Harvest Stage and Brewing Conditions Impact Mineral Content, Phenolic Compounds, and Antioxidant Capacity of Lemon Balm (Melissa officinalis L.) Herbal Tea.

Lemon balm (Melissa officinalis L.) is commonly consumed as an herbal tea for its antioxidant health benefits. Young seedlings known as microgreens are popular for their distinct flavors and can contain higher mineral content on a dry weight basis compared to their adult counterparts. However, the use of microgreens for herbal teas has not been previously investigated. In this study, lemon balm was grown to adult and microgreen harvest stages and prepared as herbal teas by brewing with boiled (100 °C) water for 5 minutes and room temperature water (22 °C) for 2 hours. The effects of harvest time and brewing method on the mineral content, phenolic compounds, and antioxidant capacity of lemon balm herbal teas were assessed. Results showed that adult lemon balm tea contained higher total phenolics, total flavonoids, rosmarinic acid, and antioxidant capacity than microgreen teas, with hot preparations containing the highest amounts (p ≤ 0.05). In contrast, microgreen lemon balm teas contained higher amounts of minerals (p ≤ 0.05), including calcium, potassium, magnesium, sodium, phosphorus, copper, and zinc. In general, brewing conditions did not impact the content of most minerals. Overall, the results support the potential of using dried microgreens as herbal teas. Microgreen lemon balm teas prepared hot and cold offer antioxidant compounds and are richer sources of minerals than adult teas. The ease of growth for microgreens offers consumers the opportunity for home preparation of a novel herbal tea beverage.

Improving the uptake: Barriers and facilitators to pulmonary rehabilitation.

BACKGROUND: Pulmonary rehabilitation (PR) is a well-established therapeutic management programme for patients with chronic lung disease. Despite good clinical evidence, patient engagement can be poor. AIM: The aim of the study was to determine the number of patients who are referred to PR at a District General Hospital, explore barriers and facilitators to attending and completing and identify strategies for improvement. METHODS: All patients invited to attend PR in the calendar year 2016 were included in an analysis (N = 281). A structured questionnaire composed of barriers and facilitators was administered to patients that did not attend (non-attenders, N = 20) and those that attended but did not complete the programme (non-completers/"drop-outs," N = 13). Improvement strategies were identified and implemented followed by analysis of patients invited to attend in 2017 and 2018. RESULTS: Age, sex and smoking status are factors that affect both attendance and completion rates of patients attending PR. In our analysis, we were able to demonstrate that lack of awareness and low perceived benefits were important barriers for non-attendance. In addition, overall uptake rate was improved but at the expense of completion rate. CONCLUSION: Our local non-attendance rate in 2016 was 42%, with strategies aimed at improving patient and physician information, this was reduced to 11% (2018), below the national United Kingdom average. Unexpectedly, there was a worsening of completion rates and this raises questions about both appropriateness of referrals and whether completion rate rather than non-attendance rate should be used as a performance indicator and standard.

A retrospective analysis comparing the use of ProCore® with standard fine needle aspiration in endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA).

Endobronchial ultrasound has become first line in the investigation of mediastinal lesions suspicious for malignancy in keeping with National Institute for Health and Care Excellence (NICE) guidelines; however, needle size and type required to maximise diagnostic sensitivity remains unclear. Previous meta-analyses have compared the use of ProCore with standard fine needle aspiration in the assessment of pancreatic masses with differences noted only in the number of passes required. We aim to assess whether a ProCore needle improves diagnostic sensitivity in EBUS-TBNA. Complete follow-up data regarding all 235 patients undergoing EBUS-TBNA in a district general hospital has been collected since the service's inception in 2012. Results were collated and retrospectively analysed allowing for calculation of test sensitivity and specificity. Comparison was then made between procedures where standard fine needle aspiration was performed and those using a ProCore needle. Overall sensitivity of EBUS-TBNA was shown to be 85% with a specificity of 100% in keeping with quoted figures from other centres. Standard fine needle aspiration produced a sensitivity of 77% (85/110) versus ProCore sensitivity of 92% (115/125) with a p value of 0.0016. Thirty percent (33/110) of patients undergoing standard fine needle aspiration required an appropriate crossover technique such as mediastinoscopy or CT-guided FNA in order to either obtain or confirm the diagnosis compared with 15% (19/125) of the ProCore group with a p value of 0.0064. Our retrospective analysis shows a statistically significant difference in the diagnostic sensitivity of sampling mediastinal lymphadenopathy using a ProCore needle compared with standard fine needle aspiration. It also shows that a significantly fewer number of patients required further procedures in order to obtain or confirm the diagnosis. This could potentially be confounded by the retrospective nature of the study design; however, due to the statistical significance demonstrated, further study is required.

Clinician-Facilitated Physical Activity Intervention Versus Pulmonary Rehabilitation for Improving Physical Activity in COPD: A Feasibility Study.

Pulmonary rehabilitation (PR) may not suit all individuals with chronic obstructive pulmonary disease (COPD) and may not result in increased physical activity. Higher levels of physical activity are associated with reduced mortality and morbidity. The aim of this study was to assess the feasibility of conducting a trial to investigate the effectiveness of a clinician-facilitated physical activity intervention (PAI) versus PR in improving physical activity in patients with COPD referred to PR. In this randomised controlled mixed methods feasibility study, all patients referred to PR who were eligible and willing were assessed at baseline and then randomised to the PAI or to PR. The assessments were repeated post-intervention and at 3-month follow-up. The main outcome was step count measured by Actigraph. Semi-structured interviews were conducted post-intervention. The N = 50 patients; mean (SD) age, 64.1(8.6) years, 24M were recruited and randomised; N = 23 (PAI) and n = 26 (PR): one patient was excluded from the analysis as that person did not meet the GOLD diagnostic criteria. Key feasibility criteria were met; recruitment was 11%, dropouts in PAI were 26% (n = 6) and 50% (n = 13/26) PR. Participants in both groups experienced a range of health benefits from their respective programmes. The PAI appears to be effective in increasing step counts in people with COPD: mean change (standard deviation) [confidence interval] for the PAI group was 972.0(3230.3)[-1080.3 to 3024.4], n = 12 and 4.3(662.7)[-440.9 to 449.5], n = 11 for the PR group. The PAI met all domains of fidelity. This study provides key information to inform a future-randomised controlled trial in physical activity.

Clinical pharmacology of an atrasentan and docetaxel regimen in men with hormone-refractory prostate cancer.

PURPOSE: This study was conducted to evaluate potential pharmacokinetic interactions between docetaxel and atrasentan as part of a phase I/II clinical trial. METHODS: Patients with prostate cancer were treated with intravenous docetaxel (60-75 mg/m(2)) every 3 weeks and oral atrasentan (10 mg) daily starting on day 3 of cycle 1 and then given continuously. The pharmacokinetics of both drugs were evaluated individually (cycle 1, day 1 for docetaxel; day 21 for atrasentan) and in combination (cycle 2, day 1 for both drugs). Pharmacogenomics of alpha-1-acid glycoprotein (AAG) were also explored. RESULTS: Paired pharmacokinetic data sets for both drugs were evaluable in 21 patients. Atrasentan was rapidly absorbed and plasma concentrations varied over a fourfold range at steady state within a typical patient. The median apparent oral clearance of atrasentan was 17.4 L/h in cycle 1 and was not affected by docetaxel administration (p = 0.9). Median systemic clearance of docetaxel was 51.1 L/h on the first cycle and significantly slower (p = 0.01) compared with that obtained during co-administration of atrasentan, 61.6 L/h. Docetaxel systemic clearance in cycle 1 was 70.0 L/h in patients homozygous for a variant allele in AAG compared with 44.5 L/h in those with at least one wild-type allele (p = 0.03). CONCLUSION: Genetic polymorphism in AAG may explain some inter-patient variability in docetaxel pharmacokinetics. The systemic clearance of docetaxel is increased by approximately 21 % when given concomitantly with atrasentan; however, atrasentan pharmacokinetics does not appear to be influenced by docetaxel administration.

Development and utilization of a combined LC-UV and LC-MS/MS method for the simultaneous analysis of tegafur and 5-fluorouracil in human plasma to support a phase I clinical study of oral UFT®/leucovorin.

Tegafur is a 5-fluorouracil (5-FU) prodrug widely used outside the United States to treat colorectal cancer as well as cancers of the head and neck. The resulting plasma concentrations of tegafur are much higher than those of 5-FU; thus, analytical methods are needed that are sensitive enough to detect low plasma concentrations of 5-FU and robust enough to simultaneously analyze tegafur. Previous LC-MS/MS methods have either failed to demonstrate the ability to simultaneously measure low 5-FU and high tegafur plasma levels, or failed to be applicable in clinical studies. Our goal was to develop a method capable of measuring low concentrations of 5-FU (8-200 ng/ml) and high concentrations of tegafur (800-20,000 ng/ml) in human plasma and to subsequently evaluate the utility of the method in patient samples collected during a phase I clinical study where oral doses of either 200mg or 300 mg UF®/LV (uracil and tegafur in a 4:1 molar ratio plus leucovorin) were administered. A combined LC-MS/MS and LC-UV method was developed utilizing negative ion atmospheric pressure ionization (API). The method provides an accuracy and precision of <10% and <6%, respectively, for both analytes. Material recoveries from the liquid-liquid extraction technique were 97-110% and 86-91% for tegafur and 5-FU, respectively. Utilization of this method to determine tegafur and 5-FU plasma concentrations followed by noncompartmental pharmacokinetic analyses successfully estimated pharmacokinetic parameters (C(MAX), t(MAX) and AUC(0-10h)) in the clinical study patients. Overall, this method is ideal for the simultaneous bioanalysis of low levels of 5-FU and relatively higher levels of its prodrug, tegafur, in human plasma for clinical pharmacokinetic analysis.

Identification of isobaric product ions in electrospray ionization mass spectra of fentanyl using multistage mass spectrometry and deuterium labeling.

Isobaric product ions cannot be differentiated by exact mass determinations, although in some cases deuterium labeling can provide useful structural information for identifying isobaric ions. Proposed fragmentation pathways of fentanyl were investigated by electrospray ionization ion trap mass spectrometry coupled with deuterium labeling experiments and spectra of regiospecific deuterium labeled analogs. The major product ion of fentanyl under tandem mass spectrometry (MS/MS) conditions (m/z 188) was accounted for by a neutral loss of N-phenylpropanamide. 1-(2-Phenylethyl)-1,2,3,6-tetrahydropyridine (1) was proposed as the structure of the product ion. However, further fragmentation (MS(3)) of the fentanyl m/z 188 ion gave product ions that were different from the product ion in the MS/MS fragmentation of synthesized 1, suggesting that the m/z 188 product ion from fentanyl includes an isobaric structure different from the structure of 1. MS/MS fragmentation of fentanyl in deuterium oxide moved one of the isobars to 1 Da higher mass, and left the other isobar unchanged in mass. Multistage mass spectral data from deuterium-labeled proposed isobaric structures provided support for two fragmentation pathways. The results illustrate the utility of multistage mass spectrometry and deuterium labeling in structural assignment of isobaric product ions.

Development of a novel DNA microarray to detect bacterial pathogens in patients with chronic obstructive pulmonary disease (COPD).

A novel microarray was constructed with DNA PCR product probes targeting species specific functional genes of nine clinically significant respiratory pathogens, including the Gram-positive organisms (Streptococcus pneumoniae, Streptococcus pyogenes), the Gram-negative organisms (Chlamydia pneumoniae, Coxiella burnetii Haemophilus spp., Legionella pneumophila, Moraxella catarrhalis, and Pseudomonas aeruginosa), as well as the atypical bacterium, Mycoplasma pneumoniae. In a "proof-of-concept" evaluation of the developed microarray, the microarray was compared with real-time PCR from 14 sputum specimens from COPD patients. All of the samples positive for bacterial species in real-time PCR were also positive for the same bacterial species using the microarray. This study shows that a microarray using PCR probes is a potentially useful method to monitor the populations of bacteria in respiratory specimens and can be tailored to specific clinical needs such as respiratory infections of particular patient populations, including patients with cystic fibrosis and bronchiectasis.

Epstein - Barr virus Pneumonitis.

Epstein-Barr virus is an unusual pathogen in the aetiology of alveolitis. We describe a case of Epstein-Barr virus induced pneumonitis and its successful treatment with Aciclovir.

British thoracic society winter meeting 2008.

The 26th annual British Thoracic Society Winter Meeting was held in London from 2 to 5 December 2008. It was attended by over 2000 participants from around the world. The program presented to attendees topics of interest across respiratory clinical and scientific fields.

Isolated mediastinal adenopathy: the case for mediastinoscopy.

BACKGROUND: We report our experience with mediastinoscopy at Auckland city hospital, a tertiary referral centre. We wished to examine correlations between clinical diagnosis and that made by histological sampling of enlarged mediastinal nodes particularly in patients with isolated mediastinal adenopathy. METHODS: We retrospectively reviewed clinical records of all patients who underwent mediastinoscopy in a five year period, mediastinoscopy was performed in the presence of enlarged lymph nodes (short axis > 1cm) found at CT. Mediastinoscopy was indicated for diagnostic staging of mediastinal adenopathy related to a parenchymal lung mass, diagnosis of isolated mediastinal adenopathy and diagnosis of mediastinal adenopathy with other CT findings. Data relating to indication, pre-test diagnosis, node stations sampled, histology, and operative complications were collected. RESULTS: Mediastinoscopy was performed in 137 consecutive patients. Seventy five patients had a lung mass, 47 had isolated mediastinal adenopathy and 15 had other CT findings. One operative complication occurred. In those patients with isolated adenopathy the following diagnoses were reached; sarcoidosis 23, TB 15, lymphoma 4, carcinoma 4, no diagnosis 1. Final diagnosis was significantly associated with patient's ethnicity. There was high sensitivity and specificity on comparison of clinical and histological diagnosis for both TB and sarcoidosis cases. CONCLUSIONS: Mediastinoscopy proved to be safe and effective in nodal assessment of the mediastinum. In carefully selected cases procedural morbidity and mortality may be avoided by application of features related to patient's ethnicity and radiological findings.

Respiratory viral infection in exacerbations of COPD.

BACKGROUND: Patients with COPD have frequent exacerbations. The role of respiratory viral infection is just emerging. We wished to determine prospectively the incidence of viral infection in exacerbated and stable COPD patients as well as smokers who do not have airways obstruction. METHODS: Stable and exacerbated COPD patients were recruited along with a group of patients who had smoked but who did not have any airways obstruction. Spirometry was performed and sputum specimens were tested for a range of 12 different respiratory viruses using PCR. RESULTS: One hundred and thirty-six patients with exacerbations of COPD, 68 stable COPD patients and 16 non-obstructed smokers were recruited. A respiratory virus was detected in 37% of exacerbations, 12% of stable COPD patients and 12% of non-obstructed smokers, p<0.0005. Rhinovirus was most frequently detected. The symptom of fever was associated with virus detection, p<0.05. Infection with more than one virus was only found in the exacerbated COPD patients. CONCLUSION: Respiratory viral infection is associated with exacerbations of COPD. Rhinovirus was the most common infecting agent identified and in two cases human metapneumovirus was also detected. Dual infections were only seen amongst those patients admitted to hospital with acute exacerbations of COPD. Viruses were more commonly detected in those with more severe airways disease.

Exudative bronchiolitis after lung transplantation.

BACKGROUND: After lung transplantation patients frequently develop small airways disease. Exudative bronchiolitis (EB) has not been described previously in this population. We describe a case series of patients who have developed EB after lung transplantation. METHODS: We reviewed the clinical records and radiologic data of 99 patients who underwent single and bilateral lung transplantation. Data relating to ethnicity, transplant indication, human leukocyte antigen (HLA) status, complications and survival were recorded. The EB cohort, defined by high-resolution computed tomographic (HRCT) evidence of the disease, was compared with a group of patients who had not developed EB. RESULTS: The majority of patients had chronic obstructive pulmonary disease (COPD; n = 51), followed by cystic fibrosis (CF; n = 22), pulmonary fibrosis (n = 8), pulmonary hypertension (n = 7), bronchiectasis (n = 5), lymphangioleiomyomatosis (n = 3) and Eisenmenger's syndrome (n = 3). Thirteen patients were found to have developed EB. EB was more commonly seen in Maori and Pacific Island patients (p < 0.05). EB was significantly associated with early infection post-transplant (p < 0.05) and a history of Aspergillus infection (p < 0.005) or diabetes (p < 0.05). The patients with EB were also significantly more likely to develop bronchiolitis obliterans syndrome (p < 0.0005), bronchiectasis (p < 0.0005) or small airways disease (p < 0.05). Patients with EB had a varied response to treatment, with the majority showing improvement. CONCLUSIONS: EB was noted to occur after lung transplantation in a significant proportion of patients, but will not be detected unless HRCT is used routinely. It has been associated with patients' ethnicity, donor haplotype, infection and the development of airways disease. EB may be a prominent indicator of the likelihood of developing BOS.

Acute and latent adenovirus in COPD.

INTRODUCTION: The COPD airway is infiltrated with CD8+ T cells, which has led to a virus being implicated in its pathogenesis. Some investigators have suggested a role for the persistence of the adenovirus E1A in bronchial epithelial cells. We examined respiratory tract specimens from COPD patients for the presence of E1A DNA and mRNA using real-time PCR. METHODS: Nucleic acid extraction was performed on sputum specimens from patients with COPD. Copy numbers for GAPDH, and adenovirus 5 E1A DNA and mRNA were determined using a quantitative real-time PCR assay. All samples were screened for the adenovirus hexon gene using nested PCR. RESULTS: One hundred and seventy-one patients, 80 male, aged 68.9+/-9.8 years with COPD were recruited. One hundred and thirty-six were seen during an exacerbation when admitted to hospital, 33 of whom were reviewed when clinically stable along with an additional 35 stable COPD patients. Ten patients in the exacerbation group were positive for the adenovirus hexon gene (7%), as were four in the stable group (6%). Only two patients in the exacerbation group were positive for adenovirus 5 E1A. Only one patient in the stable COPD group had detectable E1A DNA/mRNA and also tested positive for the adenovirus hexon gene. CONCLUSION: Adenovirus is detected in similar frequencies in exacerbated and stable COPD patients. Adenovirus E1A DNA is infrequently detected in respiratory secretions from patients with COPD. Our data suggest that the persistence of adenovirus 5 E1A in lung cells of sputum samples in patients with COPD occurs infrequently.

Evaluation of real-time PCR for the detection and quantification of bacteria in chronic obstructive pulmonary disease.

Chronic obstructive pulmonary disease (COPD) embraces a number of pathological processes including chronic bronchitis, chronic bronchiolitis and emphysema. The chronic and progressive course of COPD is often aggravated by short periods of increasing symptoms. Respiratory tract infections (RTIs) are the most common causes of COPD exacerbations. Detection and enumeration of respiratory bacteria are important techniques in diagnosing RTIs and in the validation of new treatment methods. We describe here the development and evaluation of real-time PCR assays for the simultaneous direct detection and quantification of a range of respiratory bacteria in individuals with COPD during stable periods and during acute exacerbations of the disease. Sputum samples from 30 subjects in a COPD study were analysed, and results compared with the current gold standard of culture. Real-time PCR assays proved highly sensitive, with no cross-reactivity with other species. The prevalence of bacteria detected by real-time PCR compared with that by culture was substantially higher for Streptococcus pneumoniae, Staphylococcus aureus, Haemophilus spp. and Moraxella catarrhalis. Multiple pathogens were also found with real-time PCR but were not detected by culture. This study demonstrates the potential of such methods in the detection and enumeration of respiratory bacteria.

Organisms isolated from adults with cystic fibrosis.

BACKGROUND: Patients with cystic fibrosis [CF] have frequent pulmonary exacerbations associated with the isolation of bacterial organisms from sputum samples. It is not clear however, if there are differences in the types of additional organisms isolated from patients who are infected with Burkholderia cepacia complex [BCC] or Pseudomonas aerugionsa [PA] in comparison to those who are not infected with either of these organisms [NI]. METHODS: Adult patients attending the regional CF unit were followed over a two year period and patients were assigned to three groups depending on whether they were known to be chronically infected with BCC, PA or NI. We compared the numbers and types of organisms which were isolated in each of these groups. RESULTS: Information was available on a total of 79 patients; BCC 23, PA 30 and NI 26. Total numbers of organisms isolated, expressed as median and IQR for each group, [P = 0.045] and numbers of co-infecting organisms [P = 0.003] were significantly higher in the BCC group compared to PA, and in the PA group [P < 0.001, p = 0.007 respectively] compared to NI patients. The pattern of co-infecting organisms was similar in all three groups. CONCLUSIONS: Total numbers of organisms isolated and numbers of co-infecting organisms were significantly higher in the BCC group compared to PA, and in the PA group compared to NI patients. Types of co-infecting organisms are similar in all groups of patients.

Intraischaemic hypothermia reduces free radical production and protects against ischaemic insults in cultured hippocampal slices.

Hypothermia has been demonstrated to be an effective neuroprotective strategy in a number of models of ischaemic and excitotoxic neurodegeneration in vitro and in vivo. Reduced glutamate release and free radical production have been postulated as potential mechanisms underlying this effect but no definitive mechanism has yet been reported. In the current study, we have used oxygen-glucose deprivation in organotypic hippocampal slice cultures as an in vitro model of cerebral ischaemia. When assessed by propidium iodide fluorescence, reducing the temperature during oxygen-glucose deprivation to 31-33 degrees C was significantly neuroprotective but this effect was lost if the initiation of hypothermia was delayed until the post-insult recovery period. The neuroprotective effects of hypothermia were associated with a significant decrease in both nitric oxide production, as assessed by 3-amino-4-aminomethyl-2',7'-difluorofluorescein fluorescence, and superoxide formation. Further, hypothermia significantly attenuated NMDA-induced nitric oxide formation in the absence of hypoxia/hypoglycaemia. We conclude that the neuroprotective effects of hypothermia are mediated through a reduction in nitric oxide and superoxide formation and that this effect is likely to be downstream of NMDA receptor activation.

L-arginyl-3,4-spermidine is neuroprotective in several in vitro models of neurodegeneration and in vivo ischaemia without suppressing synaptic transmission.

1. Stroke is the third most common cause of death in the world, and there is a clear need to develop new therapeutics for the stroke victim. To address this need, we generated a combinatorial library of polyamine compounds based on sFTX-3.3 toxin from which L-Arginyl-3,4-Spermidine (L-Arg-3,4) emerged as a lead neuroprotective compound. In the present study, we have extended earlier results to examine the compound's neuroprotective actions in greater detail. 2. In an in vitro ischaemia model, L-Arg-3,4 significantly reduced CA1 cell death when administered prior to induction of 60 min of ischaemia as well as when administered immediately after ischaemia. Surprisingly, L-Arg-3,4 continued to prevent cell death significantly when administration was delayed for as long as 60 min after ischaemia. 3. L-Arg-3,4 significantly reduced cell death in excitotoxicity models mediated by glutamate, NMDA, AMPA, or kainate. Unlike glutamate receptor antagonists, 300 microM L-Arg-3,4 did not suppress synaptic transmission as measured by evoked responses in acute hippocampal slices. 4. L-Arg-3,4 provided significant protection, in vitro, in a superoxide mediated injury model and prevented an increase of superoxide production after AMPA or NMDA stimulation. It also decreased nitric oxide production after in vitro ischaemia and NMDA stimulation, but did so without inhibiting nitric oxide synthase directly. 5. Furthermore, L-Arg-3,4 was significantly neuroprotective in an in vivo model of global forebrain ischaemia, without any apparent neurological side-effects. 6. Taken together, these results demonstrate that L-Arg-3,4 is protective in several models of neurodegeneration and may have potential as a new therapeutic compound for the treatment of stroke, trauma, and other neurodegenerative diseases.