Feeding Sheep Cobalt and Oregano Essential Oil Alone or in Combination on Ruminal Nutrient Digestibility, Fermentation, and Fiber Digestion Combined With Scanning Electron Microscopy.

The feeding of Co lactate (Co), an essential oil blend (EO; oregano), or a combination of Co and EO (EOC) may improve nutrient digestion of corn silage-based rations. In four separate studies, Co, EO, or EOC was fed at 0, 4, and 7 g/days to nine rumen fistulated rams arranged in a replicated 3 × 3 Latin square design. The fourth study evaluated the carrier at 0, 4, and 7 g/day. In each ram, fresh ensiled corn silage, leaf, and husk were placed in individual nylon bags inserted through the ruminal cannula and removed after 48 h. Rams fed increasing carrier rates demonstrated similar (P > 0.10) nutrient digestibilities and ruminal pH and volatile fatty acid concentrations. Feeding Co at 4 and 7 g/day increased (P < 0.05) digestibility of DM (59.4, 63.9, and 62.4% for 0, 4, and 7 g/day, respectively), NDF (59.4, 63.9, and 62.4%), and hemicellulose (HC; 56.2, 63.6, and 65.9%) compared with rams fed 0 g/day, while CP digestibility (46.4, 49.9, and 57.8%) was improved (P < 0.05) in rams fed 7 g/day compared with those fed 0 and 4 g/day. Rams fed 4 g/day EO digested greater (P < 0.05) HC (64.1, 71.4, and 69.1%) than rams fed 0 g/day, while rams fed 7 g/day were intermediate and similar (P > 0.10). Rams fed the EOC combination at 4 and 7 g/day demonstrated greater (P < 0.05) digestibilities of DM (57.7, 60.0, and 60.0%), NDF (21.4, 28.8, and 27.7%), and ADF (24.3, 33.3, and 34.4%) than rams fed 0 g/day. The SEM and SM techniques visually demonstrated minor evidence of husk and leaf digestibility in rams across the three experiments when fed 0 g/day of Co, EO, or EOC; rams fed 4 g/day of Co, EO, or EOC exhibited varying visual signs of leaf digestion with some palisade tissue, spongy tissue, and whole vein structure remaining, while in rams fed 7 g/day, only the vein structure remained. Results demonstrated that feeding Co, EO, or EOC at 4 or 7 g/day enhanced ruminal nutrient digestion and fermentation parameters, which was visually confirmed via SEM and SM.

Association of IFIH1 and pro-inflammatory mediators: Potential new clues in SLE-associated pathogenesis.

Antiviral defenses are inappropriately activated in systemic lupus erythematosus (SLE) and association between SLE and the antiviral helicase gene, IFIH1, is well established. We sought to extend the previously reported association of pathogenic soluble mediators and autoantibodies with mouse Mda5 to its human ortholog, IFIH1. To better understand the role this gene plays in human lupus, we assessed association of IFIH1 variants with soluble mediators and autoantibodies in 357 European-American SLE patients, first-degree relatives, and unrelated, unaffected healthy controls. Association between each of 135 genotyped SNPs in IFIH1 and four lupus-associated plasma mediators, IL-6, TNF-α, IFN-ß, and IP-10, were investigated via linear regression. No significant associations were found to SNPs orthologous to those identified in exon 13 of the mouse. However, outside of this region there were significant associations between IL-6 and rs76162067 (p = 0.008), as well as IP-10 and rs79711023 (p = 0.003), located in a region of IFIH1 previously shown to directly influence MDA-5 mediated IP-10 and IL-6 secretion. SLE patients and FDRs carrying the minor allele for rs79711023 demonstrated lower levels of IP-10, while only FDRs carrying the minor allele for rs76162067 demonstrated an increased level of IL-6. This would suggest that the change in IP-10 is genotypically driven, while the change in IL-6 may be reflective of SLE transition status. These data suggest that IFIH1 may contribute to SLE pathogenesis via altered inflammatory mechanisms.

Activity-dependent plasticity in the isolated embryonic avian brainstem following manipulations of rhythmic spontaneous neural activity.

When rhythmic spontaneous neural activity (rSNA) first appears in the embryonic chick brainstem and cranial nerve motor axons it is principally driven by nicotinic neurotransmission (NT). At this early age, the nicotinic acetylcholine receptor (nAChR) agonist nicotine is known to critically disrupt rSNA at low concentrations (0.1-0.5µM), which are levels that mimic the blood plasma levels of a fetus following maternal cigarette smoking. Thus, we quantified the effect of persistent exposure to exogenous nicotine on rSNA using an in vitro developmental model. We found that rSNA was eliminated by continuous bath application of exogenous nicotine, but rSNA recovered activity within 6-12h despite the persistent activation and desensitization of nAChRs. During the recovery period rSNA was critically driven by chloride-mediated membrane depolarization instead of nicotinic NT. To test whether this observed compensation was unique to the antagonism of nicotinic NT or whether the loss of spiking behavior also played a role, we eliminated rSNA by lowering overall excitatory drive with a low [K(+)]o superfusate. In this context, rSNA again recovered, although the recovery time was much quicker, and exhibited a lower frequency, higher duration, and an increase in the number of bursts per episode when compared to control embryos. Importantly, we show that the main compensatory response to lower overall excitatory drive, similar to nicotinergic block, is a result of potentiated chloride mediated membrane depolarization. These results support increasing evidence that early neural circuits sense spiking behavior to maintain primordial bioelectric rhythms. Understanding the nature of developmental plasticity in the nervous system, especially versions that preserve rhythmic behaviors following clinically meaningful environmental stimuli, both normal and pathological, will require similar studies to determine the consequences of feedback compensation at more mature chronological ages.

Non-destructive and selective imaging of the functionally active, pro-invasive membrane type-1 matrix metalloproteinase (MT1-MMP) enzyme in cancer cells.

Proteolytic activity of cell surface-associated MT1-matrix metalloproteinase (MMP) (MMP-14) is directly related to cell migration, invasion, and metastasis. MT1-MMP is regulated as a proteinase by activation and conversion of the latent proenzyme into the active enzyme, and also via inhibition by tissue inhibitors of MMPs (TIMPs) and self-proteolysis. MT1-MMP is also regulated as a membrane protein through its internalization and recycling. Routine immunohistochemistry, flow cytometry, reverse transcription-PCR, and immunoblotting methodologies do not allow quantitative imaging and assessment of the cell-surface levels of the active, TIMP-free MT1-MMP enzyme. Here, we developed a fluorescent reporter prototype that targets the cellular active MT1-MMP enzyme alone. The reporter (MP-3653) represents a liposome tagged with a fluorochrome and functionalized with a PEG chain spacer linked to an inhibitory hydroxamate warhead. Our studies using the MP-3653 reporter and its inactive derivative demonstrated that MP-3653 can be efficiently used not only to visualize the trafficking of MT1-MMP through the cell compartment, but also to quantify the femtomolar range amounts of the cell surface-associated active MT1-MMP enzyme in multiple cancer cell types, including breast carcinoma, fibrosarcoma, and melanoma. Thus, the levels of the naturally expressed, fully functional, active cellular MT1-MMP enzyme are roughly equal to 1 × 10(5) molecules/cell, whereas these levels are in a 1 × 10(6) range in the cells with the enforced MT1-MMP expression. We suggest that the reporter we developed will contribute to the laboratory studies of MT1-MMP and then, ultimately, to the design of novel, more efficient prognostic approaches and personalized cancer therapies.

Evaluation of sterilization methods following contamination of hamstring autograft during anterior cruciate ligament reconstruction.

PURPOSE: Inadvertent contamination of the hamstring autograft during ACL reconstruction is infrequent, but can result in significant complications. The purpose of this study is to evaluate bacterial contamination of hamstring autografts dropped onto the operating room floor and methods of graft decontamination. METHODS: Hamstring tendons were harvested from patients. Excess tendon not used in the ACL procedure was divided into 6 segments. Segments were assigned to 6 groups (A through F, N = 30 in each group): group A: uncontaminated graft immediately postharvest (control), group B: graft dropped onto the floor (5 s), group C: graft dropped onto the floor (15 s). grafts in groups D to F were dropped onto floor for 15 s then rinsed with saline (group D), bacitracin solution (group E) or chlorhexidine 4 % solution (group F) for 3 min. All grafts were sent to the microbiology laboratory for anaerobic and aerobic cultures. RESULTS: Cultures were positive in 23 % of graft segments from group A (7/30), 33 % of grafts from group B (10/30), 23 % from group C (7/30), 30 % from group D (9/30) and 3 % from both group E (1/30) and group F (1/30). Sixteen unique organisms were identified, with Staphylococcus aureus as the most common isolate. Grafts rinsed in either bacitracin solution or 4 % chlorhexidine solutions were significantly less likely to be culture positive when compared to control graft segments (p < 0.05). However, there was no significant difference between uncontaminated grafts retrieved in <5 versus 15 s from the floor. CONCLUSION: This study supports the practice of decontaminating a dropped ACL hamstring autograft using either 4 % chlorhexidine or bacitracin solution. Specimens should be retrieved sterilely and washed for at least 3 min. This study also demonstrates no advantage in retrieval time of less than 5 s as compared to 15 s for uncontaminated graft. Hamstring harvest in ACL reconstruction may result in positive cultures, thus routine soaking of the hamstring autograft in either bacitracin or 4 % chlorhexidine solution is recommended. In addition, dropped hamstring autograft can be effectively sterilized with bacitracin or 4 % chlorhexidine solution. LEVEL OF EVIDENCE: II.

Minimally invasive mitral valve replacement in a patient with previous omphalocele repair and severe scoliosis.

Mitral valve surgery in a patient with severe chest deformity and poor respiratory function can be associated with a high risk due to difficult access and postoperative respiratory failure. A 45-year-old man with scoliosis and respiratory dysfunction who had undergone previous omphalocele repairs presented with severe mitral regurgitation. Mitral valve replacement via right mini-thoracotomy was successfully performed. The minimally invasive approach was considered useful in this patient with anatomical difficulty and respiratory dysfunction.

Latent tricuspid valve rupture after motor vehicle accident and routine echocardiography in all chest-wall traumas.

Blunt chest-wall trauma is common; however, resultant tricuspid valve rupture is rare and can be subtle in its presentation. Transthoracic echocardiography plays a key role in diagnosis. Herein, we report the case of a 42-year-old woman who sustained substantial chest-wall trauma in a high-speed motor vehicle accident. She presented a week later with symptoms of right-heart failure, secondary to flail tricuspid valve leaflets and torrential tricuspid regurgitation. The case of this patient highlights the importance of early diagnosis and elicits discussion of the mechanisms that can underlie delayed tricuspid valve rupture. Because the clinical diagnosis of tricuspid valve rupture can be difficult, we believe that echocardiography should be used early and, if necessary, repeatedly in all patients who sustain blunt chest-wall trauma.

Drug-eluting stent treatment of a radiation-induced left internal mammary arterial graft stenosis.

A 62-year-old man underwent radiotherapy to the left upper chest for treatment of Pancoast syndrome on a background of previous coronary artery bypass grafting 12 years earlier. Within 1 year, he developed significant stenoses of both the left internal mammary artery (LIMA) graft and ostial left vertebral artery, presumably related to therapeutic radiation exposure. Initially diagnosed using computed tomography coronary angiography, the patient underwent percutaneous coronary intervention and insertion of a drug-eluting stent (DES) to the ostium of the LIMA graft via a left radial approach. He remains clinically well at 6-month follow-up. This is the first reported case in the literature of DES treatment of a radiation-induced vascular stenosis; however, the incidence of cardiovascular disease is elevated in such cases. In patients with a prior history of mantle radiation, consideration should be given to the routine assessment of internal mammary conduits prior to coronary artery bypass surgery.

On-line purge and trap gas chromatography for monitoring of trihalomethanes in drinking water distribution systems.

A method using an automated on-line purge and trap gas chromatograph with a dry electrolytic conductivity detector (DELCD) has been developed for monitoring four regulated trihalomethanes in drinking water distribution systems. This analyzer samples trihalomethanes from drinking water by pervaporation through a silicone capillary membrane contained within a gas extraction cell (GEC) followed by preconcentration using an adsorbent trap. Trihalomethanes are subsequently desorbed from the trap onto a capillary column, separated and detected. The analyzer operates in real-time, samples directly from the drinking water distribution system and is fully automated. The optimization, operation, and evaluation of the analyzer and method are discussed. Method detection limits (MDL) are less than 1.0 microg L(-1) with acceptable estimates for accuracy, and precision. The results from two on-line monitoring studies in chlorinated and chloraminated distribution systems are presented. The performance of the method is compared directly to United Stated Environmental Protection Agency Method 502.2 and shows a very slight, but acceptable bias.