Physicochemical and Antimicrobial Properties of Thermosensitive Chitosan Hydrogel Loaded with Fosfomycin

Thermosensitive chitosan hydrogels—renewable, biocompatible materials—have many applications as injectable biomaterials for localized drug delivery in the treatment of a variety of diseases. To combat infections such as Staphylococcus aureus osteomyelitis, localized antibiotic delivery would allow for higher doses at the site of infection without the risks associated with traditional antibiotic regimens. Fosfomycin, a small antibiotic in its own class, was loaded into a chitosan hydrogel system with varied beta-glycerol phosphate (β-GP) and fosfomycin (FOS) concentrations. The purpose of this study was to elucidate the interactions between FOS and chitosan hydrogel. The Kirby Bauer assay revealed an unexpected concentration-dependent inhibition of S. aureus, with reduced efficacy at the high FOS concentration but only at the low β-GP concentration. No effect of FOS concentration was observed for the planktonic assay. Rheological testing revealed that increasing β-GP concentration increased the storage modulus while decreasing gelation temperature. NMR showed that FOS was removed from the liquid portion of the hydrogel by reaction over 12 h. SEM and FTIR confirmed gels degraded and released organophosphates over 5 days. This work provides insight into the physicochemical interactions between fosfomycin and chitosan hydrogel systems and informs selection of biomaterial components for improving infection treatment.     

Two mosaic terminal inverted duplications arising post-zygotically: Evidence for possible formation of neo-telomeres

Objective

To elucidate the structure of terminal inverted duplications and to investigate potential mechanisms of formation in two cases where there was mosaicism with cells of apparently normal karyotype.

Results

A karyotype [46,XY,inv dup(4)(p16.3p15.1)/46,XY] performed on blood lymphocytes from a patient referred for developmental delay (case 1) demonstrated a normal karyotype in 60% of cells with a terminal inverted duplication 4p in the remainder. In case 2, referred for multiple fetal anomalies on an ultrasound scan, 33% of amniocyte colonies were karyotypically normal, with a terminal inv dup 10p in the remainder [46,XX,inv dup(10)(p15.3p11)/46,XX]. Duplicated FISH signals for GATA3 and NEBL loci (in case 2), and for the Wolf-Hirschhorn locus (case 1) confirmed the inverted structure of both duplications. In the GTL banded normal cells from both cases, there was a cryptic deletion detected by FISH of one copy of the subtelomere 4p (case 1, probe GS-36P21), and subtelomere 10p (case 2, probe GS-306F7). At pter on both inv dup chromosomes there was no FISH signal present for the specific subtelomere probe. However, a positive pantelomeric probe signal was detected at 4 pter and 10 pter in both the cryptically-deleted chromosomes and the inv dup chromosomes in the respective cell lines of both cases.

Conclusion

An inv dup structure was evident for both cases on GTL bands, and confirmed by the various FISH studies. The presence of telomere (TTAGGG repeat) sequences at pter on the inv dup chromosomes (where more proximal chromosome specific subtelomeric probes were negative) was indicated by the pantelomeric probe signals in both cases. We conclude the most likely mechanism of origin in both cases was by sub-telomeric breakage in the zygote at pter, and delayed repair/rearrangement until after one or more subsequent mitotic divisions. In these divisions, at least one breakage-fusion-bridge cycle occurred, to produce inverted duplications. It is proposed then that two differently "repaired" daughter cells proliferated in parallel. In one daughter cell line (with an overtly normal karyotype) there was deletion of the subtelomere and presumed repair through capping by a neo-telomere (i.e. "healing", as initially proposed by McClintock). This occurred in both cases presented. In the other daughter cell of each case, it is proposed that chromosome stabilization was achieved (after replication) by sister chromatid reunion to form a dicentric, which broke at a subsequent anaphase, to form an inverted duplication (with loss of the reciprocal product, and the other daughter cell line). One inv dup was repaired without an interstitial specific subtelomere (case 1) and one was repaired with a duplicated specific interstitial subtelomere (case 2). After repair TTAGGG repeats were detected by FISH at each respective new pter.

     

Detection of O,O-diethyl-O-(2,5-dichlorophenyl) phosphorothioate and O,O-diethyl-O-(3,6-dichloro-2-pyridyl) phosphorothioate in meat fat.

Two previously unreported organophosphorus residues have been detected in meat fat. The first, O,O-diethyl-O-(2,5-dichlorophenyl) phosphorothioate, apparently results from the debromination of bromophos-ethyl, while the other, O,O-diethy-O-(3,6-dichloro-2-pyridyl) phosphorothioate, presumably arises by the monodechlorination of chlorpyriphos. Mass spectral evidence is presented in support of the assigned structures and gas-liquid chromatographic retention data for the residues and their oxygen analogs are provided.     

Detection of antibiotic residues and association of cefquinome residues with the occurrence of Extended-Spectrum β-Lactamase (ESBL)-producing bacteria in waste milk samples from dairy farms in England and Wales in 2011

Waste milk samples from 103 farms in England and Wales were examined for the presence of β-lactam antibiotics and ESBL-producing Enterobacteriaceae. Approximately 10 months after the initial sampling, further waste milk, environmental and faecal samples from farms shown to be positive for CTX-M Escherichia coli were investigated further. Isolates with an ESBL phenotype were tested by PCR for the presence of bla_CTX-M, bla_OXA, bla_SHV and bla_TEM genes. Isolates positive for bla_CTX-M were sequenced to determine CTX-M type. Representative isolates were further examined by PFGE, plasmid replicon typing and serotyping. Of particular interest, 21.4% of waste milk samples contained residues of the cephalosporin cefquinome, which was significantly associated with CTX-M bacteria. Such bacteria occurred in 5.8% of the waste milk samples (including 3.9% CTX-M E. coli). CTX-M types identified were 1, 14, 14b and 15, but none of the E. coli were serotype O25, the serotype of the human pandemic strain.     

Studies on the sorption of organochlorine insecticides by flour stored on or near treated laminated timber or plywood as used in freight containers

Wood preservatives containing a number of organochlorine insecticides, including aldrin, dieldrin, chlordane and lindane, have been approved in Australia for treating timber used as structural components in cargo containers. Studies on the migration of these insecticides from treated laminated timber and plywood, using commercial containers and in the laboratory, are reported here. The highest insecticide residue levels were found in flour samples stored on newly treated laminated sawn timber. Physical pick-up of insecticide from the surface of the floor was considered to be the major source of contamination. Sorption of insecticides from the atmosphere of the container was the most likely source of contamination in samples stored on or near treated plywood.     

Doxorubicin area under the curve is an important predictor of neutropenia in dogs with naturally occurring cancers

Doxorubicin (DOX) area‐under‐the‐curve (AUC) was calculated for 40 dogs with spontaneously occurring cancers using a previously validated limited‐sampling approach. All dogs were administered a dose of 30 mg/m² by intravenous infusion and serum samples were collected at 5, 45 and 60 minutes post‐infusion. DOX and its major metabolite, doxorubicinol (doxol), were quantified in serum samples using high‐performance liquid chromatography tandem‐mass spectrometry. Wide interpatient variability was observed in the predicted DOX AUC with a coefficient of variation of 34%. A significant relationship was found between DOX AUC and absolute white blood cell count (P = 0.003), absolute neutrophil count (ANC; P = 0.002) and surviving fraction of neutrophils (P = 0.03) approximately 1 week after dosing (nadir). No changes in other hematologic parameters (red blood cells, platelets, lymphocytes, haemoglobin) were found to correlate with DOX AUC. The absolute dose (mg) and the dose per unit body weight (mg/kg) were not significantly correlated with nadir ANC. No relationships were found between maximum serum doxol concentration and myelosuppression. Baseline ANC was also significantly correlated to nadir ANC and a model was constructed using baseline ANC and DOX AUC that significantly described the nadir ANC. These findings demonstrate the important relationship between systemic DOX exposure and degree of neutropenia in dogs, and suggest a potential for individualized, pharmacokinetically‐guided DOX dosing in dogs.     

Implications of hypoxia tolerance for wetland refugia use in Lake Nabugabo,Uganda

Aquatic hypoxia can affect predator‐prey interactions by altering the success rate of the predator and/or the vulnerability of prey. For example, in the Lake Victoria basin of East Africa, native prey exploit hypoxic wetlands as refugia from predation by introduced Nile perch (Lates niloticus). Here, it is predicted that species exploitation of wetlands depends on their hypoxia tolerance relative to the heterogeneity of wetland hypoxia. In this study, we compared the hypoxia tolerance of four fish taxa that differ in their use of hypoxic wetlands in Lake Nabugabo, Uganda: the cichlid Pseudocrenilabrus multicolor victoriae that inhabits the dense swamp interior; and three taxa that inhabit wetland ecotones including Nile tilapia (Oreochromis niloticus), L. niloticus and juvenile endemic haplochromine cichlids. We characterised hypoxia tolerance by exposing fish to progressive hypoxia and quantifying variation in aquatic surface respiration (ASR). The effect of body size on tolerance was explored in L. niloticus by quantifying ASR behaviour across a range of size classes. ASR behaviour was also compared between O. niloticus groups from wetland versus open‐water habitats to detect habitat‐associated intraspecific variation. The most tolerant taxon was the swamp specialist P. multicolor, indicated by its low ASR thresholds and small percentage of fish using ASR during the final sample interval. The other three taxa did not differ in ASR behaviour, and no differences were detected between O. niloticus groups. Body size effects were present for L. niloticus suggesting a lower tolerance to hypoxia in larger‐bodied individuals, thus limiting their ability to penetrate wetlands.     

Fathead minnow reproduction: Implications for commercial culture

The fathead minnow, Pimephales promelas, is widely used as a bait, forage, and research fish in the United States. A hardy and fecund fish, this species has many attributes that are excellent for culture. Unfortunately, despite the widespread use of fathead minnows in toxicity testing and thousands of resulting publications, relatively little aquaculture‐related research has been conducted in support of commercial production. On the surface, current commercial fathead minnow production practices appear much the same as those of a decade ago. However, producers continue to refine culture methods, balancing broodfish stocking rates, spawning substrate quantity, and feeding rates in an attempt to meet both losses to predators and market demand. This review examines relevant reproduction‐related studies in the fields of ecology, behavior, toxicology, and aquaculture with the goal of advancing fathead minnow culture. In fathead minnow production, relative to the culture of food fish species, reproduction is vital, more so than rapid growth or a low feed conversion ratio. Fish numbers may exceed 1 million fish/ha, while feeding rates are relatively low and in feeder fish production, growth is deliberately restrained in order to maintain fish within market sizes year‐round. Underappreciated factors identified as affecting reproduction and with implications for commercial culture include: (a) poor egg adhesion to commonly used spawning substrates, (b) egg and fry cannibalism at high fish densities, and (c) the relatively short breeding period of individual males. Results highlight the importance of selecting an appropriate broodfish stocking rate based on fish size, anticipated pond fertility, and feeding program. New methods to improve reproductive performance will likely increase costs and will need to be accompanied by thorough economic analyses.