Efficacy of alcohol-based hand sanitizers against human norovirus using RNase-RT-qPCR with validation by human intestinal enteroid replication.

Successful human norovirus (HuNoV) cultivation in stem cell-derived human intestinal enteroids (HIE) was recently reported. The purpose of this study was to evaluate the anti-HuNoV efficacy of two alcohol-based commercial hand sanitizers and 60% ethanol by suspension assay using RNase-RT-qPCR, with subsequent validation of efficacy by HuNoV cultivation using the HIE model. In suspension, when evaluated by RNase-RT-qPCR, 60% ethanol resulted in less than one log10 reduction in HuNoV genome equivalent copies (GEC) regardless of contact time (30 or 60s) or soil load. The two commercial products outperformed 60% ethanol regardless of contact time or soil load, providing 2·2-3·2 log10 HuNoV GEC reductions by suspension assay. Product B could not be validated in the HIE model due to cytotoxicity. Following a 60s exposure, viral replication in the HIE model increased 1·9 ± 0·2 log10 HuNoV GEC for the neutralization (positive) control and increased 0·9 ± 0·2 log10 HuNoV GEC in challenged HIE after treatment with 60% ethanol. No HuNoV replication in HIE was observed after a 60 s exposure to Product A.

Cortical activations in sequential bilateral cochlear implant users.

This study aimed to examine auditory and visual cortical activations in bilateral cochlear implant recipients using [(18)F]-FDG positron emission tomography. We aimed to compare the activations from use of the first implant alone, the second implant alone, and both implants together. When both implants were activated simultaneously, summation of cortical activity did not occur. The first and second implants demonstrated evidence of developing distinct neural networks. The first implants show stronger bilateral recruitment of the auditory areas than the second implants. Visual cortical activations occur in response to stimulation of the second but not the first implants. When both implants were activated together, there were no visual activations suggesting interaction between the first- and second-implant networks. These findings add to the existing knowledge of plasticity following cochlear implantation and demonstrate a variability of these processes that was previously unreported.

Nasal fracture reduction: local versus general anaesthesia.

OBJECTIVE: To investigate the outcomes from nasal fracture reduction performed under local anaesthesia (LA) and general anaesthesia (GA). METHOD: A randomised multi-centred prospective trial and cohort analysis. Patients were randomised into two groups, 74 (53%) underwent closed reduction under LA, 65 (47%) patients underwent closed reduction under GA. MAIN OUTCOME MEASURES: Pain scores and patient toleration of local or general nasal manipulation was noted. RESULTS: The pain score ranged from 0 (no pain) to 10 (unbearable pain). The mean pain score in the LA group was 3, compared to 2 in the GA group. LA manipulation was tolerated equally well as GA manipulation by 85% of the patients in each group. The number of patients requiring a septorhinoplasty was compared between LA 19/74 (26%), and GA 21/65 (32%). This failed to demonstrate a significant difference with a p value of 0.50. The absolute risk difference was 5% with a 95% confidence interval of (20% to -10%). CONCLUSION: This trial clearly shows LA to be as effective as GA in the first line management of nasal fractures. The degree of septal displacement and presence of nasal tip deviation were associated with persistent nasal deformity following nasal fracture reduction.

Reduction of nasal fractures under local anaesthetic.

Forty five patients with fractured nasal bones underwent nasal fracture reduction under local anaesthetic. Patients used linear analog scores to assess pain associated with the procedure and satisfaction with the reduction. The surgeon also recorded his assessment of the reduction. The patients were reviewed two weeks later and the outcome scores repeated. The procedure was well tolerated and forty three patients (95.5%) stated they would undergo the same procedure again if they fractured their nose in the future. Fifteen patients (33.3%) had significant persistent nasal deformities and were listed for septorhinoplasty. The results of nasal fracture reduction under localanaesthetic are comparable to those obtained under general anaesthetic. The factors that may contribute to poor outcomes following reduction of nasal fractures are discussed.

Influence of dehydroepiandrosterone on rabbit intraocular pressure.

PURPOSE: The systemic concentration of dehydroepiandrosterone decreases with age in primates while in humans intraocular pressure (IOP) increases with aging. This study was designed to investigate if a relationship existed between dehydroepiandrosterone and IOP in pigmented rabbits. METHODS: Animals were treated unilaterally for 6 weeks with topical 3% dehydroepiandrosterone in 30% 2-hydroxypropyl-beta-cyclodextrin; the contralateral eye received vehicle alone. Drops were applied, and IOP measured, twice daily. RESULTS: Small, but statistically significant, drug-related effects were found. IOP was consistently higher in the afternoon; the afternoon minus morning difference in IOP, however, decreased with time. Topical, radioactive drug application indicated very low level penetration into aqueous humor, iris, corneal epithelium, the rest of the cornea, or bulbar conjunctiva. CONCLUSION: The small drug-related effects may be due, in large part, to poor intraocular drug penetration. The circadian rhythm of IOP appears to be time-dependent in chronic studies with a gradual loss of IOP difference between a.m. and p.m. readings.

Characterization of a phospholipase Dalpha cDNA from tomato fruit.

Phospholipase D (PLD) initiates phospholipid (PL) catabolism in plant cells and is also involved in signal transduction and retailoring of membrane PL. Total PL declines and phosphatidic acid increases in pericarp tissue during tomato fruit ripening, suggesting that increased PLD activity alters membrane structure. To assess the role of PLD in tomato ripening, we have begun a molecular genetic approach. Using a castor bean PLDalpha cDNA as a probe, a PLDalpha cDNA (LEPLD2) was isolated from our tomato fruit library. It has an open reading frame of 2421 nucleotides, encoding a polypeptide of 807 amino acids with a molecular mass of 92 kDa. The deduced LEPLD2 PLDalpha shares >75% sequence identity with PLDalphas from castor bean, tobacco and tomato. LEPLD2 transcript, detected by RNA gel-blot analysis, was very low in roots, low in stems, moderate in leaves, high in flowers, and increased in fruit during development and ripening. Expression of LEPLD2 in Escherichia coli yielded active enzyme, and a FLAG-PLDalpha fusion protein produced by transformed E. coli migrated close to the calculated 94 kDa on SDS/PAGE.

Lectins in drug delivery: a study of the acute local irritancy of the lectins from Solanum tuberosum and Helix pomatia.

Lectins are proteins or glycoproteins of non-immune origin capable of binding to one or more specific sugar residues. The potential for using lectins as a means of 'anchoring' a drug delivery system to the mucosal surfaces of the eye has been investigated in previous work, with the lectins from Solanum tuberosum and Helix pomatia showing particular promise. In this study the acute local dermal irritancy of these lectins, in terms of their potential to cause inflammation and tissue necrosis, was investigated. After an initial study in terminally anaesthetised animals (to ensure no gross toxicity was evident), five male New Zealand white rabbits from the same litter were briefly anaesthetised and Evans blue injected intravenously as a marker of inflammation. Sterile lectin solutions in normal saline at a range of concentrations from 50 to 500 microg ml(-1) were prepared and 50-microl volumes injected intradermally at 18 sites across a shaved area of each rabbit's back. The rabbits were then allowed to regain consciousness. There was no evidence of tissue necrosis, oedema or Evans blue infiltration with any of the lectin solutions administered. The rabbits did not display any signs of discomfort such as scratching or continued grooming throughout the experiment. Histological examination of the injection sites revealed little sign of any inflammation, such as heterophil migration, oedema or tissue damage. It was concluded that these lectins demonstrate minimal acute irritancy, and will, therefore, be taken forward for formulation and in vivo studies.

Marijuana smoking vs cannabinoids for glaucoma therapy.

OBJECTIVE: To discuss the clinical effects, including toxicological data, of marijuana and its many constituent compounds on the eye and the remainder of the body. A perspective is given on the use of marijuana and the cannabinoids in the treatment of glaucoma. RESULTS: Although it is undisputed that smoking of marijuana plant material causes a fall in intraocular pressure (IOP) in 60% to 65% of users, continued use at a rate needed to control glaucomatous IOP would lead to substantial systemic toxic effects revealed as pathological changes. CONCLUSIONS: Development of drugs based on the cannabinoid molecule or its agonists for use as topical or oral antiglaucoma medications seems to be worthy of further pursuit. Among the latter chemicals, some have no known adverse psychoactive side effects. Smoking of marijuana plant material for the reduction of elevated IOP in glaucoma is ill-advised, given its toxicological profile.

Structure and function of desmosomal transmembrane core and plaque molecules.

Desmosomes are intercellular junctions that function in cell-cell adhesion and attachment of intermediate filaments (IF) to the cell surface. Desmogleins and desmocollins are the major components of the transmembrane adhesion complex, whereas desmoplakins (DPs) are the most prominent components of the cytoplasmic plaque. Based on sequence similarity, desmogleins and desmocollins are related to the calcium-dependent homophilic adhesion molecules known as cadherins. Like the classical cadherins, the desmosomal cadherins contain four homologous extracellular domains bearing putative calcium-binding sites, a single transmembrane spanning domain, and a C-terminal cytoplasmic tail. Molecules in the desmoglein subclass contain a unique C-terminal extension within which is found a repeating motif that is predicted to form two beta-strands and two turns. Stable cell lines expressing desmoglein 1 have been generated from normally non-adherent L cell fibroblasts, to study the contribution of this cadherin to desmosomal adhesion. The predicted sequence of desmoplakin (DP) I suggests it will form homodimers comprising a central alpha-helical coiled-coil rod and two globular end domains. The C-terminus contains three regions with significant homology, each of which is made up of a 38-residue motif also found in two other molecules involved in organization of IF, bullous pemphigoid antigen and plectin. Ectopically expressed polypeptides including the C-terminus of DP I specifically align with keratin and vimentin IF in cultured cells, whereas those lacking this domain do not align with IF. The last 68 amino acids of DP are required for alignment along keratin but not vimentin IF, and residues 48-68 from the C-terminal end are critical for this interaction. These results suggest that the C-terminus of DP plays a role in the attachment of IF to the desmosome and that a specific site is necessary for interaction with keratin IF. A sequence at the most N-terminal end of DP appears to be required for efficient incorporation into the desmosomal plaque. Interestingly, this region has not been reported to be present in the homologous bullous pemphigoid antigen or plectin molecules and may represent a desmosomal targeting sequence.

Brain stem auditory evoked potentials from bone stimulation in dogs.

Brain stem auditory evoked potential (BAEP) testing with air-conducted click stimuli can be used to diagnose sensorineural deafness in dogs if conductive deafness can be ruled out. Detection of conductive deafness can be performed by recording BAEP elicited by a vibratory stimulus transducer placed against the skull. Air- and bone-conducted BAEP were compared in dogs, varying bone stimulator placement, click polarity, and stimulus intensity. Optimal bone stimulator placement was determined to be over the mastoid process, followed by the mandible and the zygomatic arch. Condensation polarity clicks gave responses preferable to those elicited by rarefaction or alternating polarity. Bone-conducted BAEP peak latencies were significantly longer than air-conducted latencies after correction of the latencies for the air conduction time accompanying air-conducted stimuli. Significant differences between stimulus modalities were not seen for BAEP peak amplitudes or interpeak latencies. Latency-intensity and amplitude-intensity regressions had similar effects for both modalities: latencies decreased and amplitudes increased as stimulus intensity increased.

Delayed lactogenesis in women with insulin-dependent diabetes mellitus.

Breast milk lactose, total nitrogen, conductivity, osmolality, and intake by infants of 33 women with insulin-dependent diabetes mellitus (IDDM), 33 control women without diabetes, and 11 reference women were determined in a 3-mo study of lactation. Milk of women with IDDM had significantly lower lactose and higher total nitrogen (2-3 d postpartum), and their infants had significantly less milk intake (7-14 d postpartum) than did control or reference women. Total nitrogen was negatively correlated with milk lactose for women with IDDM at all times and for control women through day 14 postpartum. The data indicate delayed lactogenesis for women with IDDM, which was more likely to occur with poor metabolic control. Differences in milk composition of women with IDDM do not preclude them from breast-feeding their infants.

Influence of vehicle and anterior chamber protein concentration on cyclosporine penetration through the isolated rabbit cornea.

The transcorneal penetration of cyclosporine A has been determined from each of three vehicles across isolated cornea into simulated aqueous humor containing either 50 mg % protein (0.5 mg/ml; as found in a normal eye) or 5000 mg % protein (50 mg/ml; as found in an inflamed eye). Cyclosporine entered the corneal epithelium and stroma/endothelium as well as passed through the cornea from an alpha cyclodextrin vehicle. Entry into the epithelium and stroma/endothelium occurred from an ointment vehicle with limited detectable anterior chamber penetration using 50 mg % protein solution in the anterior chamber. From corn oil vehicle, cyclosporine penetrated across the cornea with a permeability equal to that of alpha cyclodextrin vehicle. The concentration of cyclosporine in both corn oil and ointment vehicles is 8 times greater than that in alpha cyclodextrin vehicle resulting in a flux from corn oil vehicle about 7 or 8 times greater than that seen after alpha cyclodextrin vehicle. The amounts retained in the cornea, however, were relatively low after corn oil compared to cyclodextrin. The penetration of cyclosporine from either the cyclodextrin vehicle or ointment was at least doubled in the presence of 5000 mg % protein in the simulated aqueous humor relative to that seen in 50 mg % protein. This data indicates that the (presumed) absorption and binding of drug by the excess protein in the simulated aqueous humor may have removed free cyclosporine from the solution and sustained a high concentration gradient of free solute across the cornea.(ABSTRACT TRUNCATED AT 250 WORDS)

Role of eicosanoids in the ocular response to intracameral hydrogen peroxide.

The intracameral injection of hydrogen peroxide induces a sequence of responses in the tissues bounding the anterior chamber. These changes include intraocular pressure, corneal thickness, iris hyperemia, increased leakiness of the iris vasculature, and edema of the ciliary processes as judged from microscopic examination. Some of these responses appear to include inflammatory effects that may be the result of the local release of eicosanoids. Several antagonists of the arachidonic acid cascade, indomethacin, aspirin, dexamethasone, and nordihydroguaiaretic acid (NDGA) were used to examine their influence on the sequelae of hydrogen peroxide injection. Indomethacin, and high dose (7.5 mg/kg) NDGA were most effective in reducing the number of parameters that were altered after intracameral hydrogen peroxide. Microscopic observations supported the physiological changes and the responses to antagonists. The data indicate that a portion of the ocular tissue responses to intracameral hydrogen peroxide in the rabbit eye may be the result of eicosanoid production in these tissues.

Retention and distribution of liposome-entrapped [3H]methotrexate injected into normal or arthritic rabbit joints.

Normal or arthritic rabbits were injected intra-articularly (i.a.) with free [3H]methotrexate ([3H]MTX) or liposomes containing [3H]MTX with [14C]cholesteryl oleate as a lipid marker. The distribution of 3H and 14C in the injected joint and other tissues was determined. Free [3H]MTX was rapidly cleared from the joint, 79% being excreted in the urine within 24 h of injection. Liposome-entrapment retarded [3H]MTX clearance from the joint (P less than 0.001), 45.5% being recovered from the joint 24 h after injection. Uptake of liposomes by the inflamed synovium was lower than expected, 4% liposomal [3H]MTX injected being associated with the synovium after 24 h. Nevertheless, this was 40-fold greater than when free [3H]MTX was injected. Liposome entrapment should improve the efficacy and reduce the side effects of drugs injected directly into the joint cavity.

Regional ocular blood flow after chronic topical glaucoma drug treatment.

The effects of a chronic three times a day treatment over a 5 to 6 week period of rabbit eyes with 1 of 5 topically applied glaucoma drugs on ocular blood flow was determined using a radioactive microsphere technique. The drugs employed were timolol (0.5%), pilocarpine (4%), epinephrine (2%), norepinephrine (2%), and ecothiophate iodide 0.125%). The results showed that epinephrine statistically decreased blood flow to the iris and ciliary processes while not to the posterior uveal tissues or optic nerve head. Pilocarpine also showed this same trend, while the differences were not statistically significant. Other drugs were without effect on regional ocular blood flow.

Water soluble high molecular weight components from plants with potent intraocular pressure lowering activity [corrected and republished ariticle originally printed in Curr Eye Res 1987 May;6(5):733-4].

Previous work in our laboratory has shown that Cannabis sativa (marijuana) contains water-soluble, high molecular weight components that have extremely potent intraocular pressure (IOP) lowering activity. Aqueous extraction of other plants has now shown that a number of them also contain components with potent IOP lowering activity in rabbits. These include tobacco, cabbage, lettuce, several greens, Senecio anonymus, Erigeron philadelphicus, and several others. Not all plants tested were active, however, indicating that while more ubiquitous than originally thought, these materials are not apparently extractable from all plants. The chemical composition of the active fraction from tobacco was found to be different from that derived from Cannabis sativa.

Prostaglandin involvement in the responses of the rabbit eye to water-soluble marihuana-derived material.

Both anticoagulants (heparin and streptokinase) and non-steroidal anti-inflammatory compounds (aspirin and indomethacin) were used against a water-soluble derivative of marihuana, MDM. While the anticoagulants had no effect on the ocular effects of MDM, both aspirin and indomethacin altered the time course and effected the MDM-induced reduction of intraocular pressure. The usual initial hypertensive effect of intravenous MDM was eliminated and the later intraocular pressure fall occurred earlier as well as being inhibited by about 35 to 50%. Assay for prostaglandins revealed that intravenous MDM (3.86 micrograms) caused a marked rise in PGE2 concentration of the aqueous humor and iris-ciliary body during the first hour or two after administration of MDM, but normal values occurred at 4, 6, and 8 hours when the intraocular pressure is reduced by up to 60%. Following intravitreal MDM (0.002 microgram), however, the PGE2 levels remained unchanged over 24 hours, despite the induction of a fall in intraocular pressure between 14 and 18 hours which lasts for many hours. Prostaglandin appears to be involved in the hypertensive phase of intraocular pressure change after intravenous MDM injection; and, while the fall in intraocular pressure may contain a component partially mediated by prostaglandins, there is no evidence that intravitreal MDM induces any effect on prostaglandin levels. The involvement of prostaglandins, therefore, in the mediation of MDM-induced ocular hypotensive effects is apparently small.