Virtual Remote Pathology Education in Support of Virtual Remote Gynecologic-Oncology Training: The Open Pathology Education Network Pilot Proof of Concept Experience.

CONTEXT.­: The subspecialty workforce in pathology globally is inadequate for the demands of many modern therapies. The Open Pathology Education Network (OPEN) was formed to augment the global pathology workforce. The International Gynecologic Cancer Society (IGCS) virtual gynecologic-oncology (gyn-onc) fellowship training identified needs for higher-level pathology support. OBJECTIVE.­: To report on an OPEN-IGCS pilot project to support gyn-onc and pathology education efforts in a developing country. DESIGN.­: Curriculum with learning objectives and content from open sources was assembled. Mentoring sessions included bidirectional case sharing. Trainees received sequential curricula assignments and had options for communication outside mentoring sessions. Pretest and posttest digital slide assessments were included. Mentors attended the gynecology tumor board, allowing for the assessment of quality and accuracy of pathology diagnosis for cases discussed. RESULTS.­: Learners completing the pretest and posttest showed substantial improvement, with 2 practicing pathologists improving their diagnostic scores from 60% to an average of 95%. A third trainee-level participant also improved, but to a lesser degree. Qualitative assessments included increased confidence in presentation and an increased ability to anticipate questions, raise issues of expanded differential diagnoses, and articulate appropriate workup. Observations of clinicians who participated also noted increased confidence in participating pathologists. Secondary value included establishing an expanded network of support in other subspecialties for participants. Pathologic issues at the tumor board decreased, from more than 50% in the first 3 months of study to 0% in the last 3 months of study. The curriculum was embedded into a self-paced learning portal at courses.open-pathology.org. CONCLUSIONS.­: The OPEN-IGCS collaboration model shows the potential to provide subspecialty pathology training remotely.

Effect of extraction solvent on total phenol content, total flavonoid content, and antioxidant activity of Limnophila aromatica.

Limnophila aromatica is commonly used as a spice and a medicinal herb in Southeast Asia. In this study, water and various concentrations (50%, 75%, and 100%) of methanol, ethanol, and acetone in water were used as solvent in the extraction of L. aromatica. The antioxidant activity, total phenolic content, and total flavonoid content of the freeze-dried L. aromatica extracts were investigated using various in vitro assays. The extract obtained by 100% ethanol showed the highest total antioxidant activity, reducing power and DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging activity. The same extract also exhibited the highest phenolic content (40.5 mg gallic acid equivalent/g of defatted L. aromatica) and the highest flavonoid content (31.11 mg quercetin equivalent/g of defatted L. aromatica). The highest extraction yield was obtained by using 50% aqueous acetone. These results indicate that L. aromatica can be used in dietary applications with a potential to reduce oxidative stress.

Impact of pretreatments on morphology and enzymatic saccharification of shedding bark of Melaleuca leucadendron.

The effects of subcritical water (SCW) and dilute acid pretreatments on the shedding bark of Melaleuca leucadendron (paper bark tree, PBT) biomass morphology, crystallinity index (CrI) and enzymatic saccharification were studied. The morphology of PBT bark was characterized by X-ray diffraction, scanning electron microscopy and Fourier transform infrared spectroscopy. SCW pretreatment mainly extracted amorphous parts of the biomass hence its CrI increased, partial decrystallization of cellulose and exposing of intact nanofibers of cellulose were observed for SCW pretreatment at 180°C. On the other hand, dilute acid pretreatment at 160°C exhibited a large decrease in CrI, an increase in surface area, a decrease in lignin content and decrystallization of cellulose as well as the peel-off and degradation of some nanofiber bundles. Dilute acid and SCW pretreatments of PBT biomass resulted in about 4.5 fold enhancement in glucose release relative to the untreated one.

Bioethanol production from pretreated Melaleuca leucadendron shedding bark--simultaneous saccharification and fermentation at high solid loading.

Bioethanol production from the shedding bark of Melaleuca leucadendron (Paper-bark Tree, PBT) was studied using subcritical water (SCW) pretreatment at various severities (So). High ethanol production was attained by implementing a factorial design on three parameters (So, solid loading and enzyme loading) in simultaneous saccharification and fermentation (SSF) mode. Ethanol concentration of 63.2 g L(-1) corresponding to ethanol yield of 80.9% were achieved from pretreated biomass (So=2.37) at 0.25 g mL(-1) solid and 16 FPU g(-1) glucan enzyme loadings. Similarly at 0.15 g mL(-1) solid loadings both high ethanol concentration (43.7 g L(-1)) and high ethanol yield (91.25%) were achieved. Regression analysis of experimental results shows that all process parameters had significant role on maximum ethanol production, glucose solubility, ethanol yield and ethanol volumetric productivity. SSF of SCW treated PBT biomass is economically feasible for production of bioethanol.

Bioethanol production from Yarrowia lipolytica Po1g biomass.

Bioethanol production from the yeast Yarrowia lipolytica biomass was studied. The effects of temperature (90-150°C) and H2SO4 concentration (2-15%w/w) on the saccharification of biomass at a hydrolysis time of 1h were investigated. A maximum glucose concentration of 35.89 g/L can be produced from defatted biomass using 6% H2SO4 at 120°C. Subcritical water (SCW) pretreatment has negligible effect on maximizing glucose yield. Only 14.53 g/L glucose can be produced using 6% H2SO4 at 120°C if un-defatted biomass was used. The highest ethanol concentration achieved was 13.39 g/L with a corresponding ethanol yield of 0.084 g/g dry biomass (0.38 g ethanol/g glucose).

Catalyst-free fatty acid methyl ester production from wet activated sludge under subcritical water and methanol condition.

Wet activated sludge was converted directly into biodiesel using water as hydrolysis reagent to enhance the extraction of lipid in activated sludge, and as catalyst for the conversion of neutral lipids into biodiesel under subcritical conditions. At 175°C, 3.5MPa, a methanol to sludge ratio of 30 (mL/g) and a sludge water content of 84wt.%, about 90% conversion to fatty acid methyl esters was achieved within 24h without the need for conventional catalysts such as KOH and H(2)SO(4). Since water is employed as a catalyst, its removal is not required; therefore, the processing costs for producing biodiesel from activated sludge are reduced. The method has the potential for applications to other feedstock with high water contents such as micro-algae.

Seroprevalence of antibodies to avian influenza A (H5) and A (H9) viruses among market poultry workers, Hanoi, Vietnam, 2001.

BACKGROUND: The frequency of avian influenza A virus infections among poultry workers is not well understood. METHODS: A seroprevalence study of market poultry workers and persons without occupational poultry exposure was conducted during 2001 in Hanoi, Vietnam. Sera were tested for avian influenza H5 and H9 antibodies by microneutralization and Western blot assays. RESULTS: Seroprevalence of H5 and H9 antibodies was 4% and 3% in poultry workers and 1% and 3.5% in non-poultry workers, respectively. CONCLUSIONS: Seroprevalence of H5 and H9 antibodies was low among Hanoi market poultry workers in 2001, but can serve as a baseline for additional studies.

Oil production from Yarrowia lipolytica Po1g using rice bran hydrolysate.

The purpose of this study was to produce microbial oil from Yarrowia lipolytica Po1g grown in defatted rice bran hydrolysate. After removing oil from rice bran by Soxhlet extraction, the bran is subjected to acid hydrolysis with various sulfuric acid concentrations (1-4% v/v), reaction times (1-8 h), and reaction temperatures (60-120°C). The optimal conditions for maximum total sugar production from the hydrolysate were found to be 3% sulfuric acid at 90°C for 6 h. Glucose was the predominant sugar (43.20 ± 0.28 g/L) followed by xylose (4.93 ± 0.03 g/L) and arabinose (2.09 ± 0.01 g/L). The hydrolysate was subsequently detoxified by neutralization to reduce the amount of inhibitors such as 5-hydroxymethylfurfural (HMF) and furfural to increase its potential as a medium for culturing Y. lipolytica Po1g. Dry cell mass and lipid content of Y. lipolytica Po1g grown in detoxified defatted rice bran hydrolysate (DRBH) under optimum conditions were 10.75 g/L and 48.02%, respectively.

Maximizing biodiesel production from Yarrowia lipolytica Po1g biomass using subcritical water pretreatment.

The yeast Yarrowia lipolytica Po1g is one of the oleaginous microorganisms with a potential for biodiesel production. Sub-critical water (SCW) treatment has been known as an effective method for increasing the amount of extractable lipids in microorganisms. In this work, the amount of neutral lipids and fatty acid profiles in neutral lipids extracted from Y. lipolytica Po1g with and without SCW pre-treatment were investigated. The effects of temperature (125, 150 or 175°C), amount of water (20, 30 or 40 mL/g biomass) and time (10, 20 or 30 min) showed that maximum neutral lipid (42.69%, w/w) could be achieved at 175°C using 20 mL water for 20 min. The maximum neutral lipid from unpretreated samples was 23.21%. No difference in fatty acid profiles was observed, but long chain fatty acids were observed in higher amount in SCW pretreated samples. SCW pretreatment increased biodiesel yield twofold.

Irresolvable complex mixture of hydrocarbons in soybean oil deodorizer distillate.

Aliphatic hydrocarbons (HCs) can be used as a fingerprint of a given seed oil. Only by characterization of aliphatic HCs could contamination by mineral oil in that seed oil be confirmed. During the isolation of squalene from soybean oil deodorizer distillate, a significant amount of unknown HCs, ca. 44 wt%, was obtained. These seemingly-easy-to-identify HCs turned out to be much more difficult to elucidate due to the presence of an irresolvable complex mixture (ICM). The objective of this study was to purify and identify the unknown ICM of aliphatic HCs from soybean oil deodorizer distillate. Purification of the ICM was successfully achieved by using modified Soxhlet extraction, followed by modified preparative column chromatography, and finally by classical preparative column chromatography. FT-IR, TLC, elemental analysis, GC/FID, NMR and GC-MS analyses were then performed on the purified HCs. The GC chromatogram detected the presence of ICM peaks comprising two major peaks and a number of minor peaks. Validation methods such as IR and NMR justified that the unknowns are saturated HCs. This work succeeded in tentatively identifying the two major peaks in the ICM as cycloalkane derivatives.

Isolation and analysis of wax esters from activated sludge.

Neutral lipid from activated sludge (AS) as a potential source for biodiesel production has recently received considerable attentions. The utilization of useful compounds in AS may help reducing the cost of biodiesel production from AS. One of these compounds is the valuable wax esters (WEs) found in AS from a food processing company in Taiwan. About 4.13% (based on dry sludge weight) bleached wax was obtained after pretreatment and bleaching of crude sludge wax obtained from the dewaxing of crude sludge oil. The major WEs detected in the bleached wax were C46-C60 with small amounts of C37-C43 and C62 WEs. The fatty acids (FAs) and fatty alcohols (FALs) profiles of WEs were also investigated. Activated sludge WEs are mainly mixture of C14-C28 FAs and C24-C37 FALs, in which the predominant FAs are C16 and C18 while the predominant FALs are C32 and C34.

Extraction and analysis of neutral lipids from activated sludge with and without sub-critical water pre-treatment.

Activated sludge with its high lipid content is a potential raw material for producing biodiesel. Sub-critical water (SCW) treatment has been known to be a cheap and effective method for reducing organic compounds, especially carcinogenic aromatic compounds such as polycyclic aromatic hydrocarbons or polychlorobiphenyls in activated sludge as well as increasing the amount of extractable lipids. In this work, the amount of neutral lipids and fatty acid profiles in neutral lipids extracted from activated sludge both with and without SCW pre-treatment were investigated. Results showed that the amount of neutral lipid extracted from SCW treated activated sludge is almost four times to those without SCW treatment. However, there is no observable difference between the fatty acid profiles of neutral lipids obtained from activated sludge with and without SCW treatment. At least 14 kinds of fatty acids were identified in the neutral lipids of sludge oil.

Genetic analysis of avian influenza A viruses isolated from domestic waterfowl in live-bird markets of Hanoi, Vietnam, preceding fatal H5N1 human infections in 2004.

The first known cases of human infection with highly pathogenic avian influenza (HPAI) H5N1 viruses in Vietnam occurred in late 2003. However, HPAI H5N1 and low-pathogenic avian influenza (LPAI) H5N2 and H9N3 viruses were isolated from domestic waterfowl during live-bird market (LBM) surveillance in Vietnam in 2001 and 2003. To understand the possible role of these early viruses in the genesis of H5N1 strains infecting people, we performed sequencing and molecular characterization. Phylogenetic analysis revealed that the hemagglutinin (HA) genes of two geese HPAI H5N1 strains belonged to clade 3, and their surface glycoprotein and replication complex genes were most closely related (98.5-99.7% homologous) to A/duck/Guangxi/22/01 (H5N1) virus, detected contemporarily in southern China, whilst the M and NS genes were derived from an A/duck/Hong Kong/2986.1/00 (H5N1)-like virus. The H5 HA gene of the duck HPAI H5N1 strain belonged to clade 5 and acquired a gene constellation from A/quail/Shantou/3846/02 (H5N1), A/teal/China/2978.1/02 (H5N1) and A/partridge/Shantou/2286/03 (H5N1)-like viruses. The phylogenetic analysis further indicated that all eight gene segments of goose and duck HPAI H5N1 and LPAI H5N2 viruses were distinct from those of H5N1 clade-1 viruses known to have caused fatal human infections in Vietnam since late 2003. The duck H9N3 isolates derived genes from aquatic-bird influenza viruses, and their H9 HA belonged to the Korean lineage. The PB2 gene of A/duck/Vietnam/340/01 (H9N3) virus had lysine at position 627. Based on the molecular characterization of specific amino acid residues in the surface and relevant internal protein-coding genes, the Vietnamese H5N1 and H9N3 virus isolates indicated specificity to avian cell surface receptor and susceptibility for currently licensed anti-influenza A virus chemotherapeutics. Our findings suggest that the H5N1 and H5N2 viruses that circulated among geese and ducks in LBMs in Hanoi, Vietnam, during 2001 and 2003 were not the immediate ancestors of the clade-1 viruses associated with fatal human infections in Vietnam. The clade-1 HPAI H5N1 viruses were independently introduced into Vietnam.

Genetic analysis of influenza A/H3N2 and A/H1N1 viruses circulating in Vietnam from 2001 to 2006.

Influenza A virus has the ability to overcome immunity from previous infections through the acquisition of genetic changes. Thus, understanding the evolution of the viruses in humans is important for the surveillance and the selection of vaccine strains. A total of 30 influenza A/H3N2 viruses and 35 influenza A/H1N1 viruses that were collected in Vietnam from 2001 to 2006 were used to analyze the evolution of the hemagglutinin (HA), neuraminidase (NA), and matrix protein (M) genes. Phylogenetic analysis of individual gene segments revealed that the HA and the NA genes of the influenza A viruses evolved in a sequential way. However, the evolutionary pattern of the M gene proved to be nonlinear and was not linked with that of the HA and NA genes. Genetic drift in HA1 segments, especially in the antigenic sites of A/H3N2 viruses, occurred more frequently in A/H3N2 viruses than it did in A/H1N1 viruses. Two reassortants, one influenza A/H3N2 strain and one A/H1N1 strain, were found on the basis of the phylogenetic analysis of the three genes. While both genetic mutation and reassortment contributed to their evolution, the frequency of genetic changes and reassortment events differs between the two subtypes. As influenza viruses circulate throughout the year, we emphasize the importance of surveillance in tropical and subtropical zones, where the emergence of new strains may be detected earlier than it is in temperate zones.

Epidemiology of influenza in Hanoi, Vietnam, from 2001 to 2003.

OBJECTIVE: The aim of this study was to clarify the epidemiology of laboratory-confirmed influenza in Hanoi, Vietnam. METHODS: Influenza was detected by virus isolation from nasopharyngeal swabs of influenza-like-illness (ILI) patients who reported to outpatient clinics in Hanoi, Vietnam between 2001 and 2003, before the start of avian influenza A/H5N1 outbreaks. Influenza isolates were characterized by hemagglutinin inhibition test. RESULTS: A total of 4708 nasopharyngeal swabs were collected from patients with ILI. Influenza was positive in 119 (2.5%) samples by virus isolation. Influenza circulated throughout the year, with possible two peaks in summer and winter. Influenza B viruses and A/H3N2 predominated in 2001 and 2002, respectively, and mixed circulation of A/H1N1, A/H3N2 and B were observed in 2003. The seasonality of influenza roughly matched with clinical case reports in the North Region by National Communicable Disease Surveillance in Vietnam. CONCLUSIONS: The findings of year-round and biannual peak circulation of influenza in a subtropical area were in accordance with the results of previous studies in tropical and subtropical regions. Our observations indicated that establishment of laboratory-based surveillance in tropical and sub-tropical countries is important for taking actions for pandemic strategies, and links to the WHO global influenza network.

Isolation and characterization of avian influenza viruses, including highly pathogenic H5N1, from poultry in live bird markets in Hanoi, Vietnam, in 2001.

Since 1997, outbreaks of highly pathogenic (HP) H5N1 and circulation of H9N2 viruses among domestic poultry in Asia have posed a threat to public health. To better understand the extent of transmission of avian influenza viruses (AIV) to humans in Asia, we conducted a cross-sectional virologic study in live bird markets (LBM) in Hanoi, Vietnam, in October 2001. Specimens from 189 birds and 18 environmental samples were collected at 10 LBM. Four influenza A viruses of the H4N6 (n = 1), H5N2 (n = 1), and H9N3 (n = 2) subtypes were isolated from healthy ducks for an isolation frequency of over 30% from this species. Two H5N1 viruses were isolated from healthy geese. The hemagglutinin (HA) genes of these H5N1 viruses possessed multiple basic amino acid motifs at the cleavage site, were HP for experimentally infected chickens, and were thus characterized as HP AIV. These HA genes shared high amino acid identities with genes of other H5N1 viruses isolated in Asia during this period, but they were genetically distinct from those of H5N1 viruses isolated from poultry and humans in Vietnam during the early 2004 outbreaks. These viruses were not highly virulent for experimentally infected ducks, mice, or ferrets. These results establish that HP H5N1 viruses with properties similar to viruses isolated in Hong Kong and mainland China circulated in Vietnam as early as 2001, suggest a common source for H5N1 viruses circulating in these Asian countries, and provide a framework to better understand the recent widespread emergence of HP H5N1 viruses in Asia.

Anterior cervical discectomy and fusion with implantable titanium cage: initial impressions, patient outcomes and comparison to fusion with allograft.

BACKGROUND CONTEXT: The use of metal cage prosthetic devices in anterior cervical discectomy with fusion (ACDF) has increased rapidly. One of these devices is the titanium Rabea cage (Signus, Alzenau, Germany) [correction]. There are no peer-reviewed objective reports on the use of these cages in cervical discectomy. PURPOSE: The authors present preliminary outcomes data on the Rabea cage. This study is intended to provide adjunct data for surgeons who are using or are considering the use of these devices. STUDY DESIGN/SETTING: Patients in a private practice were studied prospectively as part of a long-term assessment of outcomes using several surgical procedures. Data were collated and analyzed by an independent researcher. PATIENT SAMPLE: Rabea cages were used in consecutive candidates (n=37) for ACDF. The results using Rabea cages were compared with two prospectively studied control groups, one historical (n=66) and one concurrent (n=28), both groups using ACDF with bone allograft and no instrumentation. OUTCOME MEASURES: Success at 6 months after surgery was determined using six major patient-reported outcome measures, including visual analog scales (VAS) for arm and neck pain, the Oswestry pain and disability scale, four-part (excellent-through-poor) scales for measurement of return to activities of daily living or to work and satisfaction with the results of surgery. Perioperative complications, number of vertebral levels fused, and worker's compensation and smoking status were also compared among the study groups. Fusion and subsidence were evaluated for the Rabea cage group. METHODS: Criteria for inclusion consisted of consecutive patients who presented with unremitting radicular arm pain, with or without neck pain, and/or a neurological deficit that correlated with appropriate level and side neural compression on magnetic resonance imaging or computed tomography. Questionnaire follow-up was at 6 months after surgery with 100% compliance. RESULTS: A total of 50 cages were implanted in the 37 patients. At 6-month follow-up, 78% of patients reported successful arm pain relief (VAS scores below 5). Patient satisfaction was successful in 78% of the cases. Other success rates included neck pain relief in 73% and Oswestry pain and disability scale in 70%. There was a median improvement in the Oswestry scores of 28 points (61% change). Worker's compensation patients fared dramatically worse than did the noncompensation patients in all outcome measurements. Combining all three study groups resulted in significantly worse outcomes for multilevel than for single-level procedures and for smokers compared with nonsmokers, but low case numbers precluded conclusive analysis for the Rabea group alone. In the Rabea group there were two complications, neither cage related, whereas none were reported for the ACDF controls. Rabea group fusion rates were 84% at 3 months and 95% at 6 months, but the clinical relevance of this radiological evidence when metal prostheses are used is questionable. The outcomes results were clinically and statistically indistinguishable from those of our control groups and were similar to published studies using other titanium cages. CONCLUSIONS: This is the first prospective, independently conducted report on Rabea cages. Results of the short-term (6 months) follow-up did not differ from outcomes results in our patients who have undergone ACDF with bone allograft. Although this is a preliminary assessment, the Rabea cage may represent an alternative to bone dowels and hip graft. As is the case for other allografts, artificial or bone, the main advantage is elimination of donor site complications, and the disadvantages include difficulty in radiographic assessment of fusion and potential for cage subsidence.