable minimum dose rate for a commercial product. No adverse events related to treatment with monepantel were detected.

Identification of the amino-acetonitrile derivative monepantel (AAD 1566) as a new anthelmintic drug development candidate.

Anthelmintic resistance has become a global phenomenon in gastro-intestinal nematodes of farm animals, including multi-drug resistance against the three major classes of anthelmintics. There is an urgent need for an anthelmintic with a new mode of action. The recently discovered amino-acetonitrile derivatives (AADs) offer a new class of synthetic chemicals with anthelmintic activity. The evaluation of AADs was pursued applying in vitro assays and efficacy and tolerability studies in rodents, sheep, and cattle. Amongst various suitable compounds, AAD 1566 eliminated many tested pathogenic nematode species, both at larval and adult stages, at a dose of 2.5 mg/kg bodyweight in sheep and 5.0 mg/kg bodyweight in cattle. The same doses were sufficient to cure animals infected with resistant or multi-drug-resistant nematode isolates. These findings, complemented by the good tolerability and low toxicity to mammals, suggest that AAD 1566, monepantel, would be a suitable anthelmintic drug development candidate.

PIV validation of blood-heart valve leaflet interaction modelling.

The aim of this study was to validate the 2D computational fluid dynamics (CFD) results of a moving heart valve based on a fluid-structure interaction (FSI) algorithm with experimental measurements. Firstly, a pulsatile laminar flow through a monoleaflet valve model with a stiff leaflet was visualized by means of Particle Image Velocimetry (PIV). The inflow data sets were applied to a CFD simulation including blood-leaflet interaction. The measurement section with a fixed leaflet was enclosed into a standard mock loop in series with a Harvard Apparatus Pulsatile Blood Pump, a compliance chamber and a reservoir. Standard 2D PIV measurements were made at a frequency of 60 bpm. Average velocity magnitude results of 36 phase-locked measurements were evaluated at every 10 degrees of the pump cycle. For the CFD flow simulation, a commercially available package from Fluent Inc. was used in combination with inhouse developed FSI code based on the Arbitrary Lagrangian-Eulerian (ALE) method. Then the CFD code was applied to the leaflet to quantify the shear stress on it. Generally, the CFD results are in agreement with the PIV evaluated data in major flow regions, thereby validating the FSI simulation of a monoleaflet valve with a flexible leaflet. The applicability of the new CFD code for quantifying the shear stress on a flexible leaflet is thus demonstrated.

Time-resolved PIV technique for high temporal resolution measurement of mechanical prosthetic aortic valve fluid dynamics.

Prosthetic heart valves (PHVs) have been used to replace diseased native valves for more than five decades. Among these, mechanical PHVs are the most frequently implanted. Unfortunately, these devices still do not achieve ideal behavior and lead to many complications, many of which are related to fluid mechanics. The fluid dynamics of mechanical PHVs are particularly complex and the fine-scale characteristics of such flows call for very accurate experimental techniques. Adequate temporal resolution can be reached by applying time-resolved PIV, a high-resolution dynamic technique which is able to capture detailed chronological changes in the velocity field. The aim of this experimental study is to investigate the evolution of the flow field in a detailed time domain of a commercial bileaflet PHV in a mock-loop mimicking unsteady conditions, by means of time-resolved 2D Particle Image Velocimetry (PIV). The investigated flow field corresponded to the region immediately downstream of the valve plane. Spatial resolution as in "standard" PIV analysis of prosthetic valve fluid dynamics was used. The combination of a Nd:YLF high-repetition-rate double-cavity laser with a high frame rate CMOS camera allowed a detailed, highly temporally resolved acquisition (up to 10000 fps depending on the resolution) of the flow downstream of the PHV. Features that were observed include the non-homogeneity and unsteadiness of the phenomenon and the presence of large-scale vortices within the field, especially in the wake of the valve leaflets. Furthermore, we observed that highly temporally cycle-resolved analysis allowed the different behaviors exhibited by the bileaflet valve at closure to be captured in different acquired cardiac cycles. By accurately capturing hemodynamically relevant time scales of motion, time-resolved PIV characterization can realistically be expected to help designers in improving PHV performance and in furnishing comprehensive validation with experimental data on fluid dynamics numeric modelling.

Evaluation of selected Sudanese medicinal plants for their in vitro activity against hemoflagellates, selected bacteria, HIV-1-RT and tyrosine kinase inhibitory, and for cytotoxicity.

Ethnobotanical investigations led to the selection of 19 plant species, used traditionally in Sudan against malaria and other similar tropical diseases, for further studies. Pamianthe peruviana (Amaryllidaceae) exhibited significant activity against a chloroquine-resistant Plasmodium falciparum strain (K1) and a chloroquine-sensitive strain (NF54) with IC(50) values of 0.6 and 1.1 microg/ml, respectively. Additionally, P. peruviana showed considerable activities against Trypanosoma brucei rhodesiense (IC(50) 1.5 microg/ml) and T. cruzi (IC(50) 11.8 microg/ml). The antiplasmodial activity of the different extracts of Salvadora persica (Salvadoraceae) against P. falciparum NF54 strain were found to be 0.6 microg/ml (stems) and 0.7 microg/ml (leaves). Extracts of different parts of Combretum hartmannianum (Combretaceae) possessed significant activity against the chloroquine-sensitive P. falciparum strain (NF54) with IC(50) values of 0.2 microg/ml (bark), 0.4 microg/ml (stem) and 4.3 microg/ml (leaves). Most interestingly, the extracts of the leaves of C. hartmannianum totally inhibited the enzyme HIV-1 reverse transcriptase (HIV-1 RT) at a concentration of 66 microg/ml. A comparably strong activity against p56(lck) tyrosine kinase was also seen for this extract.

Genetic variants of the TbAT1 adenosine transporter from African trypanosomes in relapse infections following melarsoprol therapy.

We have analyzed the TbAT1 gene, which codes for the P2 adenosine transporter, from Trypanosoma brucei field isolates to investigate a possible link between the presence of mutations in this gene and melarsoprol treatment failure. Of 65 T. b. gambiense isolates analyzed from a focus in north-western Uganda with high treatment failure rates following melarsoprol therapy, 38 had a mutated TbAT1. Unexpectedly, all individual isolates contained the same set of nine mutations in their TbAT1 genes. Of these, five point mutations resulted in amino acid substitutions, one resulted in the deletion of an entire codon, and three were silent point mutations. Eight of these mutations had previously been reported in a laboratory-derived Cymelarsan-resistant T. b. brucei clone. Identical sets of mutations were also found in a drug-resistant T.b.rhodesiense isolate from south-eastern Uganda and in a T.b.gambiense isolate from a relapsing patient from northern Angola. A deletion of the TbAT1 gene was found in a single T. b. gambiense isolate from a relapsing patient from northern Angola. The data presented demonstrate the surprising finding that trypanosomes from individual relapse patients of one area, as well as from geographically distant localities, contain an identical set of point mutations in the transporter gene TbAT1. They further demonstrate that many isolates from relapse patients contained the wild-type TbAT1 genes, suggesting that melarsoprol refractoriness is not solely due to a mutational inactivation of TbAT1.

Drug resistance in Trypanosoma brucei spp., the causative agents of sleeping sickness in man and nagana in cattle.

Drug resistance in pathogenic trypanosomes threatens successful control of fatal sleeping sickness in man and hinders economic livestock production in sub-Saharan Africa. We report on the occurrence and development of drug resistance, and discuss the genetic basis of such resistance in Trypanosoma brucei. Understanding these mechanisms at the molecular level will enable improved management of existing drugs and provide valuable clues to the development of new trypanocides.

Identification and characterization of trypanocides by functional expression of an adenosine transporter from Trypanosoma brucei in yeast.

The causative agents of sleeping sickness, Trypanosoma brucei rhodesiense and T. brucei gambiense, do not synthesize purines de novo but salvage purine bases and nucleosides from their hosts. We used yeast as an expression system for functional characterization of the trypanosomal adenosine transporter TbAT1. A selection of purine analogs and flavonoids were tested for their ability to interfere with adenosine transport, with the aims of identifying (a) trypanocidal TbAT1 substrates, and (b) inhibitors of trypanosomal purine transport. Cordycepin (3'-deoxyadenosine) was a TbAT1 substrate of high activity against T. brucei rhodesiense (IC50 0.2 nM). Inhibitors of mammalian nucleoside transport were not active, while the flavonol silibinin was a potent, noncompetitive inhibitor of TbAT1-mediated adenosine transport in yeast. Silibinin also inhibited melarsen-induced lysis of bloodstream form trypanosomes. IC50 values to T. brucei rhodesiense and to human carcinoma cells were 0.6 and 140 microM, respectively, indicating a good selectivity towards the parasites. Further studies are necessary to elucidate the effects of flavonoids on trypanosomal purine transport and their potential as trypanocides.

Melarsoprol refractory T. b. gambiense from Omugo, north-western Uganda.

Culture adapted T. b. gambiense isolated from Northwest Uganda were exposed to 0.001-0.14 microg/ml melarsoprol or 1.56-100 microg/ml DL-alpha-difluoromethylornithine (DFMO). Minimum inhibitory concentrations (MICs) of each drug were scored for each isolate after a period of 10 days drug exposure. The results indicate that T. b. gambiense isolates from Northwest Uganda had elevated MIC values for melarsoprol ranging from 0.009 to 0.072 microg/ml as compared with T. b. gambiense isolates from Cote d'Ivoire with MIC values ranging from 0.001 to 0.018 microg/ml or with T. b. rhodesiense from Southeast Uganda with MIC values from 0.001 to 0.009 microg/ml. All MIC values obtained fell below expected peak melarsoprol concentrations in serum of treated patients. However, it may not be possible to maintain constant drug concentrations in serum of patients as was the case in our in vitro experiments. Importantly, the MIC of 0.072 microg/ml exhibited by one of the isolates from Northwest Uganda was above levels attainable in CSF indicating that this isolate would probably not be eliminated from CSF of treated patients. PCR amplification of the gene encoding the P2-like adenosine transporter followed by restriction digestion with Sfa NI enzyme revealed presence of fragments previously observed in a trypanosome clone with laboratory-induced arsenic resistance. From our findings it appears that reduced drug susceptibility may be one factor for the frequent relapses of sleeping sickness after melarsoprol treatment occurring in Northwest Uganda.

Red palm oil in the maternal diet increases provitamin A carotenoids in breastmilk and serum of the mother-infant dyad.

BACKGROUND: Despite vitamin A supplementation programs, vitamin A deficiency in children remains a public health concern in Honduras. AIM OF THE STUDY: We investigated the effectiveness of short-term dietary supplementation of mothers with red palm oil as a strategy for improving the vitamin A status of the mother-infant dyad. METHODS: Lactating mothers in Colonia Los Pinos, a barrio of Tegucigalpa, Honduras, consumed a total of 90-mg beta-carotene as red palm oil (n = 32) supplements (n = 36) or placebo (n = 18) in six equal doses over 10 days. Carotenoids and retinol in maternal and infant serum, and breastmilk carotenoids and retinol were measured before and after supplementation. Maternal diet was evaluated by 24-hour recall. RESULTS: Maternal serum alpha-carotene and beta-carotene concentrations were increased 2 fold by palm oil compared with 1.2 fold by beta-carotene supplements. Changes were significantly different in infant serum alpha-carotene but not beta-carotene among the three experimental groups. Increases in breastmilk beta-carotene were greater for the palm oil group (2.5 fold) than for the beta-carotene supplement group (1.6 fold) and increases in milk alpha-carotene concentrations (3.2 fold) were slightly greater than those of beta-carotene. There were also small but significant changes among groups in breastmilk lutein and lycopene. Breastmilk retinol was not significantly different among the groups over the treatment period. CONCLUSIONS: Red palm oil in the maternal diet increases provitamin A carotenoids in breastmilk and serum of the mother-infant dyad. The use of dietary red palm oil to improve the vitamin A status of this population should be further investigated.

Ascosalipyrrolidinone A, an antimicrobial alkaloid, from the obligate marine fungus Ascochyta salicorniae.

From the green alga Ulva sp., the endophytic and obligate marine fungus Ascochyta salicorniae was isolated. A. salicorniae was mass cultivated and found to produce the unprecedented and structurally unusual tetramic acid containing metabolites ascosalipyrrolidinones A (1) and B (2). Additionally, the new natural product ascosalipyrone (3) and the known metabolites 4 and 5 were obtained. Ascosalipyrrolidinone A (1) has antiplasmodial activity toward Plasmodium falciparum strains K1 and NF 54, as well as showing antimicrobial activity and inhibiting tyrosine kinase p56lck.

Pelorol from the tropical marine sponge Dactylospongia elegans.

From the dichloromethane solubles of the tropical marine sponge Dactylospongia elegans, a new aromatic substituted sesquiterpene, pelorol (1), and the known sesquiterpene, ilimaquinone (2), were isolated. The structures of 1 and 2 were deduced from their spectroscopic data. The biological activities of compounds 1 and 2 were assessed in a variety of bioassays, and both compounds were found to have weak antitrypanosomal and antiplasmodial effects.

Lifetime sexual assault prevalence rates and reporting practices in an emergency department population.

STUDY OBJECTIVE: Studies suggest significant rates of female sexual assault (SA); the majority of SAs remain unreported, and few victims receive medical care. The purpose of this study was to determine lifetime prevalence rates of SA in an emergency department population and to assess reporting patterns to police, physicians, and social service agencies. METHODS: A verbally administered survey was given to all female patients during 4-hour randomized periods in an urban Level I trauma center. All English-speaking, noncritically ill women who presented during the study period were eligible. RESULTS: Four hundred forty-two women were eligible; 360 (81%) women agreed to participate. The lifetime prevalence rate of SA was 39% (n=139). Ninety-seven women (70%) were older than 15 years at the time of SA. Of these 97 SAs occurring in adulthood, 49 (52%) reported assault by an acquaintance, family member, or friend; 28 (30%) by a stranger; and 17 (18%) by a partner. Forty-five (46%) women reported the crime to the police, 42 (43%) sought medical care, and 23 (25%) contacted a social service agency. Reporting patterns for victims assaulted by a stranger versus those assaulted by a partner were: reported to police 79% (95% confidence interval [CI] 62 to 95) versus 18% (95% CI 0 to 38); P <.001), received medical care 70% (95% CI 46 to 95) versus 29% (95% CI 11 to 48; P<.01), contacted a social service agency 30% (95% CI 5 to 47) versus 24% (95% CI 1 to 46; P=.63). CONCLUSION: Lifetime female SA rates in ED populations are significant. Fewer than half of SA victims report the assault to the police or seek medical care. Women assaulted by a partner are significantly less likely to report the SA to police or seek medical care.

A new bioactive sesterterpene and antiplasmodial alkaloids from the marine sponge hyrtios cf. erecta.

From the CH(2)Cl(2) extract of the sponge Hyrtios cf. erecta, collected from Fiji, two new sesterterpenes, 1 and 2, and the known compounds isodehydroluffariellolide (3), homofascaplysin A (4), and fascaplysin (5) were isolated. The structures of 1-5 were established employing 1D and 2D NMR spectroscopy and mass spectrometry. All NMR resonances of fascaplysin (5) have been unambiguously assigned. Evaluation of the biological activity of the extracts and pure compounds toward Plasmodium falciparum, Trypanosoma brucei subsp. rhodesiense, Trypanosoma cruzi, hepatitis A virus (HAV), several other microbial targets, and HIV-1-RT and p56(lck) tyrosine kinase revealed new activities for homofascaplysin (4) and fascaplysin (5), both being potently active in vitro against P. falciparum.