ABSTRACT
Urticaria is induced by the histamine released from mast cells which develops wheals (edema) as a visual feature. In clinical practice, second-generation histamine H1 -receptor blockers are routinely used as the first-line symptomatic treatment for urticaria. Nevertheless, not much research has directly examined the second-generation histamine H1-receptor blockers' ability to reduce edema. In this study, we directly evaluated the anti-edematous activities of three second-generation histamine H1-receptor blockers available in the market (epinastine hydrochloride, cetirizine hydrochloride, and levocetirizine hydrochloride) using a λ-carrageenan-induced footpad edema model. One hour before the induction of edema with 1% λ -carrageenan injection, all second-generation histamine H1 -receptor blockers (5, 10, 50 and 100 mg/kg) were subcutaneously administered to rats. At 0.5 and 3 hours after λ -carrageenan administration, the edema volume was evaluated using a Plethysmometer. Epinastine hydrochloride significantly suppressed the edema growth in a dose-dependent manner. Cetirizine hydrochloride showed a slight anti-edematous effect, while levocetirizine significantly inhibited the development of edema in a dose-dependent manner. On the other hand, dextrocetirizine did not prevent edema from growing. In summary, second-generation histamine H1 -receptor blockers, at least those examined in this study, may be able to reduce the clinical symptoms of urticaria associated with edema. Levocetirizine hydrochloride is also anticipated to have stronger anti-edematous effects than cetirizine hydrochloride because levocetirizine is responsible for cetirizine's anti-edematous activity.
Subject(s)
Carrageenan , Cetirizine , Edema , Animals , Cetirizine/pharmacology , Edema/drug therapy , Edema/chemically induced , Rats , Male , Stereoisomerism , Histamine H1 Antagonists/pharmacology , Histamine H1 Antagonists, Non-Sedating/pharmacology , Dose-Response Relationship, Drug , Rats, Wistar , Imidazoles/pharmacology , Rats, Sprague-Dawley , DibenzazepinesABSTRACT
BACKGROUND: The efficacy and safety of transanal lateral pelvic lymph node dissection (TaLPLND) in rectal cancer has not yet been clarified. The aim of the present study was to evaluate the short-term results as an initial experience of TaLPLND. METHODS: This retrospective study included patients with middle to lower rectal cancer who underwent TaLPLND from July 2018 to July 2021. Our institutions targeted lymph nodes in the internal iliac area and the obturator area for lateral pelvic lymph node dissection (LPLND). RESULTS: A total of 30 consecutive patients with rectal cancer were included in this analysis. The median age was 60 years (range, 36-83 years), and the male-female ratio was 2:1. The median operative time was 362 min (IQR, 283-661 min), and the median intraoperative blood loss was 74 ml (IQR, 5-500 ml). Intraoperative blood transfusion was required in one case. No cases required conversion to laparotomy. TaLPLND was performed bilaterally in 13 patients (43.3%). Five patients (16.7%) underwent LPLND with combined resection of the internal iliac vessels. The median distance of the distal margin from the anal verge was 20 mm. The pathological radial margin (pRM) was positive in one case, and the negative pRM rate was 96.7%. Short-term postoperative complications (Clavien-Dindo classification grade ≥ II) were observed in nine cases (30.0%). There were no cases of reoperation or mortality. The median number of harvested lateral pelvic lymph nodes was 11 (range, 3-28). On pathological examination, lateral pelvic lymph nodes were positive for metastasis in seven cases (23.3%). CONCLUSIONS: TaLPLND appeared to be beneficial from an oncological point of view because it was close to the upstream lymphatic drainage from the tumor. The short-term outcomes of this initial experience indicate that this novel approach is feasible.
Subject(s)
Laparoscopy , Rectal Neoplasms , Humans , Male , Female , Middle Aged , Retrospective Studies , Laparoscopy/methods , Lymph Node Excision/methods , Lymph Nodes/surgery , Lymph Nodes/pathology , Rectal Neoplasms/surgery , Rectal Neoplasms/pathologyABSTRACT
BACKGROUND: We previously reported that indocyanine green fluorescence imaging (ICG-FI)-guided laparoscopic lateral pelvic lymph node dissection (LPLND) was able to increase the total number of harvested lateral pelvic lymph nodes without impairing functional preservation. However, the long-term outcomes of ICG-FI-guided laparoscopic LPLND have not been evaluated. The aim of the present study was to compare the long-term outcomes of ICG-FI-guided laparoscopic LPLND to conventional laparoscopic LPLND without ICG-FI. METHODS: This was a retrospective, multi-institutional study with propensity score matching. The study population included consecutive patients with middle-low rectal cancer (clinical stage II to III) who underwent laparoscopic LPLND between January 2013 and February 2018. The main evaluation items in this study were the 3-year overall survival, relapse-free survival (RFS), local recurrence rate, and lateral local recurrence (LLR) rate. RESULTS: A total of 172 patients with middle-lower rectal cancer who had undergone laparoscopic LPLND were included in this study. After propensity score matching, 58 patients were matched in each of the ICG-FI and non-ICG-FI groups. There were no substantial differences in the baseline characteristics between the two groups. The ICG-FI group and non-ICG-FI group included 40 and 38 women and had a median age of 65 (IQR 60-72) and 66 (IQR 60-73) years, respectively. The median follow-up for all patients was 63.7 (IQR 51.3-76.8) months. The estimated respective 3-year overall survival, RFS, and local recurrence rates were 93.1%, 70.7%, and 5.2% in the ICG-FI group and 85.9%, 71.7%, and 12.8% in the non-ICG-FI group (p = 0.201, 0.653, 0.391). The 3-year cumulative LLR rate was 0% in the ICG-FI group and 9.3% in the non-ICG-FI group (p = 0.048). CONCLUSIONS: This study revealed that laparoscopic LPLND combined with ICG-FI was able to decrease the LLR rate. It appears that ICG-FI could contribute to improving the quality of laparoscopic LPLND and strengthening local control of the lateral pelvis. TRIALS REGISTRATION: This study was registered with the Japanese Clinical Trials Registry as UMIN000041372 ( http://www.umin.ac.jp/ctr/index.htm ).
Subject(s)
Laparoscopy , Rectal Neoplasms , Humans , Female , Middle Aged , Aged , Indocyanine Green , Cohort Studies , Retrospective Studies , Propensity Score , Neoplasm Recurrence, Local/surgery , Lymph Node Excision/methods , Lymph Nodes/pathology , Laparoscopy/methods , Rectal Neoplasms/diagnostic imaging , Rectal Neoplasms/surgery , Rectal Neoplasms/pathology , Optical Imaging/methodsABSTRACT
We introduce the concept of a handlebody decomposition of a three-manifold, a generalization of a Heegaard splitting, or a trisection. We show that two handlebody decompositions of a closed orientable three-manifold are stably equivalent. As an application to materials science, we consider a mathematical model of polycontinuous patterns and discuss a topological study of microphase separation of a block copolymer melt.
ABSTRACT
AIMS: Given the significance of Salmonella enterica in both human and animal health, and a recent global dissemination of Salmonella 4,[5],12:i:-, changes in the prevalent serovars and antimicrobial resistance in clinical isolates of Salmonella from cattle and pigs were investigated in Japan. METHODS AND RESULTS: The serovars and antimicrobial susceptibilities of 1605 Salmonella enterica isolated from cattle (n = 894) and swine (n = 711) between 2002 and 2016 were examined. The most common serovar among all samples was Salmonella Typhimurium. However, its monophasic variant with antigenic structure S. 4,[5],12:i:-, which was first detected in cattle in 2006 and swine in 2010, has been rapidly increasing in incidence and resistance. Resistance rates to cefotaxime and ciprofloxacin were generally low (<10% in the cattle isolates and <5% in the swine isolates); however, isolates resistant to more than five antimicrobials, which often include these antimicrobials, were recently detected in Salmonella Dublin, S. 4,[5],12:i:-, S. Typhimurium, Salmonella Newport, Salmonella Choleraesuis and Salmonella 6,7:c:-. Among them, two S. 4,[5],12:i:- isolates possessed extended-spectrum ß-lactamase-encoding genes; blaSHV-12 or blaCTX-M-55 , respectively, while all the five S. Typhimurium isolates possessed AmpC-type ß-lactamase gene of blaCMY-2 . CONCLUSIONS: S. 4,[5],12:i:- has been rapidly increasing and exhibiting a remarkable change in antimicrobial resistance in Japan. Considering certain serovars are characterized by multidrug resistance including medically important antimicrobials, continuous monitoring and appropriate measures are required to protect public health and veterinary husbandry. SIGNIFICANCE AND IMPACT OF THE STUDY: This study presents a trend in the serovars and antimicrobial resistance in clinical isolates of Salmonella from cattle and pigs in Japan, and showed that there were certain types of Salmonella serovars depending on the animal origin which needs more attention.
Subject(s)
Anti-Bacterial Agents/pharmacology , Drug Resistance, Bacterial , Salmonella Infections, Animal/microbiology , Salmonella enterica/drug effects , Salmonella enterica/genetics , Animals , Cattle , Drug Resistance, Bacterial/drug effects , Drug Resistance, Bacterial/genetics , Japan/epidemiology , Microbial Sensitivity Tests , Salmonella Infections, Animal/epidemiology , Salmonella enterica/isolation & purification , Serogroup , SwineABSTRACT
BACKGROUND: Brain tumour patients see their primary care doctor on average three or more times before diagnosis, so there may be an opportunity to identify 'at risk' patients earlier. Suspecting a brain tumour diagnosis is difficult because brain tumour-related symptoms are typically non-specific. METHODS: We explored the predictive value of referral guidelines (Kernick and NICE 2005) for brain imaging where a tumour is suspected, in a population-based patient group referred for direct access CT of the head. A consensus panel reviewed whether non-tumour findings were clinically important or whether further investigation was necessary. RESULTS: Over a 5-year period, 3257 head scans were performed; 318 scans were excluded according to pre-specified criteria. 53 patients (1.8%) were reported to have intracranial tumours, of which 42 were significant (diagnostic yield of 1.43%). There were no false negative CT scans for tumour. With symptom-based referral guidelines primary care doctors can identify patients with a 3% positive predictive value (PPV). 559 patients had non-tumour findings, 31% of which were deemed clinically significant. In 34% of these 559 patients, referral for further imaging and/or specialist assessment from primary care was still thought warranted. CONCLUSION: Existing referral guidelines are insufficient to stratify patients adequately based on their symptoms, according to the likelihood that a tumour will be found on brain imaging. Identification of non-tumour findings may be significant for patients and earlier specialist input into interpretation of these images may be beneficial. Improving guidelines to better identify patients at risk of a brain tumour should be a priority, to improve speed of diagnosis, and reduce unnecessary imaging and costs. Future guidelines may incorporate groups of symptoms, clinical signs and tests to improve the predictive value.
Subject(s)
Brain Neoplasms/diagnostic imaging , Neuroimaging , Referral and Consultation , Tomography, X-Ray Computed , Adolescent , Adult , Aged , Aged, 80 and over , Brain Neoplasms/diagnosis , Female , Humans , Male , Middle Aged , Referral and Consultation/organization & administration , Referral and Consultation/standards , Young AdultABSTRACT
Aims: The aim of this study was to report the mid-term clinical outcome of cemented unlinked J-alumina ceramic elbow (JACE) arthroplasties when used in patients with rheumatoid arthritis (RA). Patients and Methods: We retrospectively reviewed 87 elbows, in 75 patients with RA, which was replaced using a cemented JACE total elbow arthroplasty (TEA) between August 2003 and December 2012, with a follow-up of 96%. There were 72 women and three men, with a mean age of 62 years (35 to 79). The mean follow-up was nine years (2 to 14). The clinical condition of each elbow before and after surgery was assessed using the Mayo Elbow Performance Index (MEPI, 0 to 100 points). Radiographic loosening was defined as a progressive radiolucent line of >1 mm that was completely circumferential around the prosthesis. Results: The mean MEPI scores significantly improved from 40 (10 to 75) points preoperatively to 95 (30 to 100) points at final follow-up (p < 0.0001). Complications were noted in ten elbows (ten patients; 11%). Two had an intraoperative humeral fracture which was treated by fixation and united. One had a postoperative fracture of the olecranon which united with conservative treatment and one had a radial neuropathy which resolved. Further surgery was required for one with a dislocation, three with an ulnar neuropathy and one with a postoperative humeral fracture. Revision with removal of the components was performed in one elbow due to deep infection. There was no radiographic evidence of loosening around the components. With any revision surgery or revision with implant removal as the endpoint, the rates of survival up to 14 years were 93% (95% confidence interval (CI), 83.9 to 96.6) and 99% (95% CI 91.9 to 99.8), respectively, as determined by Kaplan-Meier analysis. Conclusion: With the appropriate indications, the mid-term clinical performance of the cemented JACE TEA is reliable and comparable to other established TEAs in the management of the elbow in patients with RA. Cite this article: Bone Joint J 2018;100-B:1066-73.
Subject(s)
Aluminum Oxide/administration & dosage , Arthritis, Rheumatoid/surgery , Arthroplasty, Replacement, Elbow/methods , Bone Cements/adverse effects , Elbow Prosthesis , Adult , Aged , Arthritis, Rheumatoid/diagnostic imaging , Arthroplasty, Replacement, Elbow/instrumentation , Female , Humans , Male , Middle Aged , Prosthesis Design , Radiography , Suction/methods , Suture Techniques , Synovectomy/methods , Treatment OutcomeABSTRACT
Swine influenza viruses (SIVs) are important not only for pig farming, but also for public health. In fact, pandemic A(H1N1) 2009 viruses [A(H1N1)pdm09] were derived from SIVs. Therefore, timely characterization of locally circulating SIVs is necessary for understanding the global status of SIVs. To genetically characterize SIVs circulating in Japanese pig populations, we isolated 24 SIVs of three subtypes (17 H1N1, four H1N2 and three H3N2 strains) from 14 pig farms in Japan from 2013 to 2016. Genetic analyses revealed that the haemagglutinin (HA) and neuraminidase (NA) genes of the 17 H1N1 and the HA gene of one H1N2, A/swine/Aichi/02/2016 (H1N2), SIVs belonged to the A(H1N1)pdm09 lineage. More importantly, all of the remaining six gene segments (i.e., PB1, PB1, PA, NP, M and NS) of the 24 SIVs, regardless of the HA and NA subtype, were also classified as belonging to the A(H1N1)pdm09 lineage. These results indicate that gene segments of A(H1N1)pdm09 lineage are widely distributed in SIVs circulating in Japanese pig populations In addition, the NA gene of A/swine/Aichi/02/2016 (H1N2) shared less than 88.5% nucleotide identity with that of the closest relative A/swine/Miyagi/5/2003 (H1N2), which was isolated in Japan in 2003. These results indicate the sustained circulation of classical H1N2-derived SIVs with remarkable diversity in the NA genes in Japanese pig populations. These findings highlight the necessity of both intensive biosecurity systems and active SIV surveillance in pig populations worldwide for both animal and public health.
Subject(s)
Influenza A Virus, H1N1 Subtype/genetics , Influenza A Virus, H1N2 Subtype/isolation & purification , Influenza A Virus, H3N2 Subtype/isolation & purification , Orthomyxoviridae Infections/veterinary , Pandemics , Swine Diseases/epidemiology , Animals , Dogs , Genome, Viral/genetics , Humans , Japan , Madin Darby Canine Kidney Cells , Neuraminidase/genetics , Orthomyxoviridae Infections/epidemiology , Orthomyxoviridae Infections/virology , Phylogeny , Swine , Swine Diseases/virology , Viral Proteins/geneticsABSTRACT
We isolated eight highly pathogenic H5N8 avian influenza viruses (H5N8 HPAIVs) in the 2014/15 winter season at an overwintering site of migratory birds in Japan. Genetic analyses revealed that these isolates were divided into three groups, indicating the co-circulation of three genetic groups of H5N8 HPAIV among these migratory birds. These results also imply the possibility of global redistribution of the H5N8 HPAIVs via the migration of these birds next winter.
Subject(s)
Animal Migration , Birds/virology , Genetic Variation , Influenza A virus/genetics , Influenza A virus/isolation & purification , Animals , Influenza A virus/classification , Influenza A virus/pathogenicity , Influenza in Birds/virology , Japan , Phylogeny , Seasons , Sequence Analysis, DNAABSTRACT
BACKGROUND: CD133 and CD44 are putative cancer stem cell (CSC) markers in colorectal cancer (CRC). However, their clinical significance is currently unclear. Here, we evaluated primary CRC cell isolates to determine the significance of several CSC markers, including CD133 and CD44, as predictors of tumourigenesis and prognosis. METHODS: CD133- and CD44-positive cells from fresh clinical samples of 77 CRCs were selected by flow cytometric sorting and evaluated for tumourigenicity following subcutaneous transplantation into NOD/SCID mice. Cancer stem cell marker expression was examined in both xenografts and a complementary DNA library compiled from 167 CRC patient samples. RESULTS: CD44(+), CD133(+) and CD133(+)CD44(+) sub-populations were significantly more tumourigenic than the total cell population. The clinical samples expressed several transcript variants of CD44. Variant 2 was specifically overexpressed in both primary tumours and xenografts in comparison with the normal mucosa. A prognostic assay using qRT-PCR showed that the CD44v2(high) group (n=84, 5-year survival rate (5-OS): 0.74) had a significantly worse prognosis (P=0.041) than the CD44v2(low) group (n=83, 5-OS: 0.88). CONCLUSIONS: CD44 is an important CSC marker in CRC patients. Furthermore, CRC patients with high expression of CD44v2 have a poorer prognosis than patients with other CD44 variants.
Subject(s)
Colorectal Neoplasms/metabolism , Hyaluronan Receptors/metabolism , AC133 Antigen , Adult , Aged , Aged, 80 and over , Animals , Antigens, CD/genetics , Antigens, CD/metabolism , Colorectal Neoplasms/genetics , Colorectal Neoplasms/pathology , Gene Expression , Glycoproteins/genetics , Glycoproteins/metabolism , Humans , Hyaluronan Receptors/genetics , Male , Mice , Mice, Inbred NOD , Mice, SCID , Middle Aged , Neoplastic Stem Cells/pathology , Peptides/genetics , Peptides/metabolism , Prognosis , Up-Regulation , Xenograft Model Antitumor AssaysABSTRACT
We developed a 2(nd) generation suprachoroidal transretinal stimulation (STS) system with a 49 channel electrode array and implanted in 2 dogs. One month after surgery, all electrodes were functioning and the ocular fundus was normal in both dogs. The results indicate the 2(nd) generation STS retinal prosthesis is feasible and can be considered for clinical use.
Subject(s)
Choroid/physiology , Photic Stimulation , Retina/physiology , Visual Prosthesis , Animals , Artifacts , Dogs , Electrodes, Implanted , Electroretinography , Evoked Potentials, Visual , Feasibility Studies , Fundus Oculi , Male , Prosthesis ImplantationABSTRACT
AIMS: Early studies have shown that magnesium intake decreases the risk of Type 2 diabetes, but the results are still inconsistent. We prospectively examined the association between magnesium intake and incidence of Type 2 diabetes in a general Japanese population. METHODS: A total of 1999 subjects without diabetes aged 40-79 years who underwent a 75-g oral glucose tolerance test were followed up prospectively for a mean of 15.6 years. RESULTS: During the follow-up, 417 subjects developed Type 2 diabetes. The age- and sex-adjusted incidence of Type 2 diabetes significantly decreased with increasing magnesium intake quartile levels (≤ 148.5, 148.6-171.5, 171.6-195.5 and ≥ 195.6 mg/day, P for trend = 0.01). In multivariate analyses, after adjusting for comprehensive risk factors and other dietary factors, the hazard ratio of Type 2 diabetes was 0.67 (95% CI 0.49-0.92; P = 0.01) in the third quartile and 0.63 (95% CI 0.44-0.90; P = 0.01) in the highest quartile compared with the first quartile. In addition, the risk of Type 2 diabetes was 14% lower (P = 0.04) for a 1-sd increment of log-transformed magnesium intake in the multivariate-adjusted model. In stratified analysis, there were statistically significant interactions between magnesium intake and levels of homeostasis model assessment of insulin resistance, high-sensitivity C-reactive protein or alcohol intake on the risk of Type 2 diabetes (all P < 0.05). CONCLUSIONS: Our findings suggest that increased magnesium intake was a significant protective factor for the incidence of Type 2 diabetes in the general Japanese population, especially among subjects with insulin resistance, low-grade inflammation and a drinking habit.
Subject(s)
Diabetes Mellitus, Type 2/prevention & control , Inflammation/metabolism , Insulin Resistance , Magnesium Deficiency/drug therapy , Magnesium/therapeutic use , Adult , Aged , Diabetes Mellitus, Type 2/blood , Female , Follow-Up Studies , Glucose Tolerance Test , Humans , Incidence , Inflammation/blood , Japan , Magnesium/blood , Magnesium Deficiency/blood , Magnesium Deficiency/complications , Male , Middle Aged , Nutrition Surveys , Prevalence , Prospective Studies , Risk Factors , Time FactorsABSTRACT
An accurate method using image sharpness to determine the best focusing is proposed for ultra-high-voltage electron microscopy. This method maximizes image sharpness for adjusting the focus. Five images with different defocus values are used to calculate the image sharpness. To obtain the best focus value that produces greatest image sharpness, fitting the quasi-Gaussian function to five image sharpness is a suitable alternative. This method, which maximizes image sharpness, gives better accuracy than the wobbler method for the ultra-high-voltage electron microscope. The focusing area can be selected without moving the field of view, because the focusing area can be selected at almost any area in the image.
Subject(s)
Image Enhancement/methods , Microscopy, Electron, Transmission/methods , AlgorithmsABSTRACT
OBJECTIVE: To investigate the inhibitory effects and the regulatory mechanisms of histone deacetylase (HDAC) inhibitors on mechanical stress-induced gene expression of runt-related transcription factor (RUNX)-2 and adisintegrin and metalloproteinase with thrombospondin motif (ADAMTS)-5 in human chondrocytes. METHODS: Human chondrocytes were seeded in stretch chambers at a concentration of 5 × 10(4)cells/chamber. Cells were pre-incubated with or without HDAC inhibitors (MS-275 or trichostatin A; TSA) for 12h, followed by uniaxial cyclic tensile strain (CTS) (0.5Hz, 10% elongation), which was applied for 30 min using the ST-140-10 system (STREX, Osaka, Japan). Total RNA was extracted and the expression of RUNX-2, ADAMTS-5, matrix metalloproteinase (MMP)-3, and MMP-13 at the mRNA and protein levels were examined by real-time polymerase chain reaction (PCR) and immunocytochemistry, respectively. The activation of diverse mitogen-activated protein kinase (MAPK) pathways with or without HDAC inhibitors during CTS was examined by western blotting. RESULTS: HDAC inhibitors (TSA: 10 nM, MS-275: 100 nM) suppressed CTS-induced expression of RUNX-2, ADAMTS-5, and MMP-3 at both the mRNA and protein levels within 1h. CTS-induced activation of p38 MAPK (p38), extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK) MAPKs was downregulated by both HDAC inhibitors. CONCLUSION: The CTS-induced expression of RUNX-2 and ADAMTS-5 was suppressed by HDAC inhibitors via the inhibition of the MAPK pathway activation in human chondrocytes. The results of the current study suggested a novel therapeutic role for HDAC inhibitors against degenerative joint disease such as osteoarthritis.
Subject(s)
ADAM Proteins/metabolism , Chondrocytes/metabolism , Core Binding Factor Alpha 1 Subunit/metabolism , Histone Deacetylase Inhibitors/pharmacology , Mitogen-Activated Protein Kinases/antagonists & inhibitors , ADAM Proteins/genetics , ADAMTS5 Protein , Blotting, Western , Core Binding Factor Alpha 1 Subunit/genetics , Gene Expression , Humans , JNK Mitogen-Activated Protein Kinases/metabolism , MAP Kinase Signaling System/physiology , Male , Matrix Metalloproteinase 13/metabolism , Matrix Metalloproteinase 3/metabolism , Middle Aged , Real-Time Polymerase Chain Reaction , Stress, Mechanical , p38 Mitogen-Activated Protein Kinases/metabolismABSTRACT
The hypothesis was tested that the pluripotency of the inner cell mass (ICM) of the bovine embryo is enhanced by the glycogen synthase kinase-3ß inhibitor CHIR99021 and the MAPK1 and MAPK3 inhibitor PD032591. Treatment with the two inhibitors from Days 6 to 8 after insemination increased blastocyst steady state concentrations of mRNA for NANOG (P < 0.05) and SOX2 (P = 0.055) and tended to decrease (P = 0.09) expression of GATA6. To evaluate pluripotency, the inner cell mass was isolated by immunosurgery at Day 8, seeded on a feeder layer of bovine embryonic fibroblasts, and cultured in the presence of the inhibitors. Ten of 52 (19%) ICM from control embryos had primary outgrowth formation vs. 23 of 50 (46%) of the ICM from embryos cultured with inhibitors (P < 0.01). For ICM outgrowths from embryos cultured without inhibitors, colonies either did not persist through Passage 2 or became differentiated. In contrast, for the inhibitor group, four colonies survived beyond Passage 2, and one line persisted for 19 passages. This cell line possessed alkaline phosphatase activity, expressed several genes characteristically expressed in pluripotent cells, and differentiated into embryoid bodies when cultured in the absence of the signal transduction inhibitors and the feeder layer. Propagation of the cells was difficult due to slow growth and inefficiency in survival through each passage. In conclusion, exposure to inhibitors during the morula-blastocyst transition facilitated formation of self-renewing pluripotent cell lines from bovine blastocysts.
Subject(s)
Blastocyst Inner Cell Mass/cytology , Blastocyst/physiology , Cattle/embryology , Embryo Culture Techniques/veterinary , Morula/physiology , Pluripotent Stem Cells/cytology , Animals , Blastocyst/chemistry , Cell Separation/veterinary , Colforsin/pharmacology , Enzyme Inhibitors/pharmacology , Glycogen Synthase Kinase 3/antagonists & inhibitors , Glycogen Synthase Kinase 3 beta , Homeodomain Proteins/genetics , Mitogen-Activated Protein Kinase 1/antagonists & inhibitors , Mitogen-Activated Protein Kinase 3/antagonists & inhibitors , Pyridines/pharmacology , Pyrimidines/pharmacology , RNA, Messenger/analysis , SOXB1 Transcription Factors/geneticsABSTRACT
To determine associations between antimicrobial use and antimicrobial resistance in Campylobacter coli, 155 isolates were obtained from the feces of apparently healthy grow-finish pigs in Japan. In addition, data on the use of antibiotics collected through the national antimicrobial resistance monitoring system in Japan were used for the analysis. Logistic regression was used to identify risk factors to antimicrobial resistance in C. coli in pigs for the following antimicrobials: ampicillin, dihydrostreptomycin, erythromycin, oxytetracycline, chloramphenicol, and enrofloxacin. The data suggested the involvement of several different mechanisms of resistance selection. The statistical relationships were suggestive of co-selection; use of macrolides was associated with enrofloxacin resistance (OR=2.94; CI(95%): 0.997, 8.68) and use of tetracyclines was associated with chloramphenicol resistance (OR=2.37; CI(95%): 1.08, 5.19). The statistical relationships were suggestive of cross-resistance: use of macrolides was associated with erythromycin resistance (OR=9.36; CI(95%): 2.96, 29.62) and the use of phenicols was associated with chloramphenicol resistance (OR=11.83; CI(95%): 1.41, 99.44). These data showed that the use of antimicrobials in pigs selects for resistance in C. coli within and between classes of antimicrobials.
Subject(s)
Anti-Bacterial Agents/pharmacology , Campylobacter coli/drug effects , Drug Resistance, Multiple, Bacterial/drug effects , Swine/microbiology , Agriculture , Animals , Campylobacter coli/classification , Campylobacter coli/genetics , Campylobacter coli/isolation & purification , Cross-Sectional Studies , Feces/microbiology , Japan/epidemiology , Logistic Models , Microbial Sensitivity Tests/veterinary , Polymerase Chain Reaction/veterinary , Risk Factors , Selection, GeneticABSTRACT
In the present study, we performed quantitative reverse-transcription PCR (qPCR) to examine changes in gene expression of prolactin receptor (long form: l-PRLR; short form: s-PRLR) and 20α-hydroxysteroid dehydrogenase (20α-HSD; EC 1.1.1.149) in the bovine corpus luteum (CL) throughout the estrous cycle and pregnancy. Western blotting was used to determine protein abundance. Bovine CL were collected and luteal stages (n = 6/stage) were classified by macroscopic observation as early (d 1 to 4 after ovulation), mid (d 5 to 10), late (d 11 to 17), and regressing (d 18 to 20). A CL of pregnancy (n = 6) was determined by the presence of conceptus (d 28 to term). The mRNA for both forms of PRLR were expressed at all the luteal stages. Expression of s-PRLR and l-PRLR mRNA was less (P < 0.01) during early and regressing luteal stages compared with mid and late stages. Expression of s-PRLR mRNA in CL of pregnancy was greater (P < 0.01) than early, mid, and regressing CL and did not differ from late luteal stage expression. A greater (P < 0.01) expression of l-PRLR mRNA was observed in pregnant vs. early and regressing CL. In addition, qPCR showed the presence of 20α-HSD mRNA during all luteal stages of the estrous cycle, with the greatest (P < 0.01) expression observed in the regressing luteal stage. Western blotting revealed protein abundance of both PRLR isoforms during all luteal stages and pregnancy, with a predominance of the s-PRLR protein. Densitometry analysis indicated that protein abundances of s-PRLR were greater (P < 0.05) than l-PRLR during early, mid, and late luteal stages and did not differ during the regressing luteal stage. Protein abundances of 20α-HSD were least (P < 0.05) during the early luteal stage. In conclusion, results of the current study suggest a possible involvement of PRLR, especially s-PRLR, in the regulation of progesterone secretion and metabolism during the bovine estrous cycle and pregnancy.
Subject(s)
Cattle/physiology , Corpus Luteum/metabolism , Estrous Cycle/physiology , Progesterone/metabolism , Receptors, Prolactin/biosynthesis , 20-alpha-Hydroxysteroid Dehydrogenase/biosynthesis , 20-alpha-Hydroxysteroid Dehydrogenase/genetics , Animals , Blotting, Western/veterinary , Corpus Luteum/enzymology , Female , Gene Expression Regulation, Enzymologic , Pregnancy , Protein Isoforms , RNA, Messenger/biosynthesis , RNA, Messenger/genetics , Receptors, Prolactin/genetics , Reverse Transcriptase Polymerase Chain Reaction/veterinaryABSTRACT
Insulin like growth factor-1 (IGF1) increases the proportion of embryos that develop to the blastocyst stage. The objective of the present study was to determine whether the pro-developmental actions of IGF1 are exerted before or after Day 4 of development (i.e., on events occurring through the period of genomic activation versus events coincident with compaction and blastocoel formation) and whether mitogen-activated protein kinase (MAPK) signaling pathways mediate effects of IGF1. Treatment with IGF1 increased the proportion becoming blastocysts at concentrations of 10, 100 and 200 ng/mL, with 100 ng/mL being more effective than 10 or 200 ng/mL. At Day 8, the percent of oocytes that became blastocysts was 30, 34, 43, and 36%, respectively (SEM=2.6). As compared to controls (30.4%), IGF1 increased the percent of oocytes that were blastocysts at Day 8 when added from Days 4 to 8 (42%) or Days 0 to 8 post-insemination (40%) but there was no significant effect when IGF1 was added from Days 0 to 4 (37%; SEM=2.2). Actions of IGF1 to increase blastocyst development were reduced when embryos were co-treated with the MAPK inhibitor PD98059. The percentage of oocytes becoming a blastocyst at Day 8 was 21 versus 37% for 0 and 100 ng/mL in the absence of inhibitor and 24 versus 29% in the presence of inhibitor (IGF1×inhibitor interaction, P<0.05; pooled SEM=1.3). In conclusion, IGF1 promotes development to the blastocyst stage by regulating MAPK-dependent events at Day 4 or later of development.
Subject(s)
Blastocyst/enzymology , Insulin-Like Growth Factor I/pharmacology , MAP Kinase Signaling System , Mitogen-Activated Protein Kinases/metabolism , Animals , Blastocyst/drug effects , Blastocyst/metabolism , Cattle , Embryo, Mammalian/drug effects , Embryo, Mammalian/metabolism , Oocytes/metabolism , Time FactorsABSTRACT
Preformed donor HLA-specific antibodies are a known indicator for poor patient survival after cardiac transplantation. The role of de novo donor-specific antibodies (DSA) formed after cardiac transplantation is less clear. Here we have retrospectively analyzed 243 cardiac transplant recipients, measuring HLA antibody production every year after transplantation up to 13 years post-transplant. Production of de novo DSA was analyzed in patients who had been negative for DSA prior to their transplant. DSA including transient antibodies were associated with poor patient survival (p = 0.0018, HR = 3.198). However, de novo and persistent DSA was strongly associated with poor patient survival (p = 0.0001 HR = 4.351). Although complement fixing persistent DSA correlated with poor patient survival, this was not increased compared to noncomplement fixing persistent DSA. Multivariable analysis indicated de novo persistent DSA to be an independent predictor of poor patient survival along with HLA-DR mismatch and donor age. Only increasing donor age was found to be an independent risk factor for earlier development of CAV. In conclusion, patients who are transplanted in the absence of pre-existing DSA make de novo DSA after transplantation which are associated with poor survival. Early and regular monitoring of post-transplant DSA is required to identify patients at risk of allograft failure.
Subject(s)
HLA Antigens/immunology , Heart Transplantation/adverse effects , Heart Transplantation/immunology , Isoantibodies/biosynthesis , Adult , Antibody Specificity , Complement Fixation Tests , Female , Graft Rejection/etiology , Graft Rejection/immunology , HLA-DR Antigens/immunology , Heart Transplantation/mortality , Histocompatibility Testing , Humans , Male , Middle Aged , Multivariate Analysis , Proportional Hazards Models , Retrospective Studies , Risk Factors , Tissue DonorsABSTRACT
We treated 20 patients with cubital tunnel syndrome by anterior transposition of the ulnar nerve with endoscopic assistance. Five elbows were classified preoperatively as McGowan's stage 1, 11 as stage 2 and four as stage 3. Excellent outcomes were obtained in nine and good in eight patients. Three patients had fair results. Improvement of symptoms occurred in all patients. There were no serious complications. All ulnar nerves remained anteriorly transposed.