Different Methods Of Preservation Research Articles

The Challenge to Stabilize, Extract and Analyze Urinary Cell-Free DNA (ucfDNA) during Clinical Routine.

The "Liquid Biopsy" has become a powerful tool for cancer research during the last decade. Circulating cell-free DNA (cfDNA) that originates from tumors has emerged as one of the most promising analytes. In contrast to plasma-derived cfDNA, only a few studies have investigated urinary cfDNA. One reason might be rapid degradation and hence inadequate concentrations for downstream analysis. In this study, we examined the stability of cfDNA in urine using different methods of preservation under various storage conditions. To mimic patient samples, a pool of healthy male and female urine donors was spiked with a synthetic cfDNA reference standard (fragment size 170 bp) containing the T790M mutation in the EGFR gene. Spiked samples were preserved with three different buffers and with no buffer over four different storage periods (0 h; 4 h; 12 h; 24 h) at room temperature vs. 4 °C. The preservatives used were Urinary Analyte Stabilizer (UAS, Novosanis, Wijnegem, Belgium), Urine Conditioning Buffer (UCB, Zymo, Freiburg, Germany) and a self-prepared buffer called "AlloU". CfDNA was extracted using the QIAamp MinElute ccfDNA Mini Kit (Qiagen, Hilden, Germany). CfDNA concentration was measured using the Qubit™ 4 fluorometer (Thermo Fisher Scientific, Waltham, MA, USA). Droplet digital PCR (ddPCR) was used for detection and quantification of the T790M mutation. Almost no spiked cfDNA was recoverable from samples with no preservation buffer and the T790M variant was not detectable in these samples. These findings indicate that cfDNA was degraded below the detection limit by urinary nucleases. Stabilizing buffers showed varying efficiency in preventing this degradation. The most effective stabilizing buffer under all storage conditions was the UAS, enabling adequate recovery of the T790M variant using ddPCR. From a technical point of view, stabilizing buffers and adequate storage conditions are a prerequisite for translation of urinary cfDNA diagnostics into clinical routine.

Proximate Composition and Antioxidant Potential of Moringa oleifera Leaves as Influenced by Different Methods of Preservation

Moringa oleifera leaves are recognized as nutritionally and medicinally important for life. Maintaining the freshness of leaves is a challenge due to their very low shelf life. The aim of this study is to examine the effect of the nutritional quality of Moringa oleifera leaves by cost-effective preservation methods. 
 Samples of raw, sundried, oven-dried, and brine-preserved Moringa oleifera leaves were investigated for % proximate compositions based on AOAC methods. The antioxidant potential was analyzed via DPPH assay.
 The moisture content was significantly (p<0.05) reduced from 40.37±0.08% (fresh) to 27.48±0.20% for Moringa oleifera in brine. Ash content was significantly (p<0.05) reduced from 9.12±0.03% (fresh) to 8.63±0.15% (sun-dried), 8.17±0.08% (oven dried), 7.60±0.08% (in brine). Crude protein content of Moringa oleifera in brine (in bottles) significantly (p<0.05) increased to 26.53±0.06% from 26.50±0.05% (fresh), 24.17±0.13% (sundried), 23.34±0.24% (oven dried). Crude fat content was significantly (p<0.05) reduced from 6.33±0.10% (fresh) to 5.26±0.09% (sun-dried), 4.30±0.15% (oven dried), 3.55 ±0.12% (in brine). Crude fiber content was significantly (p<0.05) reduced from 9.63±0.09% (fresh) to 8.47±0.08% (sun-dried), 8.17±0.03% (oven dried), 6.83±0.14% (in brine). Total carbohydrate content was significantly (p<0.05) increased from 8.20±0.29% (fresh) to 45.90±0.32% (sun-dried), 52.60±0.36% (oven dried), 26.01±0.17% (in brine).
 The highest antioxidant potential was recorded in Moringa oleifera leaves in brine. (IC50 value is 54.6).
 Therefore, Moringa oleifera leaves in brine are a product that enhances the quality of nutrients while increasing its shelf life. Sun-dried and oven-dried Moringa oleifera leaf powder is also a cost-effective way of preservation.

The Impact of Preservation Techniques on Methane-Arrested Anaerobic Digestion of Nutrient-Rich Feedstocks.

The carboxylate platform is a promising biomass-to-energy pathway that uses methane-arrested anaerobic digestion (MAAD) to convert biomass to carboxylic acids, which can be chemically converted to industrial chemicals and liquid fuels. Lignocellulose is an energy-rich carbon source, but lacks nutrients necessary for microbial growth. Chicken manure (rural waste) and sewage sludge (urban waste) are rich in nitrogen and useful macronutrients; therefore, co-digesting these wastes with lignocellulose improves MAAD performance. However, waste nutrients must be digested immediately, or preserved. This study investigated the effects of various preservation techniques - frozen (fresh), air-dried, and baked - on chicken manure and sewage sludge. Batch experiments were performed with office paper (carbon source) and chicken manure or sewage sludge (nutrient source) with different methods of preservation. Fresh substrates produced higher acid yields and biomass conversion (the amount of biomass consumed during digestion) than dried substrates. Baked chicken manure showed reduced conversion and total acid production, which suggests that oven-drying reduces digestibility. From the batch data, the Continuum Particle Distribution Model (CPDM) predicted results of a four-stage countercurrent digestion. The data are displayed on maps showing the impact of liquid residence time (LRT) and volatile solids loading rate (VSLR) on conversion and product concentration. Co-digesting office paper and wet chicken manure at a non-acid volatile solid (NAVS) concentration of 300g/Lliq, the model predicted a high total acid concentration of 52.8g/L and conversion of 0.89g NAVSdigested/NAVSfed at a volatile solid loading rate of 4g/(Lliq·day) and liquid retention time of 35days.

Effects of preservation on chemical composition and sensory analysis of skipjack tuna (Katsuwonus pelamis) in Tema, Ghana

Although fish is an integral part of Ghanaian diets and play important nutritional role, much research has not been conducted with respect to the nutritional composition and sensory analysis of marine fishes in Ghana. This study evaluated chemical composition and sensory analysis of Skipjack tuna (Katsuwonus pelamis) sampled from Tema in the Greater Accra region of Ghana. Four different samples; very dry smoked (Sample A), dry smoked (sample B), moist smoked (sample C) and fresh (sample D) skipjack tuna were used for this study. Chemical composition (moisture, ash and protein) was analysed using standard laboratory methods whilst sensory parameters (colour, odour, flavour, texture and overall acceptance) were assessed using 20 trained panelists. There were significant differences in moisture and crude protein contents (P < 0.05). Ash content was however non-significantly different. Moisture content for very dry smoked skipjack tuna, dry smoked skipjack tuna, moist smoked skipjack tuna and fresh skipjack tuna were 49.79 ± 1.73 %, 60.22 ± 1.80 %, 59.32 ± 0.01 % and 68.65 ± 0.16 %, respectively whilst crude protein content were 90.83 ± 1.09 %, 73.32 ± 1.09 %, 88.15 ± 1.19 % and 91.72 ± 0.19 %, respectively for very dry smoked skipjack tuna, dry smoked skipjack tuna, moist smoked skipjack tuna and fresh skipjack tuna. It can be concluded that different methods of preservation (smoking) have significant influence on proximate composition as well as sensory analysis. It is suggested that further studies be conducted on sensory analysis and proximate composition of fish sold on Ghanaian markets. In addition, we recommend that the Food and Drugs Authority (FDA) conduct regular studies on fish to ascertain the quality.

Batuan (Garciniabinucao) Fruit Preservation

This experimental research aimed to determine the most acceptableBatuan (Garciniabinucao) Fruit Preservation.Specifically, it sought to answer if there is no significant difference in the acceptability of Batuan Fruits Preserved using different methods of preservation as to appearance, aroma, taste, texture and general acceptability. Batuanfruits weresubjected into different solutions of plain water (Product A), brine solution (Product B) and light syrup solution (Product C). It was determined through organoleptic evaluation using Five-point Hedonic Scaleby thirty (30) respondents at Iloilo Science and Technology University, La Paz, Iloilo City last April, 2018. The statistical tools used were the Mean, Analysis of Variance (ANOVA) tested at 0.01 level of significance and post hoc test.Product C is the most accepted product in terms of appearance, aroma, taste, texture and general acceptability as evaluated by the respondents. Findings also revealed that there is significant difference in Product A as to taste. The general result of the study revealed that there is no significant difference between three products in terms of appearance, aroma, taste, texture becauseit does not affect the general acceptability of the products using the three methods of preservation after post hoc test.

Methods for the determination of heavy metals in indocalamus leaves after different preservation treatment using inductively-coupled plasma mass spectrometry

The microwave digestion method was used to determine the concentrations of Cu, Mn, Pb, Cr, Cd, and Hg in indocalamus leaves by inductively-coupled plasma mass spectrometry (ICP-MS) and collision/reaction cell technology after different methods of preservation. Two digestion methods, HNO3-H2O2 and HNO3-HF, were used to digest and quantify weighed indocalamus leaves. The standard addition method was used to compare the two digestion methods' influence on the determination of heavy metal concentrations. The accuracy of the method was successfully validated using two standard reference materials, tea leaves (GBW 08513) and peach leaves (SRM 1547), through recovery experiments using standard addition. The results suggested that correlation coefficients of standard curve of the ten tested elements were between 0.9975–0.9999. The Instrument Detection Limit (IDL) was between 0.003ng·L−1–0.516ng·mL−1, the Method Detection Limit (MDL) was between 12.0ng·g−1–96.0ng·g−1, and the recovery rates of both methods were 80–115%. Both digestion methods had a high accuracy and a high recovery rate. HNO3-HF resulted in more complete digestion, and thus more accurate results. Both dry and wet preservation methods decreased the concentrations of Mn, Pb, Cr, and Cd in indocalamus leaves. The wet-preserved indocalamus leaves had an abnormally high concentration of Cu. Therefore, people should choose natural or dry-preserved indocalamus leaves.

Current situation on fertility preservation in cancer patients in Spain: Level of knowledge, information, and professional involvement

Abstract Introduction The estimated risks of infertility in childhood cancer due to radiation, chemotherapy and surgery are well known. The involvement of professionals and advances in the different methods of preservation are increasing. However, many patients do not receive information or perform any method of preservation. Material Questionnaires to paediatric onco-haematology institutions throughout Spain. The questionnaire consisted of 22 questions assessing their usual practices and knowledge about fertility preservation. Results Fifty members of the Spanish Society of Paediatric Haematology and Oncology, representing 24 of 43 centres, responded. These represented 82% of centres that treated higher numbers of patients. The effect of treatment on fertility was known by 78% of those who responded, with 76% admitting not knowing any guideline on fertility in children or adolescents. As for the ideal time and place to inform the patient and/or family, only 14% thought it should be done in the same cancer diagnosis interview. In clinical practice, 12% of those surveyed never referred patients to Human Reproduction Units, another 12% only did so if the patients showed interest, and 38% only refer patients in puberty. Just over one-third (34%) of those referrals were going to receive highly gonadotoxic treatment. Conclusions There are clear differences between pre-puberty and puberty patients. The frequency with which some method of fertility preservation is performed in patients is low. All respondents believe that the existence of national guidelines on the matter would be of interest.

ALTERNATIVE METHODS OF TECHNOLOGICAL PROCESSING TO REDUCE SALT IN MEAT PRODUCTS

The world trends in table salt reduction in meat products contemplate the use of different methods for preservation of taste and consistency in finished products as well as shelf life prolongation. There are several approaches to a sodium chloride reduction in meat products. The paper presents a review of the foreign studies that give evidence of the possibility to maintain quality of traditional meat products produced with the reduced salt content. The studies in the field of salty taste perception established that a decrease in a salt crystal size to 20 µm enabled reducing an amount of added table salt due to an increase in the salty taste intensity in food products. Investigation of the compatibility of different taste directions is also interesting as one of the approaches to a sodium chloride reduction in food products. The use of water-in-oil-in-water (w/o/w) double emulsions allows controlling a release of encapsulated ingredients (salt), which enables enhancement of salty taste. The other alternative method of technological processing of meat raw material for reducing salt in meat products is the use of high pressure processing. This method has several advantages and allows not only an increase in the salty taste intensity, but also formation of a stable emulsion, an increase in water binding capacity of minced meat and extension of shelf-life.

Situación de la preservación de fertilidad en pacientes con cáncer en nuestro medio: grado de conocimiento, información e implicación de los profesionales

IntroductionThe estimated risks of infertility in childhood cancer due to radiation, chemotherapy and surgery are well known. The involvement of professionals and advances in the different methods of preservation are increasing. However, many patients do not receive information or perform any method of preservation. MaterialQuestionnaires to paediatric onco-haematology institutions throughout Spain. The questionnaire consisted of 22 questions assessing their usual practices and knowledge about fertility preservation. ResultsFifty members of the Spanish Society of Paediatric Haematology and Oncology, representing 24 of 43 centres, responded. These represented 82% of centres that treated higher numbers of patients. The effect of treatment on fertility was known by 78% of those who responded, with 76% admitting not knowing any guideline on fertility in children or adolescents. As for the ideal time and place to inform the patient and/or family, only 14% thought it should be done in the same cancer diagnosis interview. In clinical practice, 12% of those surveyed never referred patients to Human Reproduction Units, another 12% only did so if the patients showed interest, and 38% only refer patients in puberty. Just over one-third (34%) of those referrals were going to receive highly gonadotoxic treatment. ConclusionsThere are clear differences between pre-puberty and puberty patients. The frequency with which some method of fertility preservation is performed in patients is low. All respondents believe that the existence of national guidelines on the matter would be of interest.

Applicability of the real-time PCR assay in the amplification of cyanobacterial DNA from preserved samples

Applic ability of the real-time PCR assay in the amplification of cyanobacterial DNA from preserved samples The study and monitoring of cyanobacterial blooms often involves the use of preserved samples to avoid cellular degradation. However, preserved samples may not be suitable for molecular biology studies because preservation methods can interfere with DNA quality/quantity. Real-time quantitative PCR analysis (qPCR) has been widely applied in molecular analysis and is considered a promising method for monitoring purposes. This study intended to evaluate the applicability of the real-time qPCR technique in samples that were subjected to different methods of preservation: (1) 15% Lugol’s iodine solution (2) 4% formaldehyde and (3) 25% glutaraldehyde. The ability to amplify and quantify DNA extracted from Planktothrix agardhii was assessed using the rpoC1 gene as the target fragment in both raw water samples and in vitro cultures. No reliable DNA amplification was obtained from glutaraldehyde-preserved samples. Successful amplification was obtained from Lugol’s and formaldehyde-preserved samples. In this case, however, the quantification that was achieved by real-time PCR cannot be used to infer cell numbers, because the Ct values that were obtained from the Lugol’s and formaldehydepreserved samples were significantly higher than the Ct values that were obtained from the unpreserved samples. Therefore real-time PCR can be used for the detection and identification of cyanobacteria in preserved samples but no reliable cell quantification can be performed using this method.

Evaluation of different methods for preservation of fungal cultures

O contínuo isolamento de culturas e a necessidade de sua manutenção para estudos acerca do cultivo, produtividade, caracterização nutricional, biotecnológica, tanto para objetivos acadêmicos como para fins comerciais e industriais são de fundamental importância. Assim, o objetivo deste trabalho foi avaliar diferentes métodos de preservação de culturas fúngicas, com o propósito de pesquisas acerca do cultivo de cogumelos e estudos afins. Para verificar a eficiência dos métodos, foi feita uma avaliação após seis meses de preservação. Para isso, foram utilizados dois tubos das culturas mantidas em diferentes métodos (água destilada, em óleo mineral, repicagens periódicas e sílica gel). Fragmentos dessas culturas foram novamente inoculadas em placas de Petri contendo meio malte agar (3%). Foram feitas quatro repetições (placas) por linhagem, das quais duas foram adicionadas de serragem de marupá, com o fim de testar a eficácia dos referidos métodos. Lentinus strigosu s foi o fungo que apresentou crescimento mais rápido e, nos demais métodos, cresceu em cinco dias. O período de colonização das linhagens de Pleurotus ostreatus , provenientes dos métodos de preservação em água destilada, óleo mineral e repicagem periódica foi de seis dias, sem diferença do tempo de colonização entre os métodos. A linhagem de Pleurotus sajor-caju e as linhagens de P. ostreatus colonizaram o meio em sete dias de incubação em todos os métodos, exceto em sílica gel. O tempo de colonização das linhagens de Lentinula edodes LED 25, LED 27, CHICO II, Flamulina sp . e de Pleurotus ostreatus POS 09/99 e POS 09/100 foi de nove dias de incubação. As linhagens de L. edodes LED 12, LED 20, LED 33, LED 35, LED 51, LED 55, LED 58, LED 75, LED 96/13, LED SALTO, LED JABL, LED SIII e a linhagem de Coprinus comatus CCO 01/01 obtiveram maior delonga no tempo de colonização (10 dias). Abstract The continual isolation of cultures and the need for their maintenance for studies on the cultivation, yield, nutritional and biotechnological characterization, both for academic and for commercial and industrial purposes are of fundamental importance. Thus, the objective of this study was to evaluate different methods of preserving fungal cultures for the purpose of research on the cultivation of mushrooms and correlated studies. To verify the efficiency of the methods, an evaluation was carried out after six months of preservation. Two test tubes were used from cultures maintained at different methods (distilled water, mineral oil and periodical transplanting and silica gel). Fragments of these cultures were once again inoculated in Petri plates containing malt agar (3%). Four replications were performed (plaques) per strain, two of which were added marupá sawdust in order to test the effectiveness of those methods. Lentinus strigosus was the fungus that experienced the fastest growth, and in other methods, it has grown in five days. The colonization period of Pleurotus ostreatus strains, using the methods of preservation in distilled water, mineral oil and periodical transference was six days, without difference in the colonization time between the methods. The P. sajor-caju strain and the P. ostreatus strains colonized the medium in seven days of incubation in all the methods, except on silica gel. The colonization time of Lentinula edodes LED 25, LED 27, CHICO II, Flamulina sp. and Pleurotus ostreatus POS 09/99 e POS 09/100 strains was nine days of incubation. However, the L. edodes LED 12, LED 20, LED 33, LED 35, LED 51, LED 55, LED 58, LED 75, LED 96/13, LED SALTO, LED JABL, LED SIII strains and Coprinus comatus CCO 01/01 strain had greater delay in the olonization of time (10 days).

Effect of Pre-treatments on Physical and Sensory Qualities of Dehydrated Carrot Slices

Carrot is a popular cool season crop grown throughout India. It is used as raw as well as cooked form. The preservation methods such as dehydration, canning, and pickling can be successfully adopted to preserve carrot for off-season. Dehydrated carrot in the form of grating can be used in the preparation of halwa, discs made in to chips. Carrot is rich source of vitamin A and is a seasonal crop. To make available in the off season and to exploit the nutritive value of carrot it is necessary to convert into various value added products. Among the different methods of preservation, dehydration is one of the best and cheapest method of preservation. In this regard, a study was conducted to prepare dehydrated carrot slices. The experiment was conducted in completely randomized block design with seven treatments and three replications. The various physical parameters like recovery, time taken for drying, rehydration ratio, dehydration ratio, reconstitution ratio and organoleptic evaluation were carried out. Among the different treatments carrot slices dehydrated without any pretreatment taken minimum time for dehydration (15.10 hrs) whereas carrot slices dehydrated with blanching for 2 minutes + steeping in 40° Brix syrup + 6% salt for 2 hours had taken maximum time for dehydration (18.45 hrs). The highest recovery and lowest dehydration ratio were noticed in carrot slices dehydrated with blanching for 2 minutes + steeping in 40° Brix syrup + 6% salt for 2 hours. The rehydration ratio was found higher (5.04) in the slices steeped 0.75 percent KMS for 2 hours whereas the lowest rehydration ratio (2.59) was found in carrot slices blanched for 2 minutes + steeped in 40° brix syrup + 6% salt for 2 hours. The highest reconstitution ratio (0.57) was found in slices blanched for 2 minutes + steeped in 40° brix syrup+ 6% salt for 2 hours. Higher scores for colour and appearance (4.55), taste (4.43) were recorded in the slices blanched for 2 minutes + steeped in 40° brix syrup + 6% salt for 2 hours (T6) whereas highest score for texture (4.46) and overall acceptability (4.55) were recorded in the slices blanched for 2 minutes + steeped in 300 brix syrup + 6% salt for 2 hours which was on par with T6. It indicated that sugar and salt treatment blends improved the organoleptic qualities (colour, taste and overall acceptability) of the product.

Investigation on Microstructure of CrN-Based Solid Self-Lubricant Composite Coating

The CrN-based solid self-lubricant composite coating was obtained by magnetron sputtering and low-temperature ion sulfuring. The XPS, AES and SEM equipped with EDS were utilized to observe and analyze the microstructure of the composite coating. The results show that there are Cr, Mo, N, S in the composite coating, and the phase structure is CrN, MoS2 and a few oxide. The element S was implanted 500nm in depth. For the sulfurized layer is extremely apt to be oxidized, in this paper, the source of element O is analyzed in detail through comparative studies on different methods for preservation of specimens, and the film-forming mechanism of CrN-based solid self-lubricant composite coating is discussed.

Vitality and genetic fidelity of white-rot fungi mycelia following different methods of preservation

Basidiomycetes present specific problems with regard to their preservation, because most of them do not form resistant propagules in culture but exist only as mycelium. Usually these fungi can only be preserved by serial transfer on agar (labour-intensive procedures that can increase the danger of variation or loss of physiological or morphological features), or cryopreserved in liquid nitrogen (expensive). Cryopreservation at −80 °C and lyophilisation could be good alternatives. In this work we set up and tested six protocols of cryopreservation at −80 °C, and 12 protocols of lyophilisation on 15 isolates of white-rot fungi (WRF) belonging to 10 species. The tested protocols were mainly characterized by the use of different growth media, protectants, time and number of perfusion with protectants and finally by the typology and origin of the samples to be cryopreserved (mycelium/agar plug, whole colony) or to lyophilise (mycelium/agar plug, mycelium fragment, whole colony). Cryopreservation and lyophilisation outcomes were checked, at morphological (macro- and microscopic features), physiological (growth rate and laccase, Mn-independent and Mn-dependent peroxidases activities) and genetic level (Amplified Fragment Length Polymorphisms analysis – AFLP). Vitality of all fungi was successfully preserved by all cryopreservation protocols at −80 °C, and by two lyophilisation methods. Our results showed that cryopreservation at −80 °C did not produce morphological changes in any isolate, while two isolates were affected by lyophilisation. None of the physiological features were lost, even though growth rate and enzyme activities were somehow influenced by all preservation methods. AFLP analysis showed that only the two isolates that varied in their morphology after lyophilisation produced a different DNA fingerprint pattern in comparison with that obtained before lyophilisation. These findings provide evidence that cryopreservation at −80 °C and lyophilisation are suitable alternatives to liquid nitrogen cryopreservation for preservation of some WRF strains.

Evaluation of phenotypic and genotypic alterations induced by long periods of subculturing of Cryptococcus neoformans strains

Cryptococcus neoformans is an encapsulated fungal organism that can cause disease in apparently immunocompetent, as well as immunocompromised, hosts. Since 1930, successive subculture has been used to preserve C. neoformans isolates in our Fungus Collection. In the 1970s, some of these Fungus Collection samples were selected to be subjected to a different methods of maintenance--that of lyophilized. Our objective was to analyze C. neoformans isolates in order to make a comparative evaluation between these two methods of preservation. The overall aim of this study was to qualify the preservation technique used in our mycology laboratory since the technique used might affect the survival, stability and purity of the primary isolates in culture. The samples were analyzed using classical mycology methods and using the randomly amplified polymorphic DNA technique In the analysis of phenotypes and genotypes, the typical characteristics of C. neoformans were found to differ in relation to the different methods of preservation employed. The aim of this study was to demonstrate the importance of selecting the appropriate method of preservation for fungus collections. This selection can affect the survival and purity of the cultures, and preserve the stability of their physiological, biochemical, and genetic characteristics.

Museum fish specimens and molecular taxonomy: A comparative study on DNA extraction protocols and preservation techniques

Summary Museum fish specimens are invaluable resources for genetic studies, but extraction of high quality DNA is often problematic. In this study, hairtail fishes of the genera Trichiurus and Lepturacanthus (family: Trichiuridae) representing a wide range of preservation histories and three different methods of preservation were analyzed for mitochondrial DNA (mtDNA) extraction, amplification and sequencing of marker genes. A total of six protocols, including a commercially available kit, were compared in this study. Amplification of conserved genes such as16S rRNA and 12S rRNA were done using polymerase chain reaction with sequence analyses using automated capillary sequencing techniques. The results show that mtDNA extraction, amplification and sequencing of conserved genes could be obtained successfully from frozen (−20°C) preserved specimens (1–5 years) and also from ethanol (95%) fixed specimens (2–5 years) but not from any of the formalin (10%) fixed specimens (3–4 years). However, specimens that have been fixed for only 7 days in buffered formalin (10% formalin with phosphate buffer containing 173 mm salt) and ethanol (95%) could yield successful mtDNA extraction, amplification and sequence information of both 16S rRNA and 12S rRNA.

Sporulation, radial growth and biomass production of A. robusta and M. thaumasium submitted to different methods of preservation

Biological control is based on the use of microorganisms to combat populations of a specific pathogen. Nematophagous fungi have shown to be promising agents to combat nematode parasites of plants and animals. These organisms produce special structures along the hyphae, named traps, which are responsible for capturing and killing the nematode pre-parasitic stages. The growth of the isolates I31 of Arthrobotrys robusta and NF34a of Monacrosporium thaumasium was studied after submission to four different methods of preservation during eighteen months. The isolates were stored at 4oC and cryopreserved with or without addition of cryoprotectants and stored in silica gel. The isolates did not present variation in radial growth when stored at 4oC and frozen with or without addition of cryoprotectants. There was no significant difference between the dry mycelial mass of the isolates stored at 4oC and those frozen after the addition of cryoprotectants. Storage in silica gel and freezing without cryoprotection interfered negatively in the capacity of the fungi to produce mycelial mass. This characteristic also was observed for sporulation of the two isolates. The behaviour of the isolates when maintained for prolonged periods under laboratory conditions is an important factor for selection of the most efficient method of preservation.

Molecular genetic and chemical assessment of radix Angelica (Danggui) in China.

The roots of Angelica sinensis (Danggui), a traditional Chinese medicine, have been used for invigorating blood circulation for over 2000 years in China. Three common species of Angelica roots are found in Asia: A. sinensis from China, A. acutiloba from Japan, and A. gigas from Korea. By using a molecular genetic approach, the 5S-rRNA spacer domains of the three species of Angelica were amplified, and their nucleotide sequences were determined. Diversity in DNA sequences among various species was found in their 5S-rRNA spacer domains, which could serve as markers for authentic identification of Angelica roots. In chemical analyses, the main constituents of Angelica roots including ferulic acid and Z-ligustilide were determined by HPLC; roots of A. sinensis were clearly distinct in that they contained approximately 10-fold higher levels of ferulic acid and Z-ligustilide as compared to roots of A. acutiloba and A. gigas. In addition, the amounts of main constituents in roots of A. sinensis varied according to different regions of cultivation and different methods of preservation. The chemical profile determined by HPLC therefore could serve as a fingerprint for quality control of Angelica roots.

DREI MITTELALTERLICHE SCHLŰSSELDENKMÄLER DER KUNSTGESCHICHTE UNGARNS - RESTAURIERT

Summary The study is devoted to a critical analysis of the recent conservation work made in the course of the millennary festivities on three important monuments of Hungarian art history. These important sources of art historical knowledge were treated by different methods of preservation (from conservation to reconstruction or new building). The adequacy of these methods is discussed in this paper. The monuments in question are as follows: the ruins of the royal Provostry Church of the Blessed Virgin in Székesfehérvár (11th to 15th centuries; see also P. Biczó below), the royal (later archiepiscopal) Castle in Esztergom (earlier buildings: 10th to 12th century, existant buildings: late 12th to 16th century with later alterations), the Royal Residence in Visegrád (14–15th centuries, see also L. Varga below).

The effects of different methods of preservation on the 15N concentration in Folsomia candida (Collembola)

The aim of this experiment was to study the effect of the most frequently used preservation media for soil invertebrates on the 15 N concentration in Folsomia candida (Collembola) during extraction and storage. Two stocks of F. candida were fed with food containing either a low or a high quantity of 15 N for at least 2 months. Specimens from both stocks were stored in different preservatives for 1, 5 and 27 weeks. Specimens were preserved in eleven different solutions during the first week: 70% ethyl alcohol, 70% ethyl alcohol with 5% glycerin, 1% picric acid, distilled water, tap water, 70% isopropanol, 0.5% salicylic acid, saturated NaCl, 0.5% sodium benzoate, 25% ethyl glycol, and 10% formalin. Some specimens were killed immediately using heat (60°C). Another group of animals was kept frozen at −18°C. The specimens for the 5- and 27-week experiments were kept in 70% ethyl alcohol for the first week and subsequently one group was transferred into 90% ethyl alcohol, a second into 4% formalin and a third group was frozen. The results show that the different preservatives influenced the 15 N concentration. The alcohols did not change the 15 N concentration while picric acid and sodium benzoate led to reduced amounts of the isotope. The other liquids had modest effects. Freezing or preservation in ethyl alcohol or formalin proved to be reliable methods of storage for the purpose of a 15 N analysis.