Biomimetic Chemical Reactions with Natural Products Using Metalloporphyrins and Salen Complexes as Catalysts: a Brief Review.

The cytochrome P450 is a superfamily of hemoproteins mainly present in the liver and are versatile biocatalysts. They participate in the primary metabolism and biosynthesis of various secondary metabolites. Chemical catalysts are utilized to replicate the activities of enzymes. Metalloporphyrins and Salen complexes can contribute to the products' characterization and elucidate biotransformation processes, which are investigated during pre-clinical trials. These catalysts also help discover biologically active compounds and get better yields of products of industrial interest. This review aims to investigate which natural product classes are being investigated by biomimetic chemical models and the functionalities applied in the use of these catalysts. A limited number of studies were observed, with terpenes and alkaloids being the most investigated natural product classes. The research also revealed that Metalloporphyrins are still the most popular in the studies, and the identity and yield of the products obtained depend on the reaction system conditions.

Nanoemulsions and Solid Microparticles Containing Pentyl Cinnamate to Control Aedes aegypti.

The Aedes aegypti mosquito is a vector of severe diseases with high morbidity and mortality rates. The most commonly used industrial larvicides have considerable toxicity for non-target organisms. This study aimed to develop and evaluate liquid and solid carrier systems to use pentyl cinnamate (PC), derived from natural sources, to control Ae. aegypti larvae. The liquid systems consisting of nanoemulsions with different lecithins systems were obtained and evaluated for stability over 30 days. Microparticles (MPs) were obtained by the spray drying of the nanoemulsions using maltodextrin as an adjuvant. Thermal, NMR and FTIR analysis indicated the presence of PC in microparticles. Indeed, the best nanoemulsion system was also the most stable and generated the highest MP yield. The PC larvicidal activity was increased in the PC nanoemulsion system. Therefore, it was possible to develop, characterize and obtain PC carrier systems active against Ae. aegypti larvae.

The fly liquid-food electroshock assay (FLEA) suggests opposite roles for neuropeptide F in avoidance of bitterness and shock.

BACKGROUND: Proper regulation of feeding is important for an organism's well-being and survival and involves a motivational component directing the search for food. Dissecting the molecular and neural mechanisms of motivated feeding behavior requires assays that allow quantification of both motivation and food intake. Measurements of motivated behavior usually involve assessing physical effort or overcoming an aversive stimulus. Food intake in Drosophila can be determined in a number of ways, including by measuring the time a fly's proboscis interacts with a food source associated with an electrical current in the fly liquid-food interaction counter (FLIC). Here, we show that electrical current flowing through flies during this interaction is aversive, and we describe a modified assay to measure motivation in Drosophila. RESULTS: Food intake is reduced during the interaction with FLIC when the electrical current is turned on, which provides a confounding variable in studies of motivated behavior. Based on the FLIC, we engineer a novel assay, the fly liquid-food electroshock assay (FLEA), which allows for current adjustments for each feeding well. Using the FLEA, we show that both external incentives and internal motivational state can serve as drivers for flies to overcome higher current (electric shock) to obtain superior food. Unlike similar assays in which bitterness is the aversive stimulus for the fly to overcome, we show that current perception is not discounted as flies become more food-deprived. Finally, we use genetically manipulated flies to show that neuropeptide F, an orthologue of mammalian NPY previously implicated in regulation of feeding motivation, is required for sensory processing of electrical current. CONCLUSION: The FLEA is therefore a novel assay to accurately measure incentive motivation in Drosophila. Using the FLEA, we also show that neuropeptide F is required for proper perception or processing of an electroshock, a novel function for this neuropeptide involved in the processing of external and internal stimuli.

Temporal Trends in Polybrominated Diphenylethers (PBDEs) in Blubber of Ringed Seals (Pusa hispida) from Ulukhaktok, NT, Canada Between 1981 and 2015.

Blubber taken from ringed seals (Pusa hispida) during a subsistence hunt at Ulukhaktok, NT (formerly Holman, NWT) at intervals between 2002 and 2015 was analysed for polybrominated diphenyl ether (PBDE) congeners. Results from these analyses were combined with others previously published to yield a data set of 18 tri- to hepta-substituted PBDE congeners in 102 animals sampled over a span of 19 year (females) and 34 year (males). In females, mean total PBDE concentrations increased between 1996 and 2015 by approximately 50%, from 1940 to 2780 pg/g wet wt., although not significantly so (p > 0.05) by one-way ANOVA. In males, concentrations ranged from 376 to 6470 pg/g wet wt. between 1981 and 2015 (p < 0.05). In males, the most rapid increase in PBDE concentrations occurred before 2000, but between 2002 and 2015 mean total PBDE concentrations increased by a further 50%. ANCOVA showed PBDE concentrations in females to be correlated (p < 0.05) with sampling year but not with age or condition (as measured by blubber thickness); in males, PBDE concentrations were strongly correlated (p < 0.01) with year, age and condition. The relative proportions of tetra-bromo- congeners declined weakly in both sexes over the sampling period, with a compensatory increase in penta-bromo-congener distribution. Overall, the results show no evidence yet of a decline in PBDE concentrations in western Arctic ringed seals in response to voluntary and regulated restrictions on PBDE use in the early 2000s.

Kinematic Analysis of Combined Suture-Button and Suture Anchor Augment Constructs for Ankle Syndesmosis Injuries.

BACKGROUND: Syndesmosis injuries are common, with up to 25% of all ankle injuries being reported to involve an associated syndesmosis injury. These injuries are typically treated with cortical screw fixation or suture-button implants when indicated, but the addition of a suture anchor augment implant has yet to be evaluated. The purpose of this study was to evaluate the ability of a suture anchor augment to add sagittal plane translational and transverse plane rotational constraint to suture-button constructs with syndesmosis injuries. We hypothesized that the suture anchor augment oriented in parallel with the fibers of an injured anterior-inferior tibiofibular ligament (AITFL) in addition to a suture-button construct would achieve physiological motion and stability at the syndesmosis through increased rotational and translational constraint of the fibula. METHODS: Eleven fresh-frozen cadaver ankles were stressed in external rotation using a custom-made ankle rig. Each ankle had simultaneous recording of ultrasound video, 6 degrees-of-freedom kinematics of the fibula and tibia, and torque as the ankle was stressed by an examiner. The ankles were tested in 6 different states: native uninjured; injured with interosseous ligament and AITFL sectioned; 1× suture button; 2× suture buttons, divergent; 1× suture anchor augment with 2× suture buttons, divergent; and 1× suture anchor augment with 1× suture buttons. RESULTS: Only the suture anchor augment + 2× suture buttons and suture anchor augment + 1× suture-button constructs were found to be significantly different from the injured state (P = .0003, P = .002) with mean external rotation of the fibula. CONCLUSION: Overall, the most important finding of this study was that the addition of a suture anchor augment to suture-button constructs provided a mechanism to increase external rotational constraint of the fibula. CLINICAL RELEVANCE: This study provides a mechanistic understanding of how the combined suture-button and suture anchor augment construct provides an anatomically similar reconstruction of constraints found in the native ankle. However, none of the constructs examined in this study were able to fully restore physiologic motion.

Trends of persistent organic pollutants in ringed seals (Phoca hispida) from the Canadian Arctic.

Ringed seals (Phoca hispida) have been used as bioindicator species of environmental contamination in Canada since the 1970s. In the present study, seals were harvested during subsistence hunts in four regions of the Canadian Arctic: Beaufort Sea, Arctic Archipelago, Hudson Bay, and coastal Labrador. An extensive suite of persistent organic pollutants (POPs) was determined in seal blubber collected for multiple years between 1972 and 2016. Results from this long-term study indicate geographical differences in the contaminant concentrations in seals and the significant general decrease of most POPs, including polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and related compounds, chlordanes (CHL), and hexachlorocyclohexanes (HCH) over time in ringed seals. The highest decrease rates (up to -9.1%/year for α-HCH) were found in seals from the Hudson Bay region where all chemicals investigated have significantly decreased since 1986. Significant increases in concentrations of hexachlorobenzene (HCB) in seals from Labrador and ß-HCH in Sachs Harbour, NT and Arctic Archipelago were observed. Site-specific and contaminant-specific associations between climate pattern (i.e., Arctic Oscillation, North Atlantic Oscillation, and Pacific/North American pattern) and mean ice-coverage (total, first-year ice, and old-ice) were found at sites with the longest time trend data (i.e., Arviat, Sachs Harbour/Ulukhaktok and Resolute Bay). Overall, results suggest that North American and international regulations have led to the long-term reduction of most POPs in Canadian Arctic ringed seals by reducing emissions from primary sources. However, other sources of legacy compounds (e.g., environmental reservoirs) as well changes in food web composition and structure in relation to climate changes could also be influencing the very slow rates of decline, or stable levels, of contaminants found in seals at some sites. Further work is warranted to discern between co-variation of climate changes and contaminant concentrations and cause-and-effect relationships.

Protecting the PCL During Total Knee Arthroplasty Using a Bone Island Technique.

BACKGROUND: Prior studies have shown that the posterior cruciate ligament (PCL) may be partially resected during cruciate retaining (CR) total knee arthroplasty (TKA) using highly experienced hands and standard surgical technique; therefore, proper surgical technique is aimed at preservation and balance of the PCL during CR TKA. The central objective of this study is to evaluate the effectiveness of a simple surgical technique to prevent PCL damage during performance of a CR TKA. METHODS: Sixty embalmed cadaver specimens were randomized into 2 groups, experimental and control. The control group consisted of standard tibial resection without the use of an osteotome. The experimental group utilized an osteotome in addition to standard technique to preserve a bone island anterior to the tibial attachment of the PCL. RESULTS: In the control group, PCL damage was noted in 73% (22/30) of specimens. In the experimental group, where an osteotome was used, PCL damage was found in 23% (7/30) of specimens. The use of an osteotome was found to have an absolute risk reduction of 50% when compared to the control group which did not use an osteotome to protect the PCL. CONCLUSION: In the setting of minimal surgical experience, the use of an osteotome to preserve the PCL during CR TKA by forming a bone island was found to be an effective means of protecting the PCL over standard technique. In addition, standard technique with the use of a Y-shaped PCL retractor was found to provide questionable protection to the PCL.

Determining molecular orientation via single molecule SERS in a plasmonic nano-gap.

In this work, plasmonic nano-gaps consisting of a silver nanoparticle coupled to an extended silver film have been fully optimized for single molecule Surface-Enhanced Raman Scattering (SERS) spectroscopy. The SERS signal was found to be strongly dependent on the particle size and the molecule orientation with respect to the field inside the nano-gap. Using Finite Difference Time Domain (FDTD) simulations to complement the experimental measurements, the complex interplay between the excitation enhancement and the emission enhancement of the system as a function of particle size were highlighted. Additionally, in conjunction with Density Functional Theory (DFT), the well-defined field direction in the nano-gap enables to recover the orientation of individual molecules.

BBSome function is required for both the morphogenesis and maintenance of the photoreceptor outer segment.

Genetic mutations disrupting the structure and function of primary cilia cause various inherited retinal diseases in humans. Bardet-Biedl syndrome (BBS) is a genetically heterogeneous, pleiotropic ciliopathy characterized by retinal degeneration, obesity, postaxial polydactyly, intellectual disability, and genital and renal abnormalities. To gain insight into the mechanisms of retinal degeneration in BBS, we developed a congenital knockout mouse of Bbs8, as well as conditional mouse models in which function of the BBSome (a protein complex that mediates ciliary trafficking) can be temporally inactivated or restored. We demonstrate that BBS mutant mice have defects in retinal outer segment morphogenesis. We further demonstrate that removal of Bbs8 in adult mice affects photoreceptor function and disrupts the structural integrity of the outer segment. Notably, using a mouse model in which a gene trap inhibiting Bbs8 gene expression can be removed by an inducible FLP recombinase, we show that when BBS8 is restored in immature retinas with malformed outer segments, outer segment extension can resume normally and malformed outer segment discs are displaced distally by normal outer segment structures. Over time, the retinas of the rescued mice become morphologically and functionally normal, indicating that there is a window of plasticity when initial retinal outer segment morphogenesis defects can be ameliorated.

Alcohol-Induced Behaviors Require a Subset of Drosophila JmjC-Domain Histone Demethylases in the Nervous System.

BACKGROUND: Long-lasting transcriptional changes underlie a number of adaptations that contribute to alcohol use disorders (AUD). Chromatin remodeling, including histone methylation, can confer distinct, long-lasting transcriptional changes, and histone methylases are known to play a role in the development of addiction. Conversely, little is known about the relevance of Jumonji (JmjC) domain-containing demethylases in AUDs. We systematically surveyed the alcohol-induced phenotypes of null mutations in all 13 Drosophila JmjC genes. METHODS: We used a collection of JmjC mutants, the majority of which we generated by homologous recombination, and assayed them in the Booze-o-mat to determine their naïve sensitivity to sedation and their tolerance (change in sensitivity upon repeat exposure). Mutants with reproducible phenotypes had their phenotypes rescued with tagged genomic transgenes, and/or phenocopied by nervous system-specific knockdown using RNA interference (RNAi). RESULTS: Four of the 13 JmjC genes (KDM3, lid, NO66, and HSPBAP1) showed reproducible ethanol (EtOH) sensitivity phenotypes. Some of the phenotypes were observed across doses, for example, the enhanced EtOH sensitivity of KDM3KO and NO66KO , but others were dose dependent, such as the reduced EtOH sensitivity of HSPBAP1KO , or the enhanced EtOH tolerance of NO66KO . These phenotypes were rescued by their respective genomic transgenes in KDM3KO and NO66KO mutants. While we were unable to rescue lidk mutants, knockdown of lid in the nervous system recapitulated the lidk phenotype, as was observed for KDM3KO and NO66KO RNAi-mediated knockdown. CONCLUSIONS: Our study reveals that the Drosophila JmjC-domain histone demethylases Lid, KDM3, NO66, and HSPBAP1 are required for normal EtOH-induced sedation and tolerance. Three of 3 tested of those 4 JmjC genes are required in the nervous system for normal alcohol-induced behavioral responses, suggesting that this gene family is an intriguing avenue for future research.

A Dual-Modal SERS/Fluorescence Gold Nanoparticle Probe for Mitochondrial Imaging.

A novel SERS/fluorescent multimodal imaging probe for mitochondria has been synthesised using 12 nm diameter gold nanoparticles (AuNP) surface functionalised with a rhodamine thiol derivative ligand. The normal pH-dependent fluorescence of the rhodamine-based ligand is inversed when it is conjugated with the AuNP and higher emission intensity is observed at basic pH. This switch correlates to a pKa at pH 6.62, which makes it an ideal candidate for a pH-sensitive imaging probe in the biological range (pH 6.5-7.4). The observed pH sensitivity of the ligand when attached to the AuNP is thought to be due to the formation of a spirolactam ring, going from positively charged (+18 mV) to negatively charged (-60 mV) as the pH is changed from acidic to basic. Additionally, conjugation of the ligand to the AuNP serves to enhance the Raman signal of the rhodamine ligand through surface-enhanced Raman scattering (SERS). Confocal microscopy has shown that the probe enters HEK293 (kidney), A2780 (ovarian cancer) and Min6 (pancreatic beta) cells within an hour and a half incubation time. The probe was shown to localise in the mitochondria, thus providing a novel pH-dependent SERS/fluorescent multimodal imaging probe for mitochondria.

Effect of Posterior Tibial Slope on Flexion and Anterior-Posterior Tibial Translation in Posterior Cruciate-Retaining Total Knee Arthroplasty.

Reduced posterior tibial slope (PTS) and posterior tibiofemoral translation (PTFT) in posterior cruciate-retaining (PCR) total knee arthroplasty (TKA) may result in suboptimal flexion. We evaluated the relationship between PTS, PTFT, and total knee flexion after PCR TKA in a cadaveric model. We performed a balanced PCR TKA using 9 transfemoral cadaver specimens and changed postoperative PTS in 1° increments. We measured maximal flexion and relative PTFT at maximal flexion. We determined significant changes in flexion and PTFT as a function of PTS. Findings showed an average increase in flexion of 2.3° and average PTFT increase of 1mm per degree of PTS increase when increasing PTS from 1° to 4° (P<.05). Small initial increases in PTS appear to significantly increase knee flexion and PTFT.

Temporal trends in "legacy" organochlorine contaminants in blubber of ringed seals (Phoca hispida) from Ulukhaktok, NT, Canada between 1972 and 2010.

In blubber of ringed seals (Phoca hispida) from Ulukhaktok, NT, residues of DDT, other organochlorine pesticides, and PCBs declined between 1972 and 2010. The rate of decline varied: concentrations of the DDT-group began to fall after 1981, whereas those of PCBs fell rapidly between 1972 and 1981, and then slowed. Concentrations of cis- and trans-chlordane and of HCB in both sexes, and of cis-nonachlor in males, declined slowly between 1978 and 2010; those of other organochlorine pesticides remained steady. Exponential half-lives of p,p'-DDT and p,p'-DDE in female seals are about 9 and 36 y (corresponding to initial declines of 7.8% and 1.9% per year, respectively) and those of PCB congeners from about 20 to 60 y (declines of 3.2% to <1.5% per year); the more refractory residues may be detectable for centuries to come. Exploratory PCA of PCB congener distribution identified temporal changes apparently not related to molecular structure.

Acute toxic responses of the rockfish (Sebastes schlegeli) to Iranian heavy crude oil: feeding disrupts the biotransformation and innate immune systems.

To clarify the toxic effects of Iranian Heavy Crude Oil (IHCO) from the "Hebei spirit" oil spill, innate immune toxic effects defending on biotransformation pathway have been investigated on fish exposed to IHCO. Juvenile rockfish were exposed to IHCO in gelatin capsules by feeding. The effects on multiple fish biotransformation enzymes (Cytochrome P4501A and glutathione-S-transferase) and the expression level of the several immune response genes, including interleukin-1beta, granulocyte colony-stimulating factor and Cathepsin L, were measured in the liver, spleen and kidney. The tissue-specific expression patterns of these genes demonstrated that the highest expression levels of Cytochrome P4501A, glutathione-S-transferase, interleukin-1beta, granulocyte colony-stimulating factor, interferon stimulated gene 15 and Cathepsin L were found in the liver and that the TNF receptor was high in spleen. The oil-fed fish had significantly higher concentrations of biliary fluorescent metabolites and Cytochrome P4501A expression during the initial stage (12 ∼ 48 h after exposure) than those in the liver and kidney of the sham group. Similarly, the highest mRNA expression levels of interleukin-1beta and granulocyte colony-stimulating factor were detected in the liver at the early stages of exposure (12 h after exposure). Following exposure, the levels of interferon stimulated gene 15 and granulocyte colony-stimulating factor mRNA remained high at 120 h after exposure in the liver but the levels of interleukin-1beta and Cathepsin L gradually decreased to an expression level equal to or less than the sham group. Our data suggest that the innate immune and hepatodetoxification responses in oil-fed fish were induced at the initial stage of exposure to the IHCO at the same time but several immune-related genes decreased to less than that of the sham group after the initial stage of response. Therefore, immune disturbances in fish exposed to IHCO may allow the pathogens, including the infectious diseases, to more easily affect the oil exposed fish.

An Unusual Presentation of Endocarditis Caused by Staphylococcus warneri.

Staphylococcus warneri does not generally cause serious infections in humans. We report a case of endocarditis in a healthy individual with no known past medical history. S. warneri was identified in her blood cultures and echocardiographic evidence confirmed the diagnosis of bacterial endocarditis. There was no apparent cause for her infection, and risk factors such as invasive treatment or medical implant were not present. This rare clinical presentation illustrates the importance of not overlooking low virulence species of Staphylococcus, as they can potentially serve as opportunistic etiological agents for endocarditis, especially among the elderly population.

One-bead, one-compound peptide library sequencing via high-pressure ammonia cleavage coupled to nanomanipulation/nanoelectrospray ionization mass spectrometry.

Biological screening of one-bead, one-compound (OBOC) combinatorial peptide libraries is routinely carried out with the peptide remaining bound to the resin bead during screening. After a hit is identified, the bead is isolated, the peptide is cleaved from the bead, and its sequence is determined. We have developed a new technique for cleavage of peptides from resin beads whereby exposure of a 4-hydroxymethyl benzoic acid (HMBA)-linked peptide to high-pressure ammonia gas led to efficient cleavage in as little as 5min. Here we also report a new method of extracting peptide from individual library beads for its introduction into a mass spectrometer that uses nanomanipulation combined with nanoelectrospray ionization mass spectrometry (NSI MS). Single beads analyzed by nanomanipulation/NSI MS were found to give identical MS results to those of bulk samples. Detection of 18 unique cleaved peptides 1 to 8 amino acids in length, and sequencing of 14 different peptide sequences 4 to 8 amino acids in length, was demonstrated on a combination of bulk samples and ones from individual beads of an OBOC library. The method was highly reproducible, with 100% of attempts to extract peptide resulting in high-quality MS data. This new collection of techniques allows rapid, reliable, environmentally responsible sequencing of hit beads from combinatorial peptide libraries.

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As a coagulase negative Staphylococcus species, S. caprae is not considered as a clinically-significant member, unlike S. epidermidis. In this report, we describe a case of sepsis resulting from S. caprae infection. This relatively young woman was in generally good health and contracted S. caprae most probably during her treatment of an acute pulmonary embolism. The purpose of this report is to raise awareness of this otherwise innocuous staphylococcal species in clinical settings.

Hexachlorocyclohexanes (HCH) in ringed seal (Phoca hispida) from Ulukhaktok (Holman), NT: trends from 1978 to 2006.

Trends in alpha-, beta-, and gamma-hexachlorocyclohexane (HCH) concentrations were examined in blubber lipid of ringed seals (Phoca hispida) from Ulukhaktok (Holman), NT (Canada) sampled at intervals between 1978 and 2006. alpha-HCH usually represented approximately 90% of the total HCH isomers. alpha-HCH and gamma-HCH concentrations showed no change over the sampling interval, but beta-HCH concentrations increased significantly, about 8-10-fold in females and 4-5-fold in males. Residue concentrations showed no dependence on age. Concentrations (all data as ng/g lipid, GM (range)) of alpha-HCH were significantly higher (P<0.001 by t-test) in males (217 (93.9-517), n=37) than those in females (138 (40.9-402), n=38). beta-HCH concentrations did not differ between the sexes. Concentrations of gamma-HCH were significantly higher (P<0.05) in males (6.74 (0-46.7)) than in females (4.35 (0-19.0)). Although global emissions of both alpha-HCH and beta-HCH have declined since the early 1980's, the "signal" of HCH emission changes has not yet resulted in a "response" in ringed seal residue concentrations. In the light of our current understanding of the dynamics of HCH in the Arctic, we conclude that any such response may not be detected by retrospective analyses of the sort describe here at least for another decade or so, because of the longevity of the seals.

Parenteral polymyxin B use in patients with multidrug-resistant gram-negative bacteremia and urinary tract infections: a retrospective case series.

BACKGROUND: Parenteral polymyxin use declined after the 1960s, due to safety concerns. An increase in multidrug-resistant (MDR) gram-negative infections and a shortage of new agents have prompted increased use of parenteral polymyxin. OBJECTIVE: To describe our clinical experience with parenteral polymyxin B for MDR gram-negative bacteremia and urinary tract infection (UTI). METHODS: Paper pharmacy records were used to identify patients aged 18 years or older, presence of MDR gram-negative bacteremia or UTI, and use of parenteral polymyxin B for at least 48 hours. Electronic and paper patient records were then retrospectively reviewed. Polymyxin B susceptibility was evaluated using the Kirby-Bauer method. MDR isolates were defined as resistant to at least 3 antimicrobial classes, excluding polymyxin B. Microbiologic clearance was defined by 1 repeat urine or 2 repeat blood cultures that were sterile or growing different organisms. Secondary outcomes included hospital mortality and nephrotoxicity, defined as an increase in serum creatinine of 0.5 mg/dL or more, or a 50% reduction in creatinine clearance. RESULTS: Seventeen infections in 16 patients were treated with polymyxin B (1 pt. had 2 infections that were analyzed separately). Microbiologic clearance occurred in 14 of 16 (88%) cases of MDR gram-negative bacteremia or UTI in which repeat cultures were done. Ten of 16 patients died (all-cause mortality 63%). Five patients required hemodialysis prior to polymyxin B use. Six (55%) of the remaining 11 patients with baseline renal insufficiency developed nephrotoxicity, and none of them required hemodialysis. The mean +/- SD number of days from the initiation of therapy to the onset of nephrotoxicity was 7.5 +/- 2.3 (range 4-10) days. Three (50%) of 6 patients with nephrotoxicity died. CONCLUSIONS: Our data suggest that polymyxin B may be effective for MDR gram-negative infections in patients with limited therapeutic options, but precautions should be taken to avoid toxicity.