Development and validation of a novel nomogram to predict chronic total occlusion before coronary angiography.

OBJECTIVE: Some previous studies have analyzed potential predictors related to the high incidence rate of coronary artery disease (CAD) and established a relevant nomogram for CAD in patients before coronary angiography (CAG). Nevertheless, there are still few models to predict chronic total occlusion (CTO). In this study, we aimed to construct a risk model and nomogram that could effectively predict the probability of CTO before CAG. PATIENTS AND METHODS: In total, the derivation set (n=1,105) and the validation set (n=368), which included patients with CAG diagnosis of CTO, were collected. A statistical difference test was performed for clinical, demography, echocardiography, medication history, laboratory indexes, and angiography. Univariate and multivariate logistic regression analysis were performed to determine the independent risk factors that affect the diagnosis of CTO. A nomogram was established and validated based on the independent predictors. The area under the curve (AUC), the calibration curve, and the decision curve analysis (DCA) were used to evaluate the nomogram. RESULTS: The incidence of CTO within CAD was 21.5%. Univariate and multivariate logistic regression analysis revealed that risk factors for gender (male), neutrophil percentage (NE%), hematocrit (HCT), total cholesterol (TC), high-density lipoprotein cholesterol (HDL), ejection fraction (EF), troponin I (TnI), and N-terminal pro-B-type natriuretic peptide (NT-proBNP) were independent predictors of CTO. A nomogram was constructed incorporating these independent predictors with good discrimination (0.746 in the C-index) and external validation (0.741 in the C-index). The calibration curves and the DCA showed the reliability and accuracy of this clinical prediction model. CONCLUSIONS: The nomogram, composed of gender, NE%, HCT, TC, HDL, EF, TnI, and NT-proBNP, can be used for the prediction of CTO in CAD patients, which opens a great possibility of enriching the means to predict the prognosis of these patients in clinical practice. More studies are needed to validate the effectiveness of this nomogram in other populations.

[Transcriptomic analysis of tuberculosis peptide-based vaccine MP3RT in humanized mice].

Objective: To identify the differentially expressed genes (DEGs) induced by tuberculosis peptide-based vaccine MP3RT in a humanized mouse model using transcriptomics technology. Methods: This study was conducted from August 2019 to February 2022. We used edgeR software to screen DEGs with a fold change greater than or equal to 1.5 and a P value less than 0.05 as screening conditions. Gene ontology (GO), Kyoto encyclopedia of genes and genomes (KEGG), and protein interaction network analyses were performed on the screened DEGs. Then, these DEGs were verified by RT-qPCR and statistically analyzed by GraphPad Prism 8 software. Results: A total of 367 DEGs (214 up-regulated and 153 down-regulated) were identified by transcriptomics. Bioinformatics analysis showed that the GO enrichment of the DEGs mentioned above significantly focused on cell metabolism, growth, apoptosis, inflammation, and other terms. In contrast, the KEGG enrichment significantly focused on inflammatory pathways such as the MAPK signaling pathway. Protein interaction network analysis showed that protein Abl1 had the highest aggregation, the highest aggregation coefficient, and the best connectivity. RT-qPCR results showed that gene expressions of cpne4 (t=2.48, P=0.048 0), h2-q10 (t=2.95, P=0.025 6), mef2c (t=2.87, P=0.028 4), cr2 (t=3.23, P=0.178), ablim1 (t=2.91, P=0.033 5), dll1 (t=2.70, P=0.027 3) and ms4a2 (t=3.03, P=0.019 2) genes in the MP3RT group were significantly up-regulated than those in the PBS group, while gene expressions of cd163l1 (t=2.56, P=0.043 0), il1r1 (t=2.91, P=0.022 7) and cd34 (t=2.42, P=0.046 2) genes in the MP3RT group were significantly down-regulated than those in the PBS group. Conclusions: The MP3RT vaccine induced 367 DEGs in humanized mice, which were associated with metabolic and immune responses. Furthermore, we found that p38 MAPK and JNK/MAPK signaling pathways played an important role in the molecular mechanism of the MP3RT vaccine.

[The robot-assisted system YunSRobot for soft endoscopy: a trial of remote manipulation on simulation models].

Objective: To evaluate the feasibility and safety of the robot-assisted system YunSRobot for remote manipulation endoscopy. Methods: When the master of YunSRobot was installed in the gastroenterology office in Chinese PLA General Hospital, the robot slave and upper gastrointestinal simulation model (Takahashi Lm103, Japan) were installed at the same time in the State Key Laboratory of Robotics, Shenyang Institute of Automation. Three physicians were trained to operate the master robotics and performed gastroscopy on the simulation model based on network cloud. Each physician performed 3 procedures of oesophagogastroduodenoscopy (EGD) by YunSRobot using traditional manual endoscopy, on-site operating mode, and remote manipulation mode, respectively. The operating time, lumenal anatomic exposure, man-machine interaction and other parameters were recorded. Results: The number of standard pictures obtained by traditional manual endoscopy group, on-site operating group and remote manipulation group were 39.9±0.3, 39.8±0.4, 39.9±0.3, respectively. The images of all five lesions could be obtained by each operation. The operating time in the duodenum of remote group was longer than that of on-site group, with average time (78.2±16.0)s vs. (68.9±15.8)s (P=0.021) respectively. As to the operating time on other parts or total time, all three groups were comparable. Although there was a mean delay of (572.1±48.5) ms in remote operation group, the operation was still smooth. However, compared with on-site group, the percentage of clear view time in the duodenum was significantly shortened in remote group: [(77.8±8.2)% vs. (83.9±6.4)%, P=0.024]. Statistically significant difference was detected in percentage of clear view time neither in other sites, nor was in the total operating time between two groups. The operating time in each part of remote group was obviously longer than that of manual group as followings, pharyngeal (27.3±4.2) s vs. (9.2±1.3)s (P<0.001), esophageal (29.7±6.4)s vs. (19.3±1.6)s (P=0.004), stomach (56.7±17.0)s vs. (40.3±7.0)s (P=0.003), pylorus (20.2±5.5)s vs. (9.3±1.3)s (P<0.001), duodenum (78.2±16.0)s vs. (29.3±5.6)s (P<0.001). Thus the total operating time was also longer in remote group as (559.0±87.2)s vs. (253.1±16.6)s (P<0.001). The respective time in pharynx, esophagus, stomach, pylorus, duodenum, or the overall time was all longer in remote group than that in manual group. Conclusions: The soft endoscopy robot YunSRobot has satisfactory safety and stability. Remote upper gastrointestinal endoscopy can be completed based on common network and an endoscope simulation model with smooth operation. The inspection time by YunSRobot robot per part and the overall time are longer than those of manual operation on site, still, remote operating time meets the standard of upper gastrointestinal endoscopy.

[A robot-assisted system YunSRobot for soft endoscopy: the first trial of upper gastrointestinal endoscopy on human volunteers].

Objective: To evaluate robot-assisted system YunSRobot to perform oesophagogastroduodenoscopy(EGD) in simulation model and human volunteers. Methods: The YunSRobot was invented for soft endoscopy by our team. In this trial, gastroenterologists operated the double handles of YunSRobot to perform EGD endoscopy with Olympus GIF-H 260 in simulation model Lm103 of Koken Japan and human volunteers. The operating time, lumenal anatomic exposure, man-machine interaction and other parameters were recorded and compared with manual endoscopy. Results: In the endoscopy on model, each of four doctors performed 5 procedures of EGD by YunSRobot and traditional manual endoscopy, respectively. The average time of one procedure was (626.4±120.7) seconds in the robot group and (241.5±24.7) seconds in the manual group, the operating time in robot group was more than that in manual endoscopy group. (t=14.0, P<0.001). However, the robotic manipulation time of the four endoscopists was significantly shortened from the first case to the fifth case with ((783.5±154.8)s, (667.75±85.1)s, (582.0±74.7)s, (555.0±28.9)s, (543.8±29.7)s, F=15.353, P=0.03). In the endoscopy on human, three physicians operated the EGD endoscopy in 21 volunteers, each physician performed seven volunteers by YunSRobot and traditional manual endoscopy respectively. The average time of each procedure in the robot group and the manual endoscopy group was (875.6±179.8) vs (378.8±80.4)s, the operating time in robot group was also more than that in manual endoscopy group (t=10.278, P<0.001). Like the endoscopy on model, the robotic manipulation time by the three operators was significantly reduced from the first case to the seventh cases ((954.7±62.1)s, (936.7±116.9)s, (968.7±227.1)s, (1 008.0±229.4)s, (876.7±110.5)s, (735.0±149.2)s, (649.3±81.0)s, F=3.79, P=0.024). All endoscopic anatomy of EGD were clearly and skillfully observed including esophgus, cardia, fundus, body, gastric angle, antrum, pylorus, first and second parts of duodenum, and papilla in both groups. In all procedures, there were no complications. Conclusions: The soft endoscopy robot YunSRobot is competent to conduct endoscopy of EGD on human. Presently the initial learning time of YunSRobot for EGD is longer than that of experienced manual endoscopy. The learning curve shows that the time of robot manipulation reduces rapidly in the initial procedures. It indicates that YunSRobot system has user-friendly functions and is easy to master.

Y5 receptor signalling counteracts the anorectic effects of PYY3-36 in diet-induced obese mice.

Peptide YY 3-36 (PYY3-36) is known as a critical satiety factor that reduces food intake both in rodents and humans. Although the anorexic effect of PYY3-36 is assumed to be mediated mainly by the Y2 receptor, the involvement of other Y-receptors in this process has never been conclusively resolved. Amongst them, the Y5 receptor (Y5R) is the most likely candidate to also be a target for PYY3-36, which is considered to counteract the anorectic effects of Y2R activation. In the present study, we show that short-term treatment of diet-induced obese wild-type (WT) and Y5R knockout mice (Y5KO) with PYY3-36 leads to a significantly reduced food intake in both genotypes, which is more pronounced in Y5R KO mice. Interestingly, chronic PYY3-36 infusion via minipumps to WT mice causes an increased cumulative food intake, which is associated with increased body weight gain. By contrast, lack of Y5R reversed this effect. Consistent with the observed increased body weight and fat mass in WT-treated mice, glucose tolerance was also impaired by chronic PYY3-36 treatment. Again, this was less affected in Y5KO mice, suggestive of a role of Y5R in the regulation of glucose homeostasis. Taken together, our data suggest that PYY3-36 mediated signalling via Y5 receptors may counteract the anorectic effects that it mediates via the Y2 receptor (Y2R), consequently lowering bodyweight in the absence of Y5 signalling. These findings open the potential of combination therapy using PYY3-36 and Y5R antagonists to enhance the food intake reducing effects of PYY3-36.

[Clinical application of Acutegastrointestinal injury grading system assocaited with clinical severity outcome in critically ill patients: a multi-center prospective, observational study].

Objective: To investigate the feasibility of utilizing the current acute gastrointestinal injury(AGI) grading system, and explore the association of severity of AGI grade with clinical outcome in critically ill patients. Methods: The adult patients from 14 general ICUs in Zhejiang Province with an expected admission to ICU for at least 24 h were recruited, and all clinical, laboratory, and survival data were prospectively collected. The AGI grade was daily assessed based on GIsymptoms, feeding details and organ dysfunctionon the first week of admission to ICU.The intra-abdominal pressures(IAP) was measured using AbViser device. Results: Of 550 patients enrolled, mean values for age and APACHE Ⅱ score were (64.9±17.2) years and (19.5±7.4), respectively. 456 patients(82.9%) took mechanical ventilation, and 470 patients were identified for AGI. The distribution of AGI grade on the frist day of ICU admission were 50.6%(Ⅰ grade, n=238), 34.2%(Ⅱ grade, n=161), 12.4%(Ⅲ grade, n=58) and 2.8%(Ⅳ, n=13), respectively, while the distribution of the global AGI grade based on the 7-day AGI assessment of ICU admission were 24.5%(Ⅰ grade, n=115), 49.4%(Ⅱ grade, n=232), 20.6%(Ⅲ grade, n=97) and 5.5%(Ⅳ, n=26), respectively. 28- and 60-day mortality rate was 29.3%(n=161) and 32.5%(n=179), respectively. The patients with AGI had a higher 28-(31.1% vs 18.8%, P=0.025) and 60-day survival rate(34.7% vs 20.0%, P=0.01) than those with non-AGI, and also there were positive correlations between AGI grade and 28- and 60-day mortality(P<0.001). Univariate Cox regression analysis showed that age, the source of medicial admission, diabetes mellitus, coronary heart disease, the use of vasoactive drugs, serum creatinine and lactate, mechanical ventilation, APACHE Ⅱ score, the AGI grade in the first day of ICU admission and feeding intolerance within the first week of ICU stay were significantly(P≤0.02) associated with mortality. In multivariate analysis including all these variables, the source of medical admission(χ(2)=4.34, P=0.04), diabete mellitus(χ(2)=3.96, P=0.05), the use of vasoactive drugs(χ(2)=6.55, P=0.01), serum lactate(χ(2)=4.73, P=0.03), the global AGI grade in the 7-day of ICU admission(χ(2)=7.10, P=0.008), and APACHE Ⅱ score(χ(2)=12.1, P<0.001) remained independent predictors for 60-day mortality.In the further subgroup analysis including 402 patients with 7-day survival, the feeding intolerance within the first week of ICU stay could provide independent and incremental prognostic value of 60-day mortality wtih increased χ(2)value of Cox regression model(χ(2)=52.2 vs 41.9, P=0.007) . Conclusion: The AGI grading system is useful for identifying the severity of gastrointestinal dysfunction, and could be used as a strong predictor of impaired outcome. The results provide evidence to support that feeding intolerance within 7 days of admission to ICU was an independent determinant of mortality.

[Morphological characteristics and phenotypic analysis of multiple morphological abnormalities in sperm flagella].

Objective: To analyze the sperm morphological characteristics of multiple morphological abnormalities of the sperm flagella (MMAF), and to analyze their common features and subtypes. Methods: Twenty-eight patients with abnormal morphology of flagella were analyzed by semen analysis. The morphological characteristics were analyzed by scanning electron microscopy and transmission electron microscopy. Histological observation of one case of testicular tissue was performed. Results: Of the 28 patients, only 13 patients (46.4%) had motile spermatozoa, 12 of which had a sperm motility rate of <10% and a sperm survival rate of 9.0%-80.0%. Under light and scanning electron microscope, sperm with absent, short, coiled, bent and irregular width flagella or their combinations were observed. Transmission electron microscopy showed structural abnormalities of sperm fibrous sheath, mitochondrial sheath. Loss rate of central microtubule was 41.4%-84.6%. The semen of the 2 patients with the absence or presence of the kinetic protein arm and both the inner and lateral motilin arms missing had no motile spermatozoa. There was no statistically significant difference in the proportion of flagellar malformations between the two groups of patients (without motile sperm vs with motile spermatozoa). Conclusion: MMAF is a kind of sperm flagella specific abnormalities. Initially diagnosis can be carried out using light microscopy. Clear diagnosis could be conduct using transmission electron microscopy, and the central microtubule loss of the sperm could be seen as the main feature of the flagella abnormalities. Through the morphological analysis and research, MMAF could be precisely classified, which provide a strong basis for the diagnosis.

[Study on the kidney impairment and expressions of FGF21 from a rat model of vascular calcification].

Objective: To investigate the injury and pathological changes of kidney in a rat model of aortic vascular calcification and to explore the expressions of fibroblast growth factor 21 (FGF21). Methods: A total of 14 Spraugue Dawley (SD) rats were randomly divided into two groups: control group and vitamin D3+ nicotine (VDN) group, with 7 rats in each group.The rats in VDN group received vitamin D3 and nicotine to induce vascular calcification.The content of serum creatinine was determined by sarcosine oxidase method.Alkaline phosphatases (ALP) activity was detected by ALP detection kit.The protein levels of FGF21 were measured by radioimmunoassay (RIA). The structure of kidney was observed by hematoxylin and eosin staining. Results: The serum concentration of creatinine in VDN group was significantly higher than control group[(34.00±4.69) vs (27.17±5.38) µmol/L, P<0.05], and the renal pathological changes in VDN group were more apparent. ALP activity in VDN group was significantly higher than that in control group[(62.59±22.62) vs (29.89±11.78) U/g, P<0.05]. Expression of FGF21 in VDN group increased obviously, compared with that in control group[(0.583±0.340) vs (0.207±0.105) ng/mg, P<0.05]. Meanwhile, the elevated levels of FGF21 were positively correlated with up-regulation of ALP in calcified kidneys (r=0.878, P<0.05). Conclusions: Flushing dose of vitamin D3 and nicotine can induce the change of pathology and function of the kidney.Meanwhile, the expression of FGF21 in kidney up-regulated significantly, suggesting that FGF21 may be involved in the occurrence and development of vascular calcification and subsequent kidney injury.

Sensitivity of tissue properties derived from MRgFUS temperature data to input errors and data inclusion criteria: ex vivo study in porcine muscle.

This study evaluates the sensitivity of two magnetic resonance-guided focused ultrasound (MRgFUS) thermal property estimation methods to errors in required inputs and different data inclusion criteria. Using ex vivo pork muscle MRgFUS data, sensitivities to required inputs are determined by introducing errors to ultrasound beam locations (r error = -2 to 2 mm) and time vectors (t error = -2.2 to 2.2 s). In addition, the sensitivity to user-defined data inclusion criteria is evaluated by choosing different spatial (r fit = 1-10 mm) and temporal (t fit = 8.8-61.6 s) regions for fitting. Beam location errors resulted in up to 50% change in property estimates with local minima occurring at r error = 0 and estimate errors less than 10% when r error < 0.5 mm. Errors in the time vector led to property estimate errors up to 40% and without local minimum, indicating the need to trigger ultrasound sonications with the MR image acquisition. Regarding the selection of data inclusion criteria, property estimates reached stable values (less than 5% change) when r fit > 2.5 × FWHM, and were most accurate with the least variability for longer t fit. Guidelines provided by this study highlight the importance of identifying required inputs and choosing appropriate data inclusion criteria for robust and accurate thermal property estimation. Applying these guidelines will prevent the introduction of biases and avoidable errors when utilizing these property estimation techniques for MRgFUS thermal modeling applications.

Investigation of serum vitamin D levels in Chinese infertile men.

Recently, the question of whether vitamin D exerts an effect on the pathogenic process of infertility has become the centre of attention. There are some controversial conclusions on this issue. Based on previous studies, we sought to explore the difference of serum 25-hydroxyvitamin D3 , 1,25-dihydroxyvitamin D3 levels between infertile patients and fertile men, and to find the influence on semen quality. The analysis of serum 25-hydroxyvitamin D3 level showed no significant difference between infertile patients and fertile men. However, the levels of serum 1,25-dihydroxyvitamin D3 in oligospermia (P < 0.05), asthenospermia (P < 0.01), oligoasthenospermia (P < 0.05) and azoospermia (P < 0.01) patients were significantly lower than those in fertile men. Moreover, serum 1,25-dihydroxyvitamin D3 level was positively correlated with progressive motility and total sperm number in infertile patients. In addition, a positive correlation between serum prolactin and 1,25-dihydroxyvitamin D3 was observed in fertile men. Our results indicated that lower vitamin D could be a risk factor for poor semen quality in infertile men. The 1,25-dihydroxyvitamin D3 , as the biologically active form of vitamin D, may be more significant.

Electroacupuncture decreases the urinary bladder pressure in patients with acute gastrointestinal injury.

The present study investigates the effects of electroacupuncture (EA) on urinary bladder pressure (UBP) in patients with acute gastrointestinal injury (AGI). Twenty patients with AGI admitted to the First Hospital of Jiaxing intensive care unit from December 2011 to June 2013 were evaluated. Conventional group patients (n = 10) were administered moderate enteral nutritional support, and electroacupuncture group patients (n = 10) were administered enteral nutritional support followed by EA at bilateral Zusanli (ST-36), Shangjuxu (ST-37), Hegu (LI-4), and QuChi (LI-11) acupoints. UBP was then measured every 6 h and the serum creatinine once daily for 7 days. There were no statistically significant patient demographic differences in the study groups (P > 0.05). The initial UBP of both patient groups was ≥12 mmHg. On days 6 and 7, the UBP significantly decreased in the EA group compared to the conventional group (P < 0.05). The serum creatinine concentration on day 7 was significantly lower in the EA group than in the conventional group (P < 0.05). Based on these results, electroacupuncture contributed to gastrointestinal motility recovery in patients with AGI. This procedure may reduce UBP and provide organ-protective effects in AGI patients.

Development of a 96-well plate iodine binding assay for amylose content determination.

Cereal starch amylose/amylopectin (AM/AP) is critical in functional properties for food and industrial applications. Conventional methods of AM/AP are time consuming and labor intensive making it difficult to screen the large sample sets necessary for evaluating breeding samples and investigating environmental impact on starch development. The objective was to adapt and optimize the iodine binding assay in a 96-well plate format for measurement at both λ 620 nm and λ 510 nm. The standard curve for amylose content was scaled to a 96-well plate format and demonstrated R(2) values of 0.999 and 0.993 for single and dual wavelengths, respectively. The plate methods were applicable over large ranges of amylose contents: high amylose maize starch at 61.7±2.3%, normal wheat starch at 29.0±0.74%, and a waxy maize starch at 1.2±0.9%. The method exhibited slightly greater amylose content values than the Concanavalin A method for normal type starches; but is consistent with cuvette scale iodine binding assays.

Genetic diversity and differentiation of the endangered and endemic species Sauvagesia rhodoleuca in China as detected by ISSR analysis.

Sauvagesia rhodoleuca (Ochnaceae) is an endangered plant that is endemic to southern China. The levels of genetic variation and patterns of population structure in S. rhodoleuca were investigated using inter-simple sequence repeat markers. Eleven primers were used to amplify DNA samples from 117 individuals, and a total of 92 loci were detected. Our results indicated that genetic diversity was quite low both at the species level (percentage of polymorphic bands (PPB) = 41.30%, Nei's gene diversity (h) = 0.1331, and Shannon information index (I) = 0.2028) and the population level (PPB = 16.30-28.26%, h = 0.0496-0.1012, and I = 0.0756-0.1508). A high level of genetic differentiation among populations was detected based on Nei's genetic diversity analysis (0.4344) and analysis of molecular variance (47.03%). The low genetic diversity within population and high population differentiation of S. rhodoleuca were assumed to result largely from limited gene flow, genetic drift, inbreeding, and clonal growth. Conservation strategies for this endangered species are proposed based on the genetic data.

First Report of Tomato chlorosis virus in China.

In October 2012, a severe yellowing disease was found on greenhouse and plastic house tomato (Solanum lycopersicum) plants in Beijing, China. The disease incidence varied from 5 to 80% in each of six fields across Haidian and Daxing districts. The lower leaves showed symptoms of interveinal chlorosis, leaf brittleness, and limited brown necrotic flecks, similar to symptoms induced by Tomato chlorosis virus (ToCV) and Tomato infectious chlorosis virus (TICV) (two members of genus Crinivirus, family Closteroviridae) (4). A large number of whiteflies (Bemisia tabaci) were also observed. Leaf samples were taken from eight symptomatic and two asymptomatic tomato plants in two plastic houses in the Haidian district and total RNA was isolated from the 10 samples using TRIzol reagent (Tiangen, Beijing, China). Nested reverse transcription (RT)-PCR was performed to test the presence of ToCV and TICV with degenerate primers HS-11 and HS-12 and specific primers ToC-5/ToC-6 or TIC-3/TIC-4 for ToCV or TICV, respectively (1). With ToCV primers, a 463-bp specific fragment was amplified from eight symptomatic samples but not from two asymptomatic samples, and there was no amplification with TICV primers from any sample. Sequence analysis of the amplified fragment showed 99% nucleotide sequence identity with the heat shock protein 70 homolog (HSP70h) gene of ToCV isolates from Japan (GenBank Accession No. AB513442), Spain (DQ136146), Florida (AY903448), and Greece (EU284744). The presence of ToCV was confirmed by amplification of a 848-bp fragment covering the coat protein (CP) gene of ToCV with primers CP-F (5'-GAATCTTTTAGAAGCTTTGGTTTAAGG-3') and CP-R (5'-GATCCTCTTGATCCTCATAGATTTC-3') (3). The CP had 97 to 99% amino acid sequence identity to the above-mentioned four ToCV isolates. A sequence of the CP gene obtained from one isolate was deposited at GenBank (KC311375). Additionally, virions were isolated from 25 g of symptomatic samples followed Klaassen's method (2) and their lengths were estimated to be about 800 to 850 nm by transmission electronic microscopy To our knowledge, this is the first report of ToCV on tomato in mainland China. Tomato is one of the most widely cultivated crops in China and the spread of ToCV in China may cause significant economic losses. Further information on the prevalence and incidence of ToCV is required to assess the potential impact of this virus. References: (1) C. I. Dovas et al. Plant Dis. 86:1345, 2002. (2) V. A. Klaassen et al. J. Gen. Virol. 75:1525, 1994. (3) H. Tomoki et al. J. Gen. Plant Pathol. 76:168, 2010. (4) G. C. Wisler et al. Phytopathology 88:402, 1998.

Absence of SH2B3 mutation in nonobese diabetic mice.

Type 1 diabetes is a chronic progressive autoimmune disease characterized by mononuclear cell infiltration, with subsequent destruction of insulin-producing ß-cells. Studies have identified strong associations between type 1 diabetes and several chromosome regions, including 12q24. Association between type 1 diabetes and 12q24 arises from SNP rs3184504; rs3184504 is a nonsynonymous SNP in exon 3 of SH2B3 (also known as LNK). Nonobese diabetic (NOD) mice recapitulate many aspects of the pathogenesis of type 1 diabetes in humans and are therefore frequently used in studies addressing the cellular and molecular mechanisms of this disease. It is of interest to know whether there is a similar mutation of SH2B3 in NOD mice. We found that the SH2B3 mutation is absent in NOD mice. To our knowledge, this is the first report of the sequence and the protein levels of SH2B3 in NOD mice.

NPY modulates PYY function in the regulation of energy balance and glucose homeostasis.

AIMS: Both the neuronal-derived neuropeptide Y (NPY) and the gut hormone peptide YY (PYY) have been implicated in the regulation of energy balance and glucose homeostasis. However, despite similar affinities for the same Y receptors, the co-ordinated actions of these two peptides in energy and glucose homeostasis remain largely unknown. METHODS: To investigate the mechanisms and possible interactions between PYY with NPY in the regulation of these processes, we utilized NPY/PYY single and double mutant mouse models and examined parameters of energy balance and glucose homeostasis. RESULTS: PYY(-/-) mice exhibited increased fasting-induced food intake, enhanced fasting and oral glucose-induced serum insulin levels, and an impaired insulin tolerance, - changes not observed in NPY(-/-) mice. Interestingly, whereas PYY deficiency-induced impairment in insulin tolerance remained in NPY(-/-) PYY(-/-) mice, effects of PYY deficiency on fasting-induced food intake and serum insulin concentrations at baseline and after the oral glucose bolus were absent in NPY(-/-) PYY(-/-) mice, suggesting that NPY signalling may be required for PYY's action on insulin secretion and fasting-induced hyperphagia. Moreover, NPY(-/-) PYY(-/-) , but not NPY(-/-) or PYY(-/-) mice had significantly decreased daily food intake, indicating interactive control by NPY and PYY on spontaneous food intake. Furthermore, both NPY(-/-) and PYY(-/-) mice showed significantly reduced respiratory exchange ratio during the light phase, with no additive effects observed in NPY(-/-) PYY(-/-) mice, indicating that NPY and PYY may regulate oxidative fuel selection via partly shared mechanisms. Overall, physical activity and energy expenditure, however, are not significantly altered by NPY and PYY single or double deficiencies. CONCLUSIONS: These findings show significant and diverse interactions between NPY and PYY signalling in the regulation of different aspects of energy balance and glucose homeostasis.

A rare Y chromosome constitutional rearrangement: a partial AZFb deletion and duplication within chromosome Yp in an infertile man with severe oligoasthenoteratozoospermia.

We report a case of an infertile man with severe oligoasthenoteratozoospermia with a partial azoospermia factor b (AZFb) deletion and duplication region within chromosome Yp11.2. The hormonal profile was normal for serum concentrations of follicle-stimulating hormone, luteinizing hormone, testosterone and oestradiol. The patient, who showed a 46,XY karyotype, had an approximate 2.4 Mb inherited duplication region in Yp11.2 and a de novo partial AZFb deletion, which spanned 5.25 Mb including eight protein coding genes and four non-coding transcripts, but did not remove the RBMY gene family. Both proximal and distal breakpoints of the deletion were outside any palindromic region or inverted repeat sequence and intra-chromosomal non-allelic homologous recombination could not have been the deletion mechanism. The partial AZFb deletion in our case diminished sperm production, but did not completely extinguish spermatogenesis. Considering severe oligozoospermia, spermatozoa in the patient's ejaculate were used for intracytoplasmic sperm injection, resulting in two twin pregnancies.

Additive actions of the cannabinoid and neuropeptide Y systems on adiposity and lipid oxidation.

AIMS: Energy homeostasis is regulated by a complex interaction of molecules and pathways, and new antiobesity treatments are likely to require multiple pharmacological targeting of anorexigenic or orexigenic pathways to achieve effective loss of excess body weight and adiposity. Cannabinoids, acting via the cannabinoid-1 (CB1) receptor, and neuropeptide Y (NPY) are important modulators of feeding behaviour, energy metabolism and body composition. We investigated the interaction of CB1 and NPY in the regulation of energy homeostasis, hypothesizing that dual blockade of CB1 and NPY signalling will induce greater weight and/or fat loss than that induced by single blockade of either system alone. METHODS: We studied the effects of the CB1 antagonist Rimonabant on food intake, body weight, body composition, energy metabolism and bone physiology in wild-type (WT) and NPY knockout (NPY(-/-)) mice. Rimonabant was administered orally at 10 mg/kg body weight twice per day for 3 weeks. Oral Rimonabant was delivered voluntarily to mice via a novel method enabling studies to be carried out in the absence of gavage-induced stress. RESULTS: Mice with dual blockade of CB1 and NPY signalling (Rimonabant-treated NPY(-/-) mice) exhibited greater reductions in body weight and adiposity than mice with single blockade of either system alone (Rimonabant-treated WT or vehicle-treated NPY(-/-) mice). These changes occurred without loss of lean tissue mass or bone mass. Furthermore, Rimonabant-treated NPY(-/-) mice showed a lower respiratory exchange ratio than that seen in Rimonabant-treated WT or vehicle-treated NPY(-/-) mice, suggesting that this additive effect of dual blockade of CB1 and NPY involves promotion of lipid oxidation. On the other hand, energy expenditure and physical activity were comparable amongst all treatment groups. Interestingly, Rimonabant similarly and transiently reduced spontaneous and fasting-induced food intake in WT and NPY(-/-) mice in the first hour after administration only, suggesting independent regulation of feeding by CB1 and NPY signalling. In contrast, Rimonabant increased serum corticosterone levels in WT mice, but this effect was not seen in NPY(-/-) mice, indicating that NPY signalling may be required for effects of CB1 on the hypothalamo-pituitary-adrenal axis. CONCLUSIONS: Dual blockade of CB1 and NPY signalling leads to additive reductions in body weight and adiposity without concomitant loss of lean body mass or bone mass. An additive increase in lipid oxidation in dual CB1 and NPY blockade may contribute to the effect on adiposity. These findings open new avenues for more effective treatment of obesity via dual pharmacological manipulations of the CB1 and NPY systems.

Peripheral neuropeptide Y Y1 receptors regulate lipid oxidation and fat accretion.

OBJECTIVE: Neuropeptide Y and its Y receptors are important players in the regulation of energy homeostasis. However, while their functions in feeding regulation are well recognized, functions in other critical aspects of energy homeostasis are largely unknown. To investigate the function of Y1 receptors in the regulation of energy homeostasis, we examined energy expenditure, physical activity, body composition, oxidative fuel selection and mitochondrial oxidative capacity in germline Y1(-/-) mice as well as in a conditional Y1-receptor-knockdown model in which Y1 receptors were knocked down in peripheral tissues of adult mice. RESULTS: Germline Y1(-/-) mice of both genders not only exhibit a decreased respiratory exchange ratio, indicative of increased lipid oxidation, but interestingly also develop late-onset obesity. However, the increased lipid oxidation is a primary effect of Y1 deletion rather than secondary to increased adiposity, as young Y1(-/-) mice are lean and show the same effect. The mechanism behind this is likely because of increased liver and muscle protein levels of carnitine palmitoyltransferase-1 (CPT-1) and maximal activity of key enzymes involved in beta-oxidation; beta-hydroxyacyl CoA dehydrogenase (betaHAD) and medium-chain acyl-CoA dehydrogenase (MCAD), leading to increased mitochondrial capacity for fatty acid transport and oxidation. These effects are controlled by peripheral Y1-receptor signalling, as adult-onset conditional Y1 knockdown in peripheral tissues also leads to increased lipid oxidation, liver CPT-1 levels and betaHAD activity. Importantly, these mice are resistant to diet-induced obesity. CONCLUSIONS: This work shows the primary function of peripheral Y1 receptors in the regulation of oxidative fuel selection and adiposity, opening up new avenues for anti-obesity treatments by targeting energy utilization in peripheral tissues rather than suppressing appetite by central effects.