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Biomedical subjects

M R McLaughlin

Publications and source records attributed to M R McLaughlin.

At least 19 recordsLinked to original sources

Factors affecting iron sulfide-enhanced bacteriophage plaque assays in Salmonella.

Reaction of ferric ions with hydrogen sulfide (H(2)S) enhances contrast of phage plaques in H(2)S+ Salmonella, but contrast diminishes in weak H(2)S+ strains. H(2)S was affected by concentrations of peptones, glucose, ferric ammonium citrate (FAC) and sodium thiosulfate (ST), and by FAC:ST ratio, temperature, pH, air, and host strain. Increasing peptone levels was most important for improving contrast in weak H(2)S+ strains.

Culture Media↗

Isolation of salmonella bacteriophages from swine effluent lagoons.

Bacteriophages (phages) associated with Salmonella were collected from nine swine manure lagoons in Mississippi. Phages were isolated by an enrichment protocol or directly from effluent. For enrichment, chloroform-treated samples were filtered (0.22 mum) and selectively enriched by adding a cocktail of Salmonella strains in trypticase soy broth. After overnight incubation at 35 degrees C, chloroform was added and samples stored at 5 degrees C. Enriched samples were tested by double agar layer (DAL) plaque assay against individual Salmonella isolates. Phage titers of 2.9 x 10(8) to 2.1 x 10(9) plaque forming units (pfu) per mL were produced, but estimation of phage titers in lagoons was not possible. For direct isolation, effluent was clarified by centrifugation, filtered (0.22 microm), and used in DAL plaque assays to select single-plaque isolates for 15 Salmonella strains. Plaque counts varied among Salmonella strains and lagoons. The most sensitive strain for direct phage recovery was ATCC 13311. Phage titers estimated by direct isolation with ATCC 13311 ranged among lagoons from 12 to 148 pfu per mL. In limited host range tests, 66 isolates recovered by the enrichment protocol produced plaques only on Enteritidis and Typhimurium strains of Salmonella and none produced plaques on lagoon isolates of Citrobacter, Escherichia, Proteus, Providencia, or Serratia. Electron microscopy (EM) showed purified enrichment isolates had Podoviridae morphology (tailless 50-nm icosahedral heads with tail spikes). Electron microscopy of clarified concentrated effluent showed 5.5:1 tailless to tailed phages. The isolated phages have potential as typing reagents, specific indicators, and biocontrol agents of Salmonella.

Animals↗

Enhanced contrast of bacteriophage plaques in Salmonella with ferric ammonium citrate and sodium thiosulfate (FACST) and tetrazolium red (TZR).

Visualization of bacteriophage plaques may be enhanced by addition of ferric ammonium citrate and sodium thiosulfate (FACST) or 2,3,5-triphenyltetrazolium chloride (tetrazolium red, TZR) to the soft agar layer of a traditional bacteriophage plaque assay. Background color from these reagents improved contrast between clear plaques and turbid host lawns in trypticase soy agar (TSA) plates. Enhancement by FACST is based on reaction with hydrogen sulfide gas (H2S) produced by some strains of bacteria and was tested here using H2S+ and H2S- strains of Salmonella enterica subsp. enterica with a bacteriophage (Podoviridae) isolated from swine lagoon effluent. Only the H2S+ strain produced dark brown-black color in FACST-amended agar. Both strains showed bright pinkish-red color in TZR-amended agar. Color intensity for both reagents decreased with decreasing concentrations of the reagents. Contrast in FACST-amended plates appeared greater than that with TZR, but diminished after 12 h, while contrast in TZR-amended plates remained constant. At the concentrations tested, neither reagent affected plaque counts in the H2S+ strain. The FACST should be useful in bacteriophage plaque assays with H2S+ strains of Salmonella and other H2S+ bacteria.

Bacteriological Techniques↗

Evaluation of marine bacterial lysogens for development of a marine prophage induction assay.

The demonstrated relationship between carcinogenicity of a chemical compound in mammals and its tendency to cause prophage induction in bacteria provides a method for biologically based carcinogen screening. Because of the need for this type of screening and the abundance of lysogens in the marine environment, 14 isolates were evaluated for the degree of prophage induction in exponentially growing cultures in the presence of a known mutagen (0.5 microg/ml mitomycin C). Assays were performed both in liquid culture and in microtiter plates. Virus-like particles were enumerated by epifluorescence microscopy after staining with SYBR-Gold. Two isolates designated P94-4B3 (identified as Halomonas aquamarina) and P94-4S3 (identified as Pseudomonas aeruginosa) were further evaluated for potential use. Because of the rapid growth, larger size of its virus-like particles, and linear response to increasing dose of mitomycin C, the P. aeruginosa st. P94-4S3 was determined a better candidate for the marine prophage induction assay (MPIA). The Pseudomonas isolate was then used in several experiments for the development and optimization of the MPIA procedure. Initial screenings of the MPIA were also performed with selected environmental pollutants.

Journal Article↗

C1-C2 transarticular screw fixation for atlantoaxial instability: a 6-year experience.

OBJECTIVE: We review a 6-year, single-center experience using the technique of C1-C2 transarticular screw fixation for atlantoaxial instability in 75 consecutive operations. METHODS: The study group was composed of 43 men and 32 women, with a mean age of 44 years (range, 8-76 yr). Each patient had documented atlantoaxial instability. In 28 patients (37%), atlantoaxial instability was a result of trauma; in 22 patients, (29%), it was a result of rheumatoid arthritis; in 16 patients (21%), it was a result of prior surgery; and in 9 patients (12%), it was a result of congenital abnormalities. All patients underwent stabilization with C1-C2 transfacetal screws and a posterior interspinous construct. Nine patients had unilateral screws placed. Postoperatively, the patients were maintained in a rigid cervical orthosis for a mean of 11 weeks (range, 8-15 wk); five patients were immobilized with halo fixation for a mean of 13 weeks (range, 10-16 wk). The mean follow-up period was 2.4 years (range, 1-5.5 yr). RESULTS: Osseous fusion was documented in 72 patients (96%). There were no hardware failures; however, three patients developed pseudarthrosis. Two superficial wound infections (one at the graft site and one at the cervical incision site) required antibiotic therapy. Four patients had transient suboccipital hypesthesia. No instances of an errant screw, dural laceration, or injury to the vertebral artery, spinal cord, or hypoglossal nerve were noted. CONCLUSION: C1-C2 transarticular screw fixation supplemented with an interspinous construct yielded a 96% fusion rate, with a low incidence of complications. We attribute our successful outcomes to careful preoperative assessment and meticulous surgical technique.

Adolescent↗

Interaction of the PhiHSIC virus with its host: lysogeny or pseudolysogeny?

The marine phage PhiHSIC has been previously reported to enter into a lysogenic relationship with its host, HSIC, identified as Listonella pelagia. This phage produces a variety of plaques on its host, including turbid and haloed plaques, from which lysogens were previously isolated. These lysogens were unstable during long-term storage at -80( degrees ) C and were lost. When HSIC was reinfected with phage PhiHSIC, pseudolysogen-like interactions between the phage and its host were observed. The cells (termed HSIC-2 or HSIC-2e) produced high viral titers (10(11) ml(-1)) in the absence of inoculating phage and yet reached culture densities of nearly 10(9) ml(-1). Prophages were not induced by mitomycin C or the polyaromatic hydrocarbon naphthalene in cells harboring such infections. However, such cells were homoimmune to superinfection. Colonies hybridized strongly with a gene probe from a 100-bp fragment of the PhiHSIC genome, while the host did not. Analysis of chromosomal DNA preparations suggested the presence of a chromosomally integrated prophage. Phage adsorption experiments suggested that HSIC-2 was adsorption impaired. Because of the chromosomal prophage integration and homoimmunity, we interpret these results to indicate that PhiHSIC establishes a lysogenic relationship with its host that involves an extremely high level of spontaneous induction. This could be caused by a weak repressor of phage production. Additionally, poor phage adsorption of HSIC-2 compared to the wild type probably helped maintain this pseudolysogen-like relationship. In many ways, pseudolysogenic phage-host interactions may provide a paradigm for phage-host interactions in the marine environment.

Blotting, Southern↗

Threaded cortical bone dowels for lumbar interbody fusion: over 1-year mean follow up in 28 patients.

OBJECT: The authors retrospectively reviewed a series of 35 patients with mechanical low-back or one- to two-level discogenic pain; the patients underwent lumbar interbody fusion in which threaded cortical bone dowels (TCBDs) were placed to treat degenerative disc disease. The purpose of the study was to delineate fusion rates and outcome data in this series of patients. METHODS: The series was composed of 18 women and 17 men whose mean age was 46 years (range 17-76 years). There were nine active cigarette smokers. All patients presented with symptoms consistent with mechanical low-back or discogenic pain, and magnetic resonance imaging revealed degenerative changes related to disc collapse at one or two vertebral levels. For placement of the TCBDs, 23 patients underwent posterior lumbar interbody fusion (PLIF), whereas 12 patients underwent anterior lumbar interbody fusion (ALIF). In all except one patient undergoing PLIF, pedicle screw and rod constructs were used without posterolateral fusion. In all patients undergoing ALIF except one, TCBDs were used as stand-alone devices without supplemental fixation. At follow up the success of fusion was determined by static lumbar radiography and/or computerized tomography scanning. The degree of lumbar lordosis at the diseased level was measured immediately postoperatively and compared with that documented on follow-up radiological studies. Outcomes were assessed using a modified Prolo Scale. Excellent and good outcomes were considered satisfactory, and fair or poor outcomes were considered unsatisfactory. In 28 patients (eight ALIF and 20 PLIF) radiological and clinical follow-up data were considered adequate. The mean follow-up duration was 12.3 months. Overall satisfactory outcome was 60%; 70% satisfactory outcome was noted in PLIF patients and 38% in ALIF patients. Osseous fusion was present in 95% of the patients in the PLIF group and in 13% of those in the ALIF group. Complications included one L-5 nerve root injury and two postoperative wound infections, all in patients who underwent PLIF; in an ALIF patient lateral breakout of one implant occurred at 8 months postoperatively. CONCLUSIONS: Analysis of the mean 12.3 month follow-up data indicates that there is a dramatically higher fusion rate in PLIF compared with ALIF procedures when TCBDs are used. The authors believe that it is important to note that in all the PLIF procedures except one, supplemental pedicle screw/rod constructs were used, whereas in ALIF procedures no supplemental fixation was performed. The results thus suggest that TCBDs are best used in PLIF in conjunction with pedicle screw and rod constructs.

Adolescent↗

Do current outcomes data support the technique of lumbar interbody fusion?

Over the past 20 years, significant technical advances in spinal surgery have made the concept of interbody fusion a reality. The procedure, whether approached anteriorly or posteriorly, appears to improve patient satisfaction and radiographic fusion rates in selected cases. The lack of a randomized, prospective study and difficulty in comparing outcomes between studies has made it difficult to draw such a conclusion from the literature.

Humans↗

Management of acute odontoid fractures: operative techniques and complication avoidance.

In this article the authors describe the management of Type II odontoid fractures with special attention to operative technique and avoidance of complication. Anterior odontoid screw fixation is a procedure the authors have performed over the last 8 years in cases with acute Type II and rostral Type III odontoid fractures. In cases of Chronic Type II odontoid fractures and in patients with transverse ligament disruption, the authors prefer to undertake posterior transarticular facet screw fixation supplemented by bone graft and interspinous C1-2 wiring. The technical aspects of these procedures are described with a focus on operative nuances. Selection criteria and techniques that the authors have refined over the years have helped them to optimize success rates and minimize complications.

Bone Screws↗

Single-stage anterior-posterior decompression and stabilization for complex cervical spine disorders.

OBJECT: To evaluate the applicability and safety of single-stage combined anterior-posterior decompression and fusion for complex cervical spine disorders, the authors retrospectively reviewed 72 consecutive procedures of this type performed using a uniform technique at a single center. METHODS: The indications for decompression and stabilization included: postlaminectomy kyphosis (15 patients), trauma (19 patients), spondylosis and congenital stenosis (32 patients), and ossification of the posterior longitudinal ligament (six patients). All patients underwent anterior cervical corpectomies in which allograft fibula and plates were placed, with 89% of patients undergoing two- or three-level procedures (range one-four levels). Lateral mass plating with autograft (morselized iliac crest) fusion was performed in all patients while the same anesthetic agent was still in effect. A hard cervical collar was used postoperatively in all patients (mean 13 weeks). All patients were followed for a minimum of 2 years (mean 29 months). Fusion was determined to be successful in all 72 patients (100%). Although the short-term morbidity rate reached 32%, the significant long-term morbidity rate was only 5%. At the 2-year follow-up examination, anterior cervical plate dislodgment was seen in one patient, and 16 of the 516 lateral mass screws implanted were observed to have partially backed out. However, there were no cases of nerve root injury, strut graft extrusion, or anterior plate or screw fracture. There were no clinically significant hardware complications and no patient required repeated operation. CONCLUSIONS: The combined single-stage anterior-posterior decompression, reconstruction, and instrumentation procedure represents a viable option in the treatment of a select group of patients with complex cervical spinal disorders. The technique provides immediate rigid stabilization of the cervical spine, prevents anterior plate failure or strut graft extrusion, and eliminates the need for halo immobilization postoperatively. Furthermore, a higher rate of fusion is achieved with this combined approach than with the anterior approach alone.

Adult↗

Laparoscopic anterior lumbar interbody fusion. Technical note.

In recent years, there has been an unprecedented increase in the number of patients undergoing treatment with interbody fusion devices for degenerative disease of the lumbar spine. These devices can be placed either anteriorly or posteriorly. With the advent of minimally invasive surgery and the increasing ability of general surgeons to perform transperitoneal procedures laparoscopically, a new laparoscopic technique has been developed for placing lumbar interbody fusion devices. Although this procedure has some advantages over posterior lumbar interbody fusion, it is not without significant risk, and the learning curve is steep. The authors review a series of 32 consecutive patients who underwent single-level laparoscopic anterior lumbar interbody fusion at L4-5 or L5-S1 over a 2-year period for the treatment of single-level lumbar degenerative disease. In this report they review the technical aspects of the procedure and the important lessons they have learned through their early experience with this technique.

Journal Article↗

Beneficial effects of the radioprotectant 21-aminosteroid U-74389G in a radiosurgery rat malignant glioma model.

PURPOSE: To evaluate the radioprotectant effects of the 21-aminosteroid U-74389G on the rat C6 glioma model after stereotactic radiosurgery. Because radiosurgery causes both tumor cytotoxicity, as well as regional brain edema, we hypothesized that this drug might exhibit advantageous or deleterious effects on healthy and neoplastic tissue. METHODS: Rats were implanted with 10(6) C6 glioma cells into the right frontal brain and randomized to a Control Group (n = 18), radiosurgery on Day 14 (50% isodose = 35 Gy) (n = 15), or radiosurgery preceded by a single 15 mg/kg intravenous dose of 21-aminosteroid (n = 27). All animals were killed by 90 days and evaluated for survival, tumor size, the presence or absence of regional parenchymal edema, or radiation-induced vasculopathy. RESULTS: After tumor implantation, median survival in the Control Group was 23 days. Significant improvements in median survival were noted after RS alone (median, 31 days; p = 0.02), and RS plus 21-aminosteroid (median, 59 days; p < 0.0001). In the Control Group, mean tumor diameter was 5.4 mm. After RS alone, the mean diameter was 3.2 mm (p = 0.002), and after RS plus 21-aminosteroid, 2.9 mm (p = 0.0002). In the Control Group, the tumor grew as a hypercellular, compact mass. Only 3 of 18 animals had peritumoral edema. In contrast, 7 of 15 animals in the RS group had evidence of edema (p = 0.006), but rats that received 21-aminosteroid showed no increase compared to controls (p = 0.38). Similarly, 6 of 15 animals that had radiosurgery alone showed evidence of vasculopathy (p = 0.005) compared to no animals in the control group and only 2 of 27 aminosteroid-treated animals. CONCLUSIONS: The 21-aminosteroid U-74389G exhibits a radioprotectant effect on normal brain tissue, but does not appear to protect the tumor in an in vivo rat radiosurgery model. We believe that the observed beneficial effects on healthy brain led to significant prolongation of animal survival; perhaps, by limiting the adverse effects of high-dose radiosurgery. This radioprotectant should now be evaluated in randomized clinical trials in patients with malignant brain tumors.

Animals↗

Management of acute odontoid fractures with single-screw anterior fixation.

OBJECTIVE: Accepted management strategies for odontoid fractures include external immobilization and surgical stabilization using anterior or posterior approaches. Displaced Type II fractures and rostral Type III fractures are at high risk for nonunion. Anterior fixation of odontoid fractures with a single cortical lag screw is a relatively new technique that combines rigid internal stabilization with preservation of intrinsic C1-C2 motion. We retrospectively reviewed our series of 26 consecutive patients who underwent odontoid screw fixation, to further define the safety and efficacy of the technique. METHODS: During a 5-year period, 26 patients presented with acute traumatic Type II odontoid fractures. Ten patients were female and 16 were male, with a mean age of 35 years. All patients underwent anterior odontoid screw fixation by the senior surgeon (RWH), within a mean of 3 days after injury. All patients were postoperatively maintained in external orthoses, for a mean of 7.2 weeks, and were monitored with serial clinical and radiographic examinations. RESULTS: With a mean follow-up period of 30 months, radiographic fusion was documented for 25 of 26 patients (96%). No complications related to the surgical approach were identified, and all patients remained in neurologically stable condition. Two complications (8%) were related to the instrumentation; one patient required external immobilization because of suboptimal screw placement, and one patient required posterior atlantoaxial arthrodesis because of inadequate fracture reduction. CONCLUSION: Single-screw anterior odontoid fixation was associated with a relatively low complication rate and a high fusion rate in this study. We think that this should be the preferred treatment method for acute Type II odontoid fractures.

Adolescent↗

Microvascular decompression of cranial nerves: lessons learned after 4400 operations.

OBJECT: Microvascular decompression has become an accepted surgical technique for the treatment of trigeminal neuralgia, hemifacial spasm, glossopharyngeal neuralgia, and other cranial nerve rhizopathies. The senior author (P.J.J.) began performing this procedure in 1969 and has performed more than 4400 operations. The purpose of this article is to review some of the nuances of the technical aspects of this procedure. METHODS: A review of 4415 operations shows that numerous modifications to the technique of microvascular decompression have occurred during the last 29 years. Of the 2420 operations performed for trigeminal neuralgia, hemifacial spasm, and glossopharyngeal neuralgia before 1990, cerebellar injury occurred in 21 cases (0.87%), hearing loss in 48 (1.98%), and cerebrospinal fluid (CSF) leakage in 59 cases (2.44%). Of the 1995 operations performed since 1990, cerebellar injuries declined to nine cases (0.45%), hearing loss to 16 (0.8%), and CSF leakage to 37 (1.85% p < 0.01, test for equality of distributions). The authors describe slight variations made to maximize surgical exposure and minimize potential complications in each of the six principal steps of this operation. These modifications have led to decreasing complication rates in recent years. CONCLUSIONS: Using the techniques described in this report, microvascular decompression is an extremely safe and effective treatment for many cranial nerve rhizopathies.

Arteries↗