Rapid MRSA PCR on respiratory specimens from ventilated patients with suspected pneumonia: a tool to facilitate antimicrobial stewardship.

Methicillin-resistant Staphylococcus aureus (MRSA) is an important cause of pneumonia in ventilated patients. Our objective was to evaluate the GeneXpert MRSA/SA SSTI Assay (Xpert MRSA/SA) (Cepheid, Sunnyvale, CA) for use in lower respiratory tract (LRT) specimens for rapid MRSA detection and to determine the potentially saved antibiotic-days if a culture-based identification method was replaced by this assay. Remnant LRT samples from ventilated patients submitted to the microbiology laboratory for routine culture were tested using conventional culture and Xpert MRSA/SA. One hundred of 310 LRT specimens met the inclusion criteria. Ten samples were positive for MRSA by Xpert MRSA/SA, while six were positive by routine culture methods. Xpert MRSA/SA correctly identified 5/6 positive and 89/94 negative MRSA specimens, for a sensitivity of 83.3%, specificity of 94.7%, positive predictive value of 45.6%, and negative predictive value of 98.9%. The assay also correctly detected 3/3 positive and 90/97 negative methicillin-susceptible S. aureus (MSSA) specimens, for a sensitivity of 100%, specificity of 92.8%, positive predictive value of 30%, and negative predictive value of 100%. A total of 748 vancomycin and 305 linezolid antibiotic-days were associated with the enrolled specimens. Vancomycin and linezolid utilization could decrease by 68.4% and 83%, respectively, if discontinued 1 day after negative polymerase chain reaction (PCR) results. The Xpert MRSA/SA SSTI rapid MRSA PCR assay performed well in respiratory samples from ventilated patients with suspected pneumonia and has the potential to facilitate stewardship efforts such as reducing empiric vancomycin and linezolid therapy.

Comparison and optimization of two MALDI-TOF MS platforms for the identification of medically relevant yeast species.

The rapid identification of yeast is essential for the optimization of antifungal therapy. The objective of our study was to evaluate two matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) platforms, the bioMérieux VITEK MS (IVD Knowledgebase v.2.0) and Bruker Biotyper (software version 3.1), for the rapid identification of medically relevant yeast. One hundred and seventeen isolates, representing six genera and 18 species, were analyzed using multiple direct smear methods to optimize identification. Sequence analysis was the gold standard for comparison. Isolates were analyzed with VITEK MS using the direct smear method +/- a 25 % formic acid on-plate extraction. For Biotyper, isolates were analyzed using direct smear without formic acid, and with 25 % and 100 % formic acid on-plate extractions. When all methods were included, VITEK MS correctly identified 113 (96.6 %) isolates after 24 h with one misidentification, and Biotyper correctly identified 77 (65.8 %) isolates using a threshold of ≥2.0 with no misidentifications. Using a revised threshold of ≥1.7, Biotyper correctly identified 103 (88.0 %) isolates, with 3 (2.6 %) misidentifications. For both platforms, the number of identifications was significantly increased using a formic acid overlay (VITEK MS, p < 0.01; Biotyper, p < 0.001), and reducing the Biotyper threshold from ≥2.0 to ≥1.7 significantly increased the rate of identification (p < 0.001). The data in this study demonstrate that the direct smear method with on-plate formic acid extraction can be used for yeast identification on both MS platforms, and more isolates are identified using the VITEK MS system (p < 0.01).

Effects of ovariectomization and ractopamine hydrochloride inclusion on heifer feedlot performance, meat yield, and tenderness of select muscles.

Forty-eight British cross heifers were used to examine the effects of ovariectomization (OVX v. intact; INT) and ractopamine-HCl inclusion (0 or 0.41 mg/kg of BW day(-1); NORAC or RAC) for the final 31 days on performance, meat yield, and tenderness of select muscles. Ractopamine supplemented heifers had heavier BW (P≤0.05) and greater dressing percent than NORAC. Dressing percent was also greater (P<0.01) for INT v. OVX. LM area tended to be larger for RAC (P=0.07) and was larger for INT (P=0.05). Neither ractopamine inclusion nor sex class affected (P>0.08) organ weights or percent intramuscular fat. Ractopamine inclusion increased (P≤0.05) subprimal weight for the shoulder clod and bottom round. However, ractopamine inclusion and sex class had minimal effects on subprimal yields or slice shear force. Heifers receiving ractopamine had increased BW, DP, carcass weight and select subprimal weights without impacting meat quality. Ovariectomization did not influence the affects of ractopamine supplementation.

Activation of Matrix Metalloproteinase-3 is altered at the frog neuromuscular junction following changes in synaptic activity.

The extracellular matrix surrounding the neuromuscular junction is a highly specialized and dynamic structure. Matrix Metalloproteinases are enzymes that sculpt the extracellular matrix. Since synaptic activity is critical to the structure and function of this synapse, we investigated whether changes in synaptic activity levels could alter the activity of Matrix Metalloproteinases at the neuromuscular junction. In particular, we focused on Matrix Metalloproteinase 3 (MMP3), since antibodies to MMP3 recognize molecules at the frog neuromuscular junction, and MMP3 cleaves a number of synaptic basal lamina molecules, including agrin. Here we show that the fluorogenic compound (M2300) can be used to perform in vivo proteolytic imaging of the frog neuromuscular junction to directly measure the activity state of MMP3. Application of this compound reveals that active MMP3 is concentrated at the normal frog neuromuscular junction, and is tightly associated with the terminal Schwann cell. Blocking presynaptic activity via denervation, or TTX nerve blockade, results in a decreased level of active MMP3 at the neuromuscular junction. The loss of active MMP3 at the neuromuscular junction in denervated muscles can result from decreased activation of pro-MMP3, or it could result from increased inhibition of MMP3. These results support the hypothesis that changes in synaptic activity can alter the level of active MMP3 at the neuromuscular junction.

Endometritis in postparturient cattle associated with bovine herpesvirus-4 infection: 15 cases.

Suppurative, ulcerative endometritis associated with bovine herpesvirus-4 (BHV-4) infection was identified in 15 postparturient dairy cows from 5 separate dairies. Characteristic eosinophilic to amphophilic intranuclear viral inclusion bodies were identified within degenerate endometrial lining epithelium and endothelial cells. Bovine herpesvirus-4 was confirmed as the etiology by a combination of fluorescent antibody assays, viral isolation, heminested PCR, ultrastructural examination of the uterus and inoculated tissue culture cells, and negative-stain electron microscopy of tissue culture supernatant. Viral particles measuring 70-95 nm were demonstrated in uterine epithelial and endothelial cells by electron microscopy. Bacteria including Arcanobacterium pyogenes, Escherichia coli, and an alpha-Streptococcus isolate were isolated from all uteri. Bovine herpesvirus-4-associated endometritis has been previously reported in sporadic cases in Europe but has not been previously reported in the United States. Endometritis associated with BHV-4 appears to be an emerging syndrome in Georgia dairy herds.

The potential toxicity of Ilex myrtifolia in beef cattle.

Spatial and temporal patterns of death loss in a beef cowherd indicated a possible relationship between the loss of 11 cows and the consumption of Ilex myrtifolia (myrtle leaf holly). To investigate this relationship, plant material from Ilex myrtifolia was harvested and 2 feeding trials were performed. The 1st trial involved intermittent feeding of plant material to 4-mo-o calves for 2 w, and the 2nd trial was continuous plant feeding to 2-mo-o calves for 35 d. No significant clinical pathology, histological or gross lesions resulted and no clinical signs consistent with the original herd problem were observed, suggesting that berries, leaves and stems from Ilex myrtifolia were not sufficiently toxic to induce clinical effects under these experimental conditions.

Patient-focused models of care.

The structure and the process for delivering patient care will experience major changes during the next decade. Most hospitals have tried different alternatives, including restructuring, re-engineering, redesign, and the return to patient-focused care. Staffing strategies may successfully move nurses from total patient care to delegated, shared accountability. During their short stays, new parents and their neonates receive streamlined, intensely focused care from cross-trained workers in a patient-focused care environment. Each interaction becomes a meaningful and educational one, with the focal point being the mother and the family.

Upper lip flap for reconstruction of a full-thickness ala nasi defect.

Aesthetic reconstruction of full-thickness ala nasi defects has always been a challenge to the surgeon. Although several techniques are available, none has proved to be ideal. In this paper we describe a flap from the upper lip for reconstruction of defects involving the alar base and full-thickness defects of ala nasi. A flap based on the columella-labial junction is rotated upward for reconstruction of the alar base or ala nasi. The donor site of the flap can be closed primarily. When ala reconstruction is indicated, an upper lip flap forms the inner lining of the nostril, and a skin graft is applied over it for skin coverage. With this flap, an appropriately positioned ala with natural contours may be reconstructed in one stage.

Rhizobium infection and nodule development in soybean are affected by exposure of the cotyledons to light.

The initiation of Rhizobium infections and the development of nodules on the primary root of soybean Glycine max L. Merr cv Williams seedlings are strongly affected by exposure of the cotyledons/hypocotyls to light. Seedlings in plastic growth pouches were inoculated with R. japonicum in dim light and the position of the root tip of each seedling was marked on the face of the pouch. The pouches were covered and kept in the dark for various times before exposing the upper portions of the plants (cotyledons and hypocotyls) to light. Maximum nodulation occurred if the plants were kept in the dark until 1 day after inoculation. The exposure of plants to light 2 days before inoculation reduced the number of nodules by 50% while the number of nodules was reduced by 70% if the plants were kept in the dark until 7 days after inoculation. Anatomical studies revealed that exposure to light prior to inoculation reduced both the number of infection centers with visible infection threads and the number of infections which developed nodule meristems. Plants kept in the dark for 7 days after inoculation formed a normal number of infection threads above the root tip mark, but very few of these infections developed a nodule meristem. It appears that light stimulates soybean to produce substances which can both inhibit the formation of infection threads and enhance the development of nodules from established infection threads. The effects of light on nodulation appear to be expressed independently of the Rhizobium-induced suppression of nodule formation in younger regions of the root.