Prognostic and predictive significance of GITR in metastatic renal cell carcinoma.

OBJECTIVE: Renal cell carcinoma (RCC) has gradually increased in recent years. There have been significant developments in metastatic RCC in recent years with the introduction of immune control point inhibitors. Glucocorticoid-induced tumor necrosis factor (TNF) receptor-related protein (GITR) is a co-stimulatory molecule and is seen in the highest amounts in activated CD4+ T lymphocytes and CD8+ T lymphocytes, forkhead box protein 3 (FOXP3) positive regulatory T cells (Treg). GITR leads to an increase in interleukin (IL)-2 and CD25 and Interferon Gamma. It shows an anti-tumoural effect by inhibiting the suppressive functions of FOXP3+ regulatory cells (Treg). Therefore, we aimed to evaluate the prognostic and predictive effect of GITR, tumor-infiltrating lymphocytes (CD4+CD8) (TIL), and FOXP3 in patients with metastatic RCC. PATIENTS AND METHODS: Patients diagnosed with pathologically confirmed metastatic renal cancer between 2016 and 2021 were included in our study. Clinicopathological features and some laboratory tests were recorded. GITR, CD4, CD8, and FOXP3 were evaluated by immunohistochemistry (IHC) from biopsies or nephrectomy material and recorded. RESULTS: The study included 41 patients. The median progression-free survival (PFS) was 10.5 months, and the median overall survival (OS) was 13.9 months. Median PFS was 7.9 months for the GITR-low group and 18.9 months for the GITR-high group. Median PFS was statistically significant and longer for the GITR-high group than the GITR-low group (p=0.003). When patients who received nivolumab in the 2nd line were evaluated, median PFS was found to be 5.7 months in the GITR-low group and 15.7 months in the GITR-high group. Median PFS was statistically significantly higher in the GITR-high group than in the GITR-low group (p=0.026). CONCLUSIONS: In patients with metastatic RCC, higher GITR was associated with better PFS. At the same time, in patients using nivolumab, better PFS was seen in the GITR high group. If supported by prospective studies, GITR can be used as both a prognostic and predictive marker.

Comparison of cone-beam computed tomography and panoramic radiography for mandibular morphometry.

BACKGROUND: The aim of this study was to compare the morphological differences in the mandible between patients with six age groups and to detect the correlation between these parameters on panoramic radiography (PR) and cone-beam computed tomography (CBCT). MATERIALS AND METHODS: A total of 121 subjects (50 males and 71 females) were included in the study and were divided into six age groups (10-19, 20-29, 30-39, 40-49, 50-59, and 60-69) on the basis of the chronological age. CBCT and PR methods were used to record the mandibular measurements for the same 121 patients. Differences between male and female mandibular morphometric measurements, between right and left side measurements, and differences in age subgroups compared by using independent samples t-test, paired samples t-test, and one-way ANOVA test, respectively. P < 0.05 value was considered statistically significant for all analysis. RESULTS: Males mostly have higher mandibular measurement values. There were statistically significant differences between CBCT and PR measurements (p < 0.05). PR mostly showed higher values than CBCT measurements. CONCLUSIONS: Based on the fact that PRs showing significant differences from CBCT in the morphometric measurements made on mandible, it is recommended that forensic doctors and anthropologists consider this information in their age and gender prediction studies.

Determination of CEBPA mutations by next generation sequencing in pediatric acute leukemia.

OBJECTIVES: The CCAAT/enhancer-binding protein-alpha (CEBPA) is lineage-specific transcription factor in the hematopoietic system. In this study, we aimed on the clinical features and the prognostic significance associated with CEBPA mutations in 30 pediatric patients with acute leukemia. METHODS: In addition, the association between found variants and mutations of Ten-Eleven-Translocation 2 (TET2), Kirsten rat sarcoma viral oncogene homolog (KRAS), and Casitas B-cell lymphoma (CBL), FLT3 (Fms-Related Tyrosine Kinase), JAK2 (Januse Kinase-2) and Nucleophosmin 1 (NPM1) were analyzed, which are important prognostic risk factors for pediatric acute leukemia patients. The entire CEBPA coding region was screened using the NGS method. RESULTS: CEBPA mutations were detected in 16 (53.3 %) of 30 patients. In total, ten distinct of nucleotide changes were identified in 30 patients, including 6 novel and 4 known mutations by sequencing the entire CEBPA gene. We found 6 frame shift mutations, 1 missense mutation, 3 synonymous variants. The most common mutation was the c.487del G resulting p.Glu163Ser in 5 cases. Three patients carried CEBPA double mutations. CONCLUSION: The detected variants in this article seemed to be the first screening results of genes studied by NGS in pediatric acute leukemia patients. Our results also showed some degree of association between FLT3-ITD, TET2, KRAS, CBL and CEBPA mutations (Tab. 4, Fig. 1, Ref. 24).

An osteometric study on humerus / Estudio osteométrico del húmero

Humerus is the longest and thickest bone of the upper limb. As a long bone, it has two epiphysis and diaphysis. In this study, we aimed to conduct morphometric measurements belonging to human humerus. This study was conducted on 60 humerus (28 right, 32 left) in collections of Necmettin Erbakan University Meram Medicine Faculty Anatomy Laboratory. Digital calipers, osteometric board and precision scales for humerus bone measurements were used. Measurements were classified as measurements of diaphysis and proximal and distal epiphysis of humerus. Each bone weight was determined. Also nutrient foramen number and localization was determined. In this study, it was determined that mean right humerus length 30.41±1.73 mm, mean left humerus length 30.04±2.39 mm. It was identified that mean right humerus weight was 115.05±28.06 g, mean left humerus weigh twas 111.63±33.34 g. In 9 humerus (15 %), supratrochlear foramen has been observed. 6 of these were oval and 3 of them were round. Nutrient foramen has not been observed in two humerus (3.3 %). Also, medium and weak correlation was identified between many parameters. We believe that the obtained data from this study may be qualities of reference for sex determination from humerus.

El húmero es el hueso más largo y grueso del miembro superior. Como un hueso largo, tiene dos epífisis y una diáfisis. En este estudio, se pretende realizar mediciones morfométricas pertenecientes al húmero humano. Este estudio se realizó en 60 húmeros (28 derechos, 32 izquierdos) en colecciones del Laboratorio de Anatomía de la Facultad de Medicina Meram,Necmettin Erbakan University, Se utilizaron calibradores digitales, tableros osteométricos y escalas de precisión para medir el húmero. Las mediciones se clasificaron como medidas de húmero proximal, corporal y distal. Se determinó el peso de cada hueso. También se determinó el número y la localización de los forámenes nutricios. La longitud media del húmero derecho fue de 30,41 ± 1,73 mm y la del húmero izquierdo fue de 30,04 ± 2,39 mm. El peso medio del húmero derecho fue 115,05 ± 28,06 g y el izquierdo 111,63 ± 33,34 g. En 9 húmeros (15 %), se observó un foramen supratroclear, seis de ellos eran ovales y tres redondos.No se observó foramen nutricio en dos húmeros (3,3 %). Además, se identificó una correlación media y débil entre varios parámetros. Creemos que los datos obtenidos de este estudio pueden ser de referencia para la determinación del sexo de un individio a partir del húmero.

Lepidoptera (Crambidae, Noctuidae, and Pyralidae) Injury to Corn Containing Single and Pyramided Bt Traits, and Blended or Block Refuge, in the Southern United States.

Fall armyworm, Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae); corn earworm, Helicoverpa zea Boddie (Lepidoptera: Noctuidae); southwestern corn borer, Diatraea grandiosella Dyar (Lepidoptera: Crambidae); sugarcane borer, Diatraea saccharalis F. (Lepidoptera: Crambidae); and lesser cornstalk borer, Elasmopalpus lignosellus Zeller (Lepidoptera: Pyralidae), are lepidopteran pests of corn, Zea mays L., in the southern United States. Blended refuge for transgenic plants expressing the insecticidal protein derivative from Bacillus thuringiensis (Bt) has recently been approved as an alternative resistance management strategy in the northern United States. We conducted a two-year study with 39 experiments across 12 states in the southern United States to evaluate plant injury from these five species of Lepidoptera to corn expressing Cry1F and Cry1Ab, as both single and pyramided traits, a pyramid of Cry1Ab×Vip3Aa20, and a pyramid of Cry1F×Cry1Ab plus non-Bt in a blended refuge. Leaf injury and kernel damage from corn earworm and fall armyworm, and stalking tunneling by southwestern corn borer, were similar in Cry1F×Cry1Ab plants compared with the Cry1F×Cry1Ab plus non-Bt blended refuge averaged across five-plant clusters. When measured on an individual plant basis, leaf injury, kernel damage, stalk tunneling (southwestern corn borer), and dead or injured plants (lesser cornstalk borer) were greater in the blended non-Bt refuge plants compared to Cry1F×Cry1Ab plants in the non-Bt and pyramided Cry1F×Cry1Ab blended refuge treatment. When non-Bt blended refuge plants were compared to a structured refuge of non-Bt plants, no significant difference was detected in leaf injury, kernel damage, or stalk tunneling (southwestern corn borer). Plant stands in the non-Bt and pyramided Cry1F×Cry1Ab blended refuge treatment had more stalk tunneling from sugarcane borer and plant death from lesser cornstalk borer compared to a pyramided Cry1F×Cry1Ab structured refuge treatment. Hybrid plants containing Cry1F×Cry1Ab within the pyramided Cry1F×Cry1Ab blended refuge treatment had significantly less kernel damage than non-Bt structured refuge treatments. Both single and pyramided Bt traits were effective against southwestern corn borer, sugarcane borer, and lesser cornstalk borer.

Complete ossification of the superior transverse scapular ligament in an Turkish male adult / Osificación completa del ligamento escapular superior transverso en un hombre adulto de Turquía

Scapula has three margins and angles and located posterolateral of thorax. One of the anatomical structures of scapula is suprascapular notch located medial to coracoid process base. Suprascapular notch can be seen in different shapes and depths. Suprascapular notch is surrounded by transverse scapular ligament which is a short and strong ligament. Suprascapular notch creates an osteofibrosis passage with this structure within suprascapular nerve passes. It has been reported that this osteofibrosis structure can intirely or partially ossify. All lesions of nerves occured in course as result of exposure to compression, tension and bending are called entrapment neuropathy. Like other peripheral nerves, suprascapular nerve can be exposed to compression while passing suprascapular notch. As a result of this compression suprascapular entrapment neuropathy may occur. There are direct trauma, repetitive microtrauma, neurit, progressive compressive lesions in suprascapular entrapment neuropathies etiology. A suprascapular notch taken foramen shape can be a predisposing factor to entrapment neuropathy. In the retrospective screening from Necmettin Erbakan University, Meram Medical Faculty, Department of Radiology archive, a male patient at age 68 with suprascapular notch variation has been detected. This patient's right suprascapular notch had became foramen by an osseous bridge. Diameters of foramen has been measured as 5.27 mm transverse 6.48 mm vertical. We believe having detailed knowledge of suprascapular notch is significant as a possible course of back and shoulder pain is entrapment of suprascapular nerve in suprascapular notch causing nerve paralyses, and it will give a right direction to clinicians in surgical practices.

La escápula tiene tres márgenes y tres ángulos, y se encuentra en la región posterolateral del tórax. Una de las estructuras anatómicas de la escápula es la incisura supraescapular situada medial a la base del proceso coracoides. La incisura supraescapular puede tener diferentes formas y profundidad. Está rodeada por el ligamento escapular transverso, un ligamento corto y fuerte. Con esta estructura, crea un pasaje osteofibroso por el cual pasa el nervio supraescapular. Se ha reportado que esta estructura osteofibrosa se puede osificar completa o parcialmente. Todas las lesiones de los nervios ocurren en su trayecto como resultado de la exposición a compresión, tensión y flexión, lo que se denomina neuropatía por atrapamiento. El nervio supraescapular puede estar expuesto a compresión al pasar por la incisura supraescapular, generando neuropatía por atrapamiento. Su etiología puede ser por traumatismo directo, microtraumatismo repetitivo y lesiones compresivas progresivas. Una incisura supraescapular de forma oval puede ser un factor predisponente para la neuropatía por atrapamiento. En un estudio retrospectivo realizado en la Universidad Necmettin Erbakan, Facultad de Medicina de Meram, Departamento de archivo de Radiología, se observó la imagen de un varón de 68 años con variación en la incisura supraescapular derecha, la cual se transformó en un foramen por un puente óseo. Los diámetros del foramen fueron 5,27 mm transversales y 6,48 mm verticales. Creemos que el conocimiento detallado de la incisura supraescapular es necesario en el diagnóstico y posible curso de dolor del hombro, debido al atrapamiento del nervio supraescapular en la incisura y la posible parálisis nerviosa, dando una guía a los clínicos en las prácticas quirúrgicas.

Ultrasound as a diagnostic tool to differentiate acute from chronic renal failure.

AIM: Renal ultrasound (US) is the most appropriate method for imaging renal failure; however, considerable overlap in renal size and renal echogenicity exists between normally and abnormally functioning kidneys. We compared the sonographic features of kidneys in patients with renal failure to investigate the potential role of renal US to distinguish acute from chronic renal failure and assessed the diagnostic role of body surface area-corrected renal length compared to measured renal length. MATERIALS AND METHODS: We included 127 consecutive patients with serum creatinine levels higher than 3 mg/dl and 33 healthy volunteers. The subjects with acute renal failure (ARF) and chronic renal failure (CRF) were compared for renal length, parenchymal thickness, parenchymal echogenicity, distinctness of the corticomedullary junction, and the presence of stones and cysts. RESULTS: No significant differences in age, serum albumin, creatinine, weight, height, or gender distribution were found between patients with ARF and those with CRF, except in serum hemoglobin. The right and left kidney parenchymal thickness and renal length were significantly greater in ARF patients than in those with CRF (p < 0.0001). The mean parenchymal thickness and renal length were similar in ARF patients and the control group. Grade I hyperechogenicity was the most common finding during sonography. CONCLUSIONS: Renal length, parenchymal thickness, and echogenicity differed significantly between patients with acute and chronic renal failure. A renal US examination is still the most appropriate method for imaging renal failure and should be combined with other tests to distinguish acute from chronic renal failure.

Tarnished plant bug (Hemiptera: Miridae) thresholds and sampling comparisons for flowering cotton in the midsouthern United States.

The tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Hemiptera: Miridae), has become the primary target of foliar insecticides in cotton, Gossypium hirsutum L., throughout the Midsouth over the past several years. This prompted a reevaluation of existing action thresholds for flowering cotton under current production practices and economics. A trial was conducted at 19 locations throughout the Midsouth during 2006 and 2007. Threshold treatments ranged from a weekly automatic insecticide application to a very high threshold of 10 tarnished plant bugs per 1.5 row-m on a black drop cloth. Individually, all locations reached the lowest threshold, and eight locations had a significant yield loss from tarnished plant bugs. Across all locations, lint yield decreased 0.85 to 1.72% for each threshold increase of one tarnished plant bug per 1.5 row-m. Yield loss was most closely correlated to pest density during the latter half of the flowering period. The relationship between plant bug density or damage and yield was similar for drop cloth, sweep net, and dirty square sampling methods, but the correlations among these sampling methods were not high. Incorporating actual insecticide application data from the trial and average production and economic factors for Midsouth cotton, the economic threshold, if monitoring once per week, should be between 1.6 and 2.6 tarnished plant bugs per 1.5 row-m during the flowering period. More frequent monitoring or situations where insecticide applications are more efficacious may alter this threshold.

Anticardiolipin antibodies in hemodialysis patients with hepatitis C and their role in fistula failure.

BACKGROUND/AIMS: Several conflicting results are presently reported regarding raised IgG and IgM-anticardiolipin antibodies (ACA) titers in hemodialysis (HD) patients and their role in vascular access dysfunction. We aimed to determine the prevalence of both IgM and IgG-ACA titers and to analyze retrospectively their role in primary and secondary arteriovenous fistula (AVF) failure in a homogeneous group of HD patients with chronic hepatitis C. METHODS: This study included 103 adults on maintenance hemodialysis with chronic hepatitis C infection. All participants had blood samples drawn predialysis and after an overnight fast. Analysis included biochemistry, IgG and IgM ACA, Anti-HCV, HBsAg, serum HCV RNA and HCV genotyping. RESULTS: The prevalence of IgG-ACA was 14.6% (15/103). No patient had a positive value of the IgM-ACA test. HCV replication was detected in 52 of 76 patients. The most common HCV genotype was genotype 1 (90%). The percentage of females was higher in ACA(+) group (p = 0.038). There were no significant differences between subjects with and without ACA-IgG regarding other parameters studied. No difference in regard to AVF survival was detected between ACA(+) and ACA(-) groups (p > 0.05). CONCLUSION: We found no significant differences in primary or secondary AVF failure between patients with elevated and normal ACA. Therefore, we conclude that AVFF may be caused by factors other than ACA in these patients. More prospective studies are needed to confirm this observation.

Invasion of human coronary artery endothelial cells by Streptococcus mutans OMZ175.

INTRODUCTION: Dissemination of oral bacteria into the bloodstream has been associated with eating, oral hygiene, and dental procedures; including tooth extraction, endodontic treatment, and periodontal surgery. Recently, studies identified Streptococcus mutans, the primary etiological agent of dental caries, as the most prevalent bacterial species found in clinical samples from patients who underwent heart valve and atheromatous plaque surgery. METHODS: By using antibiotic protection assays, we tested the capacity of 14 strains of S. mutans to invade primary human coronary artery endothelial cells (HCAEC). RESULTS: Serotype e strain B14 and serotype f strain OMZ175 of S. mutans were able to efficiently invade HCAEC. Among the tested strains, serotype f S. mutans OMZ175 was the most invasive, whereas strains of serotype c S. mutans, the most prevalent serotype in dental plaque, were not invasive. Based on its high invasion rate, we further investigated the invasive properties of serotype f OMZ175. Using transmission electron microscopy and antibiotic protection assays we demonstrate that S. mutans OMZ175 is capable of attaching to the HCAEC surface, entering the cells and surviving in HCAEC for at least 29 h. DISCUSSION: Our findings highlight a potential role for S. mutans in the pathogenesis of certain cardiovascular diseases.

Tarnished plant bug (Hemiptera: Miridae) thresholds for cotton before bloom in the Midsouth of the United States.

Insecticide applications to control tarnished plant bug, Lygus lineolaris (Palisot de Beauvois) (Hemiptera: Miridae), during cotton, Gossypium hirsutum L., bud formation are common throughout the Midsouth of the United States. Cultivation practices and the pest complex have changed since action thresholds were established for this pest. A trial was conducted at 33 locations over 3 yr throughout the Midsouth to evaluate tarnished plant bug damage to cotton during the prebloom period. There was no consistent yield response to action thresholds, but average tarnished plant bug density and average square loss were both significant factors impacting lint yield. Based on the yield responses and application frequency of the various action thresholds, the best economic scenario occurred when tarnished plant bug density during the prebloom period averaged eight per 100 sweeps and square retention averaged 90%. The action thresholds required to achieve these averages are expected to be higher than these levels because pest pressure is not normally constant during the prebloom period. When insecticides are required, an application interval shorter than one week may be needed to obtain satisfactory control.

Does the direction of arterial needle in AV fistula cannulation affect dialysis adequacy?

AIM: The direction of arterial access needles in fistulas and grafts has been a subject of some controversy and there is no study comparing the results either direction of arterial needle placement in cannulation of arteriovenous fistula. We compared mean urea reduction rate (URR) and Kt/V in the same HD patients when dialyzed via antegrade or retrograde arterial needle cannulation. MATERIALS AND METHODS: This was a study involving 22 adults on maintenance hemodialysis for more than 6 months. Doppler US examinations of arteriovenous fistula were performed in all subjects. Pre-dialysis and post-dialysis blood samples were obtained at the patient's midweek HD treatment 4 times a month for each direction. Arterial needle was placed in retrograde direction for the first month. On the second month, the direction of arterial needle was converted to antegrade. Means were compared by paired t-test. RESULTS: Mean URR and eKt/Vof retrograde cannulation were 74.2+/-7.2% and 1.57+/-0.33. The results were indifferent statistically from those of antegrade cannulation (73.0+/-8.7% and 1.57+/-0.35 (p=0.123)). Mean fistula blood flow was 931 +/- 483 ml/min. No cannulation complication was observed during the study period for both directions. CONCLUSIONS: Both antegrade and retrograde arterial needle placement may be preferred according to center experience without concern of HD adequacy. Longterm outcomes of antegrade and retrograde arterial needle placement such as AVF failure, thrombosis, and stenosis warrant further studies.

Parathyroid hormone as a marker for the differential diagnosis of acute and chronic renal failure.

It is often difficult to distinguish acute renal failure clinically from chronic renal failure, especially in patients who do not have records of their medical history. We investigated the magnitude of iPTH increase in ARF and the potential role of iPTH as a marker for differential diagnosis of ARF and CRF in new patients referred to our renal unit. We prospectively analyzed 122 (ARF n = 64, CRF n = 58) patients referred to our renal unit with serum creatinine higher than 2 mg/dL. ROC curve analysis was performed to investigate role of iPTH for differentiating ARF from CRF. The sensitivity, specificity, and positive predictive value of iPTH in discrimination of ARF and CRF were calculated. There was no statistically significant difference regarding the means of age, sex distribution, and serum chemistry between patients with ARF or CRF. But serum iPTH (p < 0.0001) levels were lower in patients with ARF than in those with CRF. A cutoff, set at 170 pg/mL for iPTH to discriminate patients with CRF, yielded a sensitivity of 88% and a specificity of 89%. This study confirms that the iPTH measurement is of clinical value in differentiating acute from chronic renal failure.

Chemical and spectroscopic analysis of lignin in isolated flax fibers.

The chemistry of pure flax fibers, free of contaminating nonfiber components, has not been determined. Fibers from the center sections of the stem of seed and fiber flax (Linum usitatissium L.), which had been retted after soaking in water and removal of the epidermis by hand, underwent chemical and spectroscopic analysis. Wet chemical analysis showed only trace indications of aromatics and no long chain fatty acids or alcohols in fibers. Pyrolysis mass spectroscopy (PyMS) and pyrolysis gas chromatography mass spectrometry (PyGCMS) showed only trace amounts of aromatic constituents that could be attributed to the presence of lignin. Mid-infrared (Mid-IR) and Raman spectroscopy of these fibers showed no aromatic compounds present. This study suggests that earlier work reporting the presence of lignin ranging from 1 to 4% may be the result of residual shive or epidermis/cuticle material remaining after the retting process which may be responsible for the favorable properties desired by the composites industry.

Analysis of fiber content in flax stems by near-infrared spectroscopy.

The conventional means of measuring the fiber content of flax is time-consuming and laborious, and the results obtained vary with the analysis technique used. The plant tissues must first be "retted", a process by which the fibers are separated from the rest of the stem, either by indigenous organisms in the soil when the stems are left in the field or by water (anerobic bacteria) or enzymatic retting. The fiber content is then determined by mechanical or manual separation. In this study, fiber content of flax stems was measured rapidly and objectively by near-infrared spectroscopy (NIRS) using whole pieces of stem in a large cell, in reflectance mode. Compared to the conventional method, the standard error of performance of the NIRS method was between 0.96 and 1.45% (dry matter basis), depending on the model and data processing used. NIRS calibrations were generated by hand separation of fiber from water-retted specimens. The water retting procedure takes several days to complete and requires considerable trained labor to complete the hand separation step. The NIRS procedure was conducted on pieces of stem to simulate measurement in the field.

Plant factors influencing enzyme retting of fiber and seed flax.

Retting, which is the microbial activity through which bast fibers are released from nonfiber tissues, is the limiting factor in flax processing. The objective of this work is to identify chemical and structural characteristics in a variety of fiber and seed flax types that influence enzyme retting in a recently developed method. Analyses of flax retted in a series of tests, including two enzyme rettings in some cases, indicated that lignin did not limit the separation of fibers from shive and showed that pectinases in enzyme-retting mixtures could ret fiber and seed flax. However, mature stems, such as that in flax produced for seed, had greater amounts of cutin and wax in the cleaned fiber product, suggesting that the cuticle could be a greater antiquality factor in seed versus fiber flax. With seed flax, the fraction of finer fibers produced during retting was significantly lower than with fiber flax. Results indicated that enzyme retting could be used to obtain flax fibers from seed flax stem residues and add value to this agricultural material.

Enzyme-retting of flax and characterization of processed fibers.

Enzyme-retting formulations consisting of Viscozyme L, a pectinase-rich commercial enzyme product, and ethylenediaminetetraacetic acid (EDTA) were tested on Ariane fiber flax and North Dakota seed flax straw residue. Flax stems that were crimped to disrupt the outer layers were soaked with various proportions of Viscozyme-EDTA solutions, retted, and then cleaned and cottonized with commercial processing equipment. Fiber properties were determined and crude test yarns were made of raw and Shirley cleaned flax fibers and cotton in various blend levels. Cleaned fibers were obtained from both seed and fiber flax types, but with variations due to treatment. Retting formulations produced fibers having different properties, with enzyme levels of 0.3% (v/v as supplied) giving finer but weaker fibers than 0.05% regardless of EDTA level. Experimental yarns of blended flax and cotton fibers varied in mass coefficient of variation, single end strength, and nep imperfections due to sample and formulation. With cost and fiber and yarn quality as criteria, results established a range in the amounts of components comprising retting formulations as a basis for further studies to optimize enzyme-retting formulations for flax. Under conditions examined herein, Viscozyme L at 0.3% (v/v) plus 25 mM EDTA produced the best test yarns and, therefore, established a base for future studies to develop commercial-grade, short staple flax fibers for use in textiles.

Chemical composition of components comprising bast tissue in flax.

Components from the bast region of flax (Linum usitatissium) were removed by hand from seed flax and fiber flax and analyzed by gas chromatography and mass spectrometry. Stems soaked in water were separated by hand into an outer layer, which consisted of epidermis with cuticle and parenchyma cells, and fiber bundles. Each of the two fractions were evaluated for chemical markers that could be used to predict the degree of retting and possibly fiber quality. The outer layer was subsequently treated with a mixed enzyme preparation to remove the carbohydrate portion, thus providing another fraction consisting primarily of cuticle. Four main constituent groups were investigated: dihydroxy fatty acids, long-chain fatty acids and alcohols, sterols, and aromatics. Long-chain fatty acids and alcohols located in the outer layer accounted for 80-92% of the total found in both fractions. Aromatics and sterols in the outer layer accounted for 29--72 and 27--67%, respectively, of the total and do not appear to be a reliable marker for the degree of retting. The best markers for retting were the dihydroxy fatty acids, of which 98--99% were accounted for in the outer layer. The main dihydroxy fatty acids were a mixture of 8,16- and 9,16-dihydroxyhexadecanoic acids and represented 87--89% of the total dihydroxy fatty acids measured. As a constituent of cuticle, this compound may serve as an excellent marker for indicating the degree of retting as well as a possible marker for fiber quality because this compound is almost always exclusively associated with the outer layer and not the fiber.

NTF2 monomer-dimer equilibrium.

Nuclear transport factor 2 (NTF2) mediates nuclear import of RanGDP, a central component of many nuclear trafficking pathways. NTF2 is a homodimer and each chain has independent binding sites for RanGDP and nuclear pore proteins (nucleoporins) that contain FxFG sequence repeats. We show here that the monomer-dimer dissociation constant for NTF2 obtained by sedimentation equilibrium ultracentrifugation is in the micromolar range, indicating that a substantial proportion of cellular NTF2 may be monomeric. To investigate the functional significance of NTF2 dimerization, we engineered a series of point mutations at the dimerization interface and one of these (M118E) remained monomeric below concentrations of 150 microM. CD spectra and X-ray crystallography showed that M118E-NTF2 preserved the wild-type NTF2 fold, although its thermal stability was 20 deg. C lower than that of the wild-type. M118E-NTF2 bound both RanGDP and FxFG nucleoporins less strongly, suggesting that dissociation of the NTF2 dimer could facilitate RanGDP release and thus nucleotide exchange after it had been transported into the nucleus. Moreover, colloidal gold coated with M118E-NTF2 showed reduced binding to Xenopus oocyte nuclear pores. Overall, our results indicate that dimer formation is important for NTF2 function and give insight into the formation of heterodimers by mRNA export factors such as TAP1 and NXT1 that contain NTF2-homology domains.

Regulation of functional nuclear pore size in fibroblasts.

Protein-NLS-coated gold particles up to approximately 250 A in diameter are transported through the nuclear pores in normal, proliferating BALB/c 3T3 cells. This size can increase or decrease, depending on cellular activity. It has been suggested that increases in functional pore size are related to a reduction in the amount of available p53. To further test this hypothesis, we investigated the effects of cycloheximide and pifithrin-alpha, which inhibits p53-dependent transcriptional activation, on nuclear transport. After 3 hours in cycloheximide, there was a significant increase in the size of the gold particles that entered the nucleoplasm. When the incubation period was extended to 6 hours or longer, transport capacity returned to the control level. By using proteasome inhibitors, it was shown that the cycloheximide-dependent increase in functional pore size was due to the inhibition of protein synthesis, consistent with the fact that p53 is a short-lived protein, and requires the activity of at least two different factors. Although cycloheximide increases the functional diameter of the channel available for signal-mediated transport by approximately 60 A, it had no significant effect on either the import rate of small NLS-containing substrates (FITC-BSA-NLS), or passive diffusion of fluorescent-labeled proteins across the envelope. This suggests that changes in transport capacity were not caused by an increase in overall pore diameter but instead are due to a transient increase in pore size that accompanies signal-mediated transport. Pifithrin-alpha also caused an increase in functional pore diameter without altering the import rate of FITC-BSA-NLS, providing further support for the view that p53 can initiate changes in nuclear transport capacity.