Micropropagation and cryopreservation of the rare endemic Colchicum figlalii germplasm.

BACKGROUND: The natural population of Colchicum figlalii (Varol) Parolly and Eren grows in a narrow area of serpentine rock clearings at an altitude of 1900-2100 m in Southwestern Anatolia (Sandras Mountain, Mugla, Turkey). The species is regarded as endangered according to the IUCN Red List Categories. OBJECTIVE: To develop an optimum procedure for in vitro propagation and cryopreservation of germplasm of this rare endemic. MATERIALS AND METHODS: A total of 281 bulbs were used as in vitro culture starting material and after surface sterilization, clean material was obtained from 157 of them. Woody Plant Medium (WPM), Olive Medium (OM), and Murashige and Skoog medium (MS) were used for in vitro culture establishment. RESULTS: The maximum regeneration rate (~67.3%) was obtained after four weeks of incubation on OM. The calli were successfully induced by using OM supplemented with 10.7 uM NAA from leaves of in vitro grown C. figlalii bulbs. A PVS2-vitrification procedure was used for cryopreservation of C. figlalii callus tissue. After cryo-storage, the best result for regeneration (66.7%) was obtained from calli treated with PVS2 for 75 min before plunging into liquid nitrogen. All rooted seedlings derived from cryopreserved calli were successfully acclimatized to greenhouse conditions. CONCLUSION: This study is an effective reference for future long-term conservation of similar species that are difficult to cryopreserve. Doi.org/10.54680/fr24410110412.

Acquired resistance to pamidronate treated effectively with zoledronate in juvenile Paget's disease.

Juvenile Paget's disease (JPD) is a rare autosomal recessive osteopathy. There is still a question about the most effective treatment modality in long-term prognosis. A 9-month-old boy who suffered from bone pain and deformities with a very high alkaline phosphatase level was diagnosed as JPD by radiographic findings. Genetic analysis showed a homozygous large deletion in TNFRSF11B gene encoding osteoprotegerin. Clinical improvement was observed with intravenous pamidronate therapy. However, the effect of drug reduced in time so the annual dose per kilogram body weight was increased after 2 years. Despite this increment, bone fractures developed and bone pain recurred with high-ALP levels, which suggested resistance to pamidronate. Switching to zoledronate resulted a significant improvement in bone findings radiographically and ALP level. Severe hypocalcemia requiring intravenous calcium treatment complicated the first dose of zoledronate, but not recurred thereafter. Intravenous pamidronate therapy is effective in reducing bone pain, improving bone deformities and motor development in infantile onset JPD. However, this effect can be transient. Switching to another bisphosphonate like zoledronate may provide long-term clinical and biochemical improvement as an alternative treatment in case of resistance to pamidronate therapy.

First and second trimester biochemical markers in familial mediterranean fever.

OBJECTIVES: Our aim was to investigate whether the maternal serum concentrations of first and second trimester serum analytes are altered in familial Mediterranean fever (FMF) pregnancies. MATERIALS AND METHODS: The screening tests were compared in a series of 16 serum samples from FMF pregnancies and in a cohort of 48 pregnant women with normal pregnancy. Serum samples were obtained between 11 and 13 weeks; 16 and 18 weeks gestation. RESULTS: Serum pregnancy-associated plasma protein-A (PAPP-A) levels, expressed as multiples of the median (0.9 ± 0.45 MoM) in the control group, were significantly higher than FMF patients (0.6 ± 0.3 MoM) (p = 0.027). Analyses of alpha-fetoprotein, human chorionic gonadotropin and oestriol levels showed no significant differences between FMF and normal pregnancies. CONCLUSIONS: Our study revealed that low levels of PAPP-A are associated with FMF.

Genetic diversity among melon accessions (Cucumis melo) from Turkey based on SSR markers.

Melon (Cucumis melo) is an important vegetable crop in Turkey, where it is grown in many regions; the most widely planted lines are local winter types belonging to the var. inodorous. We examined 81 melon genotypes collected from different provinces of Turkey, compared with 15 reference melon genotypes obtained from INRA/France, to determine genetic diversity among Turkish melons. Twenty polymorphic primers were used to generate the SSR markers. PCR amplification was performed and electrophoresis was conducted. SSR data were used to generate a binary matrix. For cluster analysis, UPGMA was employed to construct a clustering dendrogram based on the genetic distance matrix. The cophenetic correlation was compared with the similarity matrix using the Mantel matrix correspondence test to evaluate the representativeness of the dendrogram. A total of 123 alleles were amplified using the 20 SSR primer sets. The number of alleles detected by a single primer set ranged from 2 to 12, with an average of 6.15. The similarity ranged from 0.22 to 1.00 in the dendrogram developed from microsatellite analysis. Based on this molecular data, we concluded that genetic diversity among these Turkish accessions is relatively high.

Effects of cholesterol on progesterone production by goat luteal cell subpopulations at two different stages of the luteal phase.

The aim of the present study was to evaluate the effects of cholesterol on progesterone production during long-term culturing of luteal cell subpopulations at early and late luteal stages of the goat corpora lutea. Corpora lutea were collected from Angora goats on days 5 and 15 of the oestrous cycle. Luteal cells were isolated by collagenase digestion. The cells were separated into two distinct subpopulations by Percoll density-gradient centrifugation. Both subpopulations of luteal cells staining positively for 3ß-HSD activities (5 × 10(4) cell/well) were cultured with or without 22(R)-hydroxycholesterol (22R-HC) in serum-free culture medium for periods of up to 7 days. Cells were incubated with serum (10%) for the first 18 h of incubation followed by serum-free medium. Cell treatment (10 and 20 µg/ml) was performed on days 1, 3 and 5. Treatment of cells with both concentrations of 22R-HC resulted in significant (p < 0.01) and dose-dependent stimulation (p > 0.05) on progesterone production in both fractions of cells throughout 7 days of incubation. Treatment of the cells with cholesterol resulted in 2.5- and 9.0-fold increases in progesterone accumulation on day 3 of incubation. Steroid production was maintained throughout the incubations when cells are incubated in serum-free media treated with cholesterol and ITS premix. Cells collected from higher density of percoll layers produced 2.82 and 2.32 times more progesterone, in comparison to the lover density percoll layer, on days 5 and 15 of the oestrous cycle in untreated cell groups, respectively. Progesterone accumulation was decreased as incubation time advanced in all groups of untreated cells. These results demonstrated that goat luteal cell subpopulations secrete substantial amounts of progesterone in response to cholesterol treatment at least for 7 days, and cholesterol is required as progesterone precursor for maintaining a high-level steroidogenesis during long-life culturing of both cell subpopulations.

Genetic characterization of heat tolerant tomato (Solanum lycopersicon) genotypes by SRAP and RAPD markers.

We employed RAPD and sequence-related amplified polymorphism (SRAP) markers to evaluate polymorphisms in 15 tomato (Solanum lycopersicon) genotypes that were obtained from a tomato breeding program. Four local tomato genotypes selected from the Sanliurfa province (Southeastern Anatolia Region of Turkey), 10 heat-tolerant tomato genotypes, received from the Asian Vegetable Research and Development Center, and a sample of S. pimpinellifolium were genotyped with RAPD and SRAP markers. Eleven SRAP primer combinations were used and 66 bands were scored. The number of bands scored per primer combination ranged from three to 12, with a mean of six alleles per primer combination. All fragments scored for each primer combination were polymorphic. The percentage of polymorphic products ranged from 25 to 80%. The 15 tomato genotypes were screened for RAPD markers using 50 primers in a PCR-based DNA amplification procedure; 46 primers produced clear and good amplification. Ten of these 46 primers amplified monomorphic fragments in the tomato genotypes. A dendrogram was constructed by combining data from the RAPD and SRAP analyses. Similarity ratios of genotypes ranged from 0.87 to 0.99. The dendrogram was divided into two branches; the first main branch included only genotype CL 5915, and the second main branch included all the other genotypes.

Synthesis and molecular modeling of some novel hexahydroindazole derivatives as potent monoamine oxidase inhibitors.

A novel series of 2-thiocarbamoyl-2,3,4,5,6,7-hexahydro-1H-indazole and 2-substituted thiocarbamoyl-3,3a,4,5,6,7-hexahydro-2H-indazoles derivatives were synthesized and investigated for the ability to inhibit the activity of the A and B isoforms of monoamine oxidase (MAO). The target molecules were identified on the basis of satisfactory analytical and spectra data (IR, (1)H NMR, (13)C NMR, (2)D NMR, DEPT, EI-MASS techniques and elemental analysis). Synthesized compounds showed high activity against both the MAO-A (compounds 1d, 1e, 2c, 2d, 2e) and the MAO-B (compounds 1a, 1b, 1c, 2a, 2b) isoforms. In the discussion of the results, the influence of the structure on the biological activity of the prepared compounds was delineated. It was suggested that non-substituted and N-methyl/ethyl bearing compounds (except 2c) increased the inhibitory effect and selectivity toward MAO-B. The rest of the compounds, carrying N-allyl and N-phenyl, appeared to select the MAO-A isoform. The inhibition profile was found to be competitive and reversible for all compounds. A series of experimentally tested (1a-2e) compounds was docked computationally to the active site of the MAO-A and MAO-B isoenzyme. The autodock 4.01 program was employed to perform automated molecular docking. In order to see the detailed interactions of the docked poses of the model inhibitors compounds 1a, 1d, 1e and 2e were chosen because of their ability to reversibly inhibit the MAO-B and MAO-A and the availability of experimental inhibition data. The differences in the intermolecular hydrophobic and H-bonding of ligands to the active site of each MAO isoform were correlated to their biological data. Observation of the docked positions of these ligands revealed interactions with many residues previously reported to have an effect on the inhibition of the enzyme. Excellent to good correlations between the calculated and experimental K(i) values were obtained. In the docking of the MAO-A complex, the trans configuration of compound 1e made various very close interactions with the residues lining the active site cavity these interactions were much better than those of the other compounds tested in this study. This tight binding observation may be responsible for the nanomolar inhibition of form of MAOA. However, it binds slightly weaker (experimental K(i)=1.23 microM) to MAO-B than to MAO-A (experimental K(i)=4.22 nM).

Influence of growth conditions on the nisin production of bioengineered Lactococcus lactis strains.

Nisin production of three bioengineered strains, (LAC338, LAC339 and LAC340) with immunity (nisFEG) and/or regulation (nisRK) genes of nisin biosynthesis on plasmids in the Lactococcus lactis LL27 nisin producer, was evaluated under pH-controlled and pH-uncontrolled batch fermentations. Optimization studies showed that fructose and yeast extract yielded the highest nisin activity. The strains LAC338, LAC339, and LAC340 produced 24, 45, and 44% more nisin, respectively, than wild-type L. lactis LL27 after 12-h incubation. However, sharp decreases in the yield of nisin were observed at the late phase of fermentation with LAC339 and LL27 in contrast to LAC340 and LAC338 strains for which the high level of nisin could be maintained longer. Obviously, increasing the copy number of the regulation genes together with immunity genes in the nisin producers retarded the loss of nisin in the late phase of the fermentation.

Continuous nisin production with bioengineered Lactococcus lactis strains.

Nisin production in continuous cultures of bioengineered Lactococcus lactis strains that incorporate additional immunity and regulation genes was studied. Highest nisin activities were observed at 0.2 h(-1) dilution rate and 12.5 g l(-1) fructose concentration for all strains. Recombinant strains were able to produce greater amounts of nisin at dilution rates below 0.3 h(-1) compared to the control strain. However, this significant difference disappeared at dilution rates of 0.4 and 0.5 h(-1). For the strains LL27, LAC338, LAC339, and LAC340, optimum conditions for nisin production were determined to be at 0.29, 0.26, 0.27, and 0.27 h(-1) dilution rates and 11.95, 12.01, 11.63, and 12.50 g l(-1) fructose concentrations, respectively. The highest nisin productivity, 496 IU ml(-1) h(-1), was achieved with LAC339. The results of this study suggest that low dilution rates stabilize the high specific nisin productivity of the bioengineered strains in continuous fermentation. Moreover, response surface methodology analysis showed that regulation genes yielded high nisin productivity at wide ranges of dilution rates and fructose concentrations.

Evaluation of matrix metalloproteinase, myeloperoxidase, and oxidative damage in mesenteric ischemia-reperfusion injury.

BACKGROUND: In this study, we investigated the alterations of matrix metalloproteinase (MMP) and tissue inhibitors of metalloproteinases (TIMPs), acute inflammation, and oxidative damage in the circulatory system and the intestine in response to mesenteric ischemia/reperfusion (I/R). METHODS: Twenty-one rats were divided randomly into the following three groups (n = 7 in each group): a sham group (CG), an ischemic group (IG), and an I/R group (I/RG). MMP-9, TIMP-1, and myeloperoxidase (MPO) were measured using the enzyme-linked immunosorbent assay method, and lipid peroxidation (quantified as thiobarbituric acid reactive substances (TBARS) content), ischemia-modified albumin, the prooxidant-antioxidant balance (PAB), and ferric-reducing antioxidant power (FRAP) were measured spectrophotometrically. The degree of intestinal injury was evaluated according to the Chiu scoring system. RESULTS: A significant difference between the mean serum TIMP-1 and MMP-9 levels and the alanine transaminase activity was found among the groups. Compared with the I/RG group a significant difference in the mean tissue MMP-9, MPO, and TBARS levels in addition to the PAB and FRAP was found between the CG and IG groups. The level of MMP-9 also demonstrated a strong, positive, and valid correlation with the TBA-RS levels. A significant morphological change was observed in both the IG and the I/RG groups. The degree of intestinal injury was more severe in the I/R group and was characterized by either villous denudation or villous loss. CONCLUSIONS: These results suggest that MMP-9, TIMP-1, MPO, and oxidative stress may be important in the intestinal injury development that is induced by acute mesenteric I/R in a rat model. MMP-9 overexpression may increase the extent of intestinal villous loss, particularly when MMP-9 is upregulated by the TBARS present in the intestinal injury.

Decreased preoperative serum fibulin-3 levels in colon cancer patients.

OBJECTIVE: Fibulin-3 is known to play a role in tumor cell malignancy, invasion and metastasis, as well as in the clinical progression of tumors. This study aimed to assess serum fibulin-3 levels in patients with colon cancer compared with healthy controls and its relationship to demographics and tumor pathology. PATIENTS AND METHODS: A total of 80 patients (mean age, 58.99 years; 42% males) with colon cancer and 50 controls (mean age, 57.75; 55% males) were included. Serum levels of fibulin-3 were determined using a commercially available sandwich ELISA (Enzyme-Linked ImmunoSorbent Assay). RESULTS: Preoperative serum fibulin-3 levels were significantly lower in the group of patients with colon cancer (mean, 35.91 ng/mL; range, 10-73 ng/mL) compared with the control group (mean, 96.68 ng/mL; range, 57-168 ng/mL). CONCLUSIONS: It was concluded that fibulin-3 is expressed at a lower level in colon cancer, and it can serve as a marker for advanced colon cancer.

Fatty acid composition and quality characteristics of low-fat cooked sausages made with beef and chicken meat, tomato juice and sunflower oil.

Low-fat (5.9-10.3% fat) cooked sausages were produced with seven different formulations. Sausages produced with total replacement of fat with sunflower oil had significantly lower oleic acid (C18:1) and higher linoleic (C18:2) and behenic (C22:0) fatty acid contents. Their ratio of TUFA/TS was 3.65 compared to 0.95-1.14 for the other sausages. Also these sausages had the lowest moisture content, highest overall palatability and were less firm. Sausages with tomato juice had the lowest pH value, total aerobic count and nitrite content, but were firmer. Sausages produced with reduced beef contents had lower fat contents, lower stearic (C18:0) and higher oleic (C18:1) fatty acid contents than sausages of high beef content, their texture was very soft and had the lowest score for juiciness. Finally the sausages with chicken meat had the lowest fat and highest salt contents, and lower stearic (C18:0) and higher linoleic (C18:3) fatty acid contents than those made with beef . Also their colour was lighter, less red and more yellow and they had the lowest flavor intensity and overall acceptability.

Motives for risk-taking in adolescence: a cross-cultural study.

Most research on adolescent risk-taking has been conducted in Western societies, but it is as yet unknown whether motives to engage in risk behaviours show cultural variety. This study sets out to investigate differences in perceived motives to engage in perceived risks in Turkish and Welsh samples of young people (N=922) between 14 and 20 years of age. For this, a measurement scale to assess motives for risk-taking was constructed and validated cross-culturally. The scale was based on Kloep and Hendry's [(1999). Challenges, risks and coping in adolescence. In D. Messer, & S. Millar (Eds.), Exploring developmental psychology (pp. 400-416). London: Arnold] theoretical framework and the results of a study by Güney and Cok [(2006). Adolescent risk-taking: Calculated risks, Turkish experience. In Paper presented at the 10th Bi-annual conference of the European Association for Research on Adolescence, Antalya, Turkey]. Results show that different motives are associated with different risk behaviours, confirming Kloep and Hendry's expanded model. There were small, but significant, national differences, implying that motives to take risks-as opposed to actual risks taken-could be similar across adolescent populations, independent of culture.

Factors affecting the outcome of decompressive craniectomy for large hemispheric infarctions: a prospective cohort study.

BACKGROUND: Although surgical decompression of large hemispheric infarction is often a life-saving procedure, many patients remain functionally dependent. The aims of this study were to identify specific factors that can be used to predict functional outcome, thus establish predictive criteria to reduce poor surgical results. METHOD: In this non-randomized prospective study, we performed decompressive craniectomy in 32 patients (age range, 27 to 77 years) with large hemispheric infarctions. Based on their modified Rankin Score (RS), which was calculated 6 months postoperatively, patients were divided into two functional groups: good (RS 0-3, n = 7) and poor (RS 4-6, n = 25). The characteristics of the two groups were compared using statistical analysis. FINDINGS: One-month mortality was 31%. However, most of the surviving patients were severely disabled (RS 4 or 5), and 6-month total mortality reached 50%. Increased age (> or = 60 years) (P = 0.010), preoperative midline shift greater than 10 mm (P = 0.008), low preoperative Glasgow Coma Score (GCS < or = 7) (P = 0.002), presence of preoperative anisocoria (P = 0.032), early (within the first three days of the stroke) clinical deterioration (P = 0.032), and an internal carotid artery infarct (P = 0.069) were the positive predictors of a poor outcome. INTERPRETATION: We view decompressive craniectomy for space-occupying large hemispheric infarction as a life-sparing procedure that sometimes yields good functional outcomes. A dominant hemispheric infarction should not be an exclusion criterion when deciding to perform this operation. Early operation and careful patient selection based on the above-mentioned factors may improve the functional outcome of surgical management for large hemispheric infarction.

Mycoflora of hazelnut (Corylus avellana L.) and aflatoxin content in hazelnut kernels artificially infected with Aspergillus parasiticus.

In this research, 30 hazelnut samples were used to determine their internal mould populations and their aflatoxin contents. The results showed that Aspergillus was the most common genus in the experimental samples (96.6% of all samples). Thirty-one% of isolates were classified as Aspergillus flavus and none of the samples contained aflatoxins. In the second part of the experiment, the kernels were inoculated with conidia of Aspergillus parasiticus and incubated at three different humidified conditions and two different temperatures for 45 days. Aflatoxin contents of the samples kept in 98% relative humidity and at 28 degrees C were higher (904.6 micrograms/kg) than that of the other samples. On the other hand, no aflatoxin was detected in the control samples that were not inoculated but kept at the same conditions.

The effect of environmental pollution on the heavy metal content of raw milk.

This investigation was conducted to determine the contents of some heavy metals in milk samples collected from three different regions, an industrial region, a rural and heavy traffic intensity region around Bursa, a province of Turkey. A total of 75 raw milk samples were collected from these different regions, and the contents of lead, arsenic, copper, zinc, mercury and iron were determined. The average amounts in the samples from these three regions were for Pb 0.032, 0.049, 0.018 mg/kg; for As 0.05, 0.009, 0.0002 mg/kg; for Zu 4.49, 5.01, 3.77 mg/kg; for Cu 0.58, 0.96, 0.39 mg/kg; for Fe 1.78, 4.27, 1.01 mg/kg, while no mercury was detected in the samples. The highest heavy metal content was found in the milk samples collected from industrial region followed by traffic intensive region and rural region.

The effect of aprotinin on extraneural scarring in peripheral nerve surgery: an experimental study.

Extraneural scarring is one of the factors negatively influencing the result of peripheral nerve surgery. Many organic materials have been used to prevent fibrosis. The effect of aprotinin on peripheral nerve scarring in rats was investigated in this study. Three types of surgical intervention were carried out; namely external neurolysis (I), abrasive injury (II), and anastomosis (III). The coded samples which consisted of pure collagen fibers soaked with aprotinin or phosphate-buffered saline were applied around the left sciatic nerves of rats whereas only sham operations were performed on the right sciatic nerves. Animals were sacrificed after 4 or 6 weeks. Neurological examination, gross evaluation of extraneural fibrosis, and histological study were undertaken. The results have demonstrated that aprotinin is a promising agent in the prevention of extraneural scarring.