Targeting cyclin-dependent kinase 9 in cancer therapy.

Cyclin-dependent kinase (CDK) 9 associates mainly with cyclin T1 and forms the positive transcription elongation factor b (p-TEFb) complex responsible for transcriptional regulation. It has been shown that CDK9 modulates the expression and activity of oncogenes, such as MYC and murine double minute 4 (MDM4), and it also plays an important role in development and/or maintenance of the malignant cell phenotype. Malfunction of CDK9 is frequently observed in numerous cancers. Recent studies have highlighted the function of CDK9 through a variety of mechanisms in cancers, including the formation of new complexes and epigenetic alterations. Due to the importance of CDK9 activation in cancer cells, CDK9 inhibitors have emerged as promising candidates for cancer therapy. Natural product-derived and chemically synthesized CDK9 inhibitors are being examined in preclinical and clinical research. In this review, we summarize the current knowledge on the role of CDK9 in transcriptional regulation, epigenetic regulation, and different cellular factor interactions, focusing on new advances. We show the importance of CDK9 in mediating tumorigenesis and tumor progression. Then, we provide an overview of some CDK9 inhibitors supported by multiple oncologic preclinical and clinical investigations. Finally, we discuss the perspective and challenge of CDK9 modulation in cancer.

Identification and Validation of Novel Metastasis-Related Immune Gene Signature in Breast Cancer.

Background: Distant metastasis remains the leading cause of death among patients with breast cancer (BRCA). The process of cancer metastasis involves multiple mechanisms, including compromised immune system. However, not all genes involved in immune function have been comprehensively identified. Methods: Firstly 1623 BRCA samples, including transcriptome sequencing and clinical information, were acquired from Gene Expression Omnibus (GSE102818, GSE45255, GSE86166) and The Cancer Genome Atlas-BRCA (TCGA-BRCA) dataset. Subsequently, weighted gene co-expression network analysis (WGCNA) was performed using the GSE102818 dataset to identify the most relevant module to the metastasis of BRCA. Besides, ConsensusClusterPlus was applied to divide TCGA-BRCA patients into two subgroups (G1 and G2). In the meantime, the least absolute shrinkage and selection operator (LASSO) regression analysis was used to construct a metastasis-related immune genes (MRIGs)_score to predict the metastasis and progression of cancer. Importantly, the expression of vital genes was validated through reverse transcription quantitative polymerase chain reaction (RT-qPCR) and immunohistochemistry (IHC). Results: The expression pattern of 76 MRIGs screened by WGCNA divided TCGA-BRCA patients into two subgroups (G1 and G2), and the prognosis of G1 group was worse. Also, G1 exhibited a higher mRNA expression level based on stemness index score and Tumor Immune Dysfunction and Exclusion score. In addition, higher MRIGs_score represented the higher probability of progression in BRCA patients. It was worth mentioning that the patients in the G1 group had a high MRIGs_score than those in the G2 group. Importantly, the results of RT-qPCR and IHC demonstrated that fasciculation and elongation protein zeta 1 (FEZ1) and insulin-like growth factor 2 receptor (IGF2R) were risk factors, while interleukin (IL)-1 receptor antagonist (IL1RN) was a protective factor. Conclusion: Our study revealed a prognostic model composed of eight immune related genes that could predict the metastasis and progression of BRCA. Higher score represented higher metastasis probability. Besides, the consistency of key genes in BRCA tissue and bioinformatics analysis results from mRNA and protein levels was verified.

Aboveground Biomass Mapping and Analysis of Spatial Drivers in the Qinghai-Xizang Plateau Permafrost Zone: A Case Study of the Beilu River Basin.

Aboveground biomass (AGB) serves as a crucial measure of ecosystem productivity and carbon storage in alpine grasslands, playing a pivotal role in understanding the dynamics of the carbon cycle and the impacts of climate change on the Qinghai-Xizang Plateau. This study utilized Google Earth Engine to amalgamate Landsat 8 and Sentinel-2 satellite imagery and applied the Random Forest algorithm to estimate the spatial distribution of AGB in the alpine grasslands of the Beiliu River Basin in the Qinghai-Xizang Plateau permafrost zone during the 2022 growing season. Additionally, the geodetector technique was employed to identify the primary drivers of AGB distribution. The results indicated that the random forest model, which incorporated the normalized vegetation index (NDVI), the enhanced vegetation index (EVI), the soil-adjusted vegetation index (SAVI), and the normalized burn ratio index (NBR2), demonstrated robust performance in regards to AGB estimation, achieving an average coefficient of determination (R2) of 0.76 and a root mean square error (RMSE) of 70 g/m2. The average AGB for alpine meadows was determined to be 285 g/m2, while for alpine steppes, it was 204 g/m2, both surpassing the regional averages in the Qinghai-Xizang Plateau. The spatial pattern of AGB was primarily driven by grassland type and soil moisture, with q-values of 0.63 and 0.52, and the active layer thickness (ALT) also played a important role in AGB change, with a q-value of 0.38, demonstrating that the influences of ALT should not be neglected in regards to grassland change.

Tissue anchor use in arthroscopic glenohumeral surgery.

Arthroscopic surgery has become the mainstay of treatment of several common glenohumeral pathologies such as tears of the rotator cuff and labrum. Arthroscopic rotator cuff and labral repair provide outcomes comparable to those achieved with traditional open techniques, with the benefits of smaller incisions and less soft-tissue disruption. Development and improvement of tissue anchors and arthroscopic instrumentation has been integral to the increased popularity of arthroscopic glenohumeral repairs. Current anchors can be categorized by design and material composition. Awareness of the advantages and limitations of these implants may influence anchor selection.

Development of a self-etching primer with higher shelf life and greater dentin bond stability.

To examine the effects from the chemical structure of acidic methacrylate monomers on the primer's shelf life, N-methacryloyl-2aminoethylphosphonic acid (NMEP)- N-methacryloyl glycine (NMGly), 2-methacryloyloxyethyl dihydrogen phosphate (MEP)-NMGly and 10-methacryloyloxydecyl dihydrogen phosphate (MDP)-NMGly primers were designed. Immediately after preparation, these primers were stored at 40 degrees C for 0, 6 and 14 weeks. At the end of each storage period, (13)C NMR observations were performed. Shear bond strengths of resin to dentin, conditioned by non-stored or stored primers, were measured. Alteration rates of these primers were strongly dependent on the chemical structure of acidic methacrylate monomers. The NMEP-NMGly primer exhibited noticeably higher hydrolytic stability and greater bond strength stability than MEP-NMGly and MDP-NMGly primers. These results demonstrated that methacryl amide monomers, NMEP and NMGly, are more advantageous than methacryl ester monomers. To develop self-etching primers with longer shelf life, it is essential to utilize acidic and hydrophilic methacryl amide monomers.

Comparative study of the key enzymes and biochemical substances involved in the energy metabolism of Pacific white shrimp, Litopenaeus vannamei, with different ammonia-N tolerances.

Pacific white shrimp, Litopenaeus vannamei (9.38 ±â€¯0.17 cm, 10.08 ±â€¯0.35 g), with different ammonia-N tolerances were exposed to NH3 (1.61 mg/L) for 192 h, and the levels of key enzymes and biochemical substances involved in energy metabolism were compared to assess the role of the regulation of energy metabolism on the shrimp's adaptation to ammonia-N stress. Higher ammonia-N tolerance in the shrimp (Tolerance group) was achieved through nutritional fortification, whereas shrimp that were not nutritionally fortified comprised the Control group. The mortality rates in the Control and Tolerance groups at the end of the period of ammonia-N stress exposure were 64.44% and 40.00%, respectively. Within 1 h of exposure to ammonia-N stress, the glucose concentration in both groups declined rapidly, and no significant difference was detected between the two groups. In general, the triglyceride and cholesterol concentrations in the Control group were higher than those in the Tolerance group, and accumulations and/or fluctuations in these metabolites to varying degrees were observed. The Tolerance group presented higher phosphofructokinase (PFK) and pyruvate kinase (PK) activity compared with the Control group from 1 to 48 h of exposure to ammonia-N stress, whereas the opposite result was observed from 96 to 192 h. Similarly, during exposure to ammonia-N stress, the Tolerance group showed higher and lower lactate dehydrogenase (LDH) activity than the Control group from 1 to 24 h and from 48 to 92 h, respectively. In addition, compared with the Control group, the shrimp in the Tolerance group exhibited higher succinate dehydrogenase (SDH) activity, especially from 48 to 192 h of exposure to ammonia-N stress. The results of this study suggest that anaerobic carbohydrate (in the early stage) and aerobic metabolism (in the late stage) plays an important role in the shrimp's response to ammonia-N stress. In addition, maintenance of the normal operation of lipid metabolism is equally important for improving the tolerance of L. vannamei to ammonia-N stress.

Effect of solvent type on the degradation of 4-MET.

This study sought to investigate the degradation mechanism of 4-methacryloyloxy ethyl trimellitic acid, 4-MET, which is commonly used as an acidic monomer in solvated self-etching primers or one-step bonding agents. To this end, we examined the effects of solvent type used--such as ethanol, methanol, and acetone--on the degradation mechanism of 4-MET by using the 13C NMR technique. The degradation mechanism of 4-MET was strongly dependent on the type of solvent used. When an alcohol-based solution was used for 4-MET, the esterification of 1- or 2-carboxylic acid in 4-MET occurred. However, when an acetone solvent was used for 4-MET, the esterification reaction did not occur. Increases in the aging period of 4-MET solvated solutions resulted in the hydrolysis of the benzoyl ester portion in 4-MET. The 2-hydroxyethyl methacrylate, produced as a subproduct, also became hydrolyzed. In addition, methacrylic acid, non-esterified and esterified trimellitic acid, as well as ethylene glycol were produced as subproducts. In particular, the production of trimellitic acid and ethylene glycol affected the bonding efficacy and durability of the resin to the tooth created by self-etching primers or one-step bonding agents that contained the altered 4-MET.

Liver targeting of catalase by cationization for prevention of acute liver failure in mice.

To achieve hepatic delivery of CAT for the prevention of CCl4-induced acute liver failure in mice, two types of cationized CAT derivatives, HMD- and ED-conjugated CAT, were developed. Slight structural changes occurred during cationization and the number of increased free amino groups was 3.1 in HMD-CAT and 13.6 in ED-CAT. 111In-cationized CAT derivatives showed an increased binding to HepG2 cells, and were rapidly taken up by the liver. H2O2-induced cytotoxicity in HepG2 cells was significantly prevented by preincubation of the cells with cationized CAT derivatives. A bolus intravenous injection of the cationized CAT derivatives reduced the hepatotoxicity induced by CCl4 in mice. The ED-CAT, which showed more rapid and greater binding to the liver than the HMD-CAT, exhibited more beneficial effects as far as all the parameters examined (serum GOT, GPT, LDH and hepatic GSH) were concerned, suggesting that a high degree of cationization is effective in delivering CAT to the liver to prevent CCl4-induced hepatotoxicity. These results suggest that cationized CAT derivatives are effective in preventing acute liver failure, and ED-based cationization is a suitable method for developing liver-targetable cationized CAT derivatives, because it provides CAT with a high degree of cationization and a high remaining enzymatic activity.

Primary total knee arthroplasty with condylar allograft and MCL reconstruction for a comminuted medial condyle fracture in an arthritic knee--a case report.

Fractures of the distal femur in the elderly are usually due to low energy ground level fall onto a flexed knee. Pre-existing osteoarthritis and juxta-articular osteopenia in this age group result in high levels of comminution and articular damage at the time of injury, which challenges the management and treatment outcome. Preservation of knee function and early weight bearing should be the objectives of management in the geriatric population. We present in this case report of an elderly patient with comminuted medial condyle fracture with arthritic changes who had primary total knee arthroplasty utilizing condylar allograft and MCL reconstruction as an alternative to internal fixation.

[Phase I study of docetaxel combined with carboplatin in patients with non-small cell lung cancer].

OBJECTIVE: To determine the maximum tolerated dose (MTU) and dose-limiting toxicity (DLT) of Docetaxel and Carboplatin in patients with non-small cell lung cancer. METHODS: Docetaxel was given at escalating doses until MTD was determined from the initial dose of 65 mg/m2 to 75 mg/m2, 85 mg/ m2 on dl. Carboplatin was targeted to an area under the plasma concentration curve of 5 using Calver's equation on dl. The treatment cycle was repeated every 3 weeks. RESULTS: 16 patients received TXT and CBP for total of 54 courses (median four courses). Neutropenia was the dose-limiting toxicity. The MTD of TXT is 85 mg/m2. CONCLUSION: We recommend TXT 75 mg/m2 on d1 and CBP with a target AUC of 5 on d1, 3weeks repeated for chemotherapy in naïve patients with NSCLC.

Low-photon-number optical switch and AND/OR logic gates based on quantum dot-bimodal cavity coupling system.

We propose a new scheme based on quantum dot-bimodal cavity coupling system to realize all-optical switch and logic gates in low-photon-number regime. Suppression of mode transmission due to the destructive interference effect is theoretically demonstrated by driving the cavity with two orthogonally polarized pulsed lasers at certain pulse delay. The transmitted mode can be selected by designing laser pulse sequence. The optical switch with high on-off ratio emerges when considering one driving laser as the control. Moreover, the AND/OR logic gates based on photon polarization are achieved by cascading the coupling system. Both proposed optical switch and logic gates work well in ultra-low energy magnitude. Our work may enable various applications of all-optical computing and quantum information processing.

Cationic charge-dependent hepatic delivery of amidated serum albumin.

To obtain a quantitative correlation between the physicochemical properties of amidated bovine serum albumin (BSA) and their tissue distribution characteristics for the development of targeted delivery of proteins, BSA was amidated with hexamethylenediamine (HMD) or ethylenediamine (ED) to obtain cationized BSAs. Their structural changes were examined by spectroscopic and electrophoretic techniques then their tissue distribution was studied in mice. Circular dichroism (CD) and fluorescence measurements showed that spectroscopic changes occurred as the number of free NH2 groups increased. Capillary electrophoresis revealed a linear relationship between the mobility and the increased number of free NH2 groups. 111In-cationized BSAs were rapidly taken up by liver, but HMD-BSA showed a faster uptake than ED-BSA with a similar number of free NH2 groups, suggesting that the diamine reagent with a longer carboxyl side chain results in more efficient hepatic targeting. The hepatic uptake clearance (CL(liver)) of both derivatives increased significantly with a decrease in electrophoretic mobility (mu(ep)) towards the anode and reached a plateau at low electrophoretic mobility. The electrophoretic mobility is an appropriate indicator of the degree of amidation, which was closely correlated with the hepatic uptake clearance. The correlation between the mobility and the clearance shows that a low degree of amidation is sufficient for efficient hepatic targeting of proteins.

Physicochemical, tissue distribution, and vasodilation characteristics of nitrosated serum albumin: delivery of nitric oxide in vivo.

Conjugates of nitric oxide (NO) to serum albumins are candidates for controlled delivery of NO in vivo, but their physicochemical and tissue distribution characteristics have hardly been examined yet. In this study, to achieve its in vivo delivery, bovine serum albumin (BSA) was reacted with sodium nitrite to obtain NO-BSA, which had 0.25-0.28 molecules of S-nitrosothiol/BSA. In addition to cystein, other amino acid residues were modified by the reaction. The conjugation had no significant effect on the molecular weight, but reduced the electric charge and induced reversible changes in the secondary structure of BSA. After intravenous injection in mice at a dose of 1 mg/kg, 111In-NO-BSA slowly disappeared from plasma in a similar manner to 111In-BSA, but showed greater accumulation in the liver and kidney. NO-BSA induced a transient decrease in arterial pressure after intravenous injection in rats at a dose of 100 mg/kg, and significantly increased the distribution of 111In-BSA to the lung in mice. These results indicate that NO is released from NO-BSA shortly after injection, and this NO decreases blood pressure and increases the distribution of macromolecules to the lung. These findings provide useful basic information for designing macromolecular NO donors able to achieve controlled delivery of NO.

Transition to synchronization in a Kuramoto model with the first- and second-order interaction terms.

We investigate a Kuramoto model incorporated with the first-order and the second-order interaction terms. We show that the model displays the coexistence of multiattractors and different attractors may be characterized by the phase distributions of oscillators. By investigating the transition diagrams in both forward continuation and backward continuation, we find that the synchronous state with unimodal phase distribution is the most stable one while the state in cluster synchrony with evenly distributed bimodal phase distribution is the least stable one. We also present the phase diagram of the model in the parameter space.

Bone replacement of fast-absorbing biocomposite anchors in arthroscopic shoulder labral repairs.

BACKGROUND: Newer generation biocomposite anchors are hypothesized to resorb more reliably and faster, while allowing for bone ingrowth and replacement. PURPOSE: The purposes of this study were to (1) assess anchor resorption and bone ingrowth over time, (2) identify tunnel widening or potential reactions to the implants, (3) compare imaging findings for different sites of labral repair, and (4) determine patient subjective outcomes with the use of biocomposite anchors in glenoid labral repair. STUDY DESIGN: Case series; Level of evidence, 4. METHODS: We enrolled 22 patients to participate in a 24-month outcomes study that included subjective and objective outcome assessments after glenoid labrum repair surgery. Magnetic resonance imaging (MRI) was performed at 6 and 12 months to identify any potential reactions to implants. Computed tomography (CT) scans were performed at 12 and 24 months to determine anchor resorption and bone ingrowth. Sixteen patients and 47 anchors were available for follow-up at 24 months. An independent, fellowship-trained musculoskeletal radiologist read the scans. Subjective outcome scores measured at 24 months postoperatively included Simple Shoulder Test, Tegner activity scale, American Shoulder and Elbow Surgeons (ASES), and University of California, Los Angeles (UCLA) shoulder scores. RESULTS: No adverse events were reported with the use of biocomposite anchors at the end of the study period. At 12 and 24 months, respectively, CT scans demonstrated that an estimated 68% and 98% of combined anchor material had been absorbed, 56% and 78% of the anchor material had been replaced by soft tissue of variable density, and 9% and 20% of total anchor volume was replaced by bone. No obvious mechanical failure of the labral repairs was detected on nonarthrogram MRI. Three of the 47 anchors showed bone cyst formation. Tunnel widening (expansion beyond tunnel diameter of 3 mm; 2.9-mm drill hole utilized) was seen in 55% of the anchors but decreased between 12 and 24 months, consistent with bone replacement. Tunnel widening was seen more in anteroinferior and posterior glenoid anchor locations (84% and 57%, respectively) than in superior labral anchors (13%). Subjective outcome scores at 24 months for ASES and UCLA shoulder scores averaged 88 and 30, respectively. All but one patient were satisfied with their outcome at 24 months. CONCLUSION: Our imaging evaluation indicates resorption of newer generation biocomposite anchors with progressive bone replacement at 12 and 24 months while maintaining acceptable subjective outcomes.

The reaction of bone to tumor growth from human breast cancer cells in a rat spine single metastasis model.

STUDY DESIGN: In vivo experiments to develop a rat spine single metastasis model by using human breast cancer cells. OBJECTIVE: To study the survival and tumorigenesis of the human breast cancer cells after transplantation to vertebral body (VB) by intraosseous injection as a model for therapeutic studies of spine metastatic tumor. SUMMARY OF BACKGROUND DATA: VBs are the most common bones involved in the metastases of breast cancer. To develop experimental therapeutics requires an appropriate animal model. Moreover, it is also important to establish accurate and sensitive detection methods for the evaluation. METHODS: MDA-MB-231 human breast cancer cells were injected into 3-week-old female athymic rats. The tumorigenesis was assayed with quantitative in vivo bioluminescence (IVIS), microcomputed tomography (micro-CT), quantitative CT (qCT), micro position emission tomography (micro-PET), and histologic studies. RESULTS: A spine single metastasis model of human breast cancer was successfully developed in rats. The IVIS signal intensity from the cancer cells increased after 2 weeks. Signal from the tumor in spine can be detected by micro-PET at day 1. The signal intensity decreased after 1 week and then recovered and continually increased afterwards. Bone destruction was demonstrated in the qCT and micro-CT images. However, both qCT and micro-CT found that the bone density in the cancer cell-injected VB increased before the appearance of osteolysis. The growth of tumor and the reaction of bone in the VB were observed simultaneously by histology. CONCLUSION: A spine single metastasis model was developed by injection of human breast cancer cells into the VB of athymic rats. This is the first report of quantitative evaluation with micro-PET in a spine metastasis model. In addition, the detection of osteogenesis after the introduction of MDA-MB-231 cells in vivo is a novel observation.

Therapeutic effects of adenovirus-mediated growth and differentiation factor-5 in a mice disc degeneration model induced by annulus needle puncture.

BACKGROUND CONTEXT: The therapeutic strategies that have thus far been used for the treatment of intervertebral disc degeneration (IDD) have focused on relieving the symptoms, although reversal of the degeneration remains an important challenge for the effective treatment of IDD. Growth and differentiation factor-5 (GDF5), of which deficiency leads to early disc degeneration changes, has the potential to increase proliferation of disc cells and expression of extracellular matrix proteins. PURPOSE: The purpose of the study was to develop a lumbar disc degeneration model in mice and determine the effect of adenoviral GDF5 gene therapy. STUDY DESIGN: The study design was to compare the degeneration changes of discs punctured by different-size needles to develop a mice lumbar disc degeneration model and to evaluate the effects of in vivo gene therapy for the mice disc degeneration model by an adenoviral vector carrying GDF5 gene. METHODS: A lumbar disc degeneration model was developed by needle punctures to the discs in Balb/c mice. Afterward, a gene therapy treatment to disc degeneration was evaluated. Two of the mice lumbar discs were randomly chosen to be punctured by a 30-gauge needle and then injected with adenovirus that had been engineered to express either the luciferase gene (Ad-Luc) or the GDF5 gene (Ad-GDF5). Animals were analyzed by bioluminescent imaging, radiographic, and magnetic resonance imaging (MRI) scanning, then sacrificed at 1, 2, 4, or 8 weeks after operation, and subjected to histological and biochemical assays. RESULTS: By the detection of T2-weighted MRI scanning and histological study, the degeneration was found in all of the discs punctured by different-size needles. But the development of the degeneration in the discs injured by the 30-gauge needle was more reliable and moderate compared with that in other groups. The detection of luciferase activity by bioluminescent imaging revealed that adenovirus survived and the introduced genes were expressed over 6 weeks after injection. There were no T2-weighted MRI signals in the mice injected with either Ad-Luc or Ad-GDF5 up to 4 weeks after operation. At 6 and 8 weeks, T2-weighted signals were detected in the Ad-GDF5 group but none in the Ad-Luc control group. The percent disc height index (%DHI) was significantly decreased (approximately 20%) by 1 week after injury in both groups, indicating the development of disc degeneration. At 2 weeks, the %DHI in the mice injected with Ad-GDF5 increased significantly compared with that of the mice injected with Ad-Luc; the increase was sustained for the rest of the experiment period. The disc histology treated with Ad-GDF5 was improved compared with that in the control group. Glycosaminoglycan (GAG) levels were significantly decreased in the Ad-Luc injection group since 2 weeks after injury, and the DNA content had diminished by 4 weeks after the operation. In contrast, in the discs injected with Ad-GDF5, there was no decrease in the GAG and DNA levels after injury throughout the 8-week treatment period. CONCLUSIONS: Disc degeneration animal model can be developed by using needle puncture to the discs in mice. The adenovirus is an effective vehicle for gene delivery with rapid and prolonged expression of target protein and resulting improvement in markers of disc degeneration. Ad-GDF5 gene therapy could restore the functions of injured discs and has the potential to be an effective treatment.

Single nucleotide polymorphisms within the estrogen receptor beta gene are linked with reproductive indices in Japanese flounder, Paralichthys olivaceus.

The objectives of this study were to characterize polymorphisms within the coding region of estrogen receptor beta (ERbeta) gene in a population of 57 female Japanese flounder (Paralichthys olivaceus) and to analyze the association of ERbeta polymorphisms with reproductive indices by polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP). Two single nucleotide polymorphisms (SNPs), SNP1 (c.577delC) and SNP2 [c.A891T (p.Gln114Leu)], were identified in the ERbeta gene. A one-way ANOVA revealed that SNP1 was significantly associated with the gonadosomatic index (GSI) in female Japanese flounder (P < 0.05). And SNP2 was significantly associated with the serum 17beta-estradiol (E2) level and GSI (P < 0.05). Individuals with genotype AB of SNP2 had significantly higher serum E2 level and GSI than those of genotype AA (P < 0.05). Moreover, the hepatosomatic index (HSI), a marker for genetic effects, was significantly higher for diplotype D2 compared with the other three diplotypes (P < 0.05). These results obtained in this study suggested that SNP2 could influence reproductive endocrinology of female Japanese flounder and be useful as a potential candidate genetic marker for the selection of reproductive indices in female Japanese flounder.

Association of reproductive performance with SNPs of FOXL2 gene by SSCP in Japanese flounder (Paralichthys olivaceus).

FOXL2 which is a putative winged helix/forkhead transcription factor gene and a sexually dimorphic marker of ovarian differentiation plays an important role in ovarian development, granulosa cell differentiation, and thus the proper maintenance of ovarian function. The aims of this study were to characterize polymorphisms within the FOXL2 gene in a population of 52 female Japanese flounder and analyze the association of FOXL2 polymorphisms with reproductive performance by polymerase chain reaction-single stranded conformational polymorphism (PCR-SSCP). Results indicated that five single nucleotide polymorphisms (SNPs), which were SNP1 [c.540A>C (p.Asn102His) and c.591A>G (p.Asn119Asp)], SNP2 [c.864G>A (p.Lys210Glu)and c.875G>A] and SNP3 (c.1169C>A), were identified in the FOXL2 gene. General Linear Model (GLM) analysis showed that SNP1 in the forkhead domain was significantly associated with gonadosomatic index (GSI) (P<0.05). SNP2 in the downstream of forkhead domain was significantly associated with serum 17beta-estradiol (E(2)) level (P<0.05). And SNP3 in the 3'-UTR was significantly associated with hepatosomatic index (HSI) (P<0.05). Moreover, the evaluation of the genetic effects for both Testosterone (T) level of diplotype D3 and GSI of diplotype D5 suggested they were significantly higher than those of other four diplotypes (P<0.05), respectively. These results implied that these SNPs could influence reproductive endocrinology of female Japanese flounder and be also used in marker-assisted selection (MAS) program to reproductive performance in female Japanese flounder in the future.

Role of tyrosine and tryptophan in chemically modified serum albumin on its tissue distribution.

To investigate the effect of functional groups in bovine serum albumin (BSA) on its tissue distribution characteristics, tyrosine (Tyr) or tryptophan (Trp) residues of BSA were chemically modified by tetranitromethane (TNM) and 2-hydroxy-5-nitrobenzyl bromide (HNB), respectively. BSA was successfully modified with each reagent depending on the amount of the reagent added to the reaction mixture, and TNM- and HNB-modified BSA derivatives with different degrees of modification were obtained. Circular dichroism measurements showed that slight secondary and large tertiary changes were detectable as the degree of modification increased. After intravenous injection into mice, all synthetic BSA derivatives were eliminated very slowly from the systemic circulation. However, (111)In-TNM(6.6)- and (111)In-HNB(2.0)-BSA, derivatives with a high degree of modification, showed a slightly faster disappearance from the systemic circulation and slightly higher accumulation in the liver than (111)In-unmodified BSA. Pharmacokinetic analyses also demonstrated that the modification of Tyr or Trp residues on BSA had only marginal effects on tissue distribution. These results indicate that the Tyr and Trp residues have little effect on the tissue distribution characteristics of serum albumins, and that the specific modification of these residues may be a promising approach to designing sustained drug delivery systems using serum albumins.