Laminar Chamber Research Articles

INFLUENCE OF CLAY FRACTION ON VISCOSITY IN RELATION TO MUDFLOW

Mudflow is one type of natural disaster. This type of mass movement can be triggered by changing of water content due to increasing of rainfall intensity. The occurrence in Indonesia and Taiwan is relatively high. According to its definition, the material is dominant by fine grained soil. The viscosity as one of main rheology parameters governs mudflow movement. In order to overcome the conventional viscometer which could not measure around liquid limit, the authors developed a new laboratory test so called the flow box test (FBT). The governing equation is a couple of Terzaghi’s trap door and Bingham model. From this test, one of the merit is this test can measure the viscosity around liquid limit. Using the case study in Indonesia (Ciwidey and Karangayar) and Taiwan (Maokong), the result shows that increasing of clay fraction is followed by increasing of viscosity. Hence, this research has a contribution to explain the influence of clay fraction on viscosity in relation to mudflow.

Identification of Batrachochytrium dendrobatidis inhibiting isolates via challenge against salamander skin microbiota.

Rapid declines in global amphibian populations can be partially attributed to chytridiomycosis caused by the fungus Batrachochytrium dendrobatidis (Bd). Due to how quickly it has spread worldwide, and the number of amphibians affected, this fungus is of increasing concern in both the herpetological and mycological communities. We characterized the cutaneous microbiome of Appalachian salamanders, which persist in the presence of the fungus despite high rates of infection in co‐occurring frog populations. The cutaneous microbiome of vertebrates is known as the first line of defense against invading pathogens. We hypothesized that these salamanders might have resident bacteria on their skin with the ability to inhibit Bd growth. Our objectives were to 1) isolate resident bacteria from salamander skin swabs, 2) challenge these isolates against Bd, and 3) genotype all inhibitory isolates. During the summer of 2016, 82 swabs were collected from three salamander genera including Desmognathus (n=36), Eurycea (n=20), and Plethodon (n=26). To isolate cutaneous bacteria, swabs were streaked on Reasoner's 2A agar plates that we checked daily for two weeks. Colonies with unique morphology were isolated onto secondary plates until pure cultures were obtained. To challenge the isolates against Bd, zoospores were harvested from cultures that had been aged 5–7 days, counted with a hemocytometer, and diluted to approximately 1,000,000 cells/milliliter (mL). Tryptone agar plates were then flooded with 1.0 mL of standardized zoospore solution and dried for 45 minutes in a laminar flow hood. Bacterial isolates were streaked onto each plate in co‐culture with Bd. Plates were incubated at room temperature for four days, examined for noticeable zones of inhibition (ZOI), measured in millimeters. Isolates that produced a measurable ZOI were photographed and will be screened in further testing against Bd. We used direct colony PCR and Sanger sequencing to identify all inhibitory isolates. Presently, 63 of the 472 acquired isolates have been challenged, 16 of which produced a ZOI. The challenge phase is ongoing and we anticipate discovery of additional candidate probiotic species. In addition, we plan to test for possible synergistic inhibition by challenging candidates in polyculture.Support or Funding InformationGreat Smokey Mountains Conservation Association and Tennessee Herpetological SocietyThis abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.

Different mineral oils used for embryo culture microdrop overlay differentially impact media evaporation

Mineral oil is an important component of the embryo culture system; serving as a means to avoid media off gassing and stabilizing pH, providing temperature stability and helping avoid evaporation and harmful osmolality increases. Stability of media osmolality is especially important considering the increased use of non-humidified benchtop incubators and uninterrupted embryo culture media system. Several types of mineral oil are available for use during embryo culture, which can vary in viscosity and density. To compare different mineral oils during microdrop overlay in dry benchtop incubators and resulting impact on media evaporation. Microdrops of culture media (25ul) were prepared in a laminar flow hood by pipetting 25ul of media onto the surface of a 35mm dish and covering with 3.5ml of 1 of 4 types of mineral oil (Sigma Light Mineral Oil 0.84g/ml, Sigma Heavy Mineral Oil 0.88g/ml, OvOil™ paraffin oil and Hypure™ Heavy paraffin oil). All dishes were placed into a gassed, non-humidified benchtop incubator and cultured for 6 days. Dishes were removed and osmolality of microdrops measured using a vapor pressure osmometer (Wescor). Positive controls included media directly out of the bottle. The resulting 5 treatments were measured 3 times each. Data were analyzed using ANOVA and Tukey analysis and presented as the mean ± SEM, p<0.05. Mean osmolality of control media was 266 ± 0.88mOsm. Compared to control, evaporation and resulting significant increase in osmolality occurred in all microdrop treatments regardless of the type of oil used for overlay. However, the amount of osmolality change varied based on the type oil used. Use of Heavy Mineral Oil resulted in the least evaporation, while Light Mineral oil had the most. Both paraffin oils showed similar osmolality increases at levels between light and heavy mineral oils.Tabled 1Mineral Oil UsedOsmolality (6 days of Culture)Sigma Light Mineral Oil311 ± 2.3mOsmcSigma Heavy Mineral Oil289 ± 0.6mOsmaOvOil™ (100% pure paraffin oil)298 ± 1.2mOsmbHypure™ Heavy (paraffin oil)297 ± 1.5mOsmb Open table in a new tab Different minerals oils used for microdrop oil overlay result in different rates of media evaporation and resulting osmolality increase. Density of mineral oil seems to be an important factor to consider in terms of media evaporation. Differing nomenclature used to characterize oils used for embryo culture can be confusing and it may be beneficial for companies to list oil density as part of descriptive materials.

Validated chromatographic method for the simultaneous determination of eight drugs (morphine, ropivacaine, bupivacaine, baclofen, clonidine, sufentanil, fentanyl and ziconotide) for intrathecal analgesia

Intrathecal analgesia has increased over the past two decades in various indications: chronic refractory pain from cancerous or non-cancerous origins, spasticity. These different indications involve the use of different molecules alone or in combination such as morphine, ropivacaine, bupivacaine, fentanyl, sufentanil, clonidine, baclofen and ziconotide. Pump refills are prepared at the pharmacy under a laminar flow hood. An analytical control should be carried out before release of the preparation. A new method of analytical control by chromatography has been developed and validated according to the International Conference on Harmonization guideline in order to secure the production process.

Hand-Held Photometer for Instant On-Spot Quantification of Nucleic Acids, Proteins, and Cells.

This paper presents a novel hand-held photometer, termed "Photopette", for on-spot absorbance measurements of biochemical analytes. The Photopette is a multicomponent, highly portable device with an overall weight of 160 g, which fits within 202 mm × 47 mm × 42 mm. Designed in the form factor of a micropipette, Photopette integrates a photodiode detector with light emitting diodes (LEDs) to form a highly customizable photometer which supports a wide variety of applications within the wavelengths between 260 and 1050 nm. A dual-purpose disposable reflective tip was designed to act as a sample holder and a light-reflecting system, which is in stark contrast to the operation of mainstream spectrophotometers and photometers. Small volume analytes may be measured with low sample loss using this proprietary CuveTip. A user-friendly software application running on smart devices was developed to control and read the values from Photopette via a low-energy Bluetooth link. This one-step strategy allows measurements on-spot without sample transfer, minimizing cross-contamination and human error. The results reported in this paper demonstrate Photopette's great potential to quantify DNA, direct protein, and cell density directly within the laminar flow hood. Results are compared with a Nanodrop 2000c spectrophotometer, a mainstream spectrophotometer for small-volume measurements.

Cryotherapy by encapsulation-dehydration is effective for in vitro eradication of latent viruses from ‘Marubakaido’ apple rootstock

Apple stem pitting virus (ASPV), Apple chlorotic leaf spot virus (ACLSV) and Apple stem grooving virus (ASGV) are several major viral pathogens of apple trees, responsible for substantial damage to the world’s apple industry. This study aimed to evaluate the effectiveness of the encapsulation-dehydration cryopreservation technique to eradicate these viral pathogens from in vitro shoot tips excised from ‘Marubakaido’ apple rootstock cultures. Axillary shoot tips were excised from in vitro cultures, encapsulated in alginate beads, precultured in MS salts, dehydrated in a laminar flow hood, immersed in liquid nitrogen, then warmed and recovered on medium. After LN exposure, in vitro rooting and acclimatization, recovered ‘Marubakaido’ plants exhibited 52% survival and 35% regrowth without callus formation. After 8 months of regrowth, PCR analyses revealed that all the plants were free of ACLSV and ASPV, but 2 out of 20 recovered plants were still infected with ASGV. This is the first report in Brazil of the application of cryotherapy to eradicate viral complexes in Malus. Cryotherapy can facilitate the production of virus-free plants by producing high quality plant material.

Ethinomedicine Plant Based Product in Caries Management

Background: Ethnic medicinal practice has come into foucus in recent times due to increased awareness of side effects of the medicine (pharmaceutical drugs). In oral health care also the focus is shifted to herbal products (neem sticks/leaves, mango leaves) for brushing from commercially available tooth paste and brush. In our survey on oral health use of jatropha curcas instead of brushing with tooth brush and paste was observed and the dental caries prevalence was low. Objective: The observation from the survey initiated the present study to evaluate antibacterial activity of jatropha curcas leaves against tooth caries organism strepococcus mutans. Method: Collection And Identification of Plant Material was done Preparation Of The Jatropha Curcas Extract with leaves and dilution in to 1:3 ratio with ethyl acetate and methanol was done 3 Antibacterial Activity was performed with strain Streptococcus mutant by inoculating into 20ml Luria Bertini broth. 100 ml of Muller-Hinton agar was sterilized and poured into sterile petri plates in the laminar chamber and allow to solidify. 100μl of the cultures were spread onto the plates using a spreader. 5 wells were punched on each plate using a 5 mm megabore for four different sample concentrations. 50μl of the dilutions were pipetted into the wells and all the plates were incubated at 37°C overnight. The zone of inhibition was observed and the diameter of the zone was recorded. Results: The antibacterial activity of jatropha curcas extracts was observed and it varied with in the types of extract preparation. Methanolic extract preparation has shown better antibacterial effect against streptococcus mutans. Conclusion: Jatropha curcasa leaves can be a potential antibacterial agent against cariogenic microorganism streptococcus mutans.

180 Increasing GfrA1-Positive Spermatogonial Stem Cell Population of Goat

Spermatogonial stem cells (SSC) form the basis of spermatogenesis and continuous fertility in male. Their meagre population in the testis is a hindrance in the in vitro study of biological activity of these cells. The objective of the present study was to isolate and characterise goat SSC and increase their number during in vitro culture by different methods. Two goat testes (3 to 4 months of age) were collected from the slaughterhouse and transported to the laboratory. The testes were washed and seminiferous tubules were collected and minced in the laminar flow hood. The seminiferous tubules were washed twice with PBS to remove spermatozoa and subjected to double enzymatic digestion (collagenase, 1 mg mL−1, hyaluronidase, 1 mg mL−1, trypsin, 0.05%, and DNaseI, 10 µg mL−1 for 45 min and second digestion with same set of enzymes except trypsin for 30 min). The isolated cells were filtered sequentially through nylon mesh filters of pore size 70 and 40 µm. The cells were plated on DSA-lectin coated dishes for 4 h and the unattached cells were cultured on a Sertoli cell feeder layer prepared by treating with mitomycin-C for 3 h. The cells were cultured in DMEM/F-12 supplemented with human recombinant growth factors (glial cell-derived neurotrpic factor, 10 ng mL−1, fibroblast growth factor FGF, 10 ng mL−1, epidermal growth factor, 20 ng mL−1), 10% fetal bovine serum, and antibiotics. The expression of pluripotency markers (Oct4, Nanog, Sox2) and SSC-specific markers (Thy1, GfrA1, and Uchl1) in the SSC colonies was determined by RT-PCR and immunostaining, after in vitro culture of 3 weeks. The SSC population was enhanced by differential plating, Percoll density gradient (on Day 1) and SSC passaging (by passaging SSC colonies on Day 20). The cells were tagged with GfrA1 antibody and their population was tested by flow cytometry. The SSC colonies started appearing after 7 days and continued to grow in size and number until 3 weeks. The SSC colonies were positive for the pluripotency markers Oct4, Nanog, and Sox2 by RT-PCR and immunostaining. The SSC were also positive for the SSC-specific markers Thy1, GfrA1, and Uchl1 by RT-PCR and immunostaining. Flow cytometry showed that the GfrA1-positive population in the SSC enriched by the differential plating was 11.23%, Percoll density gradient was 23.57%, and by passaging of SSC colonies, after picking and trypsinising with 0.05% trypsin, was 91.23%. In vitro culture of the SSC enriched by these methods also revealed that the number of SSC colonies appearing in the cells enriched by passaging was higher than the other methods. From the results of present study, we conclude that SSC are positive for markers of pluripotency and SSC-specific markers. The SSC population can be enhanced to a very high level following SSC passaging, which is an inexpensive method and does not require expensive instruments like fluorescence- or magnetic-activated cell sorting.

Fresh and mechanical behavior of a self-compacting concrete with additions of nano-silica, silica fume and ternary mixtures

Self-compacting concrete (SCC) has experienced significant development in the light of results obtained from numerous studies due to a series of advantages that it offers. The use of mineral admixtures at microscale and recently at nanoscale has permitted high-performance SCC to be obtained. Over the past few years, micro silica (mSi) and nano silica (nSi) have been the most used admixtures in continuing research into the areas of civil and agricultural engineering. This paper examines the behavior of 10 mixtures of SCC prepared with binary and ternary dosages through use of Portland cement (CEM I 52.5 R), mSi and nSi. As a reference, a SCC was designed which used no mineral admixtures, with the rest of the dosages using different percentages of mSi and nSi. Three were made with 2.5%, 5% and 7.5% of nSi; three more with 2.5%, 5% and 7.5% of mSi, and the remaining three by using both admixtures (ternary mixtures) mSi and nSi, with percentages of 2.5%/2.5%, 5%/2.5% and 2.5%/5%. The properties studied are rheological (flow test, funnel test and box test), mechanical (compressive strength, tensile strength and modulus of elasticity), and microstructural (hydration composites). According to the results obtained, all the concretes meet the requirements to be classified as SCCs by monitoring the quantity of superplasticizer additive according to the type and quantity of mineral admixture used. The higher compressive strength value was obtained by the ternary mixture with 2.5%/2.5% of mSi and nSi both at 28 and 90 days. Ternary mixtures of 2.5%/5% and 5%/2.5% reach values similar to those obtained by the mixture with 7.5% of nSi. Based on these results, it is possible to say that the compressive strength depends on the particle size and the amount of the admixture, and the particle-size distribution of the resulting mixture. This allows the advantage of obtaining considerably high compressive strength by using lower amounts both of nSi and of mSi and, therefore, of SP.

Differences in Disinfection Protocols of Raw Material Vials Intended for the Production of Parenteral Nutrition Mixtures

AbstractBackgroundPreparation of parenteral nutrition (PN) mixtures is performed in a controlled atmosphere area (CAA) under a laminar flow hood. In 2014 and 2015 quality controls have revealed contaminations of PN bags by Bacillus. The different products used to avoid this contamination were Aniosurf premiumMethodsFive protocols have been tested by varying the disinfection step: hydrogen peroxide (H2O2) or sodium hypochlorite (immersion or spray), ethanol (non sporicidal) only by immersion. Prewash and aerosolization steps remained unchanged. For each series, a quantity of 30 vials have been prewashed, then disinfected according to the tested protocol, and finally aerosolized. In order to quantify the effectiveness of disinfection 10 vials were sampled using contact plates at every step of treatment.ResultsMicroorganisms were found after ethanol disinfection (n=4) and aerosolization (n=2). No germs were found when hydrogen peroxide or sodium hypochlorite were used. The atmospheric rates of H2O2 were 0.2 ppm by immersion and 0.8 ppm by spray, these values are below the recommended value level (1 ppm).ConclusionsThis study has validated the use and effectiveness of these disinfecting products. Taking into account the efficiency and ease of use for staff, immersion has been preferred to spray and hydrogen peroxide to sodium hypochlorite.

Implementation of a telechemotherapy center in the Peruvian jungle to improve patients' quality of life.

6547 Background: Instituto Nacional de Enfermedades Neoplásicas (INEN) is located in Lima (capital of Peru). There are no oncologists in regions like Peruvian jungle. Approximately 6% of medical attentions at INEN are people from this cities; so they must spend money and time to receive their treatment far from their cities. The purpose was to implement a module of outpatient chemotherapy in a non-specialized hospital, located in Peruvian jungle, for chemotherapy administration monitored by oncologists from INEN through the use of information and communication technologies (ICT). Methods: Two working teams were organized by formal agreement between INEN and local authorities to properly implement the infrastructure and to train local staff from December 2014 to November 2015. Questionnaire EORTC QLQ-C30 (v3) was used to assess QoL at the beginning and 3 months later. Results: A chemotherapy room with 18 armchairs, an area with a laminar flow hood to mix drugs at pharmacy, and videoconferencing equipment were implemented in Lamas, San Martin (1100 Km from Lima). Two general practitioners, 3 nurses and a pharmacist from Lamas, were trained during 3 months at INEN. Since November 2015, 31 patients were admitted. They received 181 cycles of chemotherapy in 248 rounds of administration, monitored by teleconference. Additionally 227 videoconference meetings to evaluate patients and to coordinate with local team were performed. Global health status improved from 76.67% to 83.33%. We observed benefit in functional scales, likewise symptoms related to disease did not vary, nevertheless financial difficulties decreased from 33.3% to 6.67%. In terms of money and time, this meant an average saving of $150.00 and 4 hours of travel time. Conclusions: Using ICT, we successfully implemented a tele-chemotherapy module in the heart of Peruvian jungle without harming patients quality of life. They will not need to travel to Lima for receiving their treatment.

Cryopreservation of somatic embryos from Petiveria alliacea L. by different techniques based on vitrification

Petiveria alliacea L. is a medicinal plant originating from the Amazon region. This study describes an efficient cryopreservation protocol for somatic embryos (SEs) produced from roots of P. alliacea based on the comparison of vitrification, encapsulation-dehydration, and D cryo-plate techniques. With the vitrification technique, SEs treated with PVS2 solution (0.4 M sucrose, 3.3 M glycerol, 2.4 M ethylene glycol, and 1.9 M DMSO) for 30 min displayed high viability (85%) and intermediate proliferation recovery (about 12 adventitious SEs produced from original SEs [SEs/SE] after 90 d of culture). With the encapsulation-dehydration technique, lower viability (70%) and very low proliferation recovery (about two SEs/SE) were achieved with cryopreserved SEs dehydrated for 10 min in a laminar air flow cabinet. The D cryo-plate technique led to high viability (85%) and proliferation recovery (19 SEs/SE) of cryopreserved SEs after 90 min dehydration. In the experimental conditions tested, the D cryo-plate method was the most efficient technique for cryopreservation of P. alliacea SEs.

Особливості клітинного циклу мезенхімальних стовбурових клітин з жирової тканини собаки за різних пасажів культивування

The features of the cell cycle of culture of adipose-derived mesenchymal stem cells from the for different cultivating passages were studied. Mesenchymal stem cells were obtained from the adipose tissue of the dog under a laminar flow hood by an explant method in our modification. Cell cultivation was carried out at 37 °C, 100% moisture and 5% CO2 in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) and 1% antibiotic-antimycotic. The culture medium was changed 2–3 times per week and the cells were selected by their capacity to attach to the flask surface. When culture flasks became 80% confluence, cells were detached with 0.25% trypsin containing 1 mmol/L EDTA and subsequently replayed at a concentration of 104 cells/cm2 for next passaging. A cells culture of adipose derived mesenchymal stem cells was obtained on the 2nd, 7th and 12th passages. The method of flow cytometry determined the level of aneuploid cells and the distribution in the cell cycle phases. The morphology of cells of different passages was studied using an inverted microscope Axiovert 40. It was investigated that the culture of mesenchymal stem cells from adipose tissue in the 2nd passage contains a significant number of the proliferative pool (S + G2/M) cells and it was 29.51 ± 3.56% of the total number of diploid cells. The number of aneuploid cells was 1.55 ± 0.43%. All cells had fibroblast-like morphology. It was established that in the middle passages (7th) in the culture of mesenchymal stem cells from the adipose tissue of the dog no significant changes were found in the distribution of cells in the phases of the cell cycle. The number of diploid cells of the proliferative pool S + G2/M and the G0/G1 pre-synthetic period remains unchanged. The level of aneuploidy increases only within the tendency. Morphologically, cells had fibroblast-like form. It was determined on 12th passage of cultivation, a significant decrease in the number of cells of the proliferative pool (S+G2/M), which was 18.93 ± 0.66% of the total number of diploid cells compared to the 2nd passage. The number of aneuploid cells increased and it was 3.49 ± 0.38%. Morphologically, separate cells had processes. The indicator of the effect of cells cultivation on the content of diploid cells of the proliferative pool (S+G2/M) in culture is ɳ2x = 70% (P &lt; 0.05). So, first characteristic properties of the aging of the culture of canine adipose-derived mesenchymal stem cells appear on the 12th passage of cultivation.

Low-grade infections in nonarthroplasty shoulder surgery

Recent studies have identified the diagnostic challenge of low-grade infections after shoulder arthroplasty surgery. Infections after nonarthroplasty procedures have not been reported. This study assessed patient-related risk factors, outcomes, and clinical presentation of low-grade infection after open and arthroscopic nonarthroplasty shoulder surgery. The cases of 35 patients presenting with suspected low-grade infection were reviewed. Biopsy specimens taken at revision surgery were cultured in the sterile environment of a class II laminar flow cabinet and incubated for a minimum of 14 days at a specialist orthopedic microbiology laboratory. Patient-related factors (age, occupation, injection), index surgery, and infection characteristics (onset of symptoms, duration to diagnosis, treatment) were analyzed. Positive cultures were identified in 21 cases (60.0%), of which 15 were male patients (71%). Of all patients with low-grade infection, 47.6% were male patients between 16 and 35 years of age. Propionibacterium acnes and coagulase-negative staphylococcus were the most common organisms isolated (81.1% [n = 17] and 23.8% [n = 5], respectively). Of 14 negative culture cases, 9 were treated with early empirical antibiotics (64.3%); 7 patients reported symptomatic improvement (77.8%). Of 5 patients treated with late empirical antibiotics, 4 stated improvement. Patients presented with symptoms akin to resistant postoperative frozen shoulder (persistent pain and stiffness, unresponsive to usual treatments). Young male patients are at greatest risk for low-grade infections after arthroscopic and open nonarthroplasty shoulder surgery. P. acnes was the most prevalent organism. Patients presented with classic postoperative frozen shoulder symptoms, resistant to usual treatments. Interestingly, 78.6% of patients with negative cultures responded positively to empirical treatment.

Experimental and numerical approaches for the quantification of tumble intensity in high-performance SI engines

The activity presented in the paper is focused on the development of a high-performance engine specifically dedicated to CNG fueling. The engine features a variable valve actuation system as well as different possible compression ratios ranging between 12 and 14. More specifically, the present work carries out an experimental and numerical characterization of the steady-state tumble flow from the engine head and deals with the development and assessment of a numerical model for the engine-cycle transient simulation. Experimental tests were carried out at Fiat Research Center (CRF) on a flow test rig equipped with two different devices for the tumble measurement, based on the Ricardo method and on the two-dimensional HWA acquisition of the axial velocity, respectively. It is worth highlighting that the HWA technique was purposely developed by CRF for the tumble characterization. The experimental results were also used for the calibration of a ‘virtual flow box’ numerical model, which was in turn exploited to gain complimentary information about the characteristics of the tumbling flow and to define the requirements for the transient CFD model of the real engine, both in terms of discretization mesh and turbulence modeling.The results showed that the HWA technique represents a factual alternative to the integral techniques for tumble characterization. It also provides additional information about the unevenness of the flow distribution generated by the intake system. The ‘virtual flow box’ simulations pointed out that good results can be obtained by using the Realizable k-ε model, whereas both the Realizable and the RNG k-ε models proved to be appropriate for the engine transient simulations. Finally, the CFD models showed to be fairly accurate and reliable, especially when the relative changes from one engine geometry and/or valve actuation to another need to be accounted for.

Morphological study of a mummified heart dated back to 1829: preliminary results

In May 2016, a cylindrical lead container was found in the chapel of the Seminary of Sassari. Based on the archive documents, the cylinder should have contained the heart of Tommaso Arnosio, Archbishop of Torres and Primate of Sardinia and Corsica from 1822 to 1829, who died in 1829 at 54 years, in Turin, his home city. The Archbishop died in the midst of the most atrocious sorrows caused by a large purulent pocket near the heart. After the autopsy and according to his will, the heart was shipped to Sassari, where the heart was buried in the chapel of the Seminary and forgotten for about 186 years. After the discovery, the container was brought for study to our CSAPS laboratory of the Department of Biomedical Sciences. A multidisciplinary team of experts hypothesized as cause of death a suppurative pleuritis or a para-cardiac lung abscess. To verify this hypothesis the cylinder was opened in a laminar flow hood, avoiding any contamination, and a mummified organ immersed in cotton wadding was found. The organ, typically heart shaped, appeared 13 x 9 cm sized, with a thickness of 1.5 cm, a weight of 80.5 g, a dark brown colour and a hard consistency. Few fragments of tissue were collected for analysis; then the organ, closed in the lead container, was returned to the chapel of the seminary. Histological analysis required a rehydration using the Sandison Solution (1); the samples were treated for light microscopy and stained with HE, PAS and Gram’s method. The first results have shown a morphology of cardiac and pericardial tissues partially preserved, and the banding and the intercalated disks appeared just in small portions. The Gram’s method was negative till now. Further histological, immunohistochemical, ultrastructural and, molecular investigations are in progress.

Cryopreservation of citrus seed via dehydration followedby immersion in liquid nitrogen

An important method for plant germplasm conservation is offered by a biotechnology-based approach of cryopreservation. Cryopreservation refers to the storage of plant material at ultralow temperatures in liquid nitrogen. A procedure for cryopreservation of polyembryonic seeds was improved for selected citrus cultivars from Turkey. Seed dehydration was performed at different exposure times, in sterile conditions of a laminar flow-hood. The tested cultivars showed the highest tolerance to low temperature storage when the seeds were first dehydrated to a moisture content of 21.8% for Poncirus trifoliata Raf. x Citrus sinensis Osb. and to 17.6% for Citrus limonia Osbeck. The postcryopreservation germinability ranged from 73.3% (Poncirus trifoliata Raf. x C. sinensis Osb. and Fortunella margarita (Lour.) Swingle) to 93.3% (C. jambhiri Lush.). Dehydration was beneficial for germination of seeds from all of the tested citrus species after the liquid nitrogen exposure. Seedlings derived from cryopreserved seeds had well-formed shoots and roots and were easily acclimated to greenhouse conditions.

PM 7/128 (1) Xanthomonas axonopodis pv. allii

Specific scopeThis Standard describes a diagnostic protocol for Xanthomonas axonopodis pv. allii.This Standard should be used in conjunction with PM 7/76 Use of EPPO diagnostic protocols.Specific approval and amendmentFirst approved in 2016‐09.

Comprehensive Quality Control of the ITG 68Ge/68Ga Generator and Synthesis of 68Ga-DOTATOC and 68Ga-PSMA-HBED-CC for Clinical Imaging.

A good-manufacturing-practices (GMP) (68)Ge/(68)Ga generator that uses modified dodecyl-3,4,5-trihydroxybenzoate hydrophobically bound to a octadecyl silica resin (C-18) as an adsorbent has been developed that allows for dilute HCl (0.05N) to efficiently elute metal-impurity-free (68)Ga(3+) ready for peptide labeling. We characterized the performance of this generator system over a year in conjunction with the production of (68)Ga-labeled DOTATOC and Glu-NH-CO-NH-Lys(Ahx)-HBED-CC (PSMA-HBED-CC) intended for clinical studies and established protocols for batch release. A 2,040-MBq self-shielded (68)Ge/(68)Ga generator provided metal-free (68)GaCl3 ready for peptide labeling in the fluidic labeling module after elution with 4 mL of 0.05N HCl. The compact system was readily housed in a laminar flow cabinet allowing an ISO class-5 environment. (68)Ga labeling of peptides using GMP kits was performed in 15-20 min, and the total production time was 45-50 min. Batch release quality control specifications were established to meet investigational new drug submission and institutional review board approval standards. Over a period of 12 mo, (68)Ga elution yields from the generator averaged 80% (range, 72.0%-95.1%), and (68)Ge breakthrough was less than 0.006%, initially decreasing with time to 0.001% (expressed as percentage of (68)Ge activity present in the generator at the time of elution), a unique characteristic of this generator. The radiochemical purity of both (68)Ga-DOTATOC and (68)Ga-PSMA-HBED-CC determined by high-performance liquid chromatography analysis was greater than 98%, with a minimum specific activity of 12.6 and 42 GBq/μmol, respectively. The radionuclidic ((68)Ge) impurity was 0.00001% or less (under the detection limit). Final sterile, pyrogen-free formulation was provided in physiologic saline with 5%-7% ethanol. The GMP-certified (68)Ge/(68)Ga generator system was studied for a year. The generator system is contained within the fluidic labeling module, and it is compact, self-shielded, and easy to operate using simple manual techniques. The system provides radiolabeled peptides with high (>98%) radiochemical purity and greater than 80% radiochemical yield. The (68)Ge levels in the final drug products were under the detection limits at all times. (68)Ga-DOTATOC and (68)Ga-PSMA-HBED-CC investigational radiopharmaceuticals are currently being studied clinically under investigational new drug (IND) applications submitted to the U.S. Food and Drug Administration.

Genetic associations of in-line recorded milkability traits and udder conformation with udder health

ABSTRACTMilkability and udder conformation traits of Swedish Holstein (SH) and Swedish Red (SR) cows from 93 herds with automatic milking systems or conventional milking parlors were used to study genetic relationships to lactation average somatic cell score (LSCS) and incidence of clinical mastitis (CM). Estimated genetic correlations between measures of milking speed (average flow rate, milking time and box time) and LSCS ranged between 0.29 and 0.57 and showed that high milking speed is associated with increasing LSCS. Regressions indicated a curvilinear relationship. Genetic correlations between milking speed and CM showed similar values as for LSCS in SH cows, but were inconsistent in SR cows. Shallow udder and strong fore udder attachment were consistently correlated with good udder health. The unfavorable relationships between milking speed and udder health traits should be considered together with a few udder conformation traits when selecting for better milkability.