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1.
J Forensic Sci ; 69(2): 659-668, 2024 Mar.
Article in English | MEDLINE | ID: mdl-38282520

ABSTRACT

This research project had two major objectives. The first was to successfully print and fire the Liberator, a 3D-printed firearm, to assess its feasibility as a lethal weapon. The second objective was to identify any individual characteristics that might be deposited during the firing process by the firearm. The Liberator was printed using unchanged files downloaded from the internet using PLA and ABS filament. The Liberator was fired remotely into newspapers at the Allegheny County Medical Examiner's Office. The printing of the Liberator was both inexpensive and relatively quick with only minor hand modifications made after printing. The Liberator fractured beyond repair after firing but successfully fired and penetrated three newspapers. Neither the bullet nor the cartridge case exhibited any individual characteristics that could be used for identification purposes. Suspected thermoplastic deposits were identified on both the bullet and cartridge case, but additional testing must be done for confirmation purposes. In conclusion, the Liberator can be used reliably for one shot and will not yield any evidence for Firearms and Toolmark Examination.

2.
BMC Cancer ; 23(1): 710, 2023 Jul 29.
Article in English | MEDLINE | ID: mdl-37516867

ABSTRACT

BACKGROUND: Several cancer immunotherapies that target the PD-L1/PD-1 pathway show promising clinical activity in patients with hepatocellular carcinoma (HCC). However, the standard of care in first-line treatment with atezolizumab (anti-PD-L1 therapy) in combination with bevacizumab is associated with a limited objective response rate. Telomerase reverse transcriptase (TERT) activation meets the criteria of oncogenic addiction in HCC and could be actionable therapeutic target and a relevant tumor antigen. Therefore we hypothesized that combining anti-PD-1/PD-L1 therapy with an anti-telomerase vaccine might be an attractive therapy in HCC. UCPVax is a therapeutic cancer vaccine composed of two separate peptides derived from telomerase (human TERT). UCPVax has been evaluated in a multicenter phase I/II study in non-small cell lung cancers and has demonstrated to be safe and immunogenic, and is under evaluation in combination with atezolizumab in a phase II clinical trial in tumors where telomerase reactivation contributes to an oncogene addiction (HPV+ cancers). The aim of the TERTIO study is to determine the clinical interest and immunological efficacy of a treatment combining the CD4 helper T-inducer cancer anti-telomerase vaccine (UCPVax) with atezolizumab and bevacizumab in unresectable HCC in a multicenter randomized phase II study. METHODS: Patients with locally advanced, metastatic or unresectable HCC who have not previously received systemic anti-cancer treatment are eligible. The primary end point is the objective response rate at 6 months. Patients will be allocated to a treatment arm with a randomization 2:1. In both arms, patients will receive atezolizumab at fixed dose of 1200 mg IV infusion and bevacizumab at fixed dose of 15 mg/kg IV infusion, every 3 weeks, according to the standard of care. In the experimental arm, these treatments will be combined with the UCPVax vaccine at 0.5 mg subcutaneously. DISCUSSION: Combining anti-PD-1/PD-L1 therapy with an anti-telomerase vaccine gains serious consideration in HCC, in order to extend the clinical efficacy of anti-PD-1/PD-L1. Indeed, anti-cancer vaccines can induce tumor-specific T cell expansion and activation and therefore restore the cancer-immunity cycle in patients lacking pre-existing anti-tumor responses. Thus, there is a strong rational to combine immune checkpoint blockade therapy and anticancer vaccine (UCPVax) in order to activate antitumor T cell immunity and bypass the immunosuppression in the tumor microenvironment in HCC. This pivotal proof of concept study will evaluate the efficacy and safety of the combination of a CD4 Th1-inducer cancer vaccine derived from telomerase (UCPVax) and atezolizumab plus bevacizumab in unresectable HCC, as well as confirming their synergic mechanism, and settling the basis for a new combination for future clinical trials. TRIAL REGISTRATION: NCT05528952.


Subject(s)
Cancer Vaccines , Carcinoma, Hepatocellular , Liver Neoplasms , Lung Neoplasms , Telomerase , Humans , Bevacizumab , Cancer Vaccines/adverse effects , Carcinoma, Hepatocellular/drug therapy , Liver Neoplasms/drug therapy , Tumor Microenvironment
3.
Dermatol Online J ; 29(2)2023 Apr 15.
Article in English | MEDLINE | ID: mdl-37220286

ABSTRACT

Proliferative verrucous leukoplakia (PVL) is a rare, aggressive form of oral leukoplakia with a substantial risk of malignant transformation. The slowly progressive course and the lack of a single defining histopathologic characteristic for PVL make this entity a diagnostic challenge. We report on a patient who presented with a 7-year history of worsening oral lesions.


Subject(s)
Lichen Planus , Humans
4.
Clin Exp Gastroenterol ; 15: 145-151, 2022.
Article in English | MEDLINE | ID: mdl-35983373

ABSTRACT

Introduction: Mucormycosis is a rare systemic fungal infection, mainly observed in immunocompromised patients. It is responsible for surface and deep tissue destruction leading to perforations and hemorrhage. Its pathogenesis represented by an angio-invasion is at the origin of a local infarction and a vascular thrombosis. We report a case of gastrointestinal (GI) mucormycosis-induced multiple gastric ulcers, GI bleeding and rectal perforation. Case Presentation: A 75-year-old man, with type II diabetes mellitus, was admitted to the intensive care unit for an acute abdominal pain associated with massive hematochezia. Clinical examination was that of an acute peritonitis and a hemorrhagic shock state. Abdominal and pelvic CT scan with intravenous contrast concluded to a perforation of the anterior wall of the rectum. He underwent immediate laparotomy with temporary colostomy. Several upper GI endoscopies had shown multiple gastric ulcer lesions. Lower GI endoscopy showed a fistulous orifice of the rectum on its anterior surface. Histopathology of the gastric biopsy showed acute and subacute inflammatory changes with filamentous elements suggesting mucormycosis. Histopathology of the rectal biopsy showed a subacute non-specific inflammation. Culture of the secretions from the rectal fistula orifice showed the strain Rhizopus sp. Antifungal susceptibility testing reported sensitivity to liposomal amphotericin B. The diagnosis of GI mucormycosis-induced multiple gastric ulcers, rectal perforation and pulmonary embolism in the patient with type II diabetes mellitus was retained. The outcomes were favorable after 6 weeks of treatment with liposomal amphotericin B associated with temporary colostomy and appropriate diabetes management. Conclusion: GI mucormycosis remains a multidisciplinary diagnostic challenge, less frequent in clinical practice, with a long diagnostic pathway. This opportunistic systemic mycosis can lead to numerous GI complications including perforation, massive GI bleeding and even multiple extra-GI complications. GI mucormycosis has a good prognosis if it is treated early with medical and surgical treatment.

5.
Int J Pharm ; 620: 121740, 2022 May 25.
Article in English | MEDLINE | ID: mdl-35421534

ABSTRACT

Since late 2019, concerns regarding trace levels of the probable human carcinogen N-dimethylnitrosamine (NDMA) in Metformin-containing pharmaceuticals have been an issue if they exceeded the maximum allowable intake of 96 ng/day for a medicine with long-term intake. Here, we report results from an extensive analysis of NDMA content along the active pharmaceutical ingredient (API) manufacturing process as well as two different drug product manufacturing processes. Our findings confirm that Metformin API is not a significant source of NDMA found in Metformin pharmaceuticals and that NDMA is created at those steps of the drug product manufacturing that introduce heat and nitrite. We demonstrate that reduction of nitrite from excipients is an effective means to reduce NDMA in the drug product. Limiting residual dimethylamine in the API has proven to be another important factor for NDMA control as dimethylamine leads to formation of NDMA in the drug products. Furthermore, analysis of historical batches of drug products has shown that NDMA may increase during storage, but the levels reached were not shelf-life limiting for the products under study.


Subject(s)
Dimethylnitrosamine , Metformin , Dimethylamines , Dimethylnitrosamine/analysis , Excipients , Humans , Nitrites
6.
Oncologist ; 26(9): e1656-e1659, 2021 09.
Article in English | MEDLINE | ID: mdl-34028132

ABSTRACT

We describe a large series of patients with solid tumors in an early COVID-19 cluster in the eastern part of France. From February to May 2020, this multicenter retrospective study enrolled 212 patients with cancer under treatment or on follow-up for any type of malignant solid tumor and positive for SARS-CoV-2. The mortality rate was 30%. Patients with gastrointestinal cancers were identified as a subset of more vulnerable patients; immunotherapy and radiotherapy within 3 months from COVID-19 diagnosis were risk factors for death. The reported data support the essential need to be proactive and weigh the risks of morbidity from COVID-19 against the magnitude of benefits of intended cancer therapies during this pandemic. IMPLICATIONS FOR PRACTICE: This article supports the essential need to be proactive (treatment delay or modification) in oncology in the setting of pandemic. This study identified patients with gastrointestinal cancers as a more vulnerable subset of patients with cancer and found that immunotherapy and radiotherapy within 3 months from COVID-19 diagnosis to be risk factors for death. The reported data indicate the necessity of weighing the risks of morbidity from COVID-19 against the magnitude of benefits of intended cancer therapies in any future wave of COVID-19.


Subject(s)
COVID-19 , Neoplasms , COVID-19 Testing , Humans , Neoplasms/epidemiology , Neoplasms/therapy , Pandemics , Retrospective Studies , SARS-CoV-2
7.
Dent Clin North Am ; 64(1): 25-37, 2020 01.
Article in English | MEDLINE | ID: mdl-31735231

ABSTRACT

Oral potentially malignant disorders (OPMDs) are precursor lesions that may undergo malignant transformation to oral cancer. These lesions most commonly present clinically as white patches (leukoplakia). However, they may also be red (erythroplakia), or red and white (erythroleukoplakia). There are many risk factors associated with the development of an OPMD, and with the risk of malignant transformation of the lesion. A biopsy with subsequent microscopic examination from the lesional tissue is necessary in identification of OPMD. This article reviews the clinical appearance of OPMDs, associated risk factors, diagnosis and histologic appearance, and treatment.


Subject(s)
Erythroplasia , Mouth Diseases , Mouth Neoplasms , Precancerous Conditions , Humans , Leukoplakia, Oral
8.
J Forensic Sci ; 61(4): 928-38, 2016 07.
Article in English | MEDLINE | ID: mdl-27364271

ABSTRACT

Due to possible secondary transfer of gunshot residue (GSR) onto a suspect in police custody prior to sampling, a baseline must be created for the amount of GSR present. With an increase of "lead free" ammunition, testing for both gunpowder and primer GSR is relevant. Seventy samples were collected using carbon-coated adhesive stubs from four Pittsburgh Police Stations and vehicles to investigate these locations as sources of secondary GSR contamination. These seventy samples were analyzed for primer GSR using scanning electron microscopy-energy-dispersive X-ray spectrometry. One primer GSR particle was detected; no sample was classified as positive for primer GSR. These same samples were then analyzed for gunpowder GSR using liquid chromatography coupled to tandem mass spectrometry to test for akardite II, ethylcentralite, diphenylamine, N-nitrosodiphenylamine, 2-nitrodiphenylamine, and 4-nitrodiphenylamine. Ethylcentralite was quantifiable in two test samples. These results suggest there is a negligible potential for secondary transfer of primer and gunpowder GSR.

9.
Histopathology ; 69(4): 607-13, 2016 Oct.
Article in English | MEDLINE | ID: mdl-27010880

ABSTRACT

AIMS: Ectomesenchymal chondromyxoid tumour (ECT) is a rare, benign intraoral neoplasm showing a predilection for the anterior dorsum of the tongue. The World Health Organization includes ECT in the pathological spectrum of soft tissue myoepithelioma. EWS RNA-binding protein 1 gene (EWSR1) rearrangement is found in 45% of cutaneous, soft tissue and bone myoepithelial neoplasms, and pleomorphic adenoma gene 1 (PLAG1) aberrations are found in 37% of EWSR1-negative soft tissue myoepitheliomas. The aim of this study was to evaluate the presence of EWSR1 and PLAG1 rearrangements in ECTs. METHODS AND RESULTS: Eleven formalin-fixed, paraffin-embedded ECTs were evaluated with fluorescence in-situ hybridization probes for EWSR1 (22q12) and PLAG1 (8q12). Among the 11 ECTs tested, three (27.3%) showed EWSR1 rearrangement in >15% of tumour cells, whereas eight (72.7%) cases did not show EWSR1 rearrangement. Eight of nine (89%) ECTs showed gain of EWSR1, probably representing gain of all or part of chromosome 22, in a varying proportion of neoplastic cells ranging between 1.4% and 27.9%. PLAG1 rearrangement was not detected in the successfully hybridized tissue sections (7/11). No correlation was observed between the molecular and histopathological findings, such as morphology of the neoplastic cells, the presence of atypia, and matrical type. CONCLUSIONS: We identified EWSR1 rearrangement in >25% of ECTs. These results suggest that some ECTs are at least genetically related to myoepithelioma of the soft parts. Finally, PLAG1 aberrations do not appear to be critical in the pathogenesis of ECT of the tongue.


Subject(s)
Calmodulin-Binding Proteins/genetics , Myoepithelioma/genetics , Myoepithelioma/pathology , RNA-Binding Proteins/genetics , Tongue Neoplasms/genetics , Tongue Neoplasms/pathology , Adult , Aged , Biomarkers, Tumor/genetics , DNA-Binding Proteins/genetics , Female , Gene Rearrangement , Humans , In Situ Hybridization, Fluorescence , Male , Middle Aged , RNA-Binding Protein EWS , Soft Tissue Neoplasms/genetics , Soft Tissue Neoplasms/pathology , Young Adult
11.
Article in English | MEDLINE | ID: mdl-26711711

ABSTRACT

OBJECTIVES: MYB rearrangement is observed in approximately 28% to 86% of adenoid cystic carcinomas (ACCs). Also, ACC features a p63+/p40+ immunophenotype in greater than 90% of cases, compared with p63+/p40- polymorphous low-grade adenocarcinoma (PLGA). Our aim was to investigate the incidence of (1) MYB rearrangement and (2) p63/p40 immunoreactivity in ACC and PLGA of minor salivary glands (MSGs). STUDY DESIGN: Seven cases of ACC as well as five of PLGA were evaluated by using a MYB (6 q23.3) break-apart fluorescence in situ hybridization (FISH) probe. In addition, all cases were immunohistochemically stained with p63 and p40 antibodies. RESULTS: All five successfully hybridized ACCs featured MYB rearrangement, whereas PLGAs did not show MYB rearrangement. Interestingly, one case of PLGA demonstrated a single intact copy of MYB in greater than 88% of the neoplastic cells. All ACCs exhibited consistent p63+/p40+ staining, whereas PLGAs demonstrated a p63+/p40- immunophenotype. CONCLUSIONS: (1) MYB rearrangement is encountered in ACCs but not PLGAs of MSGs; (2) MYB aberrations, for example, monosomy or deletion, can be seen in PLGAs; (3) combined p63/p40 immunostaining can be used to differentiate ACC from PLGA in incisionally biopsied specimens; and (4) performance of either FISH or p63/p40 immunohistochemistry is expected to be able to confirm the diagnosis of ACC or PLGA in small intraoral biopsies, since both techniques appeared to be diagnostically accurate in this pilot study.


Subject(s)
Carcinoma, Adenoid Cystic/genetics , Gene Rearrangement , Genes, myb , Salivary Gland Neoplasms/genetics , Adult , Aged , Biomarkers, Tumor/analysis , Biopsy , Carcinoma, Adenoid Cystic/pathology , Female , Humans , Immunodominant Epitopes/analysis , Immunohistochemistry , Immunophenotyping , In Situ Hybridization, Fluorescence , Male , Membrane Proteins/analysis , Middle Aged , Peptide Fragments/analysis , Pilot Projects , Salivary Gland Neoplasms/pathology , Salivary Glands, Minor/pathology
12.
Head Neck Pathol ; 10(2): 237-44, 2016 Jun.
Article in English | MEDLINE | ID: mdl-26156422

ABSTRACT

The most recent A.F.I.P. fascicle defines primordial odontogenic cyst (POC) as a distinct, nonkeratinized, odontogenic cyst of "undetermined origin" forming in the place of a developing normal or supernumerary tooth. However, the majority of examples reported in the literature under this term represent odontogenic keratocysts (keratocystic odontogenic tumors). In addition, there are rare reported cases of cystic odontomas. An 18-year-old Caucasian male presented with a unilocular mandibular radiolucent lesion in the place of a congenitally missing molar. Histologically, it featured nonkeratinizing, thin stratified squamous epithelial lining with areas of spongiosis and foci of vacuolization of individual basal cells without significant nuclear palisading. Focally, budding of the basal cell layer was identified. A zone of increased cellularity featuring induction-type fibroblasts was present subepithelially as well as dentinoid deposits with odontogenic epithelial nests. Immunohistochemically, the epithelial lining was negative for calretinin and the induction-like zone negative for S100 protein, smooth muscle actin, and CD34. The case was externally reviewed by five oral pathologists who provided various diagnostic interpretations including primordial cyst, odontogenic cyst not otherwise specified (NOS), cyst with ameloblastic changes, and unicystic ameloblastoma. At that time, a final diagnosis of odontogenic cyst NOS was rendered with a comment that it may represent a true example of POC or a cystic odontoma. The lesion has not recurred within a 13 year follow-up period after initial excision. An unusual cystic lesion is presented that may represent a true example of POC with dentinoid formation or an archegonous cystic odontoma.


Subject(s)
Odontogenic Cysts/diagnosis , Odontogenic Cysts/pathology , Odontoma/diagnosis , Odontoma/pathology , Adolescent , Biomarkers, Tumor/analysis , Humans , Immunohistochemistry , Male
13.
J Biomed Sci ; 22: 71, 2015 Aug 19.
Article in English | MEDLINE | ID: mdl-26282951

ABSTRACT

BACKGROUND: Many patients with nasopharyngeal carcinoma (NPC) face poor prognosis. Due to its hidden anatomical location, the tumor is usually diagnosed quite late, and despite initially successful treatment with radiation and cisplatin, many patients will relapse and succumb to the disease. New treatment options are urgently needed. We have performed preclinical studies to evaluate the potential NPC therapeutic activity of a newly developed analog of temozolomide (TMZ), an alkylating agent that is the current chemotherapeutic standard of care for patients with malignant glioma. RESULTS: TMZ was covalently conjugated to the natural monoterpene perillyl alcohol (POH), creating the novel fusion compound NEO212. Its impact on two NPC cell lines was studied through colony formation assays, cell death ELISA, immunoblots, and in vivo testing in tumor-bearing mice. In vitro, NEO212 effectively triggered tumor cell death, and its potency was significantly greater than that of its individual components, TMZ or POH alone. Intriguingly, merely mixing TMZ with POH also was unable to achieve the superior potency of the conjugated compound NEO212. Treatment of NPC cells with NEO212 inactivated the chemoprotective DNA repair protein MGMT (O6-methylguanine methyltransferase), resulting in significant chemosensitization of cells to a second round of drug treatment. When tested in vivo, NEO212 reduced tumor growth in treated animals. CONCLUSION: Our results demonstrate anticancer activity of NEO212 in preclinical NPC models, suggesting that this novel compound should be evaluated further for the treatment of patients with NPC.


Subject(s)
Dacarbazine/analogs & derivatives , Nasopharyngeal Neoplasms/drug therapy , Animals , Carcinoma , DNA Modification Methylases/antagonists & inhibitors , DNA Modification Methylases/metabolism , DNA Repair Enzymes/antagonists & inhibitors , DNA Repair Enzymes/metabolism , Dacarbazine/pharmacology , Humans , Mice , Mice, Nude , Nasopharyngeal Carcinoma , Nasopharyngeal Neoplasms/enzymology , Nasopharyngeal Neoplasms/pathology , Neoplasm Proteins/antagonists & inhibitors , Neoplasm Proteins/metabolism , Temozolomide , Tumor Suppressor Proteins/antagonists & inhibitors , Tumor Suppressor Proteins/metabolism , Xenograft Model Antitumor Assays
14.
Article in English | MEDLINE | ID: mdl-25561390

ABSTRACT

Sebaceous carcinoma of the oral cavity is extremely rare. To date, only 9 cases of primary sebaceous carcinoma involving the oral cavity have been reported in the English literature, with the majority of cases occurring on the buccal mucosa. This report describes a case of sebaceous carcinoma involving the maxillary gingiva. To our knowledge this is the first reported case of sebaceous carcinoma of the gingiva.


Subject(s)
Adenocarcinoma, Sebaceous/diagnosis , Gingival Neoplasms/diagnosis , Adenocarcinoma, Sebaceous/pathology , Aged , Biopsy , Diagnosis, Differential , Gingival Neoplasms/pathology , Humans , Male
15.
Head Neck Pathol ; 9(2): 269-72, 2015 Jun.
Article in English | MEDLINE | ID: mdl-24997629

ABSTRACT

Malignant rhabdoid tumors (MRTs) are exceedingly rare lesions. To our knowledge, only 2 cases have been reported in the oral cavity, with both examples occurring in infants. The current case is the third reported case of MRT of the oral cavity and the first reported case to occur in an adult at this location. The following report describes the clinical, histologic and immunohistochemical features of this tumor.


Subject(s)
Mouth Neoplasms/diagnosis , Mouth Neoplasms/therapy , Mouth/pathology , Rhabdoid Tumor/diagnosis , Rhabdoid Tumor/therapy , Biomarkers, Tumor/metabolism , Combined Modality Therapy , Female , Humans , Middle Aged , Mouth/metabolism , Mouth Neoplasms/pathology , Oral Surgical Procedures , Radiotherapy , Rhabdoid Tumor/pathology , Treatment Outcome , Vimentin/metabolism
16.
Article in English | MEDLINE | ID: mdl-22541168

ABSTRACT

A highly sensitive hydrophilic interaction liquid chromatography-tandem mass spectrometry (HILIC-MS/MS) method was developed and validated for the quantification of glycerophosphoinositol (GroPIns), glycerophosphocholine (GroPCho), glycerol 3-phosphate (GroP), inositol, and choline in the extracellular medium of Saccharomyces cerevisiae. The media samples were pretreated with a single two-phase liquid extraction. Chromatographic separation was achieved on a Waters Xbridge HILIC (150 mm × 4.6 mm, 5 µm) column under isocratic conditions using a mobile phase composed of acetonitrile/water, 70:30 (v/v) with 10mM ammonium acetate (pH adjusted to 4.5) at a flow-rate of 0.5 mL/min. Using a triple quadrupole tandem mass spectrometer, samples were detected in multiple reaction monitoring (MRM) mode via an electrospray ionization (ESI) source. The calibration curves were linear (r² ≥ 0.995) over the range of 0.5-150 nM, with the lower limit of quantitation validated at 0.5 nM for all analytes. The intra- and inter-day precision (calculated by coefficient of variation, CV%) ranged from 1.24 to 5.88% and 2.46 to 9.77%, respectively, and intra- and inter-day accuracy (calculated by relative error, RE%) was between -8.42 to 8.22% and -9.35 to 6.62%, respectively, at all quality control levels. The extracellular metabolites were stable throughout various storage stability studies. The fully validated method was successfully applied to determine the extracellular levels of phospholipid-related metabolites in S. cerevisiae.


Subject(s)
Chromatography, Liquid/methods , Glycerophospholipids/analysis , Metabolomics/methods , Saccharomyces cerevisiae/chemistry , Saccharomyces cerevisiae/metabolism , Tandem Mass Spectrometry/methods , Choline/analysis , Choline/metabolism , Fungal Proteins/genetics , Fungal Proteins/metabolism , Gene Deletion , Glycerophospholipids/metabolism , Hydrophobic and Hydrophilic Interactions , Inositol/analysis , Inositol/metabolism , Linear Models , Lipid Metabolism , Liquid-Liquid Extraction , Metabolome , Phospholipases/genetics , Phospholipases/metabolism , Reproducibility of Results , Saccharomyces cerevisiae/genetics , Sensitivity and Specificity
17.
J Mass Spectrom ; 47(2): 147-54, 2012 Feb.
Article in English | MEDLINE | ID: mdl-22359323

ABSTRACT

The employment of chemical weapons by rogue states and/or terrorist organizations is an ongoing concern in the United States. The quantitative analysis of nerve agents must be rapid and reliable for use in the private and public sectors. Current methods describe a tedious and time-consuming derivatization for gas chromatography-mass spectrometry and liquid chromatography in tandem with mass spectrometry. Two solid-phase extraction (SPE) techniques for the analysis of glyphosate and methylphosphonic acid are described with the utilization of isotopically enriched analytes for quantitation via atmospheric pressure chemical ionization-quadrupole time-of-flight mass spectrometry (APCI-Q-TOF-MS) that does not require derivatization. Solid-phase extraction-isotope dilution mass spectrometry (SPE-IDMS) involves pre-equilibration of a naturally occurring sample with an isotopically enriched standard. The second extraction method, i-Spike, involves loading an isotopically enriched standard onto the SPE column before the naturally occurring sample. The sample and the spike are then co-eluted from the column enabling precise and accurate quantitation via IDMS. The SPE methods in conjunction with IDMS eliminate concerns of incomplete elution, matrix and sorbent effects, and MS drift. For accurate quantitation with IDMS, the isotopic contribution of all atoms in the target molecule must be statistically taken into account. This paper describes two newly developed sample preparation techniques for the analysis of nerve agent surrogates in drinking water as well as statistical probability analysis for proper molecular IDMS. The methods described in this paper demonstrate accurate molecular IDMS using APCI-Q-TOF-MS with limits of quantitation as low as 0.400 mg/kg for glyphosate and 0.031 mg/kg for methylphosphonic acid.

18.
J Am Soc Mass Spectrom ; 17(2): 246-52, 2006 Feb.
Article in English | MEDLINE | ID: mdl-16413204

ABSTRACT

One of the most significant issues in any analytical practice is optimization. Optimization and calibration are key factors in quantitation. In matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS), instrument optimization is a limitation restricting quantitation. An understanding of the parameters that are most influential and the effects of these parameters on the mass spectrum is required for optimization. This understanding is especially important when characterizing synthetic polymers by MALDI-TOF-MS, due to the breadth of the polymer molecular mass distribution (MMD). Two considerations are important in quantitation, additivity of signal and signal-to-noise (S/N). In this study, the effects of several instrument parameters were studied using an orthogonal experimental design to understand effects on the signal-to-noise (S/N) of a polystyrene distribution. The instrument parameters examined included detector voltage, laser energy, delay time, extraction voltage, and lens voltage. Other parameters considered were polymer concentration and matrix. The results showed detector voltage and delay time were the most influential of the instrument parameters for polystyrene using all trans-retinoic acid (RA) as the matrix. These parameters, as well as laser energy, were most influential for the polystyrene with dithranol as the matrix.

19.
Anal Chem ; 77(14): 4539-48, 2005 Jul 15.
Article in English | MEDLINE | ID: mdl-16013871

ABSTRACT

A matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) interlaboratory comparison was conducted on mixtures of synthetic polymers having the same repeat unit and closely matching molecular mass distributions but with different end groups. The interlaboratory comparison was designed to see how well the results from a group of experienced laboratories would agree on the mass fraction, and molecular mass distribution, of each polymer in a series of binary mixtures. Polystyrenes of a molecular mass near 9000 u were used. Both polystyrenes were initiated with the same butyl initiator; however, one was terminated with -H (termed PSH) and the other was terminated with -CH2CH2OH (termed PSOH). End group composition of the individual polymers was checked by MALDI-TOF MS and by nuclear magnetic resonance (NMR). Five mixtures were created gravimetrically with mass ratios between 95:5 and 10:90 PSOH/PSH. Mixture compositions where measured by NMR and by Fourier transform infrared spectrometry (FT-IR). NMR and FT-IR were used to benchmark the performance of these methods in comparison to MALDI-TOF MS. Samples of these mixtures were sent to any institution requesting it. A total of 14 institutions participated. Analysis of variance was used to examine the influences of the independent parameters (participating laboratory, MALDI matrix, instrument manufacturer, TOF mass separation mode) on the measured mass fractions and molecular mass distributions for each polymer in each mixture. Two parameters, participating laboratory and instrument manufacturer, were determined to have a statistically significant influence. MALDI matrix and TOF mass separation mode (linear or reflectron) were found not to have a significant influence. Improper mass calibration, inadequate instrument optimization with respect to high signal-to-noise ratio across the entire mass range, and poor data analysis methods (e.g., baseline subtraction and peak integration) seemed to be the greatest obstacles in the correct application of MALDI-TOF MS to this problem. Each of these problems can be addressed with proper laboratory technique.

20.
Biomacromolecules ; 5(4): 1280-7, 2004.
Article in English | MEDLINE | ID: mdl-15244441

ABSTRACT

Facile synthesis and detailed characterization of photopolymerizable and biocompatible poly(ethylene glycol) dimethacrylates (PEGDM) and poly(ethylene glycol) urethane-dimethacrylates (PEGUDM) are described. Poly(ethylene glycol)s of various molecular masses (M(n) = 1000 to 8000 g/mol) were reacted with methacrylic anhydride or with 2-isocyanatoethyl methacrylate to form PEGDMs and PEGUDMs, respectively. PEGDMs were also prepared by a microwave-assisted route to achieve fast reaction conversions under solvent free conditions. Combined analyses of (1)H NMR and MALDI-TOF MS confirmed the formation of prepolymers of high purity and narrow mass distribution (PD < 1.02). Aqueous solutions of the PEGDMs and PEGUDMs (10% and 20% by mass fraction) were photopolymerized to yield hydrogels. Bovine chondrocytes, seeded in the hydrogels, were used to assess the biocompatibility. Preliminary rheology and uniaxial compression measurements showed varied mechanical response, and biocompatibility studies showed that cells are completely viable in both types of hydrogels after two weeks.


Subject(s)
Hydrogels , Methacrylates/chemistry , Methacrylates/chemical synthesis , Polyethylene Glycols/chemistry , Animals , Cattle , Cell Survival/drug effects , Hydrogels/chemical synthesis , Hydrogels/chemistry , Methacrylates/pharmacology , Molecular Structure , Molecular Weight , Polyethylene Glycols/chemical synthesis , Polyurethanes/chemical synthesis , Polyurethanes/chemistry , Polyurethanes/pharmacology
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