A Comprehensive Workflow for the Analysis of Bio-Macromolecular Supplements: Case Study of 20 Whey Protein Products.

The presence of bio-macromolecules as major ingredients is a primary factor in marketing many biologically derived macromolecular supplements. Workflows for analyzing these supplements for quality assurance, adulteration, and other supply-chain difficulties must include a qualitative assessment of small-molecule and macromolecular components; however, no such integrated protocol has been reported for these bio-macromolecular supplements. Twenty whey protein supplements were analyzed using an integrated workflow to identify protein content, protein adulteration, inorganic elemental content, and macromolecular and small-molecule profiles. Orthogonal analytical methods were employed, including NMR profiling, LC-DAD-QToF analysis of small-molecule components, ICP-MS analysis of inorganic elements, determination of total protein content by a Bradford assay, SDS-PAGE protein profiling, and bottom-up shotgun proteomic analysis using LC-MS-MS. All 20 supplements showed a reduced protein content compared to the claimed content but no evidence of adulteration with protein from an unclaimed source. Many supplements included unlabeled small-molecule additives (but nontoxic) and significant deviations in metal content, highlighting the importance of both macromolecular and small-molecule analysis in the comprehensive profiling of macromolecular supplements. An orthogonal, integrated workflow allowed the detection of crucial product characteristics that would have remained unidentified using traditional workflows involving either analysis of small-molecule nutritional supplements or protein analysis.

Intrinsic Buffer Hydroxyl Radical Dosimetry Using Tris(hydroxymethyl)aminomethane.

Fast photochemical oxidation of proteins (FPOP) is a powerful covalent labeling tool that uses hydroxyl radicals generated by laser flash photolysis of hydrogen peroxide to footprint protein surfaces. Because radical production varies with many experimental parameters, hydroxyl radical dosimeters have been introduced to track the effective radical dosage experienced by the protein analyte. FPOP experiments performed using adenine optical radical dosimetry containing protein in Tris buffer demonstrated unusual dosimetry behavior. We have investigated the behavior of Tris under oxidative conditions in detail. We find that Tris can act as a novel gain-of-signal optical hydroxyl radical dosimeter in FPOP experiments. This new dosimeter is also amenable to inline real-time monitoring, thereby allowing real-time adjustments to compensate for differences in samples for their quenching ability.

Towards high-throughput fast photochemical oxidation of proteins: Quantifying exposure in high fluence microtiter plate photolysis.

Protein structural analysis by mass spectrometry has gained significant popularity in recent years, including high-resolution protein topographical mapping by fast photochemical oxidation of proteins (FPOP). The ability to provide protein topographical information at moderate spatial resolution makes FPOP an attractive technology for the protein pharmaceutical discovery and development processes. However, current technology limits the throughput and requires significant manual sample manipulation. Similarly, as FPOP is being used on larger samples, sample flow through the capillary becomes challenging. No systematic comparison of the performance of static flash photolysis with traditional flow FPOP has been reported. Here, we evaluate a 96-well microtiter-based laser flash photolysis method for the topographical probing of proteins, which subsequently could be used to analyze higher order structure of the protein in a high-throughput fashion with minimal manual sample manipulation. We used multiple metrics to compare microtiter FPOP performance with that of traditional flow FPOP: adenine-based hydroxyl radical dosimetry, oxidation efficiency of a model peptide, and hydroxyl radical protein footprint of myoglobin. In all cases, microtiter plate FPOP performed comparably with traditional flow FPOP, requiring a small fraction of the time for exposure. This greatly reduced sample exposure time, coupled with automated sample handling in 96-well microtiter plates, makes microtiter-based FPOP an important step in achieving the throughput required to adapt hydroxyl radical protein footprinting for screening purposes.

Review - Medication errors and strategies for their prevention.

Medication errors occur every day causing injury to the patients and even deaths. The health care professionals are not fully aware of the damages done by medication errors in terms of patients' discomfort and economic burden. There is a need to provide information about medication errors to health care providers. This article reviews research done on the various aspects of medication errors. The research work done on prescribing errors, transcribing errors, dispensing errors, administration errors and discharged summaries errors have been examined. Eight strategies to reduce the occurrence of medication errors have been reviewed: (1) Electronic prescribing and computerized physician order entry (CPOE) with clinical decision support systems (CDSSs), (2) Bar Code, (3) Interventions to reduce medication errors, (4) Medication Error Reporting Systems (MERSs), (5) Alerts about medication errors, (6) Prevention of harm from high-alert drugs, (7) Smart Infusion Pumps and (8) Telemedicine or Telehealth or Telepharmacy. Statistical tests used in medication error studies have also been stated.

Signaling by TGF-betas in tubule cultures of adult rat testis.

Although signal transduction of transforming growth factor-betas (TGF-ßs) is well characterized in individual cell types, data about TGF-ß signaling in a cellular context is still scarce. In this study, we used ex vivo tubule cultures from adult rat testis to investigate TGF-ß signaling. We show for the first time in testicular tubules, that TGF-ßs also signal via the BMP type I receptors, with ALK2 used by TGF-ß1 and ALK3 and ALK6 by TGF-ß2. This signal transduction is mediated via Smad3 as well as via Smad1. In contrast, BMPs (BMP2 and BMP7) do not signal via the high-affinity type I and type II TGFß receptors, TBR1 or TBR2. Furthermore, treatment of tubule cultures with either TGF-ß1 or TGF-ß2 had profound significant stimulatory effects on secretion of plasminogen activator-1 (PAI-1) through utilization of TGF-ß and BMP receptors. Specific inhibitors for either TBR1 or BMP receptors yielded nearly complete inhibition of TGF-ß signaling. The TBR1-TBR2 signalosome was detected with Duolink upon stimulation with either TGF-ß1 or TGF-ß2, predominantly in spermatogenic cells of the adult rat testis, particularly in elongated spermatids. In summary, this examination of intact rat testicular tubules demonstrated for the first time that TGF-ßs signal mainly through TBR1 and TBR2 but also use BMP receptors, including for secretion of PAI-1. Whereas ALK2 participates in the TGF-ß1-induced TBR1-TBR2 signalosome, ALK3 and ALK6 are involved in signaling of TGF-ß2. Detection of the TBR1-TBR2 signalosome in late spermiogenic cells indicates a post-meiotic activity.

Trend over time; a structural review of articles published in Journal of Pakistan Medical Association from 1953-2009.

OBJECTIVE: To review the articles published in the Journal of Pakistan Medical Association from 1953 to 2009 and to assess the components of each article by a pretested proforma containing a checklist of items based on modified 'Strengthening the Reporting of Observational Studies in Epidemiology' statement. METHODS: The retrospective study was conducted at the office of the Journal of Pakistan Medical Association, Karachi, from February to November 2010. A checklist of items in the modified 'Strengthening the Reporting of Observational Studies in Epidemiology' statement was made after discussion among the authors of the study to finally include a revised checklist of 45 items instead of the checklist of 22 items of the statement. A total sample size of 370 was calculated. Simple randomisation was done for selection of articles from each year. For each article, major and minor items were documented. Data was fed into SPSS version 15. RESULTS: The scientific quality of reporting of most of the components of Introduction, Results and Discussion sections have improved progressively with time (p < 0.001) whereas most components of Methodology have remained consistent. CONCLUSION: A change in trend over time was observed over the study period in major and minor items of the articles in the Journal, showing improved reporting of various sub-components of articles.The modified 'Strengthening the Reporting of Observational Studies in Epidemiology' statement provides a checklist that may be used to improve the quality of articles.

Occurrence of medication errors and comparison of manual and computerized prescription systems in public sector hospitals in Lahore, Pakistan.

The knowledge of medication errors is an essential prerequisite for better healthcare delivery. The present study investigated prescribing errors in prescriptions from outpatient departments (OPDs) and emergency wards of two public sector hospitals in Lahore, Pakistan. A manual prescription system was followed in Hospital A. Hospital B was running a semi-computerised prescription system in the OPD and a fully computerised prescription system in the emergency ward. A total of 510 prescriptions from both departments of these two hospitals were evaluated for patient characteristics, demographics and medication errors. The data was analysed using a chi square test for comparison of errors between both the hospitals. The medical departments in OPDs of both hospitals were the highest prescribers at 45%-60%. The age group receiving the most treatment in emergency wards of both the hospitals was 21-30 years (21%-24%). A trend of omitting patient addresses and diagnoses was observed in almost all prescriptions from both of the hospitals. Nevertheless, patient information such as name, age, gender and legibility of the prescriber's signature were found in almost 100% of the electronic-prescriptions. In addition, no prescribing error was found pertaining to drug concentrations, quantity and rate of administration in e-prescriptions. The total prescribing errors in the OPD and emergency ward of Hospital A were found to be 44% and 60%, respectively. In hospital B, the OPD had 39% medication errors and the emergency department had 73.5% errors; this unexpected difference between the emergency ward and OPD of hospital B was mainly due to the inclusion of 69.4% omissions of route of administration in the prescriptions. The incidence of prescription overdose was approximately 7%-19% in the manual system and approximately 8% in semi and fully electronic system. The omission of information and incomplete information are contributors of prescribing errors in both manual and electronic prescriptions.

Dengue fever in the Indian Subcontinent: an overview.

The Indian Subcontinent has emerged as a scene of many mosquito-borne infectious diseases, including malaria and dengue fever. After the 1990s, the rate of malaria declined owing largely to preventive measures, but at the same time dengue fever (DF) and dengue hemorrhagic fever (DHF) were increasing in the region. Outbreaks were recorded in all countries of the Indian Subcontinent with India, Pakistan, Bangladesh and Sri Lanka on the forefront and suffering from the largest number of cases and deaths. We discuss annual cases of DF/DHF in these four countries and possible factors involved in DF outbreaks. We also discuss prevalent serotypes in this region where data suggest the emergence of DEN2 and DEN3 as the most dominant and lethal serotypes. Climate is an important factor influencing DF outbreaks, and rainfall, temperature and humidity play a pivotal role in DF outbreaks. Finally the economic impact of DF/DHF cases is discussed showing that direct and indirect economic loss due to DF/DHF reaches millions of USD each year.

Errors in opioid prescribing: a prospective survey in cancer pain.

CONTEXT: Cancer pain is debilitating and has multidimensional consequences. It can be treated adequately in up to 90% of patients by following pain management guidelines. Nevertheless, inadequate pain control remains a global problem. OBJECTIVES: We surveyed prescribing patterns in patients referred to our Palliative Medicine Program (PMP) to identify common errors in opioid use. METHODS: Consecutive cancer patients seen by our PMP were prospectively surveyed for the presence of pain and errors in opioid prescribing at the time of initial consultation. Our recommendations to correct and optimize pain management also were recorded. RESULTS: One hundred eighty-six consecutive cancer patients were screened. One hundred seventeen (63%) had cancer pain, 151 opioid prescribing errors were detected, and 147 different recommendations were made. Most common were failure to order around-the-clock opioids for constant pain, and the failure to treat or prevent opioid side effects. Multiple errors were more common in females, but the sex difference did not reach statistical significance. There was no difference in the errors by pain severity or reason for consultation. CONCLUSION: Opioid prescribing errors were common. Females may be at greater risk of multiple errors. A PM consultation program is effective in identifying and correcting a wide variety of opioid prescribing errors.

Cardiac trauma with gunshot injuries.

Penetrating cardiac injuries, secondary to gunfire, constitute the most lethal forms of cardiothoracic trauma with their potential fatality. We report our experience of managing two such cases who presented with haemorrhagic shock and cardiac tamponade, in a collapsed state. Prompt resuscitation and early surgical intervention (midline sternotomy and cardiorrhaphy) was successfully performed with a favourable outcome.