Prevalence of methicillin-resistant Staphylococcus aureus skin and nasal carriage isolates from bovines and its antibiogram.
This study was conducted to determine the prevalence of methicillin-resistant Staphylococcus aureus (MRSA) in cattle and buffalo and to study their antibiotic resistance pattern. A total of 136 samples (skin and nasal swab) from cattle and buffalo were collected. MRSA was identified by conventional bacterial culture techniques which were further confirmed by amplification of S. aureus-specific 16S rRNA by polymerase chain reaction (PCR). The isolates were further analyzed for the presence of mec A gene by PCR. The antimicrobial susceptibility profiling was performed by disc diffusion method. The prevalence of MRSA in the current study was 28.57% and 34.28% in cattle nasal and skin swab, respectively, with an overall prevalence of 31.43% MRSA among cattle. Buffalo nasal and skin sample showed MRSA prevalence of 54.55% and 39.4%, respectively, with 46.9% overall prevalence. PCR could detect mec A gene in 36.4% and 58% MRSA isolates from cattle and buffalo, respectively. Antimicrobial susceptibility test found MRSA resistant to penicillin and oxytetracycline (88% each), cefoxitin (75%), cotrimoxazole (62%), and amoxyclav (50%). 100% sensitivity was observed against ciprofloxacin, amikacin, chloramphenicol, and gentamicin. Three (16.7%) MRSA isolates from buffalo were found resistant to vancomycin. Cattle and buffalo were identified as a potential carrier of MRSA in Bihar (India). The isolation of vancomycin-resistant S. aureus (VRSA) in the current study indicates the emergence of VRSA in animal population which may be transmitted to the human beings working in close contact to the animals.


Vascularized periosteal bone graft from the supracondylar region of the femur.
Free, vascularized thin corticoperiosteal grafts and small periosteal bone grafts harvested from the supracondylar region of the femur are described. These grafts are nourished from the articular branch of the descending genicular artery and vein. Thin corticoperiosteal grafts consist of periosteum with a thin layer of outer cortical bone and include the cambium layer, which has a better osteogenic capacity. This graft is elastic and readily conforms to the recipient bed configuration. Thin corticoperiosteal grafts were used to treat 11 patients with fracture non-union of an upper extremity that had no massive bone defects. Early, rapid union occurred in all patients except three: one in which the anastomosed vessel became obstructed and two in which the internal fixation of the fracture was unsecured. The small bone grafts consist of periosteum, full thickness cortex, and the underlying cancellous bone. This graft can be successfully harvested without disturbing the vascularity, unlike the currently used vascularized bone grafts. This graft was used to treat three patients with avascular necrosis of the body of the talus and could prevent the necrotic talus body from progressive collapse in patients in early stages of the disease. One patient with an infectious bone defect of the first metatarsal bone was successfully treated by vascularized bone graft with an accompanying skin flap.


The developmentally regulated type X collagen gene contains a long open reading frame without introns.
Type X collagen is a recently discovered product of hypertrophic chondrocytes that is localized to presumptive mineralization zones of hyaline cartilage. Thus, in the epiphyseal growth plate of long bones it is present only in the zone of hypertrophic chondrocytes and absent in the resting and rapidly growing cartilage and in bone. Type X collagen represents, therefore, a transient and developmentally regulated collagen which is synthesized by a subpopulation of chondrocytes. We report here the isolation and characterization of cDNA and genomic clones specific for the chicken protein. The results demonstrate that the polypeptide chains of this collagen contain three distinct domains: a short non-collagenous, amino-terminal region, a collagenous domain of 460 amino acid residues, and a non-collagenous, carboxyl-terminal domain of 170 amino acid residues. The nucleotide sequence of the gene shows that these domains are encoded by a long open reading frame that is not interrupted by introns. Examination of the amino acid sequence derived from this nucleotide sequence reveals the presence of a hydrophobic segment localized 10 amino acid residues upstream from the translational stop codon. The length and sequence characteristics of this segment raise the intriguing possibility that Type X collagen polypeptides may contain a transmembrane segment.


An Sp1/KLF binding site is important for the activity of a Polycomb group response element from the Drosophila engrailed gene.
Polycomb-group response elements (PREs) are DNA elements through which the Polycomb-group (PcG) of transcriptional repressors act. Many of the PcG proteins are associated with two protein complexes that repress gene expression by modifying chromatin. Both of these protein complexes specifically associate with PREs in vivo, however, it is not known how they are recruited or held at the PRE. PREs are complex elements, made up of binding sites for many proteins. Our laboratory has been working to define all the sequences and DNA binding proteins required for the activity of a 181 bp PRE from the Drosophila engrailed gene. Here we show that one of the sites necessary for PRE activity, Site 2, can be bound by members of the Sp1/KLF family of zinc finger proteins. There are 10 Sp1/KLF family members in Drosophila, and nine of them bind to Site 2. We derive a consensus binding site for the Sp1/KLF Drosophila family members and show that this consensus sequence is present in most of the molecularly characterized PREs. These data suggest that one or more Sp1/KLF family members play a role in PRE function in Drosophila.


Drug-based cardiovascular prevention in patients with Marfan Syndrome: a systematic review.
Marfan Syndrome (MFS) is a rare and complex genetic disorder associated with increased aortic growth and aortic disease. The effectiveness of cardiovascular medical therapies aiming to slow down aortic growth has been tested in several trials, particularly beta-blockers and angiotensin receptor blockers, however showing conflicting results. We conducted a systematic review on PubMed (Medline), Cochrane library, Google Scholar, and Biomed Central databases between January and February 2022. We selected relevant articles focusing on patients with MFS treated with beta-blockers, angiotensin receptors blockers, or both, and reporting data on the effect of the drugs on 1) slowing down aortic dilatation; 2) the reduction of aortic complication (aortic dissection, mortality, aortic surgery); and with a 3) follow-up length of at least two years. A total of 16 studies were selected. Beta-blockers remain the mainstay of therapy as they have proven to slow aortic enlargement. Angiotensin receptor blockers are a useful alternative and with proven benefit as an add-on therapy to limit aortic growth. Neither beta-blockers, nor angiotensin receptor blockers have shown meaningful results on clinical aortic endpoints. The current evidence of pharmacological treatment for MFS patients is conflicting due to the lack of large, randomized clinical trials with adequate follow-up studies and homogeneous age grouping. Beta-blockers and angiotensin receptor blockers are the only available treatments to reduce aortic growth. A recently published patient-level meta-analysis confirmed that angiotensin receptor blockers and beta-blockers have a similar effect on reducing the rate of increase of the aortic root Z score, used singularly or as add-on therapy. Considering the current evidence on new features related with MFS (such as mitral annular disjunction - MAD) bearing a potential additional increased arrhythmic risk, it is of paramount importance to establish the role of beta-blockers and angiotensin receptor blockers in clinical endpoints of this population as well.


[Risk factors related to surgical site infection in elective surgery].
The risk factors for surgical site infections in surgery should be measured and monitored from admission to 30 days after the surgical procedure, because 30% of Surgical Site Infection is detected when the patient was discharged. Calculate the Relative Risk of associated factors to surgical site infections in adult with elective surgery. Patients were classified according to the surgery contamination degree; patient with surgery clean was defined as no exposed and patient with clean-contaminated or contaminated surgery was defined exposed. Risk factors for infection were classified as: inherent to the patient, pre-operative, intra-operative and post-operative. Statistical analysis; we realized Student t or Mann-Whitney U, chi square for Relative Risk (RR) and multivariate analysis by Cox proportional hazards. Were monitored up to 30 days after surgery 403 patients (59.8% women), 35 (8.7%) developed surgical site infections. The factors associated in multivariate analysis were: smoking, RR of 3.21, underweight 3.4 hand washing unsuitable techniques 4.61, transfusion during the procedure 3.22, contaminated surgery 60, and intensive care stay 8 to 14 days 11.64, permanence of 1 to 3 days 2.4 and use of catheter 1 to 3 days 2.27. To avoid all risk factors is almost impossible; therefore close monitoring of elective surgery patients can prevent infectious complications.


Bacterial adhesion to worn silicone hydrogel contact lenses.
The aim of this study was to, firstly, investigate whether silicone-hydrogel contact lenses (CL) are more or less susceptible to bacterial adhesion than conventional ones and, secondly, assess the influence of lens wear in the extent of bacterial adhesion. Four silicone-hydrogel CL (galyfilcon A, balafilcon A, lotrafilcon A, and lotrafilcon B) and one conventional hydrogel (etafilcon A) CL were tested. Bacterial adhesion experiments were performed on unworn and worn CL using the strain Staphylococcus epidermidis 9142. Worn lenses were obtained from a group of 31 subjects fitted with a silicone-hydrogel CL in one eye and a conventional hydrogel CL as contralateral pair. These lenses were used on a daily basis in combination with a multipurpose lens care solution. Adhesion assays were carried out in a parallel plate flow chamber, followed by image analysis. Hydrophobicity, roughness, and topography of the lenses surfaces were assessed through contact angle measurements and atomic force microscopy. Unworn conventional and silicone-hydrogel CL were equally susceptible to bacterial adhesion of S. epidermidis. Conversely, worn conventional hydrogel (etafilcon A) were more prone to bacterial adhesion than worn silicone-hydrogel materials, which exhibited similar adhesion extents among them. The results also showed that the lens surface properties such as hydrophobicity, roughness, and surface topography changed during wear. The alteration of surface hydrophobicity of silicone and conventional hydrogel CL during wear had a great impact on lens bacterial adhesion susceptibility. Accordingly, balafilcon A becomes significantly less hydrophobic and less prone to bacterial adhesion after lens wear, whereas etafilcon A becomes more hydrophobic and also more susceptible to bacterial adhesion (p < 0.05). Worn silicone-hydrogel galyfilcon A, balafilcon A, lotrafilcon A, and lotrafilcon B are equally prone to microbial adhesion of S. epidermidis and generally less susceptible than the conventional hydrogel.


Cerebellar ataxia in systemic lupus erythematosus.
Neuropsychiatric manifestations are commonly observed in systemic lupus erythematosus (SLE) patients; however, cerebellar involvement has rarely been reported. In the presence of acute cerebellar ataxia, etiologies related (focal edema and ischemia) and not related (infections, malignancy and paraneoplastic syndromes) to lupus have to be considered and they imply different treatment strategies. We report the clinical and radiological features of 3 SLE patients who presented with acute cerebellar ataxia. A review of the literature was performed by documenting cases of cerebellar ataxia in SLE and the importance of neuroimaging in the evaluation of these patients.


[Enhanced MR-angiography (CE-MRA) of the supraaortic arteries: dose reduction with a contrast agent of high relaxivity (Gd-BOPTA)].
Reducing the amount of contrast medium in contrast-enhanced MR-angiography (CE-MRA) of the cervical vessels could lead to considerable cost reduction. This study investigates whether the amount of contrast medium possessing high relaxivity can be reduced without loss of diagnostic information. In a prospective study the supraaortic vessels of 40 patients were examined on a 1.5 T MR-system using 10 or 20 ml Gd-BOPTA (MultiHance). The examinations were evaluated by measurement of the signal increase in the common carotid arteries in 38 patients. Furthermore, CE-MRA was rated by two independent observers, who were blinded to the amount of contrast medium and degree of stenosis. Rating criteria were quality of arterial contrast and image impression. Signal increase in the carotid artery was identical for 10 and 20 ml Gd-BOPTA in 12 patients with optimal bolus timing (10 ml: 454 +/- 58; 20 ml: 458 +/- 63). Signal intensities were significantly lower for 10 ml when bolus timing was suboptimal (281 +/- 80 vs 353 +/- 65; p < 0.02). The quality of the CE-MRA using 10 and 20 ml Gd- BOPTA was rated as identical by the observers, and the amount of contrast medium used could not be deduced from the images. Image quality and signal values of the carotid arteries did not differ in CE-MRA performed with either 10 ml or 20 ml Gd-BOPTA (MultiHance) as long as the examination is adequate. Thus, cost reduction can be achieved with a reduced standard dose of 10 ml Gd-BOPTA without loss of diagnostic information.


IgG receptors on the mast cells.
Antisera to the IgG subclasses, 1, 2a, 2b, and 2c, induced histamine release from mast cells obtained from the peritoneal washings of Lister hooded rats. The maximum responses obtained with anti-IgG1 and anti-IgG2a were as great as that for anti-IgE (more than 60% histamine release). Cells from unresponsive Wistar rats which did not secrete appreciable amounts of histamine in response to any of the antisera, produced on active sensitization with ovalbumin a small but significant response on challenge with anti-IgG1 and anti-IgG2b as well as with anti-IgE. Passive sensitization with rat myeloma serum of mast cells from the unresponsive rats produced a large response on challenge with anti-IgE but no release to the anti-IgG group 2 subclasses. IgE myeloma serum (1:1000) neutralized the histamine-releasing activity on anti-IgE serum (87% inhibition) and the antisera to all subclasses of IgG. When the IgE in the myeloma serum was inactivated by heating, the response to the IgG antisera remained completely inhibited except for anti-IgG2a where some reversal was observed. When purified myeloma IgE (30 microgram/ml) was used in place of whole serum, marked inhibition (86%) of the response to anti-IgE was obtained leaving the responses to the IgE subclasses unaffected (except for IgG2a, which was 65% inhibited).


Correction of a malunited vertical shear fracture : Staged sliding osteotomy of the pelvic ring.
The authors describe a posterior and anterior osteotomy of the pelvis for correction of a posttraumatic deformity in a 22-year-old female patient. As a shortening of the leg of 4 cm had occurred, surgery was performed in 2 stages. Between these 2 operations skeletal traction was employed. Main indications for this type of surgery included shortening of the leg, gait difficulties and poor posture while sitting. However, poor general health is considered to be a contraindication for such a risky surgery. During the surgical procedure all soft tissues preventing an proper correction have to be dissected or detached, particularly the sacrospinal and sacrotuberal ligaments. Screw fixation guarantees the correction. Exercises can be started as soon as the fragments become stable. Injuries to vessels and nerves are the most frequently encountered complications.The patient was symptom-free after 1 year postoperatively, mobility of both hips and her gait were normal. The author could observe a complete remodelling of the osteotomies after 1 year.


From global action against malaria to local issues: state of the art and perspectives of web platforms dealing with malaria information.
Since prehistory to present times and despite a rough combat against it, malaria remains a concern for human beings. While evolutions of science and technology through times allowed for some infectious diseases eradication in the 20th century, malaria resists. This review aims at assessing how Internet and web technologies are used in fighting malaria. Precisely, how do malaria fighting actors profit from these developments, how do they deal with ensuing phenomena, such as the increase of data volume, and did these technologies bring new opportunities for fighting malaria? Eleven web platforms linked to spatio-temporal malaria information are reviewed, focusing on data, metadata, web services and categories of users. Though the web platforms are highly heterogeneous the review reveals that the latest advances in web technologies are underused. Information are rarely updated dynamically, metadata catalogues are absent, web services are more and more used, but rarely standardized, and websites are mainly dedicated to scientific communities, essentially researchers. Improvement of systems interoperability, through standardization, is an opportunity to be seized in order to allow real time information exchange and online multisource data analysis. To facilitate multidisciplinary/multiscale studies, the web of linked data and the semantic web innovations can be used in order to formalize the different view points of actors involved in the combat against malaria. By doing so, new malaria fighting strategies could take place, to tackle the bottlenecks listed in the United Nation Millennium Development Goals reports, but also specific issues highlighted by the World Health Organization such as malaria elimination in international borders.


Variation in methods used to determine national mean DMFT scores for 12-year-old children in European countries.
The aim of this study was to investigate the methods used to identify national mean DMFT scores for 12-year-old children in all the Member States of the European Union and European Economic Area, and in 11 other European countries. The most recent national mean DMFT scores were accessed from the World Health Organisation Oral Health CAPP and the Council of European Chief Dental Officers databanks. A literature search was then performed to access the reports of the studies that had produced these DMFT scores, cited on these databanks. The reports were then analysed to determine: the year in which the survey/study that produced the score took place, the year the results were published, the geographical area (national, regional or local) covered, the number of children examined, how many examiners took part, how they were trained and calibrated, and the criteria used for the detection of caries. Data and information from 43 European countries were accessed. The years when the studies were performed ranged from 1990 to 2014. There were doubts over the representativeness of some samples. A wide range of different methods were used. Examiner training and calibration were very variable both in terms of duration and reported inter and intra-examiner consistency. There were important variations in the criteria employed for the detection of caries. These findings support the view that most of current national caries data for DMFT levels in 12-year-old children are not comparable across Europe.


Cough and spontaneous rupture of a normal spleen.
Rupture of the spleen is a relatively common complication of trauma and many systemic disorders affecting the reticuloendothelial system, including infections and neoplasias. A rare subtype of rupture occurring spontaneously and arising from a normal spleen was recognized as a distinct clinicopathologic entity. The pathogenesis is debated in the absence of external trauma or predisposing disease. It has been reported in association with apparently trivial insults such as vomiting. We report a case of a patient with spontaneous rupture of a normal spleen. Interestingly, it was observed after severe coughing.


Killing of different strains of Toxoplasma gondii tissue cysts by irradiation under defined conditions.
To study the effect of gamma-irradiation on the viability of Toxoplasma gondii tissue cysts, brains of mice inoculated with 95 newly isolated strains of T. gondii from pigs and 10 other laboratory isolates were pooled, flattened, packed in vacuum, and irradiated to absorbed doses of 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, and 0.9 kGy at 5 C (+/- 0.5 C) with a 137Cs gamma-radiation source. Treated samples were bioassayed for viable T. gondii in mice, cats, or both. Tissue cysts of all strains were rendered nonviable at 0.4 kGy. To study the effect of temperature on inactivation by irradiation, tissue cysts were irradiated at -4, 0, 4, 8, 12, and 16 C (+/- 0.5 C) at 0.25 kGy. Temperature during irradiation had no marked effect on the viability of tissue cysts.


Recent Advances and Challenges in 2D Metal-Free Electrocatalysts for N2 Fixation.
The ever-growing requirement of ammonia for industry and energy supply motivates people to find clean and cost-effective alternatives to overcome the shortcomings of the century-old Haber-Bosch process. Electrocatalytic N2 reduction (NRR) is considered a prospective way for ammonia production at ambient conditions. Recently, two-dimensional (2D) metal-free materials are emerging as highly efficient and robust electrocatalysts for NRR owing to their advanced features: highly exposed surface, abundant active sites, tunable electronic states, and long-term stability compared to metal-based and bulk catalysts. In this minireview, we briefly summarize the latest edge reports on 2D metal-free materials catalyzed NRR. Also, we discuss the challenges and perspectives on the design and synthesis of novel 2D metal-free catalysts.


Determining the level of stromal and epithelial cells activity in normal and hyperplastic endometrium of late reproductive and perimenopausal women.
Hyperplastic processes of the endometrium (HPE) are a group of benign endometrial and stromal cells that have undergone altered growth. This study aimed to investigate the potential role of hypoxia (as indicated by Hif-1α) and apoptosis markers (p53 and BCL-2) in the development of hyperplastic processes of the endometrium (HPE). Results showed that endometrial cells with atypical hyperplasia had increased levels of Hif-1ɑ, which indicates the presence of endometrial hypoxia and may trigger pathological manifestations. Though this result was not statistically significant, it could be the cause of atypia hyperplasia in the late reproductive period (Hif-1ɑ=1.89±0.09 units) and the perimenopausal period (Hif-1ɑ=2.09±0.07 units). Additionally, the study found that p53 markers were elevated in epithelial cells in the late reproductive period, and similar patterns were observed in the perimenopausal period, with the biggest expression in atypical hyperplasia. The study also found that the high expression of BCL-2 indicator (+++) was less common in late reproductive period women with atypia than those without it (χ2=7.2 p=0.01). A similar situation was observed in women in the perimenopausal period (χ2=4.2 p=0.04). These findings suggest that hypoxia may play a role in the development of HPE, as well as changes in apoptotic markers present in the endometrial tissue.


Circulating human peripheral blood granulocytes synthesize and secrete tumor necrosis factor alpha.
Circulating peripheral blood polymorphonuclear neutrophils (PMNs) have long been considered terminally differentiated cells that do not synthesize or secrete protein. However, work of others and ourselves has shown that PMNs can secrete the cytokine interleukin 1. In the present study we investigated whether circulating PMNs are capable of synthesizing and secreting another cytokine, tumor necrosis factor alpha (TNF-alpha). Highly purified (greater than 99% granulocytes) PMNs were isolated from normal human volunteer blood and cultured with or without bacterial lipopolysaccharide (LPS) for up to 24 hr. Cell culture supernatants were collected and tested for TNF-alpha, and total RNA was isolated from cells at various times after stimulation and assessed for TNF-alpha mRNA by Northern blot techniques. The results showed that message for TNF-alpha was produced after 60 min of in vitro stimulation with LPS and was maximal at about 4 hr. TNF-alpha was secreted into the supernatant of unstimulated PMNs from two different donors during 24 hr of culture (35-50 pg/ml), but significantly more (160-190 pg/ml) was secreted by PMNs when stimulated with LPS. PMNs from six other normal volunteers showed significant LPS-stimulated secretion of TNF at 60-180 min of culture. The secreted product also had biological activity against the TNF-sensitive L-M cell line, confirming that PMNs can make and secrete immunologically and biologically active TNF. Since it is also possible for monocytes to synthesize and secrete TNF, the amount of TNF secreted by a monocyte population equal to 20% of the PMNs cultured was measured. The results showed that monocytes at a concentration 20 times that potentially contaminating the PMN populations cultured could not produce as much TNF (unstimulated, 26-65 pg/ml; stimulated, 32-87 pg/ml). The PMN must now be considered a cell capable of altering the acute inflammatory response and modulating the immune response through the synthesis and release of cytokines.


Is it that difficult to find a good preference order for the incremental algorithm?
In a recent article published in this journal (van Deemter, Gatt, van der Sluis, & Power, 2012), the authors criticize the Incremental Algorithm (a well-known algorithm for the generation of referring expressions due to Dale & Reiter, 1995, also in this journal) because of its strong reliance on a pre-determined, domain-dependent Preference Order. The authors argue that there are potentially many different Preference Orders that could be considered, while often no evidence is available to determine which is a good one. In this brief note, however, we suggest (based on a learning curve experiment) that finding a Preference Order for a new domain may not be so difficult after all, as long as one has access to a handful of human-produced descriptions collected in a semantically transparent way. We argue that this is due to the fact that it is both more important and less difficult to get a good ordering of the head than of the tail of a Preference Order.


Chemical Changes Occur in Extra-Virgin Olive Oil during Fruits Ripeness of Zalmati Cultivar Planted in Warm Desert Climate.
The evaluation of the impact of olive ripening degree on the stability of extra virgin olive oils not only by determining the oxidative stability index, the 1-diphenyl-2-picrylhydrazyl (DPPH) radical test, but also by studying the variation of their sensory profiles, plays a key role in the assessment of the overall olive oil quality. To this end, Zalmati extra virgin olive oils samples were taken at three harvest dates relating to three ripening stages. The obtained results revealed that several analytical parameters, namely free fatty acids, peroxide index, K232 and K270, augment during repining. Besides, a substantial reduction was noticed for the monounsaturated fatty acid, chlorophyll, and carotenoids in oils produced at the advanced ripening stage. Principal component analysis (PCA) was employed to the data of the analytical analysis to delve into their capacity for discerning the oil quality consistently with the harvesting period of the Zalmati olives. Familiar plots have indicated that different olive oils are categorized in diverse harvest period groups, each of which could be obviously recognized.


Isolation of a retrovirus from the American bison and its relation to bovine retroviruses.
Tissue samples were removed at necropsy from five American bison (Bison bison) with clinical signs of a disease resembling malignant catarrhal fever (MCF). Using cell-associated virus techniques, attempts were made to isolate viruses from these tissues by culturing them directly or by co-culture with bovine fetal cells. Among the viruses isolated was one which was syncytiogenic and multiplied in bovine fetal spleen cells and remained highly cell-associated. The presence of reverse transcriptase activity indicated that it was a retrovirus. Also, it had antigenic cross activity with bovine syncytial virus, but not with bovine leukemia or bovine maedi-like retroviruses. We do not attribute a direct causative role of this retrovirus to MCF, but indirect relationships are possible.


N-nitroso-2-acetylaminofluorene: a direct-acting carcinogen inducing hepatocellular carcinoma in Sprague-Dawley rats.
To compare the hepatotoxicity and hepatocarcinogenicity of N-nitroso-2-acetylaminofluorene (NO-AAF) and its parent compound, 2-acetylaminofluorene (AAF), male Sprague-Dawley rats were given intraperitoneal (i.p.) or subcutaneous (s.c.) injections of AAF or NO-AAF (60 mg/kg body weight/week) for ten months. In the AAF group, morphological changes were produced which involved gross distortions of the liver with multiple nodule formations. The rat livers in the NO-AAF group appeared to be smooth with a blunt-thick superior segment of the lateral lobe. The serum gamma-glutamyl transpeptidase activity in both the AAF group and the NO-AAF group was significantly elevated (P < 0.0005). The present study shows that i.p. and s.c. injections of NO-AAF resulted in a high incidence of well-differentiated hepatocellular carcinomas (HCC) (7/9 and 4/6, respectively), while poorly differentiated HCCs were induced by i.p. or s.c. administration of AAF (6/9 or 2/6, respectively). Subcutaneous lesions consisting of an inflammatory reaction and fibroadenoma formation were observed in the NO-AAF-treated rats, whereas no such skin lesions were detected in the AAF-treated animals. These results suggest that NO-AAF is a new direct-acting carcinogen which may be useful for investigating hepatocarcinogenesis.


MicroRNA-302a inhibits osteosarcoma cell migration and invasion by directly targeting IGF-1R.
Osteosarcoma is one of the most frequent types of primary malignant bone neoplasm in children and adolescents. Despite advancements developed in therapeutic modalities, the 5-year overall survival rates for patients with metastatic osteosarcoma disease remain poor. The present study aimed to investigate the expression level of microRNA-302a (miR-302a) in osteosarcoma tissues and cell lines, and the biological roles of miR-302a in osteosarcoma cells. In addition, the molecular mechanism underlying its tumor suppressive roles was evaluated. miR-302a expression in osteosarcoma tissues and cell lines was detected using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Following transfection of miR-302a mimics or IGF-1R siRNA, transwell migration and invasion, luciferase reporter assay RT-qPCR and western blot assays were conducted in osteosarcoma cells. In the present study, the data demonstrated that miR-302a was frequently reduced in osteosarcoma tissue and cell lines. In addition, the expression of miR-302a was correlated with metastatic features of patients with osteosarcoma. Restoration of miR-302a expression significantly inhibited the migration and invasion capacity of osteosarcoma cells. Mechanistic studies indicated that insulin-like growth factor 1 receptor (IGF-1R) was a direct target gene of miR-302a. Overexpression of miR-302a resulted in decreased expression of IGF-1R at the mRNA and protein levels. Furthermore, the knockdown IGF-1R mimicked the functions of miR-302a overexpression on osteosarcoma cell migration and invasion. Collectively, the results of the current study indicate that miR-302a acts as a metastasis suppressing miRNA and could be investigated as a therapeutic target for the treatment of patients with osteosarcoma to prevent metastasis.


Movement Optimization for a Cyborg Cockroach in a Bounded Space Incorporating Machine Learning.
Cockroaches can traverse unknown obstacle-terrain, self-right on the ground and climb above the obstacle. However, they have limited motion, such as less activity in light/bright areas and lower temperatures. Therefore, the movement of the cyborg cockroaches needs to be optimized for the utilization of the cockroach as a cyborg insect. This study aims to increase the search rate and distance traveled by cockroaches and reduce the stop time by utilizing automatic stimulation from machine learning. Multiple machine learning classifiers were applied to classify the offline binary classification of the cockroach movement based on the inertial measuring unit input signals. Ten time-domain features were chosen and applied as the classifier inputs. The highest performance of the classifiers was implemented for the online motion recognition and automatic stimulation provided to the cerci to trigger the free walking motion of the cockroach. A user interface was developed to run multiple computational processes simultaneously in real time such as computer vision, data acquisition, feature extraction, automatic stimulation, and machine learning using a multithreading algorithm. On the basis of the experiment results, we successfully demonstrated that the movement performance of cockroaches was importantly improved by applying machine learning classification and automatic stimulation. This system increased the search rate and traveled distance by 68% and 70%, respectively, while the stop time was reduced by 78%.


The use of methylmethacrylate cement as an instantaneous fusion mass in posterior cervical fusions: a canine in vivo experimental model.
The authors previously predicted the failure of posterior cervical fusions utilizing methylmethacrylate cement as an instantaneous "fusion" mass, based on research using an in vitro canine experimental model. This report describes the results of in vivo canine studies on the same subject. Three groups of dogs had application of a posterior C4-C5 20-gauge cerclage wire and autologous iliac crest bone graft; application of a posterior C4-C5 20-gauge cerclage wire and methylmethacrylate cement; or application of a C4-C5 20-gauge cerclage wire only. This group represented the control group. The dogs were allowed to live for 3 months postoperatively, at which time they were killed and their spine fusions studied radiologically, mechanically, and histologically. Five of the bone fusions united solidly radiologically. Their flexion stability was statistically superior to the others. Histologic studies confirmed solid union of the fusion mass to the underlying bone. Four of the six methylmethacrylate fusions demonstrated cerclage wire fracture and methacrylate-bone separation by the second postoperative month. At the time the dogs were killed, their flexion stability was statistically inferior to the bone fusions and tended to be inferior to the controls as well. Histologically, fibrous tissue was noted to have grown between the methacrylate "fusion" mass and the underlying bone. This work provides a mechanical explanation for the well-known success of the traditional bony fusion. It further supports our original prediction regarding the failure of methylmethacrylate "fusions."


Photosynthetic response to globally increasing CO2 of co-occurring temperate seagrass species.
Photosynthesis of most seagrass species seems to be limited by present concentrations of dissolved inorganic carbon (DIC). Therefore, the ongoing increase in atmospheric CO2 could enhance seagrass photosynthesis and internal O2 supply, and potentially change species competition through differential responses to increasing CO2 availability among species. We used short-term photosynthetic responses of nine seagrass species from the south-west of Australia to test species-specific responses to enhanced CO2 and changes in HCO3 (-) . Net photosynthesis of all species except Zostera polychlamys were limited at pre-industrial compared to saturating CO2 levels at light saturation, suggesting that enhanced CO2 availability will enhance seagrass performance. Seven out of the nine species were efficient HCO3 (-) users through acidification of diffusive boundary layers, production of extracellular carbonic anhydrase, or uptake and internal conversion of HCO3 (-) . Species responded differently to near saturating CO2 implying that increasing atmospheric CO2 may change competition among seagrass species if co-occurring in mixed beds. Increasing CO2 availability also enhanced internal aeration in the one species assessed. We expect that future increases in atmospheric CO2 will have the strongest impact on seagrass recruits and sparsely vegetated beds, because densely vegetated seagrass beds are most often limited by light and not by inorganic carbon.


Effectiveness of a health education program on mothers' knowledge and practice regarding amebiasis.
This study was conducted to assess the effectiveness of a health education program on mothers' knowledge and practices related to amebiasis. Pre- and post-I and II-tests were used, along with a two-group quasi-experimental design. The study included a convenience sample of 120 mothers, including 60 from the study and 60 from the control group. Out of the entire number of PHCs in Wasit Governorate, seven were used for the study. A three-part structured questionnaire tests mothers' knowledge of the illness and attitudes about it. The results of this study indicated that there is statistically significant difference between the study and control groups during the post-test I and post-test II periods (t=14.968, P=.000 and t=13.731, P=.000, respectively). Data from the study group during the pre-test period of 31.17(9.14) revealed that (70%) of mothers had bad practices connected to Entameba Histolytica among children. According to the post-test I results, 47.4 (9.48) respondents, or 70%, identified good practices. The main finding showed that, after the educational program was implemented, mothers' knowledge and practice of amebiasis dramatically increased, with pre-post implementation differences that are statistically significant.


Impacts of temporary traffic control measures on vehicular emissions during the Asian games in Guangzhou, China.
To guarantee good traffic and air quality during the 16th Asian Games in Guangzhou, China, the government carried out two traffic control Drills before the Games and adopted traffic control measures during the Games. Vehicle activities before and during the first and second Drills, and during the Games, were surveyed. Based on the data under investigation, the impacts of control measures on traffic volumes and driving characteristics were analyzed during the first and second Drills, and the Games. The emission reduction of traffic control measures was also evaluated during the three stages using the MOBILE-China model. The results show that there were significant effects of implementing temporary traffic control measures on transportation activity and vehicular emissions. During the first and second Drills, and the Games, the average traffic volumes in monitored roads decreased, and the average speed of vehicles increased significantly The co-effects of traffic flow reduction, traffic congestion improvement, and the banning of high-emitting vehicles helped to greatly reduce the estimated emissions from motor vehicles in Guangzhou during the first and second Drills, and the Games. Estimated vehicular emissions were reduced by 38-52% during the first Drill and 28-36% for the second Drill. During the Asian Games, vehicular emissions of carbon monoxide (CO), hydrocarbon (HC), oxides of nitrogen (NO), and particulate matter with an aerodynamic diameter < 10 microm (PM10) reduced by an estimated 42%, 46%, 26%, and 30%, respectively, compared with those before the Games. Both the banning of high-emitting vehicles and the travel restrictions imposed by use of odd-even licenses had significant effects on the reduction of vehicular emissions of CO, HC, NOx, and PM10. Motor vehicles have become the most prevalent source of emissions and subsequently air pollution within Chinese cities. Understanding the impacts that different control measures have on vehicular emissions is very important in order to be able to control vehicle emissions. The results of this study will be very helpful for the further control of vehicle emissions in Guangzhou in the future. In addition, the effects of temporary transportation control measures will provide important awareness to other cities that will be hosting large-scale activities similar to the Asian Games.


High-CBD cannabis extracts inhibit the expression of proinflammatory factors via miRNA-mediated silencing in human small intestinal epithelial cells.
The incidence of chronic inflammatory disorders and autoimmune diseases is rapidly growing. To date, the COVID-19 pandemic caused by SARS-CoV-2 has killed over 6,209,000 people globally, while no drug has been proven effective for the disease. Screening natural anti-inflammatory compounds for clinical application has drawn much attention. In this study, we showed that high-CBD cannabis extracts #1, #5, #7, #169, and #317 suppressed the levels of expression of proinflammatory cyclooxygenase 2 (COX2) and increased the expression of the anti-inflammatory suppressor of cytokine signaling 3 (SOCS3) in human small intestinal epithelial cells (HSIEC) in TNFα/IFNγ-triggered inflammation. We revealed that these extracts, with the exception of extract #169, also profoundly attenuated induction of proinflammatory cytokines interleukin-6 (IL-6) and/or IL-8 proteins through miR-760- and miR-302c-3p-mediated silencing. The prevalent components in extracts #1 and #7 influenced the levels of IL-8 both individually as well as in combination with each other. However, the high-dose cannabis extracts displayed an inhibitory effect in the growth of HSIEC cells. These results show that our high-CBD cannabis extracts decrease the levels of proinflammatory molecules COX2, IL-6, and IL-8 via transcriptional suppression or miRNA-mediated silencing, highlighting their potential against COVID-19-associated cytokine storm syndrome.


Effects of rubidium, caesium, strontium, barium and lanthanum on ionic currents in myelinated nerve fibres from Xenopus laevis.
A comparative analysis of the effects of externally applied Rb+, Cs+, Ba2+ and La3+ on myelinated fibres under potential clamp conditions was made. The ions Cs+, Ba2+ and La3+ (belonging to groups 1a, 2a and 3a of period 6 in the periodic system) were found to affect the K system in specific ways. Ba2+ and La3+ reduced the outward K+ current associated with positive potential steps; the effect of La3+ being larger than that of Ba2+. The effect of Ba2+ depended on the K+ concentration. Cs+, Ba2+ and La3+ reduced the inward K+ tail current at repolarisation in solutions with high K+ concentration. Cs+ and Ba2+ caused a rectification of the K+ current, i.e. they reduced the inward tail current proportionally more than the outward steady state current. The effect of Ba2+ could not be described as a simple reduction of the permeability constant PK. Rb+ was found to affect the time course of the K system by increasing the rate constant beta n. Sr2+, Ba2+ and La3+ reduced the permeability constant PNa. These ions also shifted potential dependent parameters in general along the potential axis. The effect of La3+ was larger than that of Sr2+ and Ba2+. Cs+, Ba2+ and La3+ reduced the p-current, Ba2+ and La3+ the leak conductance.


Protective role of α-lipoic acid in hyperuricemia-induced endothelial dysfunction.
The aim of the current study was to determine the effects of α-lipoic acid (LA) on hyperuricemia and endothelial dysfunction, and to uncover the underlying mechanism of its action. A hyperuricemic rat model was established by administration of uric acid (UA) and the rats were orally fed with 2 g/kg/day LA or phosphate-buffered saline. Primary rat aortic endothelial cells were subsequently isolated, and a cell viability assay, apoptosis assay, enzyme nitric oxide synthase (eNOS) activity assay and mitochondrial function assay were all performed. For the in vitro study, human umbilical vein endothelial cells were used and western blotting was performed to assess Akt signaling activity. The results of the current study indicated that LA inhibited apoptosis, enhanced eNOS activity and production of nitric oxide (NO), and rescued mitochondrial mass and function in uric acid (UA)-treated endothelial cells. LA activated Akt signaling and inhibition of Akt signaling abolished the effects of LA on cell viability, NO production, ROS production and ATP levels in UA-treated endothelial cells. Therefore, the current study demonstrated that LA attenuated oxidant stress and inhibited apoptosis in UA-treated endothelial cells by activating Akt signaling. The results indicate that LA may serve as a therapeutic approach to treat hyperuricemia-induced endothelial dysfunction.


Spirocyclic Thiohydantoin Antagonists of F877L and Wild-Type Androgen Receptor for Castration-Resistant Prostate Cancer.
Androgen receptor (AR) transcriptional reactivation plays a key role in the development and progression of lethal castration-resistant prostate cancer (CRPC). Recurrent alterations in the AR enable persistent AR pathway signaling and drive resistance to the treatment of second-generation antiandrogens. AR F877L, a point mutation in the ligand binding domain of the AR, was identified in patients who acquired resistance to enzalutamide or apalutamide. In parallel to our previous structure-activity relationship (SAR) studies of compound 4 (JNJ-pan-AR) and clinical stage compound 5 (JNJ-63576253), we discovered additional AR antagonists that provide opportunities for future development. Here we report a highly potent series of spirocyclic thiohydantoins as AR antagonists for the treatment of the F877L mutant and wild-type CRPC.


A rare complication of infective endocarditis: left main coronary artery embolization resulting in sudden death.
Whilst systemic septic embolization is a common complication of infective endocarditis, a second, less frequently reported, complication is the embolization of vegetative material to the coronary arteries that rarely results in acute myocardial infarction and sudden death. The case is presented of an acute cardiac death in a patient with aortic valve endocarditis, caused by acute occlusion of the left main coronary artery with vegetative material.


Visceral obesity and diabetes.
The cross-sectional subcutaneous and visceral areas of adipose tissue, measured on computed tomographic scans at abdominal level and the subcutaneous/visceral fat area ratio (S/V ratio) distribution have been calculated in 155 subjects. Using the S/V ratio as discriminator, two groups of patients of both sexes, with subcutaneous or visceral type of obesity have been selected. Metabolic studies demonstrate a significant reduction of glucose tolerance at the oral glucose load and a significant increase of the insulin response in patients with visceral obesity. The size of the omental fat cells was larger in visceral obesity, while the size of subcutaneous fat cells was larger in subcutaneous obesity. A significant correlation was found between the S/V area ratios and the subcutaneous/visceral fat cell weight ratios. In vitro studies on subcutaneous and omental fat obtained during surgery from 9 patients with subcutaneous obesity and 7 patients with visceral obesity demonstrate a significantly higher isoprenaline-stimulated lipolytic response in omental fat of patients with visceral obesity. The lipoprotein lipase activity in subcutaneous fat was higher than in omental fat in both types of obesity.


Bacillus subtilis biosensor engineered to assess meat spoilage.
Here, we developed a cell-based biosensor that can assess meat freshness using the Gram-positive model bacterium Bacillus subtilis as a chassis. Using transcriptome analysis, we identified promoters that are specifically activated by volatiles released from spoiled meat. The most strongly activated promoter was PsboA, which drives expression of the genes required for the bacteriocin subtilosin. Next, we created a novel BioBrick compatible integration plasmid for B. subtilis and cloned PsboA as a BioBrick in front of the gene encoding the chromoprotein amilGFP inside this vector. We show that the newly identified promoter could efficiently drive fluorescent protein production in B. subtilis in response to spoiled meat and thus can be used as a biosensor to detect meat spoilage.


A new calcium binding glycoprotein family constitutes a major diatom cell wall component.
Diatoms possess silica-based cell walls with species-specific structures and ornamentations. Silica deposition in diatoms offers a model to study the processes involved in biomineralization. A new wall is produced in a specialized vesicle (silica deposition vesicle, SDV) and secreted. Thus proteins involved in wall biogenesis may remain associated with the mature cell wall. Here it is demonstrated that EDTA treatment removes most of the proteins present in mature cell walls of the marine diatom Cylindrotheca fusiformis. A main fraction consists of four related glycoproteins with a molecular mass of approximately 75 kDa. These glycoproteins were purified to homogeneity. They consist of repeats of Ca2+ binding domains separated by polypeptide stretches containing hydroxyproline. The proteins in the EDTA extract aggregate and precipitate in the presence of Ca2+. Immunological studies detected related proteins in the cell wall of the freshwater diatom Navicula pelliculosa, indicating that these proteins represent a new family of proteins that are involved in the biogenesis of diatom cell walls.


Cost benefit analysis of screening spina bifida.
Cost benefit analysis of establishing and running a mass serum alpha-fetoprotein (AFP) screening service for neural tube defects (NTDS) in the Auckland Hospital Board area has been undertaken and a cost benefit index derived. From a population of 14 066 pregnant women per year, assuming present booking patterns continue, 8017 women could be screened for NTDS. Such screening could reduce severe spina bifida in liveborn babies by 38.8 percent.


Age variation in the prevalence of DSM-IV disorders in cases of suicide of middle-aged and older persons in Sydney.
Data concerning 127 persons aged 35 years or above who died by suicide (as determined in consecutive cases by a Sydney coroner) were analyzed. Psychological autopsy (PA) interviews were conducted in 52 cases, and details were compared with the 75 cases where data were available only from coroner's files (CF). Most characteristics of the two groups were similar, although more CF suicide victims were of Asian background and unable to speak English fluently. Consensus diagnoses were reached following detailed discussion about PA and CF cases. Logistic regression showed no significant difference between age-groups in the proportion diagnosed with major depression, which contrasts with the results of an earlier U.S. study.


Single-particle fluorescence intensity fluctuations of carbon nanodots.
Fluorescent carbon nanodots (CNDs) were synthesized in oxidized and reduced forms and were analyzed at the single-particle level. Images of single CNDs at different excitation energies revealed significant heterogeneity in the lower energy trap sites between particles. We observed that a high percentage of reduced CND particles transitioned between multiple fluorescence intensity levels indicative of multichromophoric systems. Despite this behavior, individual CNDs exhibit single-step photobleaching and transient blinking to the background level suggesting single-molecule behavior.


Chloride channel inhibition blocks the protection of ischemic preconditioning and hypo-osmotic stress in rabbit ventricular myocardium.
The objective of this study was to examine the role of chloride (Cl-) channels in the myocardial protection of ischemic preconditioning (IP). Isolated rabbit ventricular myocytes were preconditioned with 10-minute simulated ischemia (SI) and 20-minute simulated reperfusion (SR) or not preconditioned (control). The myocytes then received 180-minute SI or 45-minute SI/120-minute SR. Indanyloxyacetic acid 94 (IAA-94, 10 micromol/L) or 5-nitro-2-(3-phenylpropylamino)benzoic acid (NPPB, 1 micromol/L) was administered before IP or before SI or SI/SR to inhibit Cl- channels. Electrophysiological studies indicate that these drugs, at the concentrations used, selectively abolished Cl- currents activated under hypo-osmotic conditions (215 versus 290 mOsm). IP significantly (P<0.001) reduced the percentage of dead myocytes after 60-minute (30.8+/-1.3%, mean+/-SEM), 90-minute (35.3+/-1.3%), and 120-minute (39.2+/-1.7%) SI compared with controls (44.7+/-1.6%, 54.5+/-1.3%, and 58.9+/-1.8%, respectively) and after 45-minute SI/120-minute SR (36.3+/-0.6%) compared with control (56.6+/-2.2%). Hypo-osmotic stress also produced protection similar to IP. IAA-94 or NPPB abolished the protection of both IP and hypo-osmotic stress. In buffer-perfused rabbit hearts preconditioned with three 5-minute ischemia/10-minute reperfusion cycles given before the 40-minute long ischemia and 60-minute reperfusion, IP significantly (P<0.0001) reduced infarct size (IP+vehicle, 4.7+/-0.9%, versus control+vehicle, 26.6+/-3.3%; mean+/-SEM). Again, IAA-94 or NPPB abolished the protection of IP. Our results implicate Cl- channels in the IP protection of the myocardium against ischemic/reperfusion injury and demonstrate that hypo-osmotic stress is capable of preconditioning cardiomyocytes.


Novel asthma-associated genes from genome-wide association studies: what is their significance?
Asthma is a complex syndrome leading to the easily recognizable clinical feature of wheezing. This is believed to be caused by inflammation, constriction of the airway smooth muscles, and extensive mucus secretion. However, the mechanisms initiating and maintaining these simple clinical features are not well understood, even after 100 years of asthma research. Still worse is our understanding of why certain individuals develop asthma and others do not.


Current Approaches for Dissolution Similarity Assessment, Requirements, and Global Expectations.
This report summarizes podium presentations and breakout sessions from the second day of the 2019 M-CERSI workshop on In Vitro Dissolution Similarity Assessment in Support of Drug Product Quality: What, How, and When? Presenters from the U.S. Food and Drug Administration (FDA), Health Canada (HC), European Medicines Agency (EMA), Brazilian Health Surveillance Agency (ANVISA), and the pharmaceutical industry shared experiences/concerns with dissolution profile similarity assessment supporting minor/moderate Chemistry, Manufacturing and Control (CMC) changes. Members from regulatory agencies explained that dissolution profile similarity testing is only part of the overall assessment of the acceptability of the proposed changes; decisions are usually made based on aggregate weight of evidence. Scientific shortcomings of f2 were highlighted but no proposal on how to replace it was made. Controlling dissolution timepoint variability and application of pairwise batch-to-batch comparisons (PBC) of dissolution profiles caused considerable debate. Several industry participants suggested increased sample sizes to raise confidence in decision-making and to avoid PBC. They proposed identification of a single mathematical method with predefined acceptance criteria and suggested that dissolution timepoint selection should follow EMA and HC guidance. A majority of meeting attendees favored applying clinically relevant dissolution specifications (CRDS) and dissolution safe space to determine the impact of minor/moderate CMC changes as opposed to dissolution profile similarity assessment via statistical methods. Day 2 of the workshop highlighted the need and opportunities for global harmonization including variability, timepoint selection, role of CRDS, and statistical methods to address the ambiguity globally operating pharmaceutical companies are currently facing.


Study of the prevalence of 5 HT-2C receptor gene polymorphisms in Egyptian patients with lifelong premature ejaculation.
We investigated the prevalence of 5HT2C receptor gene polymorphisms in Egyptian patients with lifelong premature ejaculation. A total of 350 participants were enrolled in a prospective study. Two hundred and forty-five cases with lifelong premature ejaculation joined this study, in addition to 105 controls. We instructed the partners of the cases to measure the IELT of the first intercourse only using a stopwatch for 1 month. Genotyping was carried out at the end of the study. The results showed that the majority of the patients and controls were Cys/Cys. A highly significant statistical association was found between the studied gene polymorphisms and IELT among cases (p-values = .009). The study emphasised the potential role of 5HT2C receptor gene polymorphisms in patients with lifelong premature ejaculation.


Anti-prion activity of Brilliant Blue G.
Prion diseases are fatal neurodegenerative disorders with no effective therapy currently available. Accumulating evidence has implicated over-activation of P2X7 ionotropic purinergic receptor (P2X7R) in the progression of neuronal loss in several neurodegenerative diseases. This has led to the speculation that simultaneous blockade of this receptor and prion replication can be an effective therapeutic strategy for prion diseases. We have focused on Brilliant Blue G (BBG), a well-known P2X7R antagonist, possessing a chemical structure expected to confer anti-prion activity and examined its inhibitory effect on the accumulation of pathogenic isoforms of prion protein (PrPres) in a cellular and a mouse model of prion disease in order to determine its therapeutic potential. BBG prevented PrPres accumulation in infected MG20 microglial and N2a neural cells at 50% inhibitory concentrations of 14.6 and 3.2 µM, respectively. Administration of BBG in vivo also reduced PrPres accumulation in the brains of mice with prion disease. However, it did not appear to alleviate the disease progression compared to the vehicle-treated controls, implying a complex role of P2X7R on the neuronal degeneration in prion diseases. These results provide novel insights into the pathophysiology of prion diseases and have important implications for the treatment.


Autophagic Degradation of Gasdermin D Protects against Nucleus Pulposus Cell Pyroptosis and Retards Intervertebral Disc Degeneration In Vivo.
Intervertebral disc degeneration (IDD) is the primary culprit of low back pain and renders heavy social burden worldwide. Pyroptosis is a newly discovered form of programmed cell death, which is also involved in nucleus pulposus (NP) cells during IDD progression. Moderate autophagy activity is critical for NP cell survival, but its relationship with pyroptosis remains unknown. This study is aimed at investigating the relationship between autophagy and pyroptotic cell death. The pyroptosis executor N-terminal domain of gasdermin D (GSDMD-N) and inflammation-related proteins were measured in lipopolysaccharide- (LPS-) treated human NP cells. Inhibition of autophagy by siRNA transfection and chemical drugs aggravated human NP cell pyroptosis. Importantly, we found that the autophagy-lysosome pathway and not the proteasome pathway mediated the degradation of GSDMD-N as lysosome dysfunction promoted the accumulation of cytoplasmic GSDMD-N. Besides, P62/SQSTM1 colocalized with GSDMD-N and mediated its degradation. The administration of the caspase-1 inhibitor VX-765 could reduce cell pyroptosis as confirmed in a rat disc IDD model in vivo, whereas ATG5 knockdown significantly accelerated the progression of IDD. In conclusion, our study indicated that autophagy protects against LPS-induced human NP cell pyroptosis via a P62/SQSTM1-mediated degradation mechanism and the inhibition of pyroptosis retards IDD progression in vivo. These findings deepen the understanding of IDD pathogenesis and hold implications in unraveling therapeutic targets for IDD treatment.


Green tea extract attenuates muscle loss and improves muscle function during disuse, but fails to improve muscle recovery following unloading in aged rats.
In this study we tested the hypothesis that green tea extract (GTE) would improve muscle recovery after reloading following disuse. Aged (32 mo) Fischer 344 Brown Norway rats were randomly assigned to receive either 14 days of hindlimb suspension (HLS) or 14 days of HLS followed by normal ambulatory function for 14 days (recovery). Additional animals served as cage controls. The rats were given GTE (50 mg/kg body wt) or water (vehicle) by gavage 7 days before and throughout the experimental periods. Compared with vehicle treatment, GTE significantly attenuated the loss of hindlimb plantaris muscle mass (-24.8% vs. -10.7%, P < 0.05) and tetanic force (-43.7% vs. -25.9%, P <0.05) during HLS. Although GTE failed to further improve recovery of muscle function or mass compared with vehicle treatment, animals given green tea via gavage maintained the lower losses of muscle mass that were found during HLS (-25.2% vs. -16.0%, P < 0.05) and force (-45.7 vs. -34.4%, P < 0.05) after the reloading periods. In addition, compared with vehicle treatment, GTE attenuated muscle fiber cross-sectional area loss in both plantaris (-39.9% vs. -23.9%, P < 0.05) and soleus (-37.2% vs. -17.6%) muscles after HLS. This green tea-induced difference was not transient but was maintained over the reloading period for plantaris (-45.6% vs. -21.5%, P <0.05) and soleus muscle fiber cross-sectional area (-38.7% vs. -10.9%, P <0.05). GTE increased satellite cell proliferation and differentiation in plantaris and soleus muscles during recovery from HLS compared with vehicle-treated muscles and decreased oxidative stress and abundance of the Bcl-2-associated X protein (Bax), yet this did not further improve muscle recovery in reloaded muscles. These data suggest that muscle recovery following disuse in aging is complex. Although satellite cell proliferation and differentiation are critical for muscle repair to occur, green tea-induced changes in satellite cell number is by itself insufficient to improve muscle recovery following a period of atrophy in old rats.


Systemic Steroid Treatment for Imatinib-Associated Severe Skin Rash in Patients with Gastrointestinal Stromal Tumor: A Phase II Study.
To achieve optimal clinical outcomes in patients with gastrointestinal stromal tumor (GIST), it is crucial to maintain sufficient dosing of imatinib. Skin rash is a common imatinib-associated adverse event and may affect compliance. This phase II study was conducted to evaluate whether imatinib-associated severe skin rash can be managed with systemic steroids without dose reduction or interruption of imatinib. This study is registered at ClinicalTrials.gov, number NCT03440515. Between 2014 and 2016, 29 patients with imatinib-associated severe skin rash were enrolled. Skin rash of grade 2 with grade ≥2 pruritus or of grade 3 was considered severe. Oral prednisolone was administered 30 mg/day for 3 weeks, then tapered off over 12 weeks. The primary endpoint was treatment success rate (TSR). Treatment success was defined as maintaining imatinib for more than 15 weeks after completion of the steroid administration schedule without skin rash that led to additional steroid treatment or dose reduction or interruption of imatinib. Of the 29 patients enrolled, 22 patients with skin rash were treated successfully (TSR, 75.8%), 2 (6.9%) were evaluated as treatment failures, and 5 (17.2%) were not evaluable. The 2-year rash-free and imatinib reduction-free interval rate was 67.2% with median follow-up of 22.0 months (range, 0.4-30.3). Recurrence of severe skin rash occurred in seven patients (24.1%). Systemic steroids were well tolerated except in one patient who experienced pneumocystis pneumonia. This study demonstrated that imatinib-associated severe skin rash can be effectively controlled by systemic steroid treatment without interruption or dose reduction of imatinib in patients with GIST. Imatinib has been the standard treatment of gastrointestinal stromal tumor in both adjuvant and palliative settings. It is crucial to maintain sufficient dosing of imatinib to achieve optimal clinical outcomes. Imatinib commonly causes imatinib-associated skin rash, which may worsen drug compliance. This phase II study demonstrated that systemic steroids could help maintaining the efficacy of imatinib by preventing interruption or dose reduction of imatinib. The present study provides a new administration strategy of systemic steroids and its efficacy and safety data. Thus, this study can be a cornerstone to establish treatment guidelines for imatinib-associated skin rash.


Development of Novel Monoclonal Antibodies for Evaluation of Transmembrane Prostate Androgen-Induced Protein 1 (TMEPAI) Expression Patterns in Gastric Cancer.
Transmembrane prostate androgen-induced protein 1 (TMEPAI) is a single-span membrane protein, functionally involved in transforming growth factor beta signaling pathway. The particular protein presented in cells in three isoforms, which differs in the length of the soluble N-terminal extracellular domain, making it challenging for the immunochemical recognition. By using quantitative real-time polymerase chain reaction, we identified significant upregulation of PMEPA1 gene expression in malignant tissues of patients with gastric adenocarcinoma. The main part of commercially available anti-TMEPAI antibodies are having polyclonal nature or not suitable for immunocytochemical localization of target protein in tissue specimens. Hence, we decide to generate a set of novel rat monoclonal antibodies (mAb) directed against conservative C-terminal cytoplasmic epitope. Immunoblotting analysis showed that monoclonal antibodies, 2E1, 6C6, and 10A7 were able to recognize specifically target protein in transiently transfected HEK293T and CHO-K1 cells. Especially established mAb, named 10A7, showed the excellent binding ability to target protein in immunohistochemistry. By using developed antibodies, we observed pronounced expression of TMEPAI in normal gastric epithelial cells while tumor cells from gastric adenomas, and adenocarcinoma samples were mostly negative for target protein expression. Also, we found that gastric epithelium cells lose the TMEPAI expression concurrently with severe dysplasia progression, which probably caused by a mechanism involving specific microRNA.


Magnetic CoFe1.95Y0.05O4-Decorated Ag3PO4 as Superior and Recyclable Photocatalyst for Dye Degradation.
The Ag3PO4/CoFe1.95Y0.05O4 nanocomposite with magnetic properties was simply synthesized by the hydrothermal method. The structure and morphology of the prepared material were characterized, and its photocatalytic activity for degradation of the methylene blue and rhodamine B dyes was also tested. It was revealed that the Ag3PO4 in the nanocomposite exhibited a smaller size and higher efficiency in degrading dyes than the individually synthesized Ag3PO4 when exposed to light. Furthermore, the magnetic properties of CoFe1.95Y0.05O4 enabled the nanocomposite to possess magnetic separation capabilities. The stable crystal structure and effective degradation ability of the nanocomposite were demonstrated through cyclic degradation experiments. It was shown that Ag3PO4/CoFe1.95Y0.05O4-0.2 could deliver the highest activity and stability in degrading the dyes, and 98% of the dyes could be reduced within 30 min. Additionally, the photocatalytic enhancement mechanism and cyclic degradation stability of the magnetic nanocomposites were also proposed.


Characterization of human colorectal mucosa, polyps, and cancers by means of computerized morphonuclear image analyses.
Fifty-eight Feulgen-stained imprint smears from freshly resected colorectal tissue were analyzed by means of a SAMBA 200 cell image processor to establish a quantitative grading of their evolution from normal to malignant mucosa on the basis of 15 morphonuclear parameters relative to morphometry (nuclear size), densitometry (DNA content), and texture (chromatin pattern). The colorectal samples belonged to six groups: normal mucosa from patients without (Group 1) or with (Group 2) colorectal cancer, adenomas (Group 3), and cancers corresponding to the three stages of the Dukes' classification (Groups 4 to 6). Results indicated that analysis of the morphonuclear parameters computed on 250 to 300 nuclei/cases objectively and quantitatively showed the adenoma-cancer sequence. This need for only a small number of nuclei to assess a highly efficient analysis created a preoperative investigative tool using cytologic smears during endoscopy. The authors also made preliminary data banks for objective and reproducible grading of unknown cases by comparisons (discriminant analyses) with their contents. This approach must be validated prospectively on a large series of cases to furnish a system for colorectal malignancy diagnosis.


Associations Between PET Textural Features and GLUT1 Expression, and the Prognostic Significance of Textural Features in Lung Adenocarcinoma.
We evaluated the association between positron emission tomography (PET) textural features and glucose transporter 1 (GLUT1) expression level and further investigated the prognostic significance of textural features in lung adenocarcinoma. We evaluated 105 adenocarcinoma patients. We extracted texture-based PET parameters of primary tumors. Conventional PET parameters were also measured. The relationships between PET parameters and GLUT1 expression levels were evaluated. The association between PET parameters and overall survival (OS) was assessed using Cox's proportional hazard regression models. In terms of PET textural features, tumors expressing high levels of GLUT1 exhibited significantly lower coarseness, contrast, complexity, and strength, but significantly higher busyness. On univariate analysis, the metabolic tumor volume, total lesion glycolysis, contrast, busyness, complexity, and strength were significant predictors of OS. Multivariate analysis showed that lower complexity (HR=2.017, 95%CI=1.032-3.942, p=0.040) was independently associated with poorer survival. PET textural features may aid risk stratification in lung adenocarcinoma patients.


The Impact of the ACA's Medicaid Expansion on Hospitals' Uncompensated Care Burden and the Potential Effects of Repeal.
ISSUE: By increasing health insurance coverage, the Affordable Care Act's Medicaid eligibility expansion was also expected to lessen the uncompensated care burden on hospitals. The expansion currently faces an uncertain future. GOAL: To compare the change in hospitals' uncompensated care burden in the 31 states (plus the District of Columbia) that chose to expand Medicaid to the changes in states that did not, and to estimate how these expenses would be affected by repeal or further expansion. METHODS: Analysis of uncompensated care data from Medicare Hospital Cost Reports from 2011 to 2015. FINDINGS AND CONCLUSIONS: Uncompensated care burdens fell sharply in expansion states between 2013 and 2015, from 3.9 percent to 2.3 percent of operating costs. Estimated savings across all hospitals in Medicaid expansion states totaled $6.2 billion. The largest reductions in uncompensated care were found for hospitals in expansion states that care for the highest proportion of low-income and uninsured patients. Legislation that scales back or eliminates Medicaid expansion is likely to expose these safety-net hospitals to large cost increases. Conversely, if the 19 states that chose not to expand Medicaid were to adopt expansion, their uncompensated care costs also would decrease by an estimated $6.2 billion.


Palisaded neutrophilic and granulomatous dermatitis as a novel cause of hypercalcemia: A case report.
Palisaded neutrophilic and granulomatous dermatitis (PNGD) is a benign, inflammatory dermatosis with distinct histopathological features often observed in patients with systemic diseases. There were no reports of PNGD without underlying systemic diseases as an underlying cause of hypercalcemia. Herein, we report a case of a 62-year-old man with hypercalcemia due to PNGD, but with no underlying systemic diseases, including tuberculosis, sarcoidosis, or vasculitis. Laboratory tests showed an elevated C-reactive protein level, an elevated corrected calcium level, a normal 25-hydroxyvitamin D level, and an elevated 1,25-dihydroxyvitamin D level. There were no other abnormalities to explain the hypercalcemia. Positron emission tomography-computed tomography showed abnormal uptake in his skin. Histopathological examination of the skin showed palisaded granulomatous infiltrate in the dermis. Neutrophils, degenerated collagen, and fibrin were present in the centers of the palisades without prominent mucin. There were no eosinophils, central necrosis, or necrotizing vasculitides. These features were consistent with PNGD. A diagnosis of PNGD with hypercalcemia was established. Oral prednisolone was administered to the patient. After treatment, his symptoms resolved, and his calcium, 1,25-dihydroxyvitamin D and CRP levels returned to normal. Skin specimens before and after treatment were assessed using immunohistochemistry for 1a-hydroxylase. Granuloma and epidermal cells were 1a-hydroxylase-positive before treatment. After treatment, the granuloma diminished in size and the 1ahydroxylase-positive areas of the epidermal cells decreased. This case was particularly unique because increased 1a-hydroxylase expression in the granuloma and epidermal cells seemed to result in hypercalcemia due to excessive transformation of 25-hydroxyvitamin D to 1,25-dihydroxyvitamin D. Physicians should consider PNGD as an underlying cause of hypercalcemia.


Small-angle neutron polarimetry apparatus (SANPA): Development at the NIST Center for Neutron Research.
Spherical neutron polarimetry directly measures the rotation of the neutron polarization after scattering from materials with magnetic structure. It is an under utilized measurement technique that is capable of measuring all nine elements of the polarization tensor of a material. In this article, we describe our new cryogen-free small-angle neutron polarimetry apparatus and infrastructure at the NIST Center for Neutron Research. The resulting apparatus is capable of continuous operation and is designed for measurements at low temperatures (4-8 K) using niobium Meissner shielding and mu-metal shielding to produce a zero-field (≤1 μT) cooling sample environment.


Plasma selenium concentration in healthy Japanese children and adults determined by flameless atomic absorption spectrophotometry.
This study showed a rapid and direct method for determining selenium concentration in plasma by flameless atomic absorption spectrophotometry, and differences in plasma selenium concentration in healthy children and adults. A direct method is possible, since selenium is heat stable in the presence of nickel. With this method, the recovery of selenium added to plasma was 100.3 +/- 5.7%, and the relative standard deviation in repeated determinations of pooled plasma selenium was 3.0% and 6.8%. The plasma selenium concentration in adults was 99.4 +/- 12.5 ng/ml, lower than reported concentrations from the United States and Canada, and higher than those from New Zealand. These variations may reflect dietary habits, bioavailability of selenium compounds in diet, racial difference, or different analytical methods. The mean concentration of plasma selenium at 1 to 6 months of age (51.0 +/- 13.1 ng/ml) was significantly lower than in adults (p less than 0.001); it increased gradually and steadily to the adult level with age. This age-related difference of plasma selenium level is similar to that reported previously.


Multiple enzyme approach for the characterization of glycan modifications on the C-terminus of the intestinal MUC2mucin.
The polymeric mucin MUC2 constitutes the main structural component of the mucus that covers the colon epithelium. The protein's central mucin domain is highly O-glycosylated and binds water to provide lubrication and prevent dehydration, binds bacteria, and separates the bacteria from the epithelial cells. Glycosylation outside the mucin domain is suggested to be important for proper protein folding and protection against intestinal proteases. However, glycosylation of these regions of the MUC2 has not been extensively studied. A purified 250 kDa recombinant protein containing the last 981 amino acids of human MUC2 was produced in CHO-K1 cells. The protein was analyzed before and after PNGase F treatment, followed by in-gel digestion with trypsin, chymotrypsin, subtilisin, or Asp-N. Peptides were analyzed by nLC/MS/MS using a combination of CID, ETD, and HCD fragmentation. The multiple enzyme approach increased peptide coverage from 36% when only using trypsin, to 86%. Seventeen of the 18 N-glycan consensus sites were identified as glycosylated. Fifty-six N-glycopeptides covering 10 N-glycan sites, and 14 O-glycopeptides were sequenced and characterized. The presented method of protein digestion can be used to gain better insights into the density and complexity of glycosylation of complex glycoproteins such as mucins.


What's in the Gift? Towards a Molecular Dissection of Nuptial Feeding in a Cricket.
Nuptial gifts produced by males and transferred to females during copulation are common in insects. Yet, their precise composition and subsequent physiological effects on the female recipient remain unresolved. Male decorated crickets Gryllodes sigillatus transfer a spermatophore to the female during copulation that is composed of an edible gift, the spermatophylax, and the ampulla that contains the ejaculate. After transfer of the spermatophore, the female detaches the spermatophylax and starts to eat it while sperm from the ampulla are evacuated into the female reproductive tract. When the female has finished consuming the spermatophylax, she detaches the ampulla and terminates sperm transfer. Hence, one simple function of the spermatophylax is to ensure complete sperm transfer by distracting the female from prematurely removing the ampulla. However, the majority of orally active components of the spermatophylax itself and their subsequent effects on female behavior have not been identified. Here, we report the first analysis of the proteome of the G. sigillatus spermatophylax and the transcriptome of the male accessory glands that make these proteins. The accessory gland transcriptome was assembled into 17,691 transcripts whilst about 30 proteins were detected within the mature spermatophylax itself. Of these 30 proteins, 18 were encoded by accessory gland encoded messages. Most spermatophylax proteins show no similarity to proteins with known biological functions and are therefore largely novel. A spermatophylax protein shows similarity to protease inhibitors suggesting that it may protect the biologically active components from digestion within the gut of the female recipient. Another protein shares similarity with previously characterized insect polypeptide growth factors suggesting that it may play a role in altering female reproductive physiology concurrent with fertilization. Characterization of the spermatophylax proteome provides the first step in identifying the genes encoding these proteins in males and in understanding their biological functions in the female recipient.


Diversity of bacterial isolates from commercial and homemade composts.
The diversity of heterotrophic bacterial isolates of three commercial and two homemade composts was studied. The commercial composts were produced from poultry litter (PC), sewage sludge (SC), municipal solid waste (MC), and homemade composts (thermal compost [DC] and vermicompost [VC]) from food wastes. The taxonomic and physiological diversity of the heterotrophic culturable bacteria was assessed using phenotypic and genotypic characterization and the analysis of the partial 16S rRNA gene sequence. Composts DC and SC presented the higher genotypic diversity, as could be inferred from the number of distinct genotypic patterns observed, 28 and 21, respectively. Gram-positive bacteria, mainly Firmicutes, were predominant in all the composts. Some organisms related with taxa rarely reported in composts, as Rhodanobacter spathiphylli, Moraxella osloensis, Lysobacter, Corynebacterium, Pigmentiphaga kullae, and new taxa were also isolated. The highest relative proportion of isolates able to degrade starch was found in compost SC (> 70%), to degrade gelatine in compost DC (> 70%), to degrade Tween 80 in compost PC (> 90%), and to degrade poly-epsilon-caprolactones in compost DC (> 80%). Compost MC presented the lowest relative proportions of isolates able to degrade starch (< 25%), gelatine (< 20%), and poly-epsilon-caprolactone (< 40%). When compared with the others, the homemade composts presented higher relative proportions of Gram-positive isolates able to inhibit the target organisms Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, or Pseudomonas aeruginosa. In compost MC, none of the Gram-positive isolates was able to inhibit those targets.


[Bioinformatics and tissue distribution analysis of Tripterygium wilfordii CYP450].
Cytochrome P450 family is a kind of biocatalyst widely existing in nature. It has many functions such as catalyzing the biosynthesis of plant secondary metabolites and regulating phytoremediation. Based on the analysis of proteome data of Tripterygium wilfordii,the CYP450 gene of T. wilfordii was preliminarily analyzed and predicted by various bioinformatics methods. The results showed that after the expression of T. wilfordii suspension cells was induced by methyl jasmonate,the proteomic data of T. wilfordii were obtained and analyzed,and 10 CYP450 proteins of T. wilfordii were finally screened out. By analyzing the phylogenetic tree constructed with CYP450 gene of Arabidopsis family,the 10 CYP450 proteins were clustered into 6 different CYP450 families. The physical and chemical properties of CYP450 proteins in different families were different. The secondary structure of CYP450 proteins was mainly composed of irregular curls. Eight subcellular localization results of CYP450 proteins were chloroplasts and the rest were plastids. Subsequently,the conserved domains( heme active sites) shared by CYP450 genes were found by analyzing the results of multiple sequence alignment. Finally,by analyzing the transcriptome data of T. wilfordii,the expression distribution of T. wilfordii in different tissues was preliminarily confirmed,which verified its correlation with the biosynthesis of active components of T. wilfordii,and provided important genetic resources for the analysis of biosynthesis pathway of active components of T. wilfordii.


[New Developments in CKD-MBD. Development and progression of CKD-MBD in predialysis period -when and how?].
Growing interest has been focused on the pathogenesis of chronic kidney disease-mineral and bone disorder (CKD-MBD) , which is not restricted to those with end-stage CKD and dialysis patients but also includes those with early-stage CKD. Recent evidence demonstrated that FGF23 levels increase early in CKD to maintain normal phosphate balance, but this results in suppression of renal production of 1,25 dihydroxyvitamin D and thereby triggers the early development of secondary hyperparathyroidism. Thus, it can be interpreted that elevated FGF23 in response to phosphate overload represent the beginning of CKD-MBD. Elevations in FGF23 may also suggest the presence of compensatory response to phosphate retention, underscoring the potential of FGF23 as a new biomarker of phosphate balance. It is hoped that further research in this area will establish the treatment strategy of predialysis CKD-MBD and improves the outcome of CKD patients.


[Perioperative complications in traditional surgery of the rectum].
The authors report their experience of 136 operations for rectal neoplasms, on all of 652 colo-rectal neoplasms, treated from January 1987 to May 1994. In detail the perianastomotic complications have been tested after "traditional" rectal operations: Anterior Resection (RA), Abdomino-perineal amputation (AAP) and Hartmann's procedure. A series of 24 AAP (18%), 81 RA (60%) and 7 Hartmann's procedure have been considered; conversely 18 final and palliative colostomies (13%) and 7 trans-anal operations haven't been taken in to account. Perioperative mortality was comparatively reasonable: 2.7%. No patient died after AAP. The most common complication proved to be anastomotic leak (17.3%), after RA; otherwise the rate of further complications was very moderate. It is feasible to avoid anastomotic leaks, by carrying out a temporary colostomy.


Improved metabolic stability and therapeutic efficacy of a novel molecular gemcitabine phospholipid complex.
The aim of the present research is to increase lipid solubility, metabolic stability and therapeutic efficacy of water soluble gemcitabine (GEM) via phospholipid complex (PC) formation. A novel phospholipid complex of GEM was successfully prepared and optimized. Physical interaction of GEM with phospholipid was evaluated by DSC, FT-IR, 1H NMR, 31P-NMR and P-XRD. SEM images of GEM-PC showed rough structure and TEM images of diluted aqueous dispersion of GEM-PC showed micellar structure. In silico study also revealed the significant interaction between drug and phospholipid. GEM-PC demonstrated sustained drug release pattern and high plasma stability (∼2.2 fold) in vitro as compared to GEM. Increased in vitro cytotoxicity and apoptosis were observed with GEM-PC, when incubated with human pancreas adenocarcinoma cell lines. In vivo pharmacokinetics showed the almost 2 fold increase in AUC0-∞ (area under curve) with phospholipid complex (8983.26ngh/ml) as compared with GEM (4371.18ngh/ml) and GEMITA (4689.29ngh/ml). Toxicity studies signify the safety of GEM-PC over GEMITA. Pharmacodynamics studies in pancreatic tumor model further revealed higher efficacy of GEM-PC than GEMITA. These findings suggested the higher potential of phospholipid based technology for the enhancement of metabolic stability and therapeutic efficacy of GEM.


Phthalate exposure and allergic diseases: Review of epidemiological and experimental evidence.
Phthalates are among the most ubiquitous environmental contaminants and endocrine-disrupting chemicals. Exposure to phthalates and related health effects have been extensively studied over the past four decades. An association between phthalate exposure and allergic diseases has been suggested, although the literature is far from conclusive. This article reviews and evaluates epidemiological (n = 43), animal (n = 49), and cell culture studies (n = 42), published until the end of 2019, on phthalates and allergic diseases, such as asthma, rhinoconjunctivitis, and eczema. In contrast to earlier reviews, emphasis is placed on experimental studies that use concentrations with relevance for human exposure. Epidemiological studies provide support for associations between phthalate exposures and airway, nasal, ocular, and dermal allergic disease outcomes, although the reported significant associations tend to be weak and demonstrate inconsistencies for any given phthalate. Rodent studies support that phthalates may act as adjuvants at levels likely to be relevant for environmental exposures, inducing respiratory and inflammatory effects in the presence of an allergen. Cell culture studies demonstrate that phthalates may alter the functionality of innate and adaptive immune cells. However, due to limitations of the applied exposure methods and models in experimental studies, including the diversity of phthalates, exposure routes, and allergic diseases considered, the support provided to the epidemiological findings is fragmented. Nevertheless, the current evidence points in the direction of concern. Further research is warranted to identify the most critical windows of exposure, the importance of exposure pathways, interactions with social factors, and the effects of co-exposure to phthalates and other environmental contaminants.


Ubiquitin in cerebrospinal fluid: a rapid competitive enzyme-linked immunoflow assay.
The aims of this study were to develop a rapid immunoassay to determine the levels of ubiquitin in cerebrospinal fluid and to establish the ubiquitin levels in the spinal fluid of normal aged individuals. A competitive enzyme-linked immunoflow assay was developed. In this assay, ubiquitin is bound to nitrocellulose membrane, after which the primary antibody-test sample mixture and the enzyme-labeled secondary antibody under vacuum are applied sequentially. The final reaction product is collected in a microtiter plate by suction. This competitive assay requires only approximately 4 h and is potentially useful for determining in biological fluids the levels of any antigen or its antibodies that might be present. Employing this immunoassay, cerebrospinal fluid ubiquitin levels were found to be 147.5 +/- 5.2 ng ml-1 in non-neurological aged cases.


Mathematical analysis of the influence of brain metabolism on the BOLD signal in Alzheimer's disease.
Blood oxygen level-dependent functional magnetic resonance imaging (BOLD-fMRI) is a standard clinical tool for the detection of brain activation. In Alzheimer's disease (AD), task-related and resting state fMRI have been used to detect brain dysfunction. It has been shown that the shape of the BOLD response is affected in early AD. To correctly interpret these changes, the mechanisms responsible for the observed behaviour need to be known. The parameters of the canonical hemodynamic response function (HRF) commonly used in the analysis of fMRI data have no direct biological interpretation and cannot be used to answer this question. We here present a model that allows relating AD-specific changes in the BOLD shape to changes in the underlying energy metabolism. According to our findings, the classic view that differences in the BOLD shape are only attributed to changes in strength and duration of the stimulus does not hold. Instead, peak height, peak timing and full width at half maximum are sensitive to changes in the reaction rate of several metabolic reactions. Our systems-theoretic approach allows the use of patient-specific clinical data to predict dementia-driven changes in the HRF, which can be used to improve the results of fMRI analyses in AD patients.


The complete amino acid sequences of ribosomal proteins L17, L27, and S9 from Bacillus stearothermophilus.
The complete primary structures of proteins L17, L27 and S9 extracted from the Bacillus stearothermophilus ribosomes with 1 M NaCl and purified to homogeneity by column chromatography have been determined. The amino acid sequences of these proteins are compared to those of the homologous ribosomal proteins from Escherichia coli. The number of identical amino acid residues between the homologous proteins lies between 33-55%.


Actinomycosis masquerading as malignancy.
We describe a patient who presented with neurological symptoms and radiological findings consistent with metastatic malignancy in several sites. However, no obvious primary site of tumour was identified and the patient later went on to develop clinical features of sepsis whilst using palliative dexamethasone therapy. A diagnosis of actinomycosis was eventually made on open lung biopsy and the patient recovered completely on penicillin V. This case serves to illustrate the need to obtain a formal tissue diagnosis whenever possible in all cases of suspected malignancy, in addition to exposing the weaknesses inherent in empiric diagnoses.


Intracellular drug delivery by poly(lactic-co-glycolic acid) nanoparticles, revisited.
We reexamined the cellular drug delivery mechanism by poly(lactic-co-glycolic acid) nanoparticles (PLGA NPs) to determine their utility and limitations as an intracellular drug delivery system. First, we prepared PLGA NPs which physically encapsulated Nile red (a hydrophobic fluorescent dye), in accordance with the usual procedure for labeling PLGA NPs, incubated them with mesothelial cells, and observed an increase in the intracellular fluorescence. We then prepared NPs from PLGA chemically conjugated to a fluorescent dye and observed their uptake by the mesothelial cells using confocal microscopy. We also used coherent anti-Stokes Raman scattering (CARS) microscopy to image cellular uptake of unlabeled PLGA NPs. Results of this study coherently suggest that PLGA NPs (i) are not readily taken up by cells, but (ii) deliver the payload to cells by extracellular drug release and/or direct drug transfer to contacting cells, which are contrasted with the prevalent view. From this alternative standpoint, we analyzed cytotoxicities of doxorubicin and paclitaxel delivered by PLGA NPs and compared with those of free drugs. Finally, we revisit previous findings in the literature and discuss potential strategies to achieve efficient drug delivery to the target tissues using PLGA NPs.


Combining transgenesis with paratransgenesis to fight malaria.
Malaria is among the deadliest infectious diseases, and Plasmodium, the causative agent, needs to complete a complex development cycle in its vector mosquito for transmission to occur. Two promising strategies to curb transmission are transgenesis, consisting of genetically engineering mosquitoes to express antimalarial effector molecules, and paratransgenesis, consisting of introducing into the mosquito commensal bacteria engineered to express antimalarial effector molecules. Although both approaches restrict parasite development in the mosquito, it is not known how their effectiveness compares. Here we provide an in-depth assessment of transgenesis and paratransgenesis and evaluate the combination of the two approaches. Using the Q-system to drive gene expression, we engineered mosquitoes to produce and secrete two effectors - scorpine and the MP2 peptide - into the mosquito gut and salivary glands. We also engineered Serratia, a commensal bacterium capable of spreading through mosquito populations to secrete effectors into the mosquito gut. Whereas both mosquito-based and bacteria-based approaches strongly reduced the oocyst and sporozoite intensity, a substantially stronger reduction of Plasmodium falciparum development was achieved when transgenesis and paratransgenesis were combined. Most importantly, transmission of Plasmodium berghei from infected to naïve mice was maximally inhibited by the combination of the two approaches. Combining these two strategies promises to become a powerful approach to combat malaria.


Correlation between peripheral blood α1-MG, DNMT1 expression, and the severity of diabetic nephropathy renal pathological damage.
To explore the correlation between peripheral blood α1-microglobulin (α1-MG) and monocyte DNA methyltransferase 1 (DNMT1) expression and the severity of renal pathological damage in diabetic nephropathy (DN). The study group comprised 100 patients with DN who underwent treatment at our hospital from January 2022 to January 2023, while the control group consisted of 50 patients with uncomplicated diabetes. The relative expression levels of peripheral blood α1-MG and DNMT1 were compared between the 2 groups of patients. Additionally, the levels of vascular endothelial growth factor (VEGF) were measured, and the diagnostic value of DN was explored using ROC curves. Furthermore, the correlation between the aforementioned indicators and the severity of renal pathological damage in the patients of the study group was analyzed. Compared to the patients in the control group, the patients in the study group showed increased relative expression levels of peripheral blood α1-MG and DNMT1, as well as elevated levels of VEGF (P < .05). The diagnostic value of peripheral blood α1-MG, DNMT1 relative expression levels, and VEGF levels for DN was explored using ROC curves. The AUC values were 0.907, 0.923, and 0.936, respectively (P < .05). The relative expression levels of peripheral blood α1-MG, DNMT1, and VEGF levels in DN patients increase with the elevation of the interstitial fibrosis and tubular atrophy scoring (IFTA) score, showing a positive correlation with r-values of 0.651, 0.710, and 0.628, respectively (P < .05). The relative expression levels of peripheral blood α1-MG, DNMT1, and VEGF levels in DN patients increase with the elevation of the interstitial inflammation score, showing a positive correlation with r-values of 0.771, 0.633, and 0.678, respectively (P < .05). The relative expression levels of peripheral blood α1-MG, DNMT1, and VEGF levels in DN patients increase with the elevation of the glomerular grading, showing a positive correlation with r-values of 0.714, 0.609, and 0.677, respectively (P < .05). The expression levels of peripheral blood α1-MG, DNMT1, and VEGF are significantly elevated in patients with DN. These levels show a positive correlation with the IFTA score, interstitial inflammation score, and glomerular grading, contributing to the diagnosis and assessment of DN.


Sentinel Lymph Node Evaluation: What the Radiologist Needs to Know.
Axillary lymph node status is the single most important prognostic indicator in patients with breast cancer. Axillary lymph node dissection, the traditional method of staging breast cancer, is associated with significant morbidity. Sentinel lymph node biopsy has become standard in patients being treated for breast cancer with clinically negative lymph nodes. There is considerable variation in the medical literature regarding technical approaches to sentinel lymph node biopsy in patients with breast cancer. The purpose of this article is to describe our preferred approaches to sentinel lymph node biopsy with a review of the literature.