Martini on the Rocks: Can a Coarse-Grained Force Field Model Crystals?

Computational chemistry is an important tool in numerous scientific disciplines, including drug discovery and structural biology. Coarse-grained models offer simple representations of molecular systems that enable simulations of large-scale systems. Because there has been an increase in the adoption of such models for simulations of biomolecular systems, critical evaluation is warranted. Here, the stability of the amyloid peptide and organic crystals is evaluated using the Martini 3 coarse-grained force field. The crystals change shape drastically during the simulations. Radial distribution functions show that the distance between backbone beads in ß-sheets increases by ∼1 Å, breaking the crystals. The melting points of organic compounds are much too low in the Martini force field. This suggests that Martini 3 lacks the specific interactions needed to accurately simulate peptides or organic crystals without imposing artificial restraints. The problems may be exacerbated by the use of the 12-6 potential, suggesting that a softer potential could improve this model for crystal simulations.

[Comparison of planimetric CT­based volumetry with simplified models for determining the size of intrapelvic hematomas due to pelvic fractures in emergency room diagnostics]. / Vergleich von planimetrisch CT­gestützter Volumetrie mit vereinfachten Modellen zur Größenbestimmung intrapelviner Hämatome bei Beckenfrakturen im Rahmen der Schockraumdiagnostik.

BACKGROUND: Bleeding in the pelvis can lead to a circulatory problem. The widely used whole-body computed tomography (WBCT) scan in the context of treatment in the trauma resuscitation unit (TRU) can give an idea of the source of bleeding (arterial vs. venous/osseous); however, the volume determination of an intrapelvic hematoma by volumetric planimetry cannot be used for a quick estimation of the blood loss. Simplified measurement techniques using geometric models should be used to estimate the extent of bleeding complications. OBJECTIVE: To determine whether simplified geometric models can be used to quickly and reliably determine intrapelvic hematoma volume in fractures type Tile B/C during emergency room diagnostics or whether the time-consuming planimetric method must always be used. MATERIAL AND METHODS: Retrospectively, 42 intrapelvic hemorrhages after pelvic fractures Tile B + C (n = 8:B, 34:C) at two trauma centers in Germany were selected (66% men, 33% women; mean age 42 ± 20 years) and the CT scans obtained during the initial trauma scan were analyzed in more detail. The CT datasets of the included patients with 1-5 mm slice thickness were available for analysis. By area labelling (ROIs) of the hemorrhage areas in the individual slice images, the volume was calculated by CT volumetrically. Comparatively, volumes were calculated using simplified geometric figures (cuboid, ellipsoid, Kothari). A correction factor was determined by calculating the deviation of the volumes of the geometric models from the planimetrically determined hematoma size. RESULTS AND DISCUSSION: The median planimetric bleeding volume in the total collective was 1710 ml (10-7152 ml). Relevant pelvic bleeding with a total volume > 100 ml existed in 25 patients. In 42.86% the volume was overestimated in the cuboid model and in 13 cases (30.95%) there was a significant underestimation to the planimetrically measured volume. Thus, we excluded this volume model. In the models ellipsoid and measuring method according to Kothari, an approximation to the planimetrically determined volume could be achieved with a correction factor calculated via a multiple linear regression analysis. The time-saving and approximate quantification of the hematoma volume using a modified ellipsoidal calculation according to Kothari makes it possible to assess the extent of bleeding in the pelvis after trauma if there are signs of a C-problem. This measurement method, as a simple reproducible metric, could be embedded in trauma resuscitation units (TRU) in the future.

Prophylactic clipping using the over-the-scope clip (OTSC) system after complex ESD and EMR of large colon polyps.

BACKGROUND: Delayed bleeding is the most frequent complication after endoscopic submucosal dissection (ESD) and endoscopic mucosal resection (EMR) of large colon polyps. Today, prophylactic clipping with through-the-scope clips (TTSCs) is commonly used to reduce the risk of bleeding. However, the over-the-scope clip (OTSC) system might be superior to TTSCs in achieving hemostasis. This study aims to evaluate the efficacy and safety of prophylactic clipping using the OTSC system after ESD or EMR of large colon polyps. METHODS: This is a retrospective analysis of a prospective collected database from 2009 until 2021 of three endoscopic centers. Patients with large (≥ 20 mm) colon polyps were enrolled. All polyps were removed by either ESD or EMR. After the resection, OTSCs were prophylactically applied on parts of the mucosal defect with a high risk of delayed bleeding or/and perforation. The main outcome measurement was delayed bleeding. RESULTS: A total of 75 patients underwent ESD (67%, 50/75) or EMR (33%, 25/75) in the colorectum. The mean resected specimen diameter was 57 mm ± 24.1 (range 22-98 mm). The mean number of OTSCs placed on the mucosal defect was 2 (range 1-5). None of the mucosal defects were completely closed. Intraprocedural bleeding occurred in 5.3% (ESD 2.0% vs. EMR 12.0%; P = 0.105), and intraprocedural perforation occurred in 6.7% (ESD 8% vs. EMR 4%; P = 0.659) of the patients. Hemostasis was achieved in 100% of cases of intraprocedural bleeding, whereas two patients required surgical conversion due to intraprocedural perforation. Among the remaining 73 patients who received prosphylactic clipping, delayed bleeding occurred in 1.4% (ESD 0% vs. EMR 4.2%; P = 0.329), and delayed perforation occurred in 0%. CONCLUSIONS: The prophylactic partial closure of large post-ESD/EMR mucosal defects using OTSCs could serve as an effective strategy to reduce the risk of delayed bleeding and perforation. The prophylactic partial closure of large complex post-ESD/EMR mucosal defects using OTSCs could serve as an effective strategy to reduce the risk of delayed bleeding and perforation.

Simulations of Amyloid-Forming Peptides in the Crystal State.

There still is little treatment available for amyloid diseases, despite their significant impact on individuals and the social and economic implications for society. One reason for this is that the physical nature of amyloid formation is not understood sufficiently well. Therefore, fundamental research at the molecular level remains necessary to support the development of therapeutics. A few structures of short peptides from amyloid-forming proteins have been determined. These can in principle be used as scaffolds for designing aggregation inhibitors. Attempts to this end have often used the tools of computational chemistry, in particular molecular simulation. However, few simulation studies of these peptides in the crystal state have been presented so far. Hence, to validate the capability of common force fields (AMBER19SB, CHARMM36m, and OPLS-AA/M) to yield insight into the dynamics and structural stability of amyloid peptide aggregates, we have performed molecular dynamics simulations of twelve different peptide crystals at two different temperatures. From the simulations, we evaluate the hydrogen bonding patterns, the isotropic B-factors, the change in energy, the Ramachandran plots, and the unit cell parameters and compare the results with the crystal structures. Most crystals are stable in the simulations but for all force fields there is at least one that deviates from the experimental crystal, suggesting more work is needed on these models.

Consequences for Norway from a hypothetical accident at the Sellafield reprocessing plant: Atmospheric transport of radionuclides.

The potential consequences for Norway should a nuclear accident at the Sellafield nuclear site occur, have been of concern for Norwegian authorities for several decades. Meteorological data from a 33-year period and the dispersion model 'SNAP' were used to evaluate meteorological conditions for which atmospheric transport of radionuclides from Sellafield to Norway would lead to the most severe impacts. The worst-case meteorological scenario for Norway, was found on 25th June 1989 for a low elevation (0-800 m) release and on 29th June 2001 for a higher elevation (800-1600 m) release. In both cases the western part of Norway was most affected. In general, the probability for depositions (>10 Bq/m2 of 137Cs) increased about 40% during the autumn and winter compared to the spring and summer months. An influence of climate change on the depositions was analysed, but not verified. Results from a number of simulations were also compared to identify how factors such as radioactive particle characteristics and initial release conditions could affect the predicted radionuclide deposition. The impact on predicted total depositions as well as hot-spot depositions by varying particle density and size as well as release elevation in worst-case scenario simulations amounted to about 40%-50%.

Evaluation of Heating and Irradiation Methods for Production of Purified Protein Derivative (PPD) of Mycobacterium Tuberculosis.

Tuberculin skin test, also known as the tuberculin or purified protein derivative (PPD) test, is an extensively applied diagnostic test for the detection of primary infection with Mycobacterium tuberculosis (Mtb). The production of PPD is accompanied by some difficulties that require a series of modifications in the production and purification processes. The present study aimed to determine the facilitation level of the manufacturing process by modifying evaluation methods for the production of PPD tuberculin. Mtb strains were cultured in Lowenstein-Jensen media, and the cultured strains were inoculated into the Dorset-Henley liquid medium by the biphasic medium of potato-Dorset-Henley. After incubation, flasks containing cultured strain were selected for bacterial inactivation, and the optimal gamma radiation dose(s) was determined. Tuberculoproteins were precipitated by ammonium sulfate (AS) and Trichloroacetic acid (TCA). Protein concentration was determined using the Bradford and Kjeldahl protein assay methods. Finally, the lymphocyte transformation test and potency test were performed. Based on the results, the Dorset-Henley liquid medium is suitable for the massive growth of the bacterium. The transferal of Mtb from solid to liquid medium was directly carried out without intermediate culture. It was found that during tuberculoprotein production, heating at 100°C for 3 h would be safe for killing mycobacterium. Furthermore, the simultaneous use of heating and gamma irradiation (8 kGgy) killed all of the mycobacteria, while doses of 1, 1.5, and 7 kGy decreased a significant number of bacterial cells. The results also indicated that the concentration of tuberculoprotein extracted by TCA precipitation method was higher than that obtained by AS precipitation. The tuberculoproteins which were produced by these two methods in the lymphocyte transformation test were not significantly different in terms of potency (P>0.05). Moreover, due to the high volume of produced protein, the protein measurement was more efficiently carried out by the Kjeldahl method, compared to the Bradford method. Finally, the results of the present study demonstrated that in addition to the novel approach of gamma irradiation, optimum methods are efficient and applicable in the production of PPD tuberculin.

Immunization with recombinant PlpE of ovine Mannheimia haemolytica isolate provides protection against lethal challenge in mice.

BACKGROUND: Mannheimia haemolytica primarily causes pneumonia leading to heavy morbidity and mortality in domestic livestock world-wide. Recently, outer membrane lipoproteins have emerged as targets for inducing protective immunity against the Pasteurella infection. AIMS: This study aimed to evaluate recombinant outer membrane lipoprotein E (PlpE) from the ovine M. haemolytica isolate, as a potential vaccine candidate. METHODS: Recombinant PlpE was constructed using pET26 (b) expression vector in Escherichia coli. Expressed recombinant PlpE was purified and injected subcutaneously to mice. The protection index of the vaccine was evaluated by challenge of mice intraperitoneally. RESULTS: Anti-PlpE antibody responses in the immunized mice was significantly increased in comparison with the control group which in turn, provided effective protection when challenged with strain of virulent M. haemolytica. CONCLUSION: Recombinant PlpE from ovine M. haemolytica isolate had the potential to be used as a vaccine candidate against M. haemolytica infection in sheep flocks.

Oncologic outcomes of patients with incidental prostate cancer who underwent RARC: a comparison between nerve sparing and non-nerve sparing approach.

BACKGROUND: Incidental Prostate cancer (iPCa) is a relatively common finding during histopathological evaluation of radical cystectomy (RC) specimens. To reduce the high impact of RC on erectile function, several sexual-preserving techniques have been proposed. The aim of this study was to evaluate and compare the oncologic outcomes of patients with iPCa who underwent nerve spring and no-nerve sparing robot-assisted radical cystectomy (RARC). METHODS: The clinicopathologic data of male patients who underwent RARC at our institution between 2006 and 2016 were retrospectively analysed. Patients with iPCa at definitive pathological examinations were stratified in two groups, according to the preservation of the neurovascular bundles (nerve sparing vs no nerve sparing). Significant PCa was defined as any Gleason score ≥ 3 + 4. Biochemical recurrence (BR) was defined as a sustained PSA level > 0.2 ng/mL on two or more consecutive appraisals. BR rate was assessed only in patients with incidental prostate cancer and at least 2 years of follow-up. Differences in categorical and continuous variables were analysed using the chi-squared test and the Mann-Withney U test, respectively. Biochemical recurrence curves were generated using the Kaplan-Meier method and compared with the Log-rank test. RESULTS: Overall, 343 male patients underwent RARC for bladder cancer within the study period. Nerve-sparing surgery was performed in 143 patients (41%), of these 110 had at least 2 years of follow up after surgery. Patients who underwent nerve-sparing surgery were significantly younger (p < 0.001). Clinically significant PCa was found in 24% of patients. No significant differences regarding preoperative PSA value (p = 0.3), PCa pathological stage (p = 0.5), Gleason score (p = 0.3) and positive surgical margin rates (p = 0.3) were found between the two groups. After a median follow-up of 51 months only one patient, in the no-nerve-sparing group had developed a biochemical recurrence (p = 0.4). CONCLUSIONS: In our series most of the iPca detected in RC specimens can be considered as insignificant with a low rate of BR (0.9%). Nerve-sparing RARC is a safe procedure which did not affect oncological outcomes of patients with iPCa.

Novel modified endoscopic mucosal resection of large GI lesions (> 20 mm) using an external additional working channel (AWC) may improve R0 resection rate: initial clinical experience.

BACKGROUND: En-bloc resection of large, flat dysplastic mucosal lesions of the luminal GI tract can be challenging. In order to improve the efficacy of resection for lesions ≥2 cm and to optimize R0 resection rates of lesions suspected of harboring high-grade dysplasia or early adenocarcinoma, a novel grasp and snare EMR technique utilizing a novel over the scope additional accessory channel, termed EMR Plus (EMR+), was developed. The aim of this pilot study is to describe the early safety and efficacy data from the first in human clinical cases. METHODS: A novel external over-the-scope additional working channel (AWC) (Ovesco, Tuebingen, Germany) was utilized for the EMR+ procedure, allowing a second endoscopic device to be used through the AWC while using otherwise standard endoscopic equipment. The EMR+ technique allows tissue retraction and a degree of triangulation during endoscopic resection. We performed EMR+ procedure in 6 patients between 02/2018-12/2018 for lesions in the upper and lower GI tract. RESULTS: The EMR+ technique utilizing the AWC was performed successfully in 6 resection procedures of the upper and/or lower GI tract in 6 patients in 2 endoscopy centers. All resections were performed successfully with the EMR+ technique, all achieving an R0 resection. No severe adverse events occurred in any of the procedures. CONCLUSIONS: The EMR+ technique, utilizing an additional working channel, had an acceptable safety and efficacy profile in this preliminary study demonstrating it's first use in humans. This technique may allow an additional option to providers to remove complex, large mucosal-based lesions in the GI tract using standard endoscopic equipment and a novel AWC device.

[Refixation of osteochondral fragments with resorbable polylactid implants : Long-term clincal and MRI results]. / Refixation von osteochondralen Fragmenten mit resorbierbaren Polylactid-Implantaten : Klinische und MR-morphologische Langzeitergebnisse.

BACKGROUND: Refixation with resorbable implants is a common surgical treatment in patients who suffer an injury with shearing of an osteochondral flake due to trauma of the knee or the upper ankle joint. To date there are no studies which outline long-term outcomes for this procedure. The aim of this study was to evaluate long-term clinical and magnetic resonance imaging (MRI) results after refixation with resorbable polylactide (PLLA) implants. MATERIAL AND METHODS: In this retrospective study 12 patients with 13 injuries were examined 13.9 years (±1.2 years) after refixation of an osteochondral fragment of the knee (10 patients) and the upper ankle joint (2 patients) with a mean size of 3.33 cm2 (±2.33) by resorbable polylactide (PLLA) implants (nails, pins, screws, Bionx, Tampere, Finland). To objectify the clinical results eight established clinical scores (VASS, Tegner, Lysholm, McDermott, KSS, WOMAC, AOFAS, FADI+Sports) were used. Furthermore, the morphological integration of bone and cartilage was assessed by MRI (3 T) using proton-weighted and cartilage-sensitive 3D double-echo steady-state (DESS) sequences. The morphological results were objectified with a modified MRI score according to Henderson et al. RESULTS: After 13.9 years (±1.2) the patients with an injury of the knee as well as of the upper ankle joint showed good to excellent results (knee: VASS 1.2 (±1.7), Tegner 4.4 (±1.3), Lysholm 85.7 (±12.2), McDermott 90.7 (±8.6), KSS 189 (±14.2), WOMAC (6.16% (±8.45)) (upper ankle joint: VASS 2.5 (±2.5), Tegner 5.5 (±1.5), Lysholm 87 (±13), McDermott 88 (±12); WOMAC (8.54% (±8.54), AOFAS 75.5 (±24.5), FADI+Sports 118 (±18)). In all cases there was evidence of good integration of the osteochondral fragment in MRI. In five patients there was moderate subchondral cyst formation (∅ ≤1 mm); however, mild changes of the cartilage contour were found in all patients. The mean modified Henderson score achieved was 14.4 (±2.0, best 8, worst 32), which corresponds to a good morphological result. CONCLUSION: Because of good clinical and morphological results shown by MRI, refixation through resorbable implants (PLLA) can be recommended for treatment of traumatic osteochondral flakes.

Large-scale network dysfunction in vascular cognitive disorder supports connectional diaschisis in advanced arteriosclerosis.

BACKGROUND AND PURPOSE: The interrelation of cognitive performance, cerebrovascular damage and brain functional connectivity (FC) in advanced arteriosclerosis remains unclear. Our aim was to investigate the associations between FC, white matter damage and cognitive impairment in carotid artery disease. METHODS: Seventy-one participants with a recent cerebrovascular event and with written informed consent underwent resting-state functional magnetic resonance imaging and the Addenbrooke's Cognitive Examination - Revised (ACE-R). Network and inter-hemispheric FC metrics were compared between cognitively normal and impaired subjects, and interrelated with cognition. In order to explore the nature of FC changes, their associations with microstructural damage of related white matter tracts and cognitive performance were investigated, followed by mediation analysis. RESULTS: Participants with global cognitive impairment showed reduced FC compared to the cognitively intact subjects within the central executive network (CEN), and between hemispheres. Patients with executive dysfunction had decreased CEN FC whilst patients with memory loss demonstrated low FC in both the CEN and the default mode network (DMN). Global performance correlated with connectivity metrics of the CEN hub with DMN nodes, and between hemispheres. Cingulum mean diffusivity (MD) was negatively correlated with ACE-R and CEN-DMN FC. The cingulum MD-cognition association was partially mediated by CEN-DMN FC. CONCLUSIONS: Long-range functional disconnection of the CEN with DMN nodes is the main feature of cognitive impairment in elderly subjects with symptomatic carotid artery disease. Our findings provide further support for the connectional diaschisis concept of vascular cognitive disorder, and highlight a mediation role of functional disconnection to explain associations between microstructural white matter tract damage and cognitive impairment.

Cloning and Expression of Mannheimia haemolytica PlpE Gene in Escherichia coli and its Immunogenicity Assessment.

Mannheimia haemolytica is responsible for considerable economic losses to cattle, sheep, and goat industries in many parts of the world. This bacterium isone of the causative agents of shipping fever in cattle. Current vaccines against M. haemolytica are moderately efficacious since they do not provide complete protection against the disease. Production of an economic vaccine for protecting farm animals against M. haemolytica has attracted the attention of many scientists. The outer membrane proteins (OMPs) play a major role in the pathogenesis and immunogenicity of M. haemolytica. Research on M. haemolytica OMPs has shown that antibodies to a particular OMP may be important in immune protection. In the current study, the gene for M. haemolytica OMP PlpE was cloned into the expression vector pET26-b, and then expressed in Escherichia coli BL21. The expression of the protein was carried out by the induction of cultured Escherichiacoli Bl21 cells with 1mM isopropyl-&beta;-D-thiogalactopyranoside. The recombinant PlpE was purified using Ni-NTA agarose resin, and then subjected to sodium dodecyl sulfate polyacrylamide gel electrophoresis. The identity of the expressed protein was analyzed by western blotting. It was revealed that rPlpE was expressed and produced properly. To assess the immunogenicity of the recombinant protein, the purified rPlpE was used as an antigen for antibody production in goats. The observations suggested that the produced recombinant protein can be used as a antigen for developing diagnostic tests and or as a vaccine candidate.

EVALUATING SHORT-TERM EXPOSURE TO WI-FI SIGNALS ON STUDENTS' REACTION TIME, SHORT-TERM MEMORY AND REASONING ABILITY.

This study investigated the effect of short-term exposure to Wi-Fi signals on the cognitive functions of the mind. After obtaining permission from the local Ethics Committee of Shiraz University of Medical Sciences and approval by the Iranian Registry of Clinical Trials (IRCT2017041233398N1), 45 male and female students from Shiraz University of Medical Sciences volunteered to participate in this study. They were exposed to Wi-Fi signals in two sham and exposure sessions, each for 2 hours. After completion, they took part in reaction time, short-term memory, and reasoning ability tests. After scoring, the data were analysed by SPSS software. In addition, the electric field strength and power density were calculated. The results showed no statistically significant differences between the mean scores of reaction time, short-term memory, and reasoning ability in sham and exposure. Also, the obtained values from the electric field strength and power density (E = 4.1 Vm-1, P = 0.446 Wm-2) were lower than that of threshold values by the International Commission on Non-Ionizing Radiation Protection (ICNIRP). Our results can greatly reduce concerns regarding the effects of short-term exposure to Wi-Fi waves on cognitive functions.

Treatment of third-degree burn wounds in animal specimens: acellular dermis or partial-thickness skin graft.

Several dermal products have been introduced to substitute dermal tissues. In this study we review the effects of these products on repairing third-degree burn wounds and managing complications in animal specimens. Using an interventional approach, rats were randomly assigned to four groups (G1 to G4). Two wounds were created on the back of each rat. An open wound was left on the back of rats in G1; in G2, wounds were covered with a thick rat derived-ADM product and overlying thin skin graft; on G3 rats, similar third degree ulcers were made with one ulcer covered with harvested thin skin graft. In G4, ulcers were covered with a thin rat derived-ADM product and thin graft. Factors such as take rate, histopathological score, wound contracture and graft contracture were compared on the 7th, 15th, 21st and 30th day. Mean graft take rate on the 30th day in the thick ADM, thin ADM and graft group showed a significant difference (p=0.015). Histopathological score on the 30th day in the thin ADM, thick ADM and graft group showed no considerable difference. Mean graft take rate was significantly better in the thin ADM and graft group than in the thick ADM group. Wound contracture was significantly more severe in the thick ADM and control group than in the thin ADM and graft group.

Plusieurs produits ont été introduits dans le but de substituer le derme. Dans cette étude, nous avons étudié l'effet de ces produits sur la cicatrisation et la gestion des complications après brûlure expérimentale. Nous avons étudié 4 groupes (G1 à G4) de rats ayant subi deux brûlures du dos. Celles de G1 étaient laissées à l'air, G2 recevaient un Derme Artificiel Acellulaire (DAA) dérivé de rats épais recouvert d'une greffe fine, G3 recevaient une greffe conservée, G4 recevaient DAA fin et greffe fine. Le taux de prise de greffe, le score histologique, la rétraction de la brûlure et de la greffe ont été comparés à J7, J15, J21 et J30. A J30, les taux de prise de greffe étaient significativement différents entre G2, G3 et G4 (p=0,015), étant moins bons en cas d'utilisation de DAA en couche épaisse. De même, la rétraction était plus intense après utilisation de DAA en couche épaisse qu'en couche fine ou après greffe seule. Les résultats histologiques étaient comparables.

Dietary energy level affects adipose depot mass but does not impair in vitro subcutaneous adipose tissue response to short-term insulin and tumor necrosis factor-α challenge in nonlactating, nonpregnant Holstein cows.

We assessed effects of overfeeding energy to nonlactating and nonpregnant Holstein cows during a length of time similar to a typical dry period on body lipid storage and the abundance of genes related to insulin signaling, inflammation, and ubiquitination in subcutaneous adipose tissue (SAT) in vitro challenged with insulin and recombinant bovine tumor necrosis factor-α. Fourteen cows were randomly assigned to either a high-energy (OVE; net energy for lactation = 1.60 Mcal/kg of dry matter; n = 7) or control (CON; net energy for lactation = 1.30 Mcal/kg of dry matter; n = 7) diet for 6 wk. Immediately after slaughter, liver, kidneys, and mammary gland were separated and weighed. The adipose tissue mass in the omental, mesenteric, and perirenal depots was dissected and weighed. Subcutaneous adipose tissue was collected from the tail-head region and was used as follows: control, bovine insulin (INS) at 1 µmol/L, tumor necrosis factor-α at 5 ng/mL (TNF), and their combination. Despite a lack of difference in final body condition score, OVE cows had greater energy intake and were heavier than CON cows. Furthermore, overfeeding led to greater mass of mesenteric and perirenal adipose, liver, and mammary gland. Overall, SAT incubated with INS had an upregulation of insulin receptor (INSR), interleukin-10 (IL10), small ubiquitin-like modifier 3 (SUMO3), and ubiquitin conjugating enzyme E2I (UBC9), whereas TNF upregulated peroxisome proliferator-activated receptor gamma (PPARG), diacylglycerol O-acyltransferase 2 (DGAT2), interleukin-6 (IL6), nuclear factor kappa B subunit 1 (NFKB1), small ubiquitin-like modifier 2 (SUMO2), and UBC9. Regardless of in vitro treatment, feeding OVE upregulated PPARG, fatty acid synthase (FASN), and insulin induced gene 1 (INSIG1). Abundance of PPARG was greater in SAT of OVE cows cultured individually with INS and TNF. The interaction between diet and in vitro treatment revealed that sterol regulatory element binding transcription factor 1 (SREBF1) had greater abundance in SAT from the CON group in response to culture with INS, whereas SAT from OVE cows had greater SREBF1 abundance in response to culture with TNF. The mRNA abundance of IL6 and NFKB1 was greater in response to TNF treatment and overall in CON cows. Furthermore, SAT from these cows had greater IL10 abundance when cultured with INS and TNF. Overall, data highlighted that overfeeding energy increases adipose tissue mass in part by stimulating transcription of key genes associated with insulin signaling, adipogenesis, and lipogenesis. Because SAT thickness or mass was not measured, the lack of effect of overfeeding on body condition score limits its use to predict overall body lipid storage. An overt inflammatory response in SAT after a 6-wk period of over-consumption of energy could not be discerned.

Effect of time of eCG administration on the fate of ovarian follicle in Holstein heifers.

The objective of this study was to investigate the effect of equine chorionic gonadotropin (eCG) on ovarian follicles at three stages of development (emergence, dominance and early static phases) during the first follicular wave (FFW) in Holstein heifers. Heifers (n=20) were randomly assigned into four experimental groups (n=5 in each group). Heifers received eCG (500 IU; Folligon®; Intervet, Holland; i.m) a) on the day of follicle emergence (day of ovulation; group 1), b) on the dominant phase (dominant follicle (DF): the first day in which follicle was observed at ≥10 mm; group 2, and c) on the early static phase (group 3) of the FFW. Control group heifers did not receive any treatment. Daily ultrasonography was conducted to monitor ovarian structure throughout estrous cycle. All treatment group heifers, regardless of the stage of follicle development, displayed follicle growth after eCG injection. Administration of eCG, in group 1, hastened DF detection and induced co-dominant follicles; whereas, in groups 2 and 3, it delayed DF regression, and increased cycle length compared to control. In all treatment group heifers, DF was present 84 h after eCG injection. Maximum diameter of corpus luteum was larger in eCG treated groups compared to control (P<0.05). In conclusion, depending on the time of eCG administration throughout the FFW (emergence, dominant and early static phases), co-dominancy, maintenance of DF, enhancement of follicle and corpus luteum growth and increase in estrous cycle length could be observed in Holstein heifers.

Lactation-related changes in tissue expression of PEDF in dairy cows.

Pigment epithelium-derived factor (PEDF) is evolving as metabolic regulatory protein. Albeit mostly considered in only pathological conditions related to excess energy intake resulting in obesity and insulin resistance, PEDF is likely to be involved in other physiological processes such as the homeorhetic adaptation of metabolism to lactation. We aimed to characterize the expression of PEDF and its association to the concomitant mobilization of body reserves during lactation in nonobese subjects. This mobilization is particularly distinct in dairy cows, and we therefore assessed the mRNA expression of PEDF and its putative receptors in different tissues in 2 trials with dairy cows fed with or without conjugated linoleic acids (CLAs). Conjugated linoleic acids depress milk fat synthesis and may thus reduce the drain of energy via milk. In pluriparous cows, the serum PEDF concentrations and the mRNA abundance in subcutaneous adipose tissue (scAT), as well as the hepatic and scAT mRNA abundance of the putative receptors, adipose triglyceride lipase, and laminin receptor 1, changed over time of sampling (day -21 until day 252 relative to calving). Conjugated linoleic acid treatment was associated with reduced PEDF concentrations in serum and lower PEDF mRNA abundance in scAT on day 21 postpartum. Comparing different tissues from primiparous cows, PEDF mRNA was highest in the liver, followed by scAT, visceral adipose tissue (AT), and mammary gland, and lowest in the muscle. Significant changes in PEDF expression with time of sampling were limited to AT in primiparous and pluriparous cows. Our data support a regulatory role for PEDF. The similarities between the time course of the serum concentrations of PEDF and its mRNA abundance in scAT may point to a regulatory role for AT rather than the liver for PEDF in dairy cows.

Enhancement of Th1 immune response against Leishmania cysteine peptidase A, B by PLGA nanoparticle.

Recombinant cysteine peptidase vaccine can induce protective immunity against cutaneous leishmaniasis. However, the antigenic diversity and variable immunogenicity prevents them from being approved for general vaccination. Different approaches like adjuvant application and antigen delivery systems are studied to increase their efficacy. Nanoparticles can both stimulated antigen uptakes and affect direction of immune response. In this study the effect of PLGA nanoparticles were considered to enhance the immune response against recombinant CPA (CPA) and CPB (CPB). For this purpose, L. major CPA and CPB were prepared. PLGA were conjugated to the proteins using Aldehyde/Hydrazine Reaction. Conjugation efficacy and created nanoparticle morphology were determined by FTIR and SEM methods, respectively. BALB/c mice were received intraperitoneally three boosts of 7 µg/mouse of each antigen alone (CPA/CPB/CPA + CPB) or as PLGA conjugated form in different Study groups, at 3 weeks interval. After vaccination, mice were challenged with 106L. major, subcutaneously. Time course study of lesion development demonstrated nanoparticle efficacy in parasite dissemination control that confirmed by spleen parasite burden assay. Significant induction of nitric oxide production by peritoneal macrophages and increase in splenocyte IFN-γ production showed the protective effect of PLGA-CPA/CPB vaccination in comparison to CPA and CPB alone. Current study demonstrated that the conjugation of the antigen with the PLGA can activate immune responses against L. major. However, further study is necessary to assess the long-term effect and other aspects of immune response.

Is there any correlation between sperm parameters and chromatin quality with embryo morphokinetics in patients with male infertility?

The main goal was to evaluate the correlation between sperm parameters and chromatin quality with embryo kinetics via time-lapse monitoring system (TLM). A total of 40 couples involved in the ICSI program as a result of male infertility. For assessment of sperm chromatin and DNA quality, we used aniline blue, toluidine blue, chromomycin A3, acridine orange and terminal transferase-mediated deoxyuridine triphosphate biotin end labelling assays. All mature oocytes were injected, and the generated zygotes (2PNs) were cultured in TLM. In day 3 after injection, single embryo transfer (SET) was carried out according to the morphology and morphokinetics. The patients were followed up until delivery. There were positive significant correlations between sperm count with CC2 (r = .330, p = .049), T4 (r = .329, p = .038), T6 (r = .342, p = .035) and T7 (r = .374, p = .025). Also, there were positive significant correlations between nonprogressive motility and T2 (r = 0.323, p = .042), T3 (r = .411, p = .013) and T4 (r = .418, p = .007). Regarding the sperm chromatin quality assays, there were negative significant correlations between CMA3 and CC2 (r = -.272, p = .049) and between acridine orange and T5 (r = -.221, p = .040). It seems that the abnormal sperm parameters and chromatin alteration affect the normal embryo kinetics in ICSI program.