Evaluation of non-additive genetic effects on carcass and meat quality traits in Korean Hanwoo cattle using genomic models.

The traditional genetic evaluation methods generally consider additive genetic effects only and often ignore non-additive (dominance and epistasis) effects that may have contributed to genetic variation of complex traits of livestock species. The available dense single nucleotide polymorphisms (SNPs) panels offer to investigate the potential benefits of including non-additive genetic effects in the genomic evaluation models. Data from 16 971 genotyped (Illumina Bovine 50 K SNP chip) Korean Hanwoo cattle were used to estimate genetic variance components and prediction accuracy of genomic breeding values (GEBVs) for four carcass and meat quality traits: carcass weight (CWT), eye muscle area (EMA), back fat thickness (BFT) and marbling score (MS). Five different genetic models were evaluated through including additive, dominance and epistatic interactions (additive by additive, A × A; additive by dominance, A × D and dominance by dominance, D × D) successively in the models. The estimates of additive genetic variances and narrow sense heritabilities (ha2) were found similar across the evaluated models and traits except when additive interaction (A × A) was included. The dominance variance estimates relative to phenotypic variance ranged from 1.7-3.4% for CWT and MS traits, whereas, they were close to zero for EMA and BFT traits. The magnitude of A × A epistatic heritability (haa2) ranged between 14.8 and 27.7% in all traits. However, heritability estimates for A × D and D × D epistatic interactions (had2 and hdd2) were quite low compared to haa2 and were contributed only 0.0-9.7% of the total phenotypic variation. In general, broad sense heritability (hG2) estimates were almost twice (ranging between 0.54 and 0.68) the ha2 for all of the investigated traits. The inclusion of dominance effects did not improve the prediction accuracy of GEBV but improved 2.0-3.0% when epistatic effects were included in the model. More importantly, rank correlation revealed that partitioning of variance components considering dominance and epistatic effects in the model would enable to re-rank of top animals with better prediction of GEBV. The present result suggests that dominance and epistatic effects could be included in the genomic evaluation model for better estimates of variance components and more accurate prediction of GEBV for carcass and meat quality traits in Korean Hanwoo cattle.

Nosocomial outbreak of Achromobacter spp. bacteremia due to germicide contamination: a systematic review.

OBJECTIVE: Achromobacter spp. are aerobic, non-fermentative Gram-negative bacilli that can be widely found in aquatic environments. Nosocomial outbreaks and pseudo-outbreaks of Achromobacter spp. bacteremia have been recognized for decades. Notably, commonly used germicides in hospital settings constitute important sources for these outbreaks. This review aims at summarizing the latest studies and presents the characteristics of nosocomial outbreaks of Achromobacter spp. bacteremia caused by germicide contamination. MATERIALS AND METHODS: A systematic search of the PubMed and EMBASE databases was conducted for articles published in English between January 1, 2000, and June 10, 2022. RESULTS: Overall, 170 articles were retrieved, and 7 studies were finally included in the systematic review. Whether true or pseudo-bacteremia, positive blood culture results were most commonly reported in immunosuppressed patients or those with indwelling catheters. The most commonly reported contaminated germicide was chlorhexidine solution used as both an antiseptic and disinfectant. Atomizers, dispensers, and various product containers were identified as reservoirs. The prognoses of the affected patients were generally favorable. CONCLUSIONS: Awareness about the high survival ability of Achromobacter spp. in germicides and the possible hospital reservoirs of these microbes will help to improve infection control and prevent nosocomial outbreaks or pseudo-outbreaks caused by Achromobacter spp.

Bevacizumab for recurrent glioblastoma: a systematic review and meta-analysis.

OBJECTIVE: The phenomenon is that few randomized control trials (RCTs) directly compared the effects of bevacizumab with other types of standard treatments for recurrent glioblastoma (GBM). We conducted a systematic review and meta-analysis to assess the efficacy of bevacizumab in recurrent GBM patients. MATERIALS AND METHODS: We searched electronic databases (Medline, Embase, and Web of Science) contrasting the bevacizumab with standard treatments up to May 2021. For the continuous outcomes of median progression-free survival (PFS) and median overall survival (OS), we summarized the mean difference (MD) as the effective index. We used relative risk (RR) to estimate the data with a random-effects model to get the outcomes of objective response rate (ORR), 12-month OS, 6-month PFS, and any mentioned adverse events. RESULTS: A total of 807 patients in 5 RCTs included into our systematic review and meta-analysis. The results showed bevacizumab could provide benefits of the ORR (RR, 2.67; 95% CI: 1.14-6.26, p = 0.02), median PFS (MD, 1.12 months; 95% CI: 0.35-1.90 months, p = 0.005), but not the median OS (MD, -0.19 months; 95% CI: -1.37-0.99 months, p = 0.75). Whereas the rates of the secondary outcomes of interest were similar between the bevacizumab group and control group, including 6 month-PFS (RR, 1.23; 95% CI, 0.82-1.84, p = 0.32) and 12 month-OS (RR, 0.93; 95% CI, 0.79-1.09, p = 0.36). As for adverse events, patients with bevacizumab showed higher rates of grade 3/4 and any grade hypertension compared with those with standard treatments (RR, 3.71; 95% CI: 1.17-11.76, p = 0.03; RR, 2.68; 95% CI: 1.26-5.76, p = 0.01, respectively). CONCLUSIONS: This study provides clear proof of the beneficial effects of bevacizumab treatment in recurrent GBM patients. The only observed adverse event was grade 3/4 or any grade hypertension.

[Pathogen distribution and antimicrobial resistance among lower respiratory tract infections in patients with hematological malignancies].

Objective: To investigate the pathogen distribution and antimicrobial resistance among lower respiratory tract infections in patients with hematological malignancies. Methods: Sputum samples were collected from 967 patients with hematological malignancies and lower respiratory tract infections in Department of Hematology,the Second Hospital of Shanxi Medical University from January 2017 to July 2020. The pathogens and drug sensitivity reports were carried out by automatic bacterial identification instruments. WHONET 5.6 and SPSS 20.0 softwares were used for statistical analysis. Results: A total of 961 strains of pathogens were isolated, 516 (53.7%) pathogens were Gram-negative bacteria, mainly 118 strains of Klebsiella pneumonia (12.3%), 68 strains of Pseudomonas aeruginosa (7.1%), 67 strains of Acinetobacter baumannii (7.0%),52 strains of Stenotrophomonas maltophilia (5.4%), 43 strains of Escherichia coli (4.5%), and 42 strains of Enterbacter cloacae (4.4%). There were 171 (17.8%) strains of Gram-positive bacteria and 274 (28.5%) fungi. The drug resistance rates of Pseudomonas aeruginosa and Acinetobacter baumannii to carbapenem were 22.1%-31.3%. Stenotrophomonas maltophilia was sensitive to levofloxacin, compound sulfamethoxazole and minocycline. The antimicrobial resistance rates of these three enterobacteria to carbapenems, cefoperazone/sulbactam, piperacillin/tazobactam were low (<10%). The resistant Gram-positive bacteria to ticoplanin, vancomycin and linazolamide were not detected. Conclusion: The major pathogens related to lower respiratory tract infections in patients with hematological malignancies are gram-negative bacteria in our centre. Different pathogens appear different characteristics of antimicrobial resistance.

Low-dose CT angiography of the lower extremities: a comparison study of image quality and radiation dose.

AIM: To investigate the image quality and radiation dose of ultralow-dose (ULD) and low-dose (LD) lower-extremity computed tomography (CT) angiography (LE-CTA) using the advanced modelled iterative reconstruction (ADMIRE) algorithm to detect peripheral arterial disease (PAD) in comparison with standard-dose (SD) CT. MATERIALS AND METHODS: One hundred and seven consecutive patients were examined using LE-CTA at 70 kVp and a dual-source scanner to achieve three image sets using 30% (ULD), 70% (LD), and 100% (SD) tube loads. Qualitative analysis was conducted by examining the three image sets for overall quality. The image quality of arterial segments was analysed by two independent readers. In addition, the CT dose index (CTDIvol) was measured in the three image sets. RESULTS: The mean overall quality scores were 3.4±0.6 for ULD CT, 3.9±0.3 for LD CT, and 3.9±0.2 for SD CT. Both readers scored the arterial segments as 2-4 (adequate-excellent) in the three image sets. In addition, 89.4% (93/104) and 54.8% (57/104) segments of PAD with calcified plaques were scored 4 between SD and LD CT and between SD and ULD CT, respectively, and 45.2% (47/104) segments had a lower score by one point in ULD CT compared with SD CT. The mean CTDIvol was 4.1±1.1 mGy for SD CT, 2.9±0.8 mGy for LD CT, and 1.2±0.3 mGy for ULD CT. CONCLUSIONS: LD/ULD CT at 70 kVp using ADMIRE reconstruction enables a reduction in the radiation dose while enabling adequate evaluation or follow-up of PAD based on LE-CTA.

[Prevalence and risk factors of Blastocystis infections among primary school students in Jiangjin District, Chongqing City].

OBJECTIVE: To investigate the prevalence and risk factors of Blastocystis infections among primary school students in Jiangjin District, Chongqing City. METHODS: A cross-sectional questionnaire survey was conducted among students sampled from a primary school in Jiangjin District, Chongqing City on April, 2018, and their stool samples were collected for microscopic examinations, in vitro culture and PCR assays to analyze the prevalence of Blastocystis infections and subtype of the parasite. In addition, the risk factors of Blastocystis infections among primary school students were identified using univariate analysis and multivariate logistic regression analysis. RESULTS: A total of 466 primary students were surveyed, and the subjects had a mean age of (9.81±1.66) years and included 236 males (50.64%) and 230 females (49.36%). The prevalence of Blastocystis infections was 15.24% (71/466) among the study students, and there was no significance difference in the prevalence between male and fe- male students (16.52% vs. 13.91%; χ2 = 0.616, P = 0.433). In addition, there was a significant difference in the prevalence of Blastocystis infections among grade 1 (6.35%, 4/63), grade 2 (5.17%, 3/58), grade 3 (21.74%, 15/69), grade 4 (25.30%, 21/83), grade 5 (10.19%, 11/108) and grade 6 students (20.00%, 17/85) (χ2 = 15.410, P = 0.009). There were four Blastocystis subtypes characterized (ST1, ST3, ST6 and ST7), in which ST6 was the most common subtype (45.07%, 32/71), followed by ST3 (25.35%, 18/71). Multivariate logistic regression analysis revealed that minority ethnicity [odds ratio (OR) = 4.259, 95% confidential inter- val (CI) : (1.161, 15.621)] and low maternal education level (primary school and below) [OR = 9.038, 95% CI: (1.125, 72.642)] were identified as risk factors of Blastocystis infection among primary school students in Jiangjin District, Chongqing City. CONCLUSIONS: There is a high prevalence of Blastocystis infections detected among primary school students in Jiangjin District, Chongqing City, and ST6 and ST3 are predominant subtypes. Minority ethnicity and low maternal education level (primary school and below) are risk factors for Blastocystis infections in primary school students.

Clinical outcomes of 201 neonates born to mothers with COVID-19: a systematic review.

OBJECTIVE: To evaluate the clinical manifestations and outcomes of neonates born to women who had Coronavirus Disease 2019 (COVID-19) during pregnancy. MATERIALS AND METHODS: A systematic literature search was conducted on PubMed and Embase till April 15, 2020, by combining the terms (COVID-19, Severe Acute Respiratory Syndrome Coronavirus 2, SARS-CoV-2, Novel Coronavirus, 2019-nCov, Wuhan pneumonia) and (pregnancy, pregnant women, mother, fetus, neonate, newborn, infant). RESULTS: We included 16 case series and 12 case reports describing a total of 223 pregnant women and 201 infants. Four newborns born to mothers affected by COVID-19 were reported to have laboratory-confirmed Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) infection within 48 hours after birth. However, Reverse Transcription-Polymerase Chain Reaction tests of the breast milk, placenta, amniotic fluids, and cord blood and maternal vaginal secretions were all negative for SARS-CoV-2 in the reported cases. Fetal death was reported in two cases, and 48 of 185 newborns (25.9%) were born prematurely. Infants born small for gestational age and low birth weight (< 2,500 g) accounted for 8.3% and 15.6% of reported cases, respectively. Birth asphyxia and respiratory distress syndrome were observed in 1.8% and 6.4% of neonates, respectively. There was one neonatal death due to intractable gastric bleeding among the SARS-CoV-2-negative infants. CONCLUSIONS: Current evidence suggests that COVID-19 during pregnancy rarely affects fetal and neonatal mortality, but can be associated with adverse neonatal morbidities. Vertical transmission has not been observed in the majority of the reported cases. The infants born to mothers with COVID-19 are carefully monitored for accompanying complication, and quarantine of infected mothers is warranted.

[Prevalence and risk factors of Blastocystis hominis infection in inpatients in Jiangjin District, Chongqing City].

OBJECTIVE: To understand the prevalence and risk factors of Blastocystis hominis infection in inpatients in Jiangjin District, Chongqing City. METHODS: A cross-sectional study was conducted in a community hospital in Jiangjin District, Chongqing City, and the inpatients were surveyed by questionnaires. After obtaining the informed consent from the inpatients or legal guardians, the stool and blood samples were collected and examined by microscopy and PCR from April 17 to May 1, 2018. The univariate analysis and logistic regression analysis were used to analyze the risk factors of the B. hominis infection. RESULTS: A total of 198 hospitalized patients were investigated, and the infection rate of B. hominis was 10.61% (21/198), and the infection rate of the females (12.10%) was higher than that of the males (8.11%), but the difference was not statistically significant. The highest rate of infection was 19.23% in the age group of 10 to 20 years, followed by 17.74% in the age group of 60 years and above, and the lowest rate was 2.38% in the age group of 20 to 40 years. The difference in infection rates of B. hominis among the different age groups was statistically significant (P < 0.05). The infection rate of B. hominis in the people who used dry pail latrines was 33.30%, which was higher than that of the people who used water flush toilets (9.10%) (P < 0.05). The genotypes of B. hominis were ST1, ST3, ST6 and ST7, and ST6 and ST3 being the most predominant genotypes which accounted for 47.62% (10/21) and 38.10% (8/21) respectively, and among the infected males, the genotypes were only ST3 and ST6. The multiple logistic regression analysis showed that among the factors affecting B. hominis infection, only keeping pets was a risk factor [OR = 3.798, 95% CI (1.245, 11.581), P < 0.05]. CONCLUSIONS: A high prevalence of B. hominis infection is found in the inpatients in Jiangjin District, Chongqing City, the predominant genotypes are ST6 and ST3, and keeping pets may be one of the main risk factors.

Suggested new breakpoints of anti-MERS-CoV antibody ELISA titers: performance analysis of serologic tests.

To provide optimal cut-off values of anti-Middle East respiratory syndrome coronavirus (MERS-CoV) serologic tests, we evaluated performance of ELISA IgG, ELISA IgA, IFA IgM, and IFA IgG using 138 serum samples of 49 MERS-CoV-infected patients and 219 serum samples of 219 rRT-PCR-negative MERS-CoV-exposed healthcare personnel and patients. The performance analysis was conducted for two different purposes: (1) prediction of neutralization activity in MERS-CoV-infected patients, and (2) epidemiologic surveillance of MERS-CoV infections among MERS-CoV-exposed individuals. To evaluate performance according to serum collection time, we used 'days post onset of illness (dpoi)' and 'days post exposure (dpex)' assessing neutralization activity and infection diagnosis, respectively. Performance of serologic tests improved with delayed sampling time, being maximized after a seroconversion period. In predicting neutralization activity, ELISA IgG tests showed optimal performance using sera collected after 21 dpoi at cut-off values of OD ratio 0.4 (sensitivity 100% and specificity 100%), and ELISA IgA showed optimal performance using sera collected after 14 dpoi at cut-off value of OD ratio 0.2 (sensitivity 85.2% and specificity 100%). In diagnosis of MERS-CoV infection, ELISA IgG exhibited optimal performance using sera collected after 28 dpex, at a cut-off value of OD ratio 0.2 (sensitivity 97.3% and specificity 92.9%). These new breakpoints are markedly lower than previously suggested values (ELISA IgG OD ratio 1.1, sensitivity 34.8% and specificity 100% in the present data set), and the performance data help serologic tests to be practically used in the field of MERS management.

[Construction of the pIRES2-ZsGreen1 eukaryotic expression vector of Factor â ¨ gene and expression in HEK-293 cells].

Objective: To construct pIRES2-ZsGreen1/F Ⅸ expression vector, using the pcDNA/FⅨ plasmid containing FⅨ cDNA as template, and express in HEK-293 cells. Methods: The total ORF of F Ⅸ gene was amplified from pcDNA/F Ⅸ plasmid, then the amplified fragment was clonded into the pIRES2-ZsGreen1 vector using the Infusion enzyme. The positive clones of eukaryotic expression vector of pIRES2-ZsGreen1/F Ⅸ were screened and expanded after transfection, then were constructed and confirmed by PCR and sequencing. Transient expression experiments were performed using HEK-293 cells transfected with the expression vectors and observed the expression of ZsGreen1 protein by confocal laser microscope. The relative expression levels of FⅨ mRNA, protein and FⅨ activity (FⅨ∶C) were detected by real time PCR (RT-PCR), immunofluorescence microscopy, One-Stage method, respectively. Results: The expression vector, pIRES2-ZsGreen1/F Ⅸ, was successfully constructed and expressed in HEK-293 cells. RT-PCR detected the expression of F Ⅸ mRNA in HEK-293 cells and the immunofluorescence microscopy showed FⅨ protein distributed in the surrounding of nucleus. FⅨ∶C of cell lysates and cell culture fluid transfected with the expression vectors were (92.03 ± 0.29)% and (86.89 ± 8.78)%, respectively; while both F Ⅸ∶C of cell lysates and cell culture fluid transfected with or without the expression vectors were 0. Conclusion: The experimental results showed the expression vector, pIRES2-ZsGreen1/FⅨ, was successfully constructed , which provided experiment basement for the follow study on the location, function and molecular pathology of hemophilia B.

Living Donor Liver Transplantation in a Patient With Extensive Portomesenteric Venous Thrombosis: Case Report.

Extensive portomesenteric venous thrombus preventing restoration of adequate portal venous flow used to be considered a contraindication to liver transplantation. The subject was a 49-year-old male with hepatitis B cirrhosis and extensive thrombosis of portal, splenic, and superior mesenteric veins, and two large collateral vessels; one dilated and tortuous inferior to the pancreaticoduodenal vein and relevant to splanchnic venous return and the other a dilated coronary vein relevant to splenic venous return. During operation, the portal vein was anastomosed to these large collateral vessels using cryopreserved iliac vein. In conclusion, portal reconstruction with large collateral vessels in living-donor liver transplantation could be used selectively for patients with extensive portomesenteric venous thrombosis.

Relationship between tinnitus and suicidal behaviour in Korean men and women: a cross-sectional study.

OBJECTIVES: This study investigated the prevalence of suicidal ideation and behaviour in a representative sample of South Koreans with or without tinnitus. DESIGN: A cross-sectional study. SETTING: Based on data from the 2010 to 2012 Korean National Health and Nutrition Examination Survey (KNHANES). PARTICIPANTS: The study included 17 446 Korean individuals. MAIN OUTCOME MEASURES: Participants provided demographic, socio-economic and behavioural information, as well as responses to questionnaires assessing the presence and severity of tinnitus, mental health status regarding stress, depression, and suicidal ideation and attempts. In the univariate analysis, the Rao-Scott chi-square test and logistic regression analysis were used to test the association between tinnitus and risk factors. Simple and multiple linear regression analyses were used to examine the association between tinnitus and mental status. RESULTS: A total of 20.9% and 1.2% of participants with tinnitus, and 12.2% and 0.6% of those without, reported suicidal ideation and attempts, respectively (P < 0.0001 and P = 0.001). Participants reporting suicide attempts showed a higher proportion of severe annoying (6.0%) and irritating (11.8%) tinnitus than those with suicidal ideation (1.4% and 10.2%, respectively). Risks for experiencing tinnitus were significantly associated with suicidal ideation and attempts after adjusting for confounding variables. CONCLUSION: This study has important implications for enhanced screening and evaluation of mental health status and suicidal ideation/behaviour among tinnitus patients.

Characterization of biochemical properties of a selenium-independent glutathione peroxidase of Cryptosporidium parvum.

Glutathione peroxidase (GPx; EC 1.11.1.9) is an important antioxidant enzyme that catalyses the reduction of organic and inorganic hydroperoxides to water in oxygen-consuming organisms, using glutathione as an electron donor. Here, we report the characterization of a GPx of Cryptosporidium parvum (CpGPx). CpGPx contained a standard UGU codon for cysteine instead of a UGA opal codon for seleno-cysteine (SeCys) at the active site, and no SeCys insertion sequence (SECIS) motif was identified within the 3'-untranslated region (UTR) of CpGPx, which suggested its selenium-independent nature. In silico and biochemical analyses indicated that CpGPx is a cytosolic protein with a monomeric structure. Recombinant CpGPx was active over a wide pH range and was stable under physiological conditions. It showed a substrate preference against organic hydroperoxides, such as cumene hydroperoxide and t-butyl hydroperoxide, but it also showed activity against inorganic hydroperoxide, hydrogen peroxide. Recombinant CpGPx was not inhibited by potassium cyanide or by sodium azide. The enzyme effectively protected DNA and protein from oxidative damage induced by hydrogen peroxide, and was functionally expressed in various developmental stages of C. parvum. These results collectively suggest the essential role of CpGPx for the parasite's antioxidant defence system.

PPP1R1B-STARD3 chimeric fusion transcript in human gastric cancer promotes tumorigenesis through activation of PI3K/AKT signaling.

Fusion genes act as potent oncogenes, resulting from chromosomal rearrangements or abnormal transcription in many human cancers. Although multiple gastric cancer genomes have been sequenced, the driving recurrent gene fusions have not been well characterized. Here, we used paired-end transcriptome sequencing to identify novel gene fusions in 18 human gastric cancer cell lines and 18 pairs of primary human gastric cancer tissues and their adjacent normal tissues. Multiple samples revealed expression of PPP1R1B-STARD3 fusion transcript. The presence of PPP1R1B-STARD3 correlated with elevated levels of PPP1R1B mRNA. PPP1R1B-STARD3 fusion transcript was detected in 21.3% of primary human gastric cancers but not in adjacent matched normal gastric tissues. Based on reverse transcription PCR analysis of DNA, unlike other fusions described in gastric cancer, the PPP1R1B-STARD3 appears to be generated by RNA processing without chromosomal rearrangement. Overexpression of PPP1R1B-STARD3 in MKN-28 significantly increased cell proliferation and colony formation. This increased proliferation was mediated by activation of phosphatidylinositol-3-kinase (PI3K)/AKT signaling. Furthermore, expression of PPP1R1B-STARD3 fusion transcript enhanced the tumor growth of MKN-28 cells in athymic nude mice. These findings show that PPP1R1B-STARD3 fusion transcript has a key role in subsets of gastric cancers through the activation of PI3K/AKT signaling.

Atypical upswing at initial part of phase III in capnograms due to technical cause.

We sporadically experienced three paediatric cases of atypical upswing at the initial part of phase III in the capnograms via side-stream capnometer immediately following endotracheal intubation and mechanical ventilation. No fault was found in the monitor or anaesthetic system including breathing circuits, carbon dioxide (CO2 ) sampling tube, water trap, and unidirectional valves. The upsurge of CO2 disappeared with increasing the respiratory rate; however, it reappeared with decreasing the respiratory rate and vice versa. We experimentally reproduced the phenomenon of overshooting CO2 measurement after the luer lock connection of the sample gas line at the water trap had been unscrewed a little bit.

Cryptostatin, a chagasin-family cysteine protease inhibitor of Cryptosporidium parvum.

Cysteine proteases of pathogenic protozoan parasites play pivotal roles in the life cycle of parasites, but strict regulation of their activities is also essential for maintenance of parasite physiology and interaction with hosts. In this study, we identified and characterized cryptostatin, a novel inhibitor of cysteine protease (ICP) of Cryptosporidium parvum. Cryptostatin showed low sequence identity to other chagasin-family ICPs, but 3 motifs (NPTTG, GXGG, and RPW/F motifs), which are evolutionarily conserved in chagasin-family ICPs, were found in the sequence. The overall structure of cryptostatin consisted of 8 ß-strands that progressed in parallel and closely resembled the immunoglobulin fold. Recombinant cryptostatin inhibited various cysteine proteases, including papain, human cathepsin B, human cathepsin L, and cryptopain-1, with K i's in the picomolar range. Cryptostatin was active over a wide pH range and was highly stable under physiological conditions. The protein was thermostable and retained its inhibitory activity even after incubation at 95°C. Cryptostatin formed tight complexes with cysteine proteases, so the complexes remained intact in the presence of sodium dodecyl sulfate and ß-mercaptoethanol, but they were disassembled by boiling. An immunogold electron microscopy analysis demonstrated diffused localization of cryptostatin within oocystes and meronts, but not within trophozoites, which suggests a possible role for cryptostatin in host cell invasion by C. parvum.

CBL enhances breast tumor formation by inhibiting tumor suppressive activity of TGF-ß signaling.

Casitas B-lineage lymphoma (CBL) protein family functions as multifunctional adaptor proteins and E3 ubiquitin ligases that are implicated as regulators of signaling in various cell types. Recent discovery revealed mutations of proto-oncogenic CBL in the linker region and RING finger domain in human acute myeloid neoplasm, and these transforming mutations induced carcinogenesis. However, the adaptor function of CBL mediated signaling pathway during tumorigenesis has not been well characterized. Here, we show that CBL is highly expressed in breast cancer cells and significantly inhibits transforming growth factor-ß (TGF-ß) tumor suppressive activity. Knockdown of CBL expression resulted in the increased expression of TGF-ß target genes, PAI-I and CDK inhibitors such as p15(INK4b) and p21(Cip1). Furthermore, we demonstrate that CBL is frequently overexpressed in human breast cancer tissues, and the loss of CBL decreases the tumorigenic activity of breast cancer cells in vivo. CBL directly binds to Smad3 through its proline-rich motif, thereby preventing Smad3 from interacting with Smad4 and blocking nuclear translocation of Smad3. CBL-b, one of CBL protein family, also interacted with Smad3 and knockdown of both CBL and CBL-b further enhanced TGF-ß transcriptional activity. Our findings provide evidence for a previously undescribed mechanism by which oncogenic CBL can block TGF-ß tumor suppressor activity.

Molecular cloning and characterization of a M17 leucine aminopeptidase of Cryptosporidium parvum.

Leucine aminopeptidases (LAPs) are a group of metalloexopeptidases that catalyse the sequential removal of amino acids from the N-termini of polypeptides or proteins. They play an important role in regulating the balance between catabolism and anabolism in living cells. LAPs of apicomplexa parasitic protozoa have been intensively investigated due to their crucial roles in parasite biology as well as their potentials as drug targets. In this study, we identified an M17 leucine aminopeptidase of Cryptosporidium parvum (CpLAP) and characterized the biochemical properties of the recombinant protein. Multiple sequence alignment of the deduced amino acid sequence of CpLAP with those of other organisms revealed that typical amino acid residues essential for metal binding and active-site formation in M17 LAPs were well conserved in CpLAP. Recombinant CpLAP shared similar biochemical properties such as optimal pH, stability at neutral pHs, and metal-binding characteristics with other characterized LAPs. The enzyme showed a marked preference for Leu and its activity was effectively inhibited by bestatin. These results collectively suggest that CpLAP is a typical member of the M17 LAP family and may play an important role in free amino acid regulation in the parasite.