The number of TM patients seen at each practice or clinic varied

The number of TM patients seen at each practice or clinic varied considerably; the median number was 267 per year (IQR 150–500 patients per year). Specialty vaccines used for travel were offered at a similar frequency compared with the 2005 survey (Table 3). TM consultations were most often between 11 and 20 min in length (67.3% of YFVCs). In addition to pre-travel health consultations, 72.6% of centers gave telephone advice. YFVCs were asked about TM training. Nurses had received some training in 96.7% of YFVCs compared with physicians in 32.2% of centers

(p < 0.0005). The number of physicians with TM training was less than in the baseline survey, where MAPK inhibitor 56.6% of physicians had such training. The most common type of training for nurses were study days run by vaccine manufacturers (87.0% of nurses had attended one), compared to 40.0% in the IWR-1 research buy baseline survey. Self-study was reported by 60.8% of nurses (Figure 2), and was the most common form of training for physicians (51.7%), followed by vaccine manufacturer

training days (44.6%). Forty percent of physicians attended vaccine manufacturer training days in 2005. Few nurses or physicians had membership of the Faculty of Travel Medicine (Royal College of Physicians and Surgeons, Glasgow)23 (3.6 and 3.3%, respectively), or had passed the International Society of Travel Medicine Certificate of Knowledge examination in TM (1.5 and 1.9%, respectively)24; 7 to 13% had completed a diploma level course (a year of distance learning in TM). All but one YFVC reported having internet access at their

center, and nearly all of these centers had it available during a TM consultation (98.7%). Of those who did have internet access during the consultation, 84.8% used it for each patient, compared to the 44.0% who reported using it for each patient in the baseline survey. The internet was used during a consultation for country recommendations (95.9% of YFVCs), general TM information (83.1%), information sheets on travel diseases (80.5%), and information on global disease outbreaks (65.1%). The most frequently accessed websites were the NaTHNaC website (87.8% of respondents) and Health Protection Scotland’s TRAVAX website Celecoxib (73.5%). In contrast, the NaTHNaC website was used by only 18% of YFVCs in 2005. Regarding printed resources, the Department of Health book, Immunisation against Infectious Disease, which covers immunization guidelines for the UK (92.9%), and the British National Formulary, an information source about the use of medicines (71.9%), were the most widely used resources. Vaccine charts in health professional periodicals were used by only 29.5% compared with 73.7% in the baseline survey. The NaTHNaC telephone advice line was the most commonly used telephone line (77.1%), a marked increase from the 14.4% of centers previously using it. Respondents reported that training courses on travel health topics (69.


“Institute of Biological, Environmental & Rural Sciences,


“Institute of Biological, Environmental & Rural Sciences, Aberystwyth University (IBERS), Aberystwyth, Ceredigion, Wales, UK Marlborough, Wiltshire, UK Concentrations of Na+, K+ and Ca2+ in the growth medium were varied within limits normally found in vivo to determine how cation concentrations affect the sensitivity of ruminal bacteria to the ionophores, monensin (a Na+/H+ and K+/H+ exchanger) and tetronasin (Ca2+/H+). High [Na+] (172 mM cf. 137 mM in control medium) enhanced the efficacy of monensin towards Eubacterium ruminantium 2388, Streptococcus learn more bovis C277,

Lactobacillus casei LB17 and Prevotella albensis M384. High [K+] (35 mM cf. 19 mM) alone caused a decreased potency of both ionophores, except with L. casei. Added Ca2+ (7.4

cf. 2.8 mM) increased the potency of tetronasin when [Na+] was low. High [Na+] alone also potentiated the efficacy of tetronasin. Monensin caused intracellular [Na+] and [K+] to be decreased in the most sensitive of these organisms, E. ruminantium, whereas only intracellular [Ca2+] fell with tetronasin. The changes were small; however, Δp fell by only 20 mV after 2 h when ionophores caused immediate cessation of growth. ATP concentrations fell by 77% and 75% with monensin and tetronasin, respectively. Thus, altering cation concentrations might be used to potentiate the efficacy of ionophores, by increasing the rate of energy expenditure to maintain ionic homoeostasis in sensitive bacteria. Monensin and tetronasin are feedlot ionophores that improve feed efficiency in cattle (Goodrich et al., 1984; Bartle et al., 1988). Although CDK inhibitor banned in Europe since 2006, they remain in widespread use elsewhere in the world, and research on their efficacy and mode of action continues (Dubuc et al., 2009; Felix & Loerch, 2011; Packer et al., 2011), particularly in the context of their ability to lower methane emissions

(Martin et al., 2010). Their nutritional effects are due largely to changes in the fermentation stoichiometry and the metabolism of dietary nitrogen by ruminal microorganisms (Bergen & Bates, 1984; Russell & Strobel, 1989; Duffield et al., 2012). These changes arise partly from the elimination of many MYO10 Gram-positive bacteria (Chen & Wolin, 1979; Henderson et al., 1981; Nagaraja & Taylor, 1987; Newbold et al., 1988) and partly from adaptations which resistant Gram-negative bacteria undergo when grown in the presence of ionophores (Morehead & Dawson, 1992; Newbold et al., 1992; Callaway & Russell, 1999). There has been much speculation about the molecular mode of action of feedlot ionophores, mainly by analogy with the action of ionophores on nonruminal species of bacteria (Bergen & Bates, 1984; Russell, 1987; Russell & Strobel, 1989). How ionophores affect ruminal bacteria has important implications for the possible enhancement of their potency in vivo by altering the dietary content of the cations that they translocate (Rumpler et al., 1986; Chirase et al.

Of these 61 patients, 57 with a primary infection and 4 with

Of these 61 patients, 57 with a primary infection and 4 with AZD6244 a secondary infection would otherwise be labeled as negative.[2] “
“Dengue outbreaks occur annually in Far North Queensland, Australia. Advice on topical insect repellents provided by health authorities rarely addresses the wide range of formulations and active ingredients currently registered for use in Australia. Recommendations on the use of registered products require review. Mosquito-borne disease in Australia is a major

concern.1 Since the early 1990s, there has been almost annual activity of dengue recorded from Far North Queensland, where the only species of mosquito currently present in Australia capable of transmitting dengue, Aedes aegypti (L.), is present, and culminating in one of the largest epidemics of dengue in 50 years reported during 2008 to 2009.1,2 Advice is provided to Doxorubicin clinical trial residents and tourists regarding the need to protect themselves through the use of repellents. However, there are some important differences in the personal protection advice provided

by health authorities in areas of dengue risk compared to elsewhere in the country. Australia supports a diverse mosquito fauna, but of the more than 300 species known to exist in the country relatively few pose a serious threat to public health either through nuisance-biting or transmission of disease-causing pathogens.1 The vast majority of these species are most active in host seeking at dusk and dawn with varying activity

levels during the night or in the late afternoon.1 However, the two mosquitoes capable of transmitting dengue in Australia, Ae aegypti and Aedes albopictus (Skuse) (recently introduced to the Torres Strait and may potentially spread to mainland Australia3,4), are severe nuisance-biting pests that predominantly bite humans during the day. Personal protection advice provided by local and state health authorities on websites, fact sheets, and press releases typically includes the recommended use of insect repellents, in combination with behavioral practices and physical Adenosine barriers, to prevent bites by mosquitoes. Topical repellents containing the active ingredients diethyltoluamide (DEET) and picaridin are widely recommended, represent low risk to human health, and have been demonstrated to provide effective protection from biting mosquitoes.5–7 However, the advice provided by local health authorities, with regard to both active ingredients and formulations, does not reflect the wide range of commercially available repellents currently registered with the Australian Pesticides and Veterinary Medicines Authority (APVMA). While DEET and picaridin are the most common active ingredients, botanical products containing extracts from Melaleuca spp. or Eucalyptus spp. are also widely available, but products containing botanical active ingredients and the extracts from a range of Australian native plants have been shown to provide only limited protection again A aegypti.

Because of this, the PFC can filter out distractors and up-modula

Because of this, the PFC can filter out distractors and up-modulate important sensory information before it even reaches the cortex. This type of attentional bias in the thalamus has been demonstrated in several studies

(Crick, 1984; McAlonan et al., 2006, 2008). The BF and mAChRs are also thought to influence sensory processing. click here Therefore, we tested how mAChR and BF stimulation affect between-trial correlations with and without attention applied to RF1. As indicated by comparing Fig. 11D and E (excitatory neurons), mAChR stimulation in RF1 seemed to have little effect on changing the reliability of the input. BF stimulation, however, was able to increase the reliability of both inputs to the cortex (Fig. 11, bottom). Goard & Dan (2009) also showed that stimulation of the BF leads to an increase in the reliability of neurons in the LGN and cortex. In addition, comparing Fig. 11E and F (excitatory neurons) shows that when the BF is stimulated, the reliability of RF2 increases to match that of RF1. This demonstrates that BF stimulation is able to override the attentional bias imposed onto RF1 and enhance both sensory inputs to the cortex. This happens as a result of GABAergic projections from the BF to the TRN, which have been Small Molecule Compound Library shown anatomically (Bickford et al., 1994). These projections make the BF very important for regulating the flow of information from the sensory periphery to the cortex. In

contrast to excitatory neurons, inhibitory neurons in our simulation showed hardly any increase in reliability when top-down attention was applied (Fig. 11, inhibitory neurons) and only a weak increase in reliability when the BF was stimulated (Fig. 11I and L). To see how the type of neuron affected between-trial correlations, we changed fast-spiking neurons in RF1 to regular-spiking neurons as above (Fig. 12). Comparing Fig. 12A–D with plots Fig. 11D, J, F and L, respectively, we see no significant changes. Thus, we can conclude that changing the spike waveform of inhibitory neurons appears not significantly to affect the between-trial correlations of either inhibitory or excitatory neurons.

The present model illustrates several important mechanisms underlying attention and neuronal correlations in visual cortex. First, our model accounts for the BF enhancement of both bottom-up sensory Carnitine dehydrogenase input and top-down attention through ‘local’ and ‘global’ neuromodulatory circuitry. Within the context of our model, glutamatergic projections from frontal cortex synapse onto cholinergic fibers in V1, causing local cholinergic transients, which, ultimately, lead to a local enhancement of top-down attention. In contrast, stimulation of the BF has a more global effect and can actually decrease the efficacy of top-down projections and increase sensory input by blocking top-down projections in the thalamus. Second, our model suggests an important role for mAChRs on both inhibitory and excitatory neurons.

To sum up, our results suggest that eye tracking can provide a se

To sum up, our results suggest that eye tracking can provide a sensitive, real-time, non-invasive measure of attentional fluctuations Selleck ERK inhibitor due to TOT, without

interfering with task performance or compromising safety. Decreased attentional levels can cause operators to misread or ignore incoming information, with the effect of compromising safety and job performance. Thus, there is a great need to monitor mental state in real-time in complex systems such as ATC towers, where the combination of long duty periods, insufficient sleep, monotonous tasks and high stress leads to physical and mental operator fatigue. Numerous studies have focused on assessing and/or improving ATC work conditions (McKinley et al., 2012), but fatigue-related incidents continue to occur. To address this problem, international agencies have conducted extensive research on ATC operators’ fatigue (Eurocontrol, 2012) and put in place new regulations to increase staff numbers and decrease work hours (FAA, 2012). Here we show that eye movement parameters such as (micro)saccadic and drift velocities can serve as indicators of mental fatigue. These findings are valuable because fixational eye movements occur not only during prolonged fixation but also in the intersaccadic Dapagliflozin purchase fixation periods during normal visual exploration (Otero-Millan et al., 2008; McCamy et al., 2013b). Thus, it is possible to monitor eye movement indices of mental fatigue while

operators are involved in their duty, without the need for currently used artificial oculomotor tests such as the guided saccade task (e.g. Hirvonen et al., 2010; Di Stasi et al., 2012; Ahlstrom et al., 2013). Continuous on-line eye-movement-based evaluation of ATC operators could improve safety and efficiency and reduce operational costs. This effort will require the translation of research findings and methods to ecological and complex environments to enable system

designs that maximise human–system interaction. Fixational eye movements (microsaccades and drift) and saccadic parameters can indicate mental fatigue reliably during prolonged visual search, irrespective of task complexity. These findings have potential impacts in the development of neuroergonomic tools to detect fatigue in ecological situations, and moreover suggest that fixational eye movement dynamics have heptaminol the potential to signal the nervous system’s activation state. We thank Behrooz Kousari, Peter Wettenstein and Andrew Danielson for technical assistance and Jorge Otero-Millan for his comments. We thank Dr David Riascos for his valuable advice and help with the figures. This study was supported by the Barrow Neurological Foundation (to S.L.M. and S.M.-C.), the National Science Foundation (awards 0852636 and 1153786 to S.M.-C), the Spanish Ministry of Economy and Finance (projects PSI2012 and PSI2012-39292 to J.J.C. and A.C.) and the MEC-Fulbright Postdoctoral Fellowship program (grant PS-2010-0667 to L.L.D.S.). The authors have no conflicts of interest to declare.

In addition, two flavin-binding monooxygenases were found to be u

In addition, two flavin-binding monooxygenases were found to be upregulated during growth on alkanes indicative of two novel pathways likely to be involved in alkane degradation by A. borkumensis (ABO_0282, ABO_1097, Table 1). PD0332991 cost Moreover, we detected the up-expression of two genes similar to the ones involved in the degradation of halogenated alkanes in other bacteria, namely haloacid dehalogenase-like hydrolase dhlA (ABO_1537, Table 1) and haloalkane dehalogenase dhmA (ABO_2415,

Table 1). If the first enzyme is known to convert haloalkanes to corresponding alcohols and halides, the second one catalyzes hydrolytic cleavage of carbon-halogen bonds in halogenated aliphatic compounds, leading to the formation of primary alcohols, halide ions, and protons. Alkane-induced coexpression of these enzymes mediating the breakdown of haloalkanes, alongside the induction of enzymes degrading aliphatic alkanes, signifies unspecific upregulation of expression, probably reflecting the presence of halogenated alkanes in sea water. Additionally, we found alkane-induced

expression of aldehyde reductase (ABO_2414, Table 1). This gene is predicted to be involved in the metabolic activation of polycyclic aromatic hydrocarbons (PAHs), as shown recently for human aldehyde reductase AKR1A1 (Palackal et al., 2001). However, as yet, A. borkumensis has not been shown to either degrade or transform PAHs, and thus requires further experimentation to explore what coexpression INCB018424 chemical structure of this gene alongside those mediating the degradation of aliphatic alkanes may signify for the degradation of alkanes or petroleum. These data allow us to update the list of enzymatic systems shown before by our proteomic study to be potentially involved in the initial terminal oxidation of alkanes by A. borkumensis (Figs 1 and 2). Attachment of A. borkumensis to hydrocarbons and its molecular mechanisms have not yet been studied, although such abilities are likely to form part of the specific ecological adaptation of this bacterium. EM observation of Alcanivorax SK2 indeed indicates that this organism forms biofilm-supporting structures during growth on alkanes (Fig.

selleck chemicals 3). Cells grown on alkane seem to more connect to each other rather than to the solid surface of the carrier slide, and they are shorter and rounder, and produce considerable amount of extracellular polymeric substances (EPS), which appears to support the three-dimensional structure of a biofilm. After 10 days of growth, alkane-grown cells develop a biofilm, which exhibits a pronounced three-dimensional architecture supported by extracellular matrix (Fig. 3). The argument of an alkane-induced formation of EPS is supported by alkane-induced up-expression of gmhA (ABO_0584). GmhA encodes a phosphoheptose isomerases that mediates the synthesis of heptose, a conserved component of outer membrane lipopolysaccharide, that for example in Yersinia, was shown to contribute to the formation of biofilms (Darby et al., 2005).

The only change to the method which we used for examining the pos

The only change to the method which we used for examining the posture effects within each separate experiment was that the analysis was now based on independent-samples Selleckchem JAK inhibitor t-tests which compared the Posture (Uncrossed-hands ‘UnX’ and Crossed-hands ‘X’) × Hemisphere (Contralateral ‘Con’ and Ipsilateral ‘Ipsi’) contrast waveforms observed in Experiment 1 vs. Experiment 2; such t-tests equate to the three-way interaction between Experiment,

Posture and Hemisphere. Figure 6 shows the time course of this three-way interaction according to the specific subtractive contrast: Positive values of this contrast occur when posture effects are relatively more contralaterally distributed in Experiment 1 and relatively more ipsilaterally distributed in Experiment 2. The vertical dashed line in Fig. 6 shows the onset of the significant interval. Thus, a significant effect of sight of the limbs (the variable manipulated between the two experiments) on the laterality of postural remapping started at 152 ms and was observed until the end of the interval Anti-infection Compound Library mouse tested, i.e. 200 ms (a sequence of consecutive

significant t-tests, all P < 0.05, over 38 ms in length was deemed significant by our Monte Carlo simulation). The mean first-order autocorrelation at lag 1 (estimated in our data, and used for our Monte Carlo simulations) was 0.97 for this analysis. This interaction reflects the different hemispheric distribution of postural effects observed in the two experiments reported above, and confirms that when participants have sight of the hands the first effect of posture on the SEP is observed over contralateral sites (Exp. 1), whereas when participants do not have sight of their hands the first effect of posture is observed over ipsilateral sites (Exp. 2). Keeping track of the layout of Lck one’s body and limbs is

of central importance, not just to guide action, but also in making sense of the multisensory environment (see Holmes & Spence, 2004; Bremner et al., 2008). Without processes of remapping across changes in body posture (i.e. processes which take account of movements of the limbs, the head or even the eyes in their sockets; see Pöppel, 1973), we would be hard-pressed to comprehend the spatial correspondences between stimuli which arise from the same objects, but which arrive to the brain through different sensory channels. Given the central importance of processes of postural remapping in sensory spatial representation, it is crucial to determine how and when these processes occur in the brain. To address these questions, the current study investigated how changes in body posture modulate the electrophysiological time course of somatosensory spatial processing.

Both preshaping and reaching efficiency improved with practice, w

Both preshaping and reaching efficiency improved with practice, while selective CST

lesion abrogated both. The loss of preshaping was greatest for pasta oriented vertically, suggesting loss of supination, as seen with human CST injury. The degree of preshaping loss strongly correlated with the amount of skill acquired at baseline, suggesting that the CST mediates the learned component of preshaping. Finally, the amount of preshaping lost after injury strongly correlated with reduced retrieval success, showing an important functional consequence for preshaping. We have thus demonstrated, for the first time, preshaping in the rat and dependence of this skill on the CST. Understanding the basis for this skill and measuring CP 868596 its recovery after injury will be important for studying higher-level motor control in rats. Selumetinib ic50
“Caspase 3 activation has been linked to the acute neurotoxic effects of central nervous system damage, as in traumatic brain injury or cerebral ischaemia, and also to the early events leading to long-term neurodegeneration, as in Alzheimer’s disease. However, the

precise mechanisms activating caspase 3 in neuronal injury are unclear. RhoB is a member of the Rho GTPase family that is dramatically induced by cerebral ischaemia or neurotrauma, both in preclinical models and clinically. In the current study, we tested the hypothesis that RhoB might directly modulate

caspase 3 activity and apoptotic or necrotic responses in neurons. Over-expression of RhoB in the NG108-15 neuronal cell line or in cultured corticohippocampal Methocarbamol neurons elevated caspase 3 activity without inducing overt toxicity. Cultured corticohippocampal neurons from RhoB knockout mice did not show any differences in sensitivity to a necrotic stimulus – acute calcium ionophore exposure – compared with neurons from wild-type mice. However, corticohippocampal neurons lacking RhoB exhibited a reduction in the degree of DNA fragmentation and caspase 3 activation induced by the apoptotic agent staurosporine, in parallel with increased neuronal survival. Staurosporine induction of caspase 9 activity was also suppressed. RhoB knockout mice showed reduced basal levels of caspase 3 activity in the adult brain. These data directly implicate neuronal RhoB in caspase 3 activation and the initial stages of programmed cell death, and suggest that RhoB may represent an attractive target for therapeutic intervention in conditions involving elevated caspase 3 activity in the central nervous system. “
“The effects of a GABAB agonist, baclofen, on mechanical noxious and innocuous synaptic transmission in the substantia gelatinosa (SG) were investigated in adult rats with the in vivo patch-clamp technique.

, 1999; Decker et al, 2003a, b, 2008; Hauser-Gerspach et al, 20

, 1999; Decker et al., 2003a, b, 2008; Hauser-Gerspach et al., 2007; Meier et al., 2008; Vig Slenters et al., 2008); thus only

brief descriptions of its main parts are given here. The system consists of an anaerobic flow chamber (Minucells, Bad Abbach, Germany) with (1) a test specimen mounted with its test surface not facing the flow direction; (2) a Teflon® dispenser (Multimed GmbH, Kirchheim unter Teck, Germany) containing the bacterial suspension; and (3) a peristaltic pump click here (Spetec GmbH, Erding, Germany) with an integrated speed controller. In this study, the system was modified to mimic conditions related to peri-implantitis, namely an anaerobic atmosphere and a slow-flowing, nutrient-poor environment containing three different strains of peri-implantitis-related bacteria. Specifically, the circulating bacteria were allowed to adhere to the protein-coated titanium specimens under anaerobic conditions (MACS MG; Don Whitley Scientific Ltd; atmosphere of 80% N2, 10% H2 and 10% CO2) at 37 °C for 72 h. Sterile polished disks of commercially pure titanium (Grade 2, ASTM F-67), 5 mm diameter and 1 mm thickness, with a mean surface roughness of 120 nm (Straumann AG, Basel, Switzerland), were sterilized by steam autoclaving and gamma irradiation and used as substrates. The disks were placed for 15 min in freshly mixed serum/saliva Lenvatinib mixture (1 : 10) prior

to each experiment in order to allow protein pellicle formation (Hauser-Gerspach et al., 2007). Fasting stimulated saliva of three healthy

volunteers was homogenized, filtered through a 70-μm filter (Cell Strainer; Becton Dickinson), and centrifuged at 22 000 g for 45 min at 4 °C. The supernatant was filter-sterilized (45 and 0.22 μm; Millex-HV and Millex-GV respectively; Millipore, Switzerland) and mixed with pooled serum (Blutspendezentrum, Basel, Switzerland). The protein-coated substrates were placed in the anaerobic flow chamber, 0.2% glucose was added to the bacterial suspension, and the suspension was circulated at 0.8 mL min−1 for 72 h. To compensate for the decrease in pH of the bacterial suspension (7.26 ± 0.07 to 4.84 ± 0.21), it was renewed Exoribonuclease in 24-h intervals. After 72 h, the biofilm-coated titanium disks were evaluated using SEM, CLSM, and IMC. The biofilms were fixed overnight in 2% glutaraldehyde solution (Sigma, Buchs, Switzerland), washed once with PBS, and dehydrated in stepwise increasing concentrations of ethanol – 30%, 50%, 70%, 90%, 2 × 100% for 10 min each. The samples (n = 3) were then critical-point-dried and coated with 10 nm of gold and examined (Fei Nova NanoSEM 230®, Eindhoven, the Netherlands). Oligonucleotide DNA probes, labeled at the 5′-end with Cy3 and Cy5 or with 6-carboxyfluorescein (FAM) and additionally labeled at the 3′-end (Microsynth AG, Balgach, Switzerland), are listed with their sequences and specificities in Table 1.

, 1996) Also, cystatins, a superfamily of cysteine PI in human s

, 1996). Also, cystatins, a superfamily of cysteine PI in human saliva, is known to interfere with the growth of oral bacteria such as Porphyromonas gingivalis (Blankenvoorde et al., 1998). Synthetic PIs have been developed against a number of proteolytic enzymes as potential antibiotics to retard the growth http://www.selleckchem.com/products/MG132.html and proliferation of bacterial pathogens and viruses. Based on human cysteine protease inhibitors, Björck et al. (1989) synthesized a peptide derivative known as Z-LVG-CHN2 and showed its specific inhibitory effect on the growth of group A streptococci strains, both in vivo and in vitro. Aprotinin was found to have antibacterial activity by its ability

to permeate the cell walls of Gram-positive and Gram-negative bacteria and disintegrate

the cytoplasm (Pellegrini et al., 1992). Lopes et al. (1999) pointed out the direct correlation between the action of aprotinin and inhibiting growth of the Gram-positive bacterium Streptomyces alboniger. The growth of P. gingivalis and Fusobacterium nucleatum was specifically inhibited by protease inhibitors, such as bestatin (Rogers et al., 1998; Grenier et al., 2001a). Because protease inhibitors are widely added during standard purification procedures for proteomic studies, we examined whether such a protocol might ultimately affect our ability to qualitatively and quantitatively evaluate ubiquitin-Proteasome system the growth and diversity of oral bacteria. To address this question, we used a commercially available PI cocktail that consists of the serine protease inhibitors AEBSF and aprotinin, the cysteine protease inhibitor E-64, the serine and cysteine Tolmetin protease inhibitor leupeptin, the amino-protease inhibitor bestatin, and the aspartic acid protease inhibitor pepstatin A. Based on previously published studies, we hypothesized that the cocktail would affect total cultivable bacterial growth and, therefore, would interfere

with the evaluation of bacterial composition in whole oral saliva samples. Unexpectedly, however, the results of our study showed that neither oral bacterial counts nor DGGE profiles of the bacterial composition with protease inhibitors displayed significant statistical differences when compared with the nonprotease inhibitor group, suggesting that the addition of PI in saliva samples has no effect on either the growth or the composition of oral microbiota. Pellegrini et al. (1990) tested the antibacterial properties of a wide variety of protease inhibitors and found that most did not inhibit the growth of bacteria. Their observation suggested that the concentration of aprotinin and bestatin might dictate selective antibacterial activities. For instance, bestatin was only found to completely affect the proliferation of P. gingivalis in the oral cavity at a concentration of 2.5 μg mL−1 (Grenier & Michaud, 1994).