Evaluation of omadacycline against intracellular Mycobacterium abscessus in an infection model in human macrophages.

Background: Omadacycline is an aminomethylcycline antibiotic in the tetracycline class that was approved by the US FDA in 2018 for the treatment of community-acquired bacterial pneumonia and acute bacterial skin and skin structure infections. It is available in both IV and oral formulations. Omadacycline has broad-spectrum in vitro activity and clinical efficacy against infections caused by Gram-positive and Gram-negative pathogens. Omadacycline is being evaluated in a 3 month placebo-controlled Phase 2 clinical trial of oral omadacycline versus placebo in adults with non-tuberculous mycobacteria (NTM) pulmonary disease caused by Mycobacterium abscessus (NCT04922554). Objectives: To determine if omadacycline has intracellular antimicrobial activity against NTM, bacteria that can cause chronic lung disease, in an ex vivo model of intracellular infection. Methods: Two strains of M. abscessus were used to infect THP-1 macrophages. Intracellular M. abscessus was then challenged with omadacycline and control antibiotics at multiples of the MIC over time to evaluate intracellular killing. Results: At 16 ×  the MIC at 72 h, omadacycline treatment of intracellular NTM yielded a log10 reduction in cfu of 1.1 (91.74% reduction in cfu) and 1.6 (97.65% reduction in cfu) consistent with killing observed with tigecycline, whereas amikacin and clarithromycin at 16 ×  the MIC did not show any reduction in cfu against the intracellular M. abscessus. Conclusions: Omadacycline displayed intracellular activity against M. abscessus within macrophages. The activity was similar to that of tigecycline; as expected, intracellular killing was not observed with clarithromycin and amikacin.

Effects of oil sands process water mixtures on the mayfly Hexagenia and field-collected aquatic macroinvertebrate communities.

Extraction of Canada's oil sands has created 1 billion m3 of tailings, which are stored in on-site tailings ponds. Due to limited storage capacity, the planned release of tailings into the surrounding environment may be required. This represents an environmental management challenge, as the tailings contain contaminants that are known toxins to aquatic communities. Of particular concern are naphthenic acids and their metallic counterparts, as they are the principal toxic components of tailings, are relatively soluble, and are persistent in aquatic environments. This study examines the acute toxicity of environmentally relevant 10:1 mixtures of two process water components: naphthenic acid and sodium naphthenate. We assess the effects of these simplified oil sands process water (OSPW) mixtures under planned and unplanned tailings release scenarios, using traditional and cutting-edge bioindicators for aquatic invertebrate taxa. We found that safe concentrations for mayflies and other aquatic macroinvertebrates were less than 1 mg/l, as no mayfly taxa survived repeated exposure to this dose in either the 48-h or 72-h acute toxicity test. In the 72-h test, no mayflies survived treatment levels greater than 0.5 mg sodium naphthenate/l. In the mesocosm study, even a 90% dilution of the OSPW mixture was not sufficient to protect sensitive macroinvertebrate communities. The results of this study highlight the potential environmental damage that will occur if OSPW is not carefully managed. This information will aid with the development of a management plan for oil sands tailings ponds, which will provide insight into the potential for process water release into the surrounding environment while conserving unique ecosystems downstream of development in the oil sands region.

Chronic maternal hyperglycemia induced during mid-pregnancy in rats increases RAGE expression, augments hippocampal excitability, and alters behavior of the offspring.

Maternal diabetes during pregnancy may increase the risk of neurodevelopmental disorders in the offspring by increasing inflammation. A major source of inflammatory signaling observed in diabetes is activation of the receptor for advanced glycation end-products (RAGE), and increased RAGE expression has been reported in psychiatric disorders. Thus, we sought to examine whether maternal diabetes creates a proinflammatory state, triggered largely by RAGE signaling, that alters normal brain development and behavior of the offspring. We tested this hypothesis in rats using the streptozotocin (STZ; 50mg/kg; i.p.) model of diabetes induced during mid-pregnancy. Following STZ treatment, we observed a significant increase in RAGE protein expression in the forebrain of the offspring (postnatal day 1). Data obtained from whole-cell patch clamping of hippocampal neurons in cultures from the offspring of STZ-treated dams revealed a striking increase in excitability. When tested in a battery of behavioral tasks in early adulthood, the offspring of STZ-treated dams had significantly lower prepulse inhibition, reduced anxiety-like behavior, and altered object-place preference when compared to control offspring. In an operant-based strategy set-shifting task, STZ offspring did not differ from controls on an initial visual discrimination or reversal learning but took significantly longer to shift to a new strategy (i.e., set-shift). Insulin replacement with an implantable pellet in the dams reversed the effects of maternal diabetes on RAGE expression, hippocampal excitability, prepulse inhibition and object-place memory, but not anxiety-like behavior or set-shifting. Taken together, these results suggest that chronic maternal hyperglycemia alters normal hippocampal development and behavior of the offspring, effects that may be mediated by increased RAGE signaling in the fetal brain.

Altered object-in-place recognition memory, prepulse inhibition, and locomotor activity in the offspring of rats exposed to a viral mimetic during pregnancy.

Infection during pregnancy (i.e., prenatal infection) increases the risk of psychiatric illnesses such as schizophrenia and autism in the adult offspring. The present experiments examined the effects of prenatal immune challenge on behavior in three paradigms relevant to these disorders: prepulse inhibition (PPI) of the acoustic startle response, locomotor responses to an unfamiliar environment and the N-methyl-d-aspartate antagonist MK-801, and three forms of recognition memory. Pregnant Long-Evans rats were exposed to the viral mimetic polyinosinic-polycytidylic acid (PolyI:C; 4 mg/kg, i.v.) on gestational day 15. Offspring were tested for PPI and locomotor activity before puberty (postnatal days (PNDs)35 and 36) and during young adulthood (PNDs 56 and 57). Four prepulse-pulse intervals (30, 50, 80, and 140 ms) were employed in the PPI test. Recognition memory testing was performed using three different spontaneous novelty recognition tests (object, object location, and object-in-place recognition) after PND 60. Regardless of sex, offspring of PolyI:C-treated dams showed disrupted PPI at 50-, 80-, and 140-ms prepulse-pulse intervals. In the prepubescent rats, we observed prepulse facilitation for the 30-ms prepulse-pulse interval trials that was selectively retained in the adult PolyI:C-treated offspring. Locomotor responses to MK-801 were significantly reduced before puberty, whereas responses to an unfamiliar environment were increased in young adulthood. Both male and female PolyI:C-treated offspring showed intact object and object location recognition memory, whereas male PolyI:C-treated offspring displayed significantly impaired object-in-place recognition memory. Females were unable to perform the object-in-place test. The present results demonstrate that prenatal immune challenge during mid/late gestation disrupts PPI and locomotor behavior. In addition, the selective impairment of object-in-place recognition memory suggests tasks that depend on prefrontal cortex may be particularly vulnerable following prenatal immune challenge.

Hip protectors: recommendations for conducting clinical trials--an international consensus statement (part II).

INTRODUCTION: While hip protectors are effective in some clinical trials, many, including all in community settings, have been unable to demonstrate effectiveness. This is due partly to differences in the design and analysis. The aim of this report is to develop recommendations for subsequent clinical research. METHODS: In November of 2007, the International Hip Protector Research Group met to address barriers to the clinical effectiveness of hip protectors. This paper represents a consensus statement from the group on recommended methods for conducting future clinical trials of hip protectors. RESULTS AND CONCLUSIONS: Consensus recommendations include the following: the use of a hip protector that has undergone adequate biomechanical testing, the use of sham hip protectors, the conduct of clinical trials in populations with annual hip fracture incidence of at least 3%, a run-in period with demonstration of adequate adherence, surveillance of falls and adherence, and the inclusion of economic analyses. Larger and more costly clinical trials are required to definitively investigate effectiveness of hip protectors.

Hip protectors: recommendations for biomechanical testing--an international consensus statement (part I).

INTRODUCTION: Hip protectors represent a promising strategy for preventing fall-related hip fractures. However, clinical trials have yielded conflicting results due, in part, to lack of agreement on techniques for measuring and optimizing the biomechanical performance of hip protectors as a prerequisite to clinical trials. METHODS: In November 2007, the International Hip Protector Research Group met in Copenhagen to address barriers to the clinical effectiveness of hip protectors. This paper represents an evidence-based consensus statement from the group on recommended methods for evaluating the biomechanical performance of hip protectors. RESULTS AND CONCLUSIONS: The primary outcome of testing should be the percent reduction (compared with the unpadded condition) in peak value of the axial compressive force applied to the femoral neck during a simulated fall on the greater trochanter. To provide reasonable results, the test system should accurately simulate the pelvic anatomy, and the impact velocity (3.4 m/s), pelvic stiffness (acceptable range: 39-55 kN/m), and effective mass of the body (acceptable range: 22-33 kg) during impact. Given the current lack of clear evidence regarding the clinical efficacy of specific hip protectors, the primary value of biomechanical testing at present is to compare the protective value of different products, as opposed to rejecting or accepting specific devices for market use.

Amygdaloid kindling is anxiogenic but fails to alter object recognition or spatial working memory in rats.

Kindling in rats produces enduring behavioral changes that parallel the psychobehavioral disturbances frequently accompanying temporal lobe epilepsy. Some evidence suggests that the site of kindling is an important determinant of the type of behavioral changes observed following kindling, although this variable has not been systematically investigated. In the present experiments, the effects of amygdaloid kindling were assessed on a battery of behavioral tests we used previously to assess the effects of kindling in dorsal hippocampus or perirhinal cortex. Three generalized seizures were kindled with stimulation in or near the basolateral amygdala. One week later, rats were tested successively on measures of anxiety, activity, object recognition memory, and spatial working memory over a period of 3 weeks. Amygdaloid kindling produced increased anxiety, but spared all other behaviors assessed. This pattern of results is partially distinct from the previously described effects of perirhinal cortical kindling, which increases anxiety but also impairs object recognition memory, and is completely distinct from dorsal hippocampal kindling, which selectively increases activity and impairs spatial working memory. The observations suggest that kindling of distinct highly interconnected temporal lobe sites produces distinct patterns of behavioral comorbidity. The underlying mechanisms are thus most likely localized to intrinsic circuits at the site of seizure origination.

Anterior perirhinal cortex kindling produces long-lasting effects on anxiety and object recognition memory.

Temporal lobe epilepsy (TLE) is frequently accompanied by memory impairments and, although their bases are unknown, most research has focused on the hippocampus. The present study investigated the importance of another medial temporal lobe structure, the perirhinal cortex (Prh), in changes in memory in TLE using kindling as a model. Rats were kindled twice daily with anterior Prh stimulation until three fully generalized seizures were evoked. Beginning 7 days later and on successive days, rats were tested in an elevated plus maze, a large circular open field, an open field object exploration task and a delayed-match-to-place task in a water maze in order to assess anxiety-related and exploratory behaviour, object recognition memory and spatial cognition. Kindling increased anxiety-related behaviour in both the elevated plus and open field mazes and disrupted spontaneous object recognition but spared all other behaviours tested. These results are consistent with other findings indicating a greater role for the Prh in object memory and emotional behaviour than in spatial memory and contrast with the selective disruption of spatial memory produced by dorsal hippocampal kindling. The site-selectivity of the behavioural disruptions produced by kindling indicates that such effects are probably mediated by changes particular to the site of seizure initiation rather than to changes in the characteristic circuitry activated by limbic seizure generalization. Further investigation of the behavioural effects of Prh kindling may be useful for studying the mechanisms of mnemonic and affective dysfunction associated with TLE and offer insights into bases for variability in such dysfunction across patients.

Medial prefrontal cortex is involved in spatial temporal order memory but not spatial recognition memory in tests relying on spontaneous exploration in rats.

The present study describes two novel tasks relying on spontaneous patterns of exploration in a radial-arm maze that can be used to assess spatial recognition memory and spatial temporal order memory (i.e. memory for the order in which places have been visited) in the rat. In the recognition memory task, rats were permitted to freely explore two arms in the maze on a first trial and one 'familiar' arm and one novelly located arm on a second trial 105 min later. In the temporal order memory task, rats were permitted to explore two arms in the maze on a first trial, two novel arms on a second trial 60 min later, and one 'older familiar' arm and one 'more recent familiar' arm on a third trial 45 min later. Using these tasks, we found that rats direct greater exploration at a novel than a familiar arm location, thus showing long-term spatial recognition memory, and at an older familiar arm than a more recent familiar arm, thus showing long-term spatial temporal order memory. Lidocaine inactivation of the mPFC prior to the final trial in each task disrupted performance on the temporal order but not the recognition memory task, thereby demonstrating a role for the mPFC in the retrieval and/or use of temporal order information but not in spatial memory per se. These findings highlight the specific involvement of the rat mPFC in temporal order memory and have important implications for a broader understanding of mPFC function.

Microbial survivors: thermophiles, halophiles, and other prodigies.

When biologists encounter microbes flourishing in boiling water and other extreme habitats, they often consider such creatures as merely odd, and only search for possible protective mechanisms. But it may also be that extreme habitats resemble those where life first occurred, and that such organisms provide links with earlier evolution.

Dorsal hippocampal kindling produces a selective and enduring disruption of hippocampally mediated behavior.

Kindling produces enduring neural changes that are subsequently manifest in enhanced susceptibility to seizure-evoking stimuli and alterations in some types of behavior. The present study investigated the effects of dorsal hippocampal (dHPC) kindling on a variety of behaviors to clarify the nature of previously reported effects on spatial task performance. Rats were kindled twice daily with dHPC stimulation until three fully generalized seizures were evoked. Beginning 7 d later and on successive days, rats were tested in an elevated plus maze, a large circular open field, an open field object exploration task, and a delayed-match-to-place (DMTP) task in a water maze to assess anxiety-related and activity-related behavior (tasks 1 and 2), object recognition memory (task 3), and spatial cognition (task 4). Kindling disrupted performance on the DMTP task in a manner that was not delay dependent and produced a mild enhancement of activity-related behaviors in the open field task but not the elevated plus maze. All other aspects of testing were spared. These findings indicate that dHPC kindling produces enduring and selective effects on behavior that are consistent with a restricted disruption of hippocampally mediated functions. Possible bases for these effects are changes in local NMDA receptor function and/or changes in local inhibition, which might alter the optimal conditions for experience-dependent induction of intrahippocampal plasticity. This preparation may be useful for studying the mechanisms of mnemonic dysfunction associated with temporal lobe epilepsy and may offer unique insights into the mechanisms underlying normal hippocampal function.

Susceptibility to kindling and neuronal connections of the anterior claustrum.

The claustrum has been implicated in the kindling of generalized seizures from limbic sites. We examined the susceptibility of the anterior claustrum itself to kindling and correlated this with an anatomical investigation of its afferent and efferent connections. Electrical stimulation of the anterior claustrum resulted in a pattern of rapid kindling with two distinct phases. Early kindling involved extremely rapid progression to bilaterally generalized seizures of short duration. With repeated daily kindling stimulations, early-phase generalized seizures abruptly became more elaborate and prolonged, resembling limbic-type seizures as triggered from the amygdala. We suggest that the rapid rate of kindling from the anterior claustrum is an indication that the claustrum is functionally close to the mechanisms of seizure generalization. In support of our hypothesis, we found significant afferent, efferent, and often reciprocal connections between the anterior claustrum and areas that have been implicated in the generation of generalized seizures, including frontal and motor cortex, limbic cortex, amygdala, and endopiriform nucleus. Additional connections were found with various other structures, including olfactory areas, nucleus accumbens, midline thalamus, and brainstem nuclei including the substantia nigra and the dorsal raphe nucleus. The anatomical connections of the anterior claustrum are consistent with its very high susceptibility to kindling and support the view that the claustrum is part of a forebrain network of structures participating in the generalization of seizures.

Personal values and sexual decision-making among virginal and sexually experienced urban adolescent girls.

PURPOSE: To guide the development of an intervention to reduce the incidence of sexually transmitted diseases (STDs) in urban, adolescent girls, we investigated such girls' reasons for deciding to have or not to have sexual intercourse. METHODS: Consecutive girls >or=14 years of age attending an urban adolescent clinic were invited to complete an anonymous survey about sexual decision-making. In this pilot study, girls were asked: (a) whether they agreed with a statement that they had or had not had sexual intercourse "because of my values and beliefs"; and (b) to select from a list one or more specific reasons why they had or had not had intercourse. The girls were categorized by self-report as either "virgins," "currently inactive" (no intercourse in the preceding 3 months), or "currently active" (had intercourse during the preceding 3 months). RESULTS: Usable surveys were obtained from 197 adolescents whose age (18.2 +/- 2.6 years) and race (69% black) were comparable to those of clinic attendees in general. Forty girls (20%; age 16.1 +/- 2.1 years) were virgins, 25 girls (13%; age 17.8 +/- 2.3 years) were inactive, and 132 girls (67%; age 18.9 +/- 2.5 years) were currently active. "Values and beliefs" were cited as the reason for decisions about sexual behavior by 53% of the virgins, but only by 24% of the sexually inactive and 24% of the sexually active girls (p = .002). Virgins were more likely than inactive girls to cite three specific reasons for not having sex: "not the right thing for me now" (82% vs. 50%, p = .007), "waiting until I am older" (69% vs. 8%, p = .001), and "waiting until I am married" (67% vs. 38%, p = .02). The reason "against my religious beliefs" was cited by 23% of virgins and 13% of inactive girls (p = not significant). Personal values were implicit in the two specific reasons for having sex that active girls chose most frequently, namely, "I like/love the person" (86%) and "I like having sex" (37%), although only 24% of these girls had explicitly cited "values and beliefs" as their reason for having sex. CONCLUSIONS: Our data indicate that urban girls, both those who have had sexual intercourse and those who have not, view their sexual behavior as being based on personal (although infrequently religious) values. Many of the virginal urban, adolescent girls we surveyed hold abstinence as a personal value. The sexually active adolescents perceive the decision to have sexual intercourse as being based affirmatively on their personal values rather than on the chance occurrence of opportunities to have intercourse. These data may be useful in the development of new strategies for reducing urban adolescent girls' risk of acquiring sexually transmitted diseases.

Initial sequencing and analysis of the human genome.

The human genome holds an extraordinary trove of information about human development, physiology, medicine and evolution. Here we report the results of an international collaboration to produce and make freely available a draft sequence of the human genome. We also present an initial analysis of the data, describing some of the insights that can be gleaned from the sequence.

Effects of low-dose alcohol exposure on simulated merchant ship piloting by maritime cadets.

The US Department of Transportation (DOT) regulates on-the-job alcohol use by operators of certain categories of commercial transport. For aircraft, trains, and commercial vessels, operators are subject to sanctions for having > or = 0.04 g% blood alcohol concentration (BAC). This study examines the effects of alcohol (between 0.04 and 0.05 g% BAC) on simulated merchant ship handling. A two-group randomized factorial design was used to compare beverage alcohol to placebo while controlling for baseline performance on a previous day. The study was conducted in the Maritime Simulation Center at Maine Maritime Academy, Castine, ME. Participants were 38 volunteer deck officer cadets in their junior or senior year, at least 21 years of age, with previous experience on a bridge simulator. Following a baseline trial on Day 1, on Day 2 participants were randomized to receive alcohol (0.6 g/kg for males and 0.5 g/kg for females) or placebo. After allowing time for absorption, participants completed a bridge simulator task. For baseline and performance trials, participants were randomized to one of four bridge simulator scenarios, each representing passage of a fully loaded container vessel through a channel with commercial traffic. The aggregate scenario score given by blinded maritime educators measured performance. A main effect for alcohol was found indicating that performance was significantly impaired by this low dose of alcohol relative to performance in the placebo condition. These findings are consistent with current federal regulations that limit low-dose alcohol exposure for the operators of commercial transport vehicles. Further research is required to determine effects at lower BACs.

Age of drinking onset and unintentional injury involvement after drinking.

CONTEXT: In 1997, unintentional injury was the leading cause of death for persons aged 1 to 34 years. Approximately one third of deaths due to unintentional injury in the United States are estimated to be alcohol related. Onset of drinking at an early age has been found to be associated with alcohol dependence, but whether early-onset drinking increases risk for unintentional injury while drinking is unknown. OBJECTIVE: To explore whether persons who started drinking at an early age are more likely to have experienced unintentional injuries while under the influence of alcohol. DESIGN AND SETTING: The National Longitudinal Alcohol Epidemiology Survey, a cross-sectional survey conducted in 1992 of a representative sample of the US population. PARTICIPANTS: A total of 42,862 randomly selected adults (response rate, 90%; mean age, 44 years). MAIN OUTCOME MEASURES: Unintentional injury involvement while under the influence of alcohol by age of drinking onset (categorized as <14 years, each age from 14-20 years, or >/=21 years). RESULTS: Relative to respondents who began drinking at age 21 years or older, those who started before age 14 years as well as those who started at each intervening age up to 21 years were significantly more likely to have been injured while under the influence of alcohol, even after controlling for history of alcohol dependence, heavy drinking frequency during the period that they drank most, family history of alcoholism, and other characteristics associated with earlier onset of drinking. After adjusting for these variables, odds ratios for having been injured while under the influence of alcohol were as follows: for younger than 14 years, 2.98 (95% confidence interval [CI], 2.29-3.89); age 14 years, 2.96 (95% CI, 2.26-3.88); age 15 years, 3.14 (95% CI, 2.48-3.97); age 16 years, 2.38 (95% CI, 1.90-2.98); age 17 years, 2.12 (95% CI, 1.66-2.71); age 18 years, 1. 33 (95% CI, 1.08-1.64); age 19 years, 1.42 (95% CI, 1.07-1.89); and age 20 years, 1.39 (95% CI, 1.01-1.91). CONCLUSION: Drinking onset at ages younger than 21 years is associated with having experienced alcohol-related injuries. JAMA. 2000;284:1527-1533

Effects of low-dose alcohol exposure on simulated merchant ship handling power plant operation by maritime cadets.

AIMS: Under current US Department of Transportation (DOT) regulations, some commercial vehicle operators are subject to sanctions for having > or = 00.04 g% blood alcohol concentrations (BACs) on the job. This study investigated the effects of alcohol (between 0.04 and 0.05 g% BAC) on the simulated operation of a commercial ship's power plant. DESIGN: A balanced placebo design was used in which alcohol administration was fully crossed with expectancy that alcohol was administrated. SETTING: The study was conducted at the Massachusetts Maritime Academy, Buzzards Bay, MA, USA. PARTICIPANTS: Participants were 18 volunteer engineering students in their senior year, at least 21 years of age, with previous experience on the diesel simulator. INTERVENTION: Following a baseline trial on day 1, on day 2 participants were randomized to expectancy (told alcohol or placebo) and beverage (receive alcohol or placebo). The dosage was 0.6 g/kg for males and 0.5 g/kg for females. In both baseline and performance days, participants were randomized to one of four diesel simulator scenarios, each replicating a system failure in a power plant subsystem (e.g. main propulsion, electrical generating). MEASUREMENT: Performance was measured by time required for problem identification and remediation. FINDINGS: A main effect for alcohol administration was found. In the alcohol condition, simulator performance time was almost twice as long (351 sec) as on the placebo condition (186 sec). The expectancy effect was not significant. CONCLUSION: These findings support the current federal policy on the hazards of this low alcohol level within the context of commercial shipping.

Boater training, drinking and boating, and other unsafe boating practices.

OBJECTIVES: Identify associations between lack of formal boater training, drinking and boating, and other unsafe boating practices. METHODS: A telephone survey queried respondents (age 16 or older in continental United States) about boating experience, type of boat used, and training. RESULTS: Of the 3,042 boaters surveyed (70% response), most had no formal training (73%). Boaters with formal training failed to use PFDs about as often as those without formal training and were equally or more likely to use alcohol while boating. CONCLUSIONS: The unexpected association between formal training and unsafe boating practices is probably due to reduced risk perception and inadequacies of boater training programs. Such programs seldom mention the risks of alcohol use while boating. Decisions to mandate formal training should be informed by these results; if mandated, training should address the risks of alcohol use while boating, and should be renewed frequently enough to offset reductions in risk perception.