Dual Roles for Regulatory T-cell Depletion and Costimulatory Signaling in Agonistic GITR Targeting for Tumor Immunotherapy.
Agonistic monoclonal antibodies (mAb) targeting the T-cell receptor coregulatory molecule GITR exert potent therapeutic activities in preclinical tumor models. Although anti-GITR mAb are thought to act by depleting and destabilizing the intratumoral T regulatory cell (Treg) population, the precise mechanism of action is obscure. Here, we addressed this issue using a Treg fate-mapping approach, which revealed that Treg loss was primarily due to cell depletion, with minimal evidence of Treg conversion to a non-Foxp3-expressing population. Further characterization of persisting Tregs following anti-GITR mAb treatment showed that a highly activated subpopulation of CD44hiICOShi intratumoral Tregs were preferentially targeted for elimination, with the remaining Tregs exhibiting a less suppressive phenotype. With these changes in the Treg population, intratumoral CD8+ T cells acquired a more functional phenotype characterized by downregulation of the exhaustion markers PD-1 and LAG-3. This reversal of CD8+ T-cell exhaustion was dependent on both agonistic GITR signaling and Treg depletion, as neither mechanism by itself could fully rescue the exhaustion phenotype. Tests of anti-human GITR antibody MK-4166 in a humanized mouse model of cancer mimicked many of the effects of anti-mouse GITR mAb in syngeneic tumor models, decreasing both Treg numbers and immune suppressor phenotype while enhancing effector responsiveness. Overall, our results show how anti-GITR mAb shifts Treg populations to enable immune attack on tumors, with clinical implications for molecular markers to modify emerging treatments. Cancer Res; 77(5); 1108-18. ©2016 AACR.


Efficacy of garcinia cambogia on body weight, inflammation and glucose tolerance in high fat fed male wistar rats.
Obesity leads to derangements in lipid and glucose homeostasis resulting in various metabolic complications. Plants containing vital phytochemicals are known to posses anti obesity properties and have proved to exert beneficial effects in obesity. The present study was aimed to investigate the effects of Garcinia Cambogia on body weight, glucose tolerance and inflammation in high fat diet fed male Wistar rats. Five month old male wistar rats (n=40) were divided into four groups. Two groups were fed with standard rodent diet and the remaining two with 30% high fat diet. One group in each of the two sets received the crude ethanolic extract of Garcinia Cambogia at a dose of 400mg/kg body weight/day for ten weeks. Body weight, intraperitoneal glucose tolerance test, leptin, tumour necrosis factor-α (TNF-α) and renal function (urea, creatinine, uric acid) were studied. High fat diet fed rats showed increased body weight gain, glucose intolerance, elevated levels of plasma leptin and TNF-α. Supplementation of Garcinia Cambogia extract (GE) along with high fat diet significantly decreased body weight gain, glucose intolerance, plasma leptin and TNF-α level. No significant changes were observed in the renal function parameters in any of the groups. Supplementation of the Garcinia Cambogia extract with high fat diet reduced body weight gain, inflammation and glucose intolerance.


High-definition vs. standard-definition endoscopy with indigo carmine for the in vivo diagnosis of colonic polyps.
There is growing evidence that indigo carmine chromoendoscopy is effective for the in vivo diagnosis of colonic polyps. However, the impact of colonoscope resolution on diagnostic accuracy has not been investigated. We aimed to compare the effectiveness of in vivo diagnosis of small colonic polyps using indigo carmine dye spray with standard-definition and high-definition colonoscopes. Procedures were performed using Fujinon colonoscopes and EPX 4400 processor. Fujinon standard-definition (SD) and high-definition (HD) colonoscopes were used, with the endoscopist blinded to colonoscope definition. Polyps <10 mm were assessed using 0.2% indigo carmine dye spray, with the predicted diagnosis recorded. In each case the kind of colonoscope (SD or HD) was recorded. Polyps were removed and sent for histological analysis, with the pathologist blinded to the diagnosis made by the endoscopist. The predicted diagnosis was compared with the true histology to calculate the accuracy, sensitivity and specificity of in vivo assessment using either SD or HD scopes. In total 237 polyps <10 mm in size were examined. There was no statistically significant difference for any of the measured parameters between SD and HD assessments, with an accuracy, sensitivity and specificity of 89%, 91% and 87% with SD colonoscopes and 92%, 96% and 84% with HD colonoscopes. The accuracy of in vivo assessment of small colonic polyps with indigo carmine dye spray is excellent with standard-definition colonoscopes and is not improved with high-definition colonoscopes.


Translocation domain of botulinum neurotoxin A subtype 2 potently induces entry into neuronal cells.
Botulinum neurotoxin (BoNT) is the causative agent of botulism in humans and animals. Only BoNT serotype A subtype 1 (BoNT/A1) is used clinically because of its high potency and long duration of action. BoNT/A1 and BoNT/A subtype 2 (BoNT/A2) have a high degree of amino acid sequence similarity in the light chain (LC) (96%), whereas their N-and C-terminal heavy chain (HN and HC ) differ by 13%. The LC acts as a zinc-dependent endopeptidase, HN as the translocation domain, and HC as the receptor-binding domain. BoNT/A2 and BoNT/A1 had similar potency in the mouse bioassay, but BoNT/A2 entered faster and more efficiently into neuronal cells. To identify the domains responsible for these characteristics, HN of BoNT/A1 and BoNT/A2 was exchanged to construct chimeric BoNT/A121 and BoNT/A212. After expression in Escherichia coli, chimeric and wild-type BoNT/As were purified as single-chain proteins and activated by conversion to disulfide-linked dichains. The toxicities of recombinant wild-type and chimeric BoNT/As were similar, but dropped to 60% compared with the values of native BoNT/As. The relative orders of SNAP-25 cleavage activity in neuronal cells and toxicity differed. BoNT/A121 and recombinant BoNT/A2 have similar SNAP-25 cleavage activity. BoNT/A2 HN is possibly responsible for the higher potency of BoNT/A2 than BoNT/A1.


Different diagnostic criteria for Parkinson disease: what are the pitfalls?
As there are no definite diagnostic tests or reliable biomarkers for Parkinson disease (PD), its diagnosis still relies on the presence of a combination of cardinal motor features, along with the exclusion of other causes of Parkinsonism and the presence of some of supportive features. To date, several diagnostic criteria have been developed for different purposes through expert opinions or comprehensive review of the literature. However, none of them are without limitations. In this article, we review different diagnostic criteria for PD which have been published in the English medical literature, highlighting specific limitations and pitfalls. With considerable progress in the understanding of PD, particularly in a view of diverse clinical symptomatology and its evolution, it will be difficult to establish a single criterion that is capable of capturing all cases at different disease stages. Rather, we should aim to develop a set of criteria which include a consensus on clinical gold standard or reliable biomarkers at different levels of diagnostic certainty for different purposes. Despite a more refined set of criteria that may aid in the recognition of PD, the accuracy of its diagnosis still largely depends on the observational skills and clinical sensitivity of the treating physician.


Isolating syntax in natural language: MEG evidence for an early contribution of left posterior temporal cortex.
Although the construction of syntactic structures is considered a fundamental component of language processing, insights concerning its neurobiological basis have remained elusive. This may be due, in part, to the inherent difficulty of isolating incremental syntax from other components of language, such as semantic composition. However, while pure manipulations of syntax are challenging to design, natural language grammars do have a sparse set of constructions presenting the possibility. Here we examined one such case, English post-nominal adjectives (mountain TALL enough for a strenuous hike), which were contrasted with semantically parallel but structurally simpler noun-adjective sequences in an MEG experiment. We observed a sharp activity increase in the left posterior temporal lobe (PTL) when syntactic composition was more straightforward, approximately 200 msec after adjective onset. The semantic fit between the noun and adjective was also varied, but this affected anterior temporal cortex, consistent with prior work. These findings offer a unique demonstration of the relevance of posterior temporal cortex for syntactic processing in natural language. We also present connectivity evidence that the syntax-related PTL responses were relayed to ipsilateral inferior frontal and anterior temporal regions. The combined results draw an initial picture of the rapid spatio-temporal dynamics of the syntactic and semantic composition network in sentence processing.


[Response to aggression: evaluation and therapeutic implications. Work Group on Metabolism and Nutrition of the SEMIUC].
The metabolic response to stress/aggression is a complex process which is mainly mediated by the interaction between the neuro-endocrine system and the circulating cytokines. This interaction brings about physiological and metabolic alterations--severe metabolic-nutritional deficits--of the hypermetabolic type, muscular proteolysis, lipolysis, glycogenolysis, and gluconeogenesis among others, which should be studied and understood prior to initiating or not a nutritional support. The parenteral or enteral nutritional support is usually indicated to prevent a worsening of these situations of altered metabolism frequently associated with inanition, although one should not attempt to revert to normal preexisting deficit situations. On the other hand, and is this special context, we should not forget the advances in nutrients with pharmacological effects, and in pharmacological nutrition. At this II Consensus Conference of the SEMIUC (Sociedad Española de Medicine Intensiva y Unidades Coronarias = Spanish Society of Intensive Medicine and Coronary Units), we have proposed four objectives: i) To make recommendation, based both on the scientific evidence, as on the experience of the components. ii) Define the scientific terminology to be used in this specific context. iii) Give an answer to the different and assorted clinical problems which are secondary to a situation of stress which has a multiple etiology. iv) Provide new ideas for the development of clinical trails and studies of this specific context.


Liprin-α1 regulates breast cancer cell invasion by affecting cell motility, invadopodia and extracellular matrix degradation.
Migration of cells and degradation of the extracellular matrix (ECM) are required for efficient tumor cell invasion, but the underlying molecular mechanisms are only partially known. The PPFIA1 gene for liprin-α1 is frequently amplified in human breast cancers. We recently demonstrated that liprin-α1 is an important regulator of cell edge dynamics during motility. We show, herein, that the liprin-α1 protein is highly expressed in human breast tumors. Functional analysis shows that liprin-α1 is specifically required for the migration and invasion of highly invasive human breast cancer MDA-MB-231 cells. We used time-lapse analysis to demonstrate defects in the motility of liprin-α1-depleted cells that include a striking instability of the lamellipodia. Liprin-α1 levels altered by either RNA interference or overexpression affected also cell spreading and the number of invadopodia per cell, but not the density of invadopodia per unit of surface area. On the other hand, silencing of liprin-α1 inhibited the degradation of the ECM, whereas its overexpression enhanced degradation, resulting in significant negative or positive effects, respectively, on the area of degradation per invadopodium. Transfection of fluorescent-labeled cortactin revealed that depletion of liprin-α1 also affected the assembly and disassembly of invadopodia, with decrease of their lifetime. Our results strongly support a novel important role of liprin-α1 in the regulation of human tumor cell invasion.


Significance of angina pectoris in aortic valve stenosis.
Of 60 patients aged 45 to 66 years with aortic valve stenosis, 28 (47 per cent) had angina pectoris. Significant coronary arterial obstruction was shown by selective coronary cineangiography in 14 of them. Systolic pressure gradients across the aortic valve were lower in patients with angina than in those without. In those with angina, systolic gradients were higher in those with normal coronary arteriograms than in those with demonstrable coronary arterial disease. Aortic valve replacement relieved the angina in all patients who had normal coronary arteriograms. When valve replacement was combined with coronary bypass grafting in those with coronary arterial disease, surgical mortality was higher and symptomatic relief less predictable. Incapacitating angina in patients with aortic stenosis was nearly always associated with significant coronary disease. In those with less severe angina it was impossible to predict the state of the coronary arteries. Two patients, who did not have angina and who did not undergo coronary arteriography, died after aortic valve replacement and were found at necropsy to have unsuspected severe coronary disease. We, therefore, suggest that coronary arteriography should be carried out in all patients over the age of 40 years in whom surgery is being considered for aortic stenosis.


Atomic-resolution microscopy in water.
The scanning tunneling microscope is revolutionizing the study of surfaces. In ultra-high vacuum it is capable not only of imaging individual atoms but also of determining energy states on an atom-by-atom basis. It is now possible to operate this instrument in water. Aqueous optical microscopy is confined to a lateral resolution limit of about 2000 angstroms, and aqueous x-ray microscopy has yielded a lateral resolution of 75 angstroms. With a scanning tunneling microscope, an image of a graphite surface immersed in deionized water was obtained with features less than 3 angstroms apart clearly resolved. Further, an image measured in saline solution demonstrated that the instrument can be operated under conditions useful for many biological samples.


Cyclin B1, unlike cyclin G1, increases significantly during colorectal carcinogenesis and during later metastasis to lymph nodes.
In collaboration with p53, cyclins B1 and G1 regulate the G2/M transition, a key checkpoint in the active cell cycle, which can be monitored by Ki67. However, the cyclin B1 expression remains unclear during colorectal carcinogenesis and during later metastasis to lymph nodes, and cyclin G1 expression is not clear in colorectal tumors. To clarify the variations of the two cyclins in colorectal tumors, cyclin B1, cyclin G1, p53, and Ki67 were immunohistochemically stained in 22 normal mucosa, 62 adenomas, 17 carcinomas in adenomas, 194 primary carcinomas, and 21 lymph node metastases; and the two cyclins were examined by Western blot in other 10 pairs of normal mucosa and primary carcinomas. Located in cytoplasms, nuclei or both, cyclin B1 expression increased significantly from normal mucosa through adenomas to primary carcinomas, from adenomas with mild dysplasia through those with moderate to those with severe, from peripheral adenomas to their central carcinomas, and from primary to metastatic foci. These increased expressions were confirmed by Western blot. Cyclin B1 expression, however, declined significantly in primary carcinomas showing large size, mucinous type, deep invasion, or short postoperative-patient-survival time. High cyclin B1 was linked to high p53 in adenomas, and to high Ki67 in adenomas and primary carcinomas. In contrast, found limited to nuclei, cyclin G1 expression did not vary significantly from normal mucosa through to metastatic carcinomas, and was not associated with clinicopathological parameters, p53 or Ki67. The unchanged expressions were confirmed by Western blot. Thus, increased cyclin B1, but not cyclin G1, may promote colorectal carcinogenesis and later metastasis to lymph nodes.


Dressings and ajunctive therapies: AHCPR guidelines revisited.
The Agency for Health Care Policy and Research Pressure Ulcer Treatment Guidelines released in 1994 offered 11 recommendations relating to the use of dressings and adjunctive therapies. These recommendations were based on evidence gleaned from the medical literature from 1966 to mid-1993 as well as the accumulated expertise of a multidisciplinary advisory panel. As the recommendations are now 5 years old, a review of the literature published since 1993 was undertaken in order to determine additional evidence for their continued validity or disapproval. The review focused on evidence at the level of controlled clinical trials of dressings and adjunctive therapies in the treatment of pressure ulcers.


Immunization of mice with plasmids coexpressing HPV16 E5 and the novel oncogene hWAPL.
The novel human oncogene hWAPL is associated with uterine cervical cancer. The HPV16 E5 oncoprotein could induce genomic instability in normal human cells. However, the mechanism of E5 interaction with hWAPL still awaits definition. In our present studies, the eukaryotic expression plasmids, pcDNA3-hWAPL and pcDNA3-hWAPL-E5 were constructed and carried out to vaccinate mice directly. The result that indicated the polyclonal antibody titer in immunized mice sera was increased by enzyme-linked immunosorbent assay. In addition, the proliferative responses of immunized mice spleen cells showed the optical densities values in vaccinated group remarkably higher than that in the control group. In conclusion, the recombinant plasmids could induce strong humoral and cellular immune response and exhibited great potential as therapeutic targets in the treatment of cervical cancer. However, the result didn't show significant difference in group with coexpression of HPV16 E5-hWAPL and group with only hWAPL expression. Consistent with these observations, we demonstrated that HPV16 E5 was not the optimal factor to cooperate with hWAPL in gene therapy.


Influence of cigarette smoke on cross-linking of dermal collagen.
Effect of cigarette smoke on collagen crosslinking was studied in male albino rat skins. Skin samples taken for the analysis on 120th day of exposure to cigarette smoke showed that, compared to controls, the exposed animal had a decreased tendency in the percent reversibility of neutral salt soluble collagen gel, susceptibility of insoluble collagen to denaturing agents and bacterial collagenase. Electrophoresis on SDS-polyacrylamide gel revealed a marked increase in the beta components of the collagen of the rats exposed to cigarette smoke (5.34%) and an appreciable decrease in the ratio of alpha/beta (7.78%). An increase in the aldehyde content of neutral salt soluble collagen was also noticed (32.11%). These changes collectively could indicate the increased crosslinking of dermal collagen in cigarette smoke exposed rats.


Role of Laboratory in Emerging Infectious Disease Control in Iran, Pasteur Institute of Iran, and national laboratory network.
Strengthening surveillance systems is a key aspect of outbreak response and was particularly important during the COVID-19 pandemic. Respiratory pathogens spread rapidly, and laboratory capacity is key to monitoring the spread. Prior to the pandemic, Iran had established a rapid response team and laboratory network to provide identification, monitoring, and detection of emerging infectious diseases, but did not have the laboratory capacity to respond to COVID-19. Following the announcement of the COVID-19 pandemic, the rapid response team diverted all attention to supporting COVID-19 surveillance. Iran built on the existing national laboratory infrastructure to incorporate SARS-CoV-2 surveillance into the response network. Based on existing international protocols, in-house molecular diagnosis capacity was operationalized, and commercial controls and assays were acquired and validated to national standards. The first COVID-19 laboratory was operational by January 25, less than 4 weeks before the initial detection of SARS-CoV-2 was announced. Assays and support were expanded and rolled out to form the COVID-19 National Laboratory Network, which consists of 560 multi-sectoral laboratories covering all provinces of Iran. The national laboratory network supports a wide range of operational capacities, including assay validation and protocol development, quality assurance, respiratory pathogen diagnosis and surveillance, and variant identification and assessment using multiple sequencing platforms. This network has supported the testing of over 55 million samples over the past 36 months using RT-qPCR and has sequenced approximately 2200 samples across the country, contributing the data to international databases, including GISAID.


Experiences of women who underwent induced lactation: A literature review.
This article aims to review the literature published over the past five decades related to the experiences of women who have undergone induced lactation. A comprehensive electronic search was conducted using PubMed, the Library of Congress, Google Scholar, SAGE, and ScienceDirect. The following search keywords were used: adoptive breastfeeding, induced lactation, non-puerperal lactation, extraordinary breastfeeding, and milk kinship. The search was restricted to articles written in English and published from 1956 to 2019. All study designs were included except for practice protocols. A total of 50 articles about induced lactation were retrieved. Of these, 17 articles identified the experiences of women who underwent induced lactation. The articles included original papers (n=7), reviews (n=5), and case reports (n=5). Four articles were specifically related to Malaysia, and the others were international. These 17 articles concerning the experiences of women who induced lactation will be reviewed based on four themes related to inducing lactation: (a) understanding women's perception of satisfaction, (b) emotional aspects, (c) enabling factors, and (d) challenges. Identifying a total of only 17 articles on induced lactation published over the last 53 years suggests that the subject is understudied. This review provides emerging knowledge regarding the experiences of women who have induced lactation in terms of satisfaction, emotions, enabling factors and challenges related to inducing lactation.


Healthberry 865® and Its Related, Specific, Single Anthocyanins Exert a Direct Vascular Action, Modulating Both Endothelial Function and Oxidative Stress.
In recent years, epidemiological studies have identified a relationship between diet and cerebro-cardiovascular disease (CVD). In this regard, there is a promising dietary group for cardiovascular protection are polyphenols, especially anthocyanins. Vascular reactivity studies were performed using Healthberry 865® and constituent single anthocyanins to characterize vasomotor responses; immunofluorescence analysis with dichlorofluorescein diacetate and dihydroethidium were used to evaluate nitric oxide and oxidative stress; lucigenin assay was used to measure NADPH oxidase activity; and gel electrophoresis and immunoblotting were used to dissect the molecular mechanisms involved. We demonstrated that Healthberry 865® exerts an important vasorelaxant effect of resistance artery functions in mice. Its action is mediated by nitric oxide release through the intracellular signaling PI3K/Akt. Moreover, behind its capability of modulating vascular tone, it also exerts an important antioxidant effect though the modulation of the NADPH oxidase enzyme. Interestingly, its cardiovascular properties are mediated by the selective action of different anthocyanins. Finally, the exposure of human dysfunctional vessels to Healthberry 865® significantly reduces oxidative stress and improves NO bioavailability. Although further investigations are needed, our data demonstrate the direct role of Healthberry 865® on the modulation of vasculature, both on the vasorelaxation and on oxidative stress; thus, supporting the concept that a pure mixture of anthocyanins could be helpful in preventing the onset of vascular dysfunction associated with the development of CVD.


IFN-gamma is critical to the control of murine autoimmune encephalomyelitis and regulates both in the periphery and in the target tissue: a possible role for nitric oxide.
NO and IFN-gamma have normally been considered cytotoxic and proinflammatory molecules, respectively, in the setting of the central nervous system inflammatory disease autoimmune encephalomyelitis (EAE). Using mice lacking the ligand binding chain of the IFN-gamma receptor (IFNgammaR-/-), we have previously shown that IFN-gamma is not essential for myelin oligodendrocyte glycoprotein peptide (MOG35-55) induced EAE expression but is in fact essential for its down-regulation. Here we examined the downstream molecular and cellular mechanism(s) of IFN-gamma regulation and demonstrate that neither IL-4 nor IL-10 appear to play a role in down-regulation nor do various lymphoid cell populations. Cells of the macrophage lineage are key to down-regulation as evidenced by the fact that peritoneal exudate cells from IFNgammaR+/+ mice inhibit Ag-driven proliferation of IFNgammaR-/- lymphocytes, whereas IFNgammaR-/- peritoneal exudate cells do not. High levels of reactive nitrogen intermediates are detected in the former cultures but not the latter, and the inhibition of proliferation is reversible with an inhibitor of inducible NO synthase, indicating a key role for NO in down-regulation. Studies with bone marrow chimeras indicate that down-regulation occurs not only systemically but also within the target tissue. These data suggest that IFN-gamma down-regulates EAE by inducing inducible NO synthase and subsequently NO production, both by macrophages in the periphery and, by inference, microglia and astrocytes in the target tissue.


Developing leadership and excellence with vulnerable populations in research, education, and practice.
The development of leadership and excellence in an area of nursing scholarship depends on the interactions among practice, research, theory, and education. This article describes the development of leadership at the UCLA School of Nursing through scholarship in the health-related problems of vulnerable populations. Leadership in scholarship began and continues in practice, influences education, generates theory, is tested in research, and returns to influence all of these areas. The vitality and creativity of scholarship and science with vulnerable populations depends on an ongoing infusion of knowledge from practice, research, theory, and education. While focusing on one school, there are many that have followed a similar course in developing scholarship and others that may wish to develop their scholarship based on this model.


Lactobacillus acidipiscis sp. nov. and Weissella thailandensis sp. nov., isolated from fermented fish in Thailand.
Eleven strains of homofermentative, rod-shaped lactic acid bacteria and five strains of heterofermentative, sphere-shaped lactic acid bacteria were isolated from fermented fish (pla-ra and pla-chom) in Thailand. They were identified as new species and named Lactobacillus acidipiscis sp. nov. and Weissella thailandensis sp. nov., respectively, on the basis of phylogenetic analysis of the 16S rRNA gene sequences, DNA relatedness and phenotypic characteristics. The type strain of L. acidipiscis is FS60-1T (= PCU 207T = NRIC 0300T = HSCC 1411T = JCM 10692T = TISTR 1386T) and the type strain of Weissella thailandensis is FS61-1T (= PCU 210T = NRIC 0298T = HSCC 1412T = JCM 10695T = TISTR 1384T).


CD4-independent binding of SIV gp120 to rhesus CCR5.
CCR5 and CD4 are coreceptors for immunodeficiency virus entry into target cells. The gp120 envelope glycoprotein from human immunodeficiency virus strain HIV-1(YU2) bound human CCR5 (CCR5hu) or rhesus macaque CCR5 (CCR5rh) only in the presence of CD4. The gp120 from simian immunodeficiency virus strain SIVmac239 bound CCR5rh without CD4, but CCR5hu remained CD4-dependent. The CD4-independent binding of SIVmac239 gp120 depended on a single amino acid, Asp13, in the CCR5rh amino-terminus. Thus, CCR5-binding moieties on the immunodeficiency virus envelope glycoprotein can be generated by interaction with CD4 or by direct interaction with the CCR5 amino-terminus. These results may have implications for the evolution of receptor use among lentiviruses as well as utility in the development of effective intervention.


[Pharmacological and clinical profile of linaclotide (Linzess®), a novel therapeutic agent for irritable bowel syndrome with constipation and chronic constipation].
Linaclotide (Linzess® tablets 0.25 mg) is a guanylate cyclase-C (GC-C) agonist with high selectivity and binding affinity to GC-C. In Japan, linaclotide was approved for 〝irritable bowel syndrome with constipation (IBS-C)〟 in December 2016 and 〝chronic constipation (CC) (excluding constipation due to organic disease)〟 in August 2018. Non-clinical studies demonstrated that linaclotide binding to GC-C increases intracellular cyclic guanosine monophosphate (cGMP), resulting in increased fluid secretion and gastrointestinal transit. In rats with colonic hyperalgesia, but not in normal rats, linaclotide suppressed the visceral nociceptive response, mediated by increased submucosal cGMP. In clinical studies in Japan, improvements were observed in the responder rates for global assessment of IBS symptom relief, complete spontaneous bowel movements in patients with IBS-C, and the frequency of spontaneous bowel movement in patients with CC, which were maintained during long-term treatment. Additionally, abdominal bloating, which has been associated with lower quality of life (QOL) and lower satisfaction with other approved therapies, and IBS QOL were improved throughout treatment with linaclotide. Diarrhea, a consequence of linaclotide's mechanism of action, was observed during the clinical studies, but was generally controllable by decreasing the linaclotide dose. No drug resistance was observed during the clinical studies, unlike some other approved agents. These results of non-clinical and clinical studies demonstrate that linaclotide can improve constipation, various abdominal symptoms, and QOL with a favorable safety profile in patients with IBS-C and CC.


Biosynthesized Silver Nanoparticles by Aqueous Stem Extract of Entada spiralis and Screening of Their Biomedical Activity.
Silver nanoparticles (Ag-NPs) have been established as antibacterial nanoparticles and have been innovatively developed to overcome the occurrence of antibiotic resistance in the environment. In this study, an environmentally friendly and easy method of the biosynthesis of Ag-NPs plants, mediated by aqueous extract stem extract of Entada spiralis (E. spiralis), was successfully developed. The E. spiralis/Ag-NPs samples were characterized using spectroscopy and the microscopic technique of UV-visible (UV-vis), X-ray Diffraction (XRD), Field Emission Transmission Electron Microscope (FETEM), zeta potential, and Fourier Transform Infrared (FTIR) analyses. Surface Plasmon Resonance (SPR) absorption at 400-450 nm in the UV-vis spectra established the formation of E. spiralis/Ag-NPs. The crystalline structure of E. spiralis/Ag-NPs was displayed in the XRD analysis. The small size, around 18.49 ± 4.23 nm, and spherical shape of Ag-NPs with good distribution was observed in the FETEM image. The best physicochemical parameters on Ag-NPs biosynthesis using E. spiralis extract occurred at a moderate temperature (~52.0°C), 0.100 M of silver nitrate, 2.50 g of E. spiralis dosage and 600 min of stirring reaction time. The antibacterial activity was tested against Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, and Proteus vulgaris using an antibacterial disk diffusion assay. Based on the results, it is evident that E. spiralis/Ag-NPs are susceptible to all the bacteria and has promising potential to be applied in both the industry and medical fields.


White matter microstructure in chronic moderate-to-severe traumatic brain injury: Impact of acute-phase injury-related variables and associations with outcome measures.
This study examines how injury mechanisms and early neuroimaging and clinical measures impact white matter (WM) fractional anisotropy (FA), mean diffusivity (MD), and tract volumes in the chronic phase of traumatic brain injury (TBI) and how WM integrity in the chronic phase is associated with different outcome measures obtained at the same time. Diffusion tensor imaging (DTI) at 3 T was acquired more than 1 year after TBI in 49 moderate-to-severe-TBI survivors and 50 matched controls. DTI data were analyzed with tract-based spatial statistics and automated tractography. Moderate-to-severe TBI led to widespread FA decreases, MD increases, and tract volume reductions. In severe TBI and in acceleration/deceleration injuries, a specific FA loss was detected. A particular loss of FA was also present in the thalamus and the brainstem in all grades of diffuse axonal injury. Acute-phase Glasgow Coma Scale scores, number of microhemorrhages on T2*, lesion volume on fluid-attenuated inversion recovery, and duration of posttraumatic amnesia were associated with more widespread FA loss and MD increases in chronic TBI. Episodes of cerebral perfusion pressure <70 mmHg were specifically associated with reduced MD. Neither episodes of intracranial pressure >20 mmHg nor acute-phase Rotterdam CT scores were associated with WM changes. Glasgow Outcome Scale Extended scores and performance-based cognitive control functioning were associated with FA and MD changes, but self-reported cognitive control functioning was not. In conclusion, FA loss specifically reflects the primary injury severity and mechanism, whereas FA and MD changes are associated with objective measures of general and cognitive control functioning.


Pulmonary Sarcomatoid carcinoma: a surgical diagnosis and prognostic factors.
Pulmonary Sarcomatoid Carcinoma (PSC) is a rare group of tumors accounting for about 0.4% of non-small cell lung carcinoma (NSCLC). Five subtypes were described: pleomorphic carcinoma, spindle cell carcinoma, carcinosarcoma, giant cell carcinoma and pulmonary blastoma. The diagnosis is pathological but requires a good quality sampling of the tumor. On a series of 1582 patients operated on for lung cancer from 1992 to 2016, 43 patients were retrospectively identified as having been treated surgically for pulmonary sarcomatoid carcinoma. The population consisted of 33 males and 10 females with mean age of 55 years. Imaging findings showed a peripheral mass in the majority of cases (n=29). Careful investigation failed to discover a primitive lesion elsewhere. Six patients received induction therapy for wall involvement. Lobectomy or bilobectomy was performed in 30 patients and pneumonectomy in 11 patients. A wedge resection was performed in one patient and an exploratory thoracotomy in another. In macroscopy, the mean tumor's size was 5.2 cm (1-17.5cm). The histologic diagnoses were: pleomorphic carcinoma (n=30), carcinosarcoma (n=5), spindle cell carcinoma (n=1), giant cell carcinoma (n=3) and blastoma (n=4). Two patients died within 1 month of surgical complications and 5 died of disease within 17 months. Adjuvant therapy was performed in 6 patients. Recurrence happened in 4 patients within 12 months after operation. Median survival for all patients was 8months. Resection of primary pulmonary sarcomatoid carcinoma is associated with an acceptable survival rate if the resection is complete. The size of the tumor is the most important prognosis factor. Nevertheless, a carefully follow-up is essential.


Hydrogen peroxide-induced apoptosis and necrosis in human lung fibroblasts: protective roles of glutathione.
Although reactive oxygen species (ROS)-related cell damage has been implicated in pathogenesis of fibrogenetic pulmonary disorders, features of ROS-mediated cell death in human lung fibroblasts are not completely understood. We therefore examined the effects of hydrogen peroxide (H2O2) on cell growth kinetics in human lung fibroblasts (HFL-1 cells) and tested the roles of antioxidants on the H2O2-induced cell death (i.e., necrosis and apoptosis) in HFL-1 cells. We found that the relatively low concentrations of H2O2 ranging from 10 microM to 100 microM induced predominantly apoptosis, whereas higher concentration of H2O2 ranging 1 mM-10 mM induced predominantly necrosis in HFL-1 cells. Extracellular supplementation of glutathione (GSH) in culture media significantly abolished the H2O2-induced cell death, whereas GSH-depleted cells by pretreatment with buthionine sulfoxime (BSO) were likely to undergo cell death caused by a lower concentration of H2O2 than normal HFL-1 cells without BSO treatment. These results indicate that H2O2 induces both necrosis and apoptosis of human lung fibroblasts at least in part through the action of ROS and that modulation of the ROS production inside and outside of cells may influence the cell survival during oxidative insults.


Knowledge and attitudes towards mpox and effect of intervention among College of Applied Medical Sciences students.
Objectives: This research aims to assess the base level of knowledge and attitude of Applied Medical Sciences Students regarding mpox and whether an educational intervention could improve their knowledge and attitude. Methods: A quasi-experimental research was used, involving 960 medical students from Applied Medical Sciences College at Prince Sattam Bin Abdulaziz University, KSA. They were recruited from the beginning of November 2022 till the mid of January 2023 based on the non-randomized sampling method. A standardized, anonymous, and closed-ended questionnaire was used, compromising three main sections: participants' demographics, knowledge, and attitudes toward the mpox epidemic. Results: Total knowledge scores of the studied sample in the pretest phase were 45.43 ± 6.29 compared to 65.03 ± 2.93 in the post-test phase. Besides, total attitude scores were 48.62 ± 4.78 before program implementation, while after conducting the program reached 70.65 ± 5.13. There was a notable improvement in the total knowledge score of the sample studied after the intervention was implemented, particularly for neurological manifestations. Conclusion: After the program's implementation, there was an obvious improvement in the medical students' total knowledge and attitude scores concerning the mpox epidemic. It is necessary to initiate well-organized training initiatives for all other medical faculties, paramedics, and applied health institutions in Saudi Arabia.


Increased tumor uptake of chemotherapeutics and improved chemoresponse by novel non-anticoagulant low molecular weight heparin.
Recent prospective clinical trials of low molecular weight heparins (LMWHs) have demonstrated that these agents may provide significant advantages in terms of progression-free and overall survival in certain subgroups of cancer patients. The mechanisms of improved survival associated with LMWHs are not known, and may involve direct and/or indirect effects on tumor growth. The purpose of this study was to investigate the effects of LMWH and a sulfated non-anticoagulant LMWH (S-NACH) on tumor chemotherapeutic uptake and chemoresponse. LMWH and S-NACH were tested for their ability to reduce tumor growth and tumor-associated angiogenesis using three different in vivo models. Biodistribution studies were undertaken to determine the effect of these agents on uptake of paclitaxel (PACL) and doxorubicin (Dox) by breast cancer tumor xenografts. LMWH and S-NACH (10 mg/kg s.c. daily) effectively limited tumor growth of human A549 lung adenocarcinoma xenografts in the nude mouse. In an MDA453/LCC6 breast tumor xenograft model, PACL plus S-NACH showed significant (p < 0.01) tumor growth suppression and improved survival when compared to PACL alone. LMWH increased [(124-)I]-PACL uptake into MDA453/LCC6 tumors, with tumor:muscle ratios several fold greater than that of [(124-)I]-PACL alone 24 h post-injection. Similarly, LMWH and S-NACH significantly (p < 0.01) increased the uptake of Dox by 1.5-2 fold in MCF7 Dox-resistant tumor xenografts. Protocols utilizing adjuvant or neo-adjuvant therapy with LMWH or S-NACH could lead to increased tumor chemo responsiveness, potentially overcoming tumor chemoresistance.


Factors associated with the likelihood of Giardia spp. and Cryptosporidium spp. in soil from dairy farms.
A study was conducted to identify factors associated with the likelihood of detecting Giardia spp. and Cryptosporidium spp. in the soil of dairy farms in a watershed area. A total of 37 farms were visited, and 782 soil samples were collected from targeted areas on these farms. The samples were analyzed for the presence of Cryptosporidium spp. oocysts, Giardia spp. cysts, percent moisture content, and pH. Logistic regression analysis was used to identify risk factors associated with the likelihood of the presence of these organisms. The use of the land at the sampling site was associated with the likelihood of environmental contamination with Cryptosporidium spp. Barn cleaner equipment area and agricultural fields were associated with increased likelihood of environmental contamination with Cryptosporidium spp. The risk of environmental contamination decreased with the pH of the soil and with the score of the potential likelihood of Cryptosporidium spp. The size of the sampling site, as determined by the sampling design, in square feet, was associated nonlinearly with the risk of detecting Cryptosporidium spp. The likelihood of the Giardia cyst in the soil increased with the prevalence of Giardia spp. in animals (i.e., 18 to 39%). As the size of the farm increased, there was decreased risk of Giardia spp. in the soil, and sampling sites which were covered with brush or bare soil showed a decrease in likelihood of detecting Giardia spp. when compared to land which had managed grass. The number of cattle on the farm less than 6 mo of age was negatively associated with the risk of detecting Giardia spp. in the soil, and the percent moisture content was positively associated with the risk of detecting Giardia spp. Our study showed that these two protozoan exist in dairy farm soil at different rates, and this risk could be modified by manipulating the pH of the soil.


Identification and characterization of foot-and-mouth disease virus O1 Burgwedel/1987 as an intertypic recombinant.
The foot-and-mouth disease virus field isolate Burgwedel/1987 subtype O1 was found to differ genetically from the antigenically related strain O1 Kaufbeuren within the region encoding the non-structural proteins. This genetic difference was indicated by the RNase mismatch cleavage method and confirmed by nucleotide sequencing. An alignment of sequences encoding proteinase 3C of the Burgwedel isolate and several other virus strains identified this isolate as an intertypic recombinant; the parent strains were O1 Kaufbeuren and a subtype C1 strain. Recombination occurred between nucleotide positions 5493 and 5521, within the region encoding peptide 3B1. Thus, the 5' three-quarters of the O1 genome were fused to the 3'-terminal quarter of the C1 genome. Other contemporary isolates from the same district are not recombinants. Sequence alignment distinguished four patterns of proteinase 3C-coding sequences among the virus strains analysed: subtypes A12, C1 and O1 exhibit one pattern each, and another pattern is common to subtypes A5, A10 and O2.


Role of common genetic variants in ovarian cancer susceptibility and outcome: progress to date from the Ovarian Cancer Association Consortium (OCAC).
In this article, we review the current knowledge of the inherited genetics of epithelial ovarian cancer (EOC) susceptibility and clinical outcome. We focus on recent developments in identifying low-penetrance susceptibility genes and the role of the Ovarian Cancer Association Consortium (OCAC) in these discoveries. The OCAC was established to facilitate large-scale replication analyses for reported genetic associations for EOC. Since its inception, the OCAC has conducted both candidate gene and genome-wide association studies (GWAS); the latter has identified six established loci for EOC susceptibility, most of which showed stronger association with the serous histological subtype. Future GWAS and sequencing studies are likely to result in the discovery of additional susceptibility loci and may result in established associations with clinical outcome. Additional rare and uncommon ovarian cancer loci will likely be uncovered from high-throughput next-generation sequencing studies. Applying these novel findings to establish improved preventative and clinical intervention strategies will be one of the major challenges of future work.


Perspectives and Experiences of Patient-Led Melanoma Surveillance Using Digital Technologies From Clinicians Involved in the MEL-SELF Pilot Randomized Controlled Trial: Qualitative Interview Study.
The growing number of melanoma patients who need long-term surveillance increasingly exceeds the capacity of the dermatology workforce, particularly outside of metropolitan areas. Digital technologies that enable patients to perform skin self-examination and send dermoscopic images of lesions of concern to a dermatologist (mobile teledermoscopy) are a potential solution. If these technologies and the remote delivery of melanoma surveillance are to be incorporated into routine clinical practice, they need to be accepted by clinicians providing melanoma care, such as dermatologists and general practitioners (GPs). This study aimed to explore perceptions of potential benefits and harms of mobile teledermoscopy, as well as experiences with this technology, among clinicians participating in a pilot randomized controlled trial (RCT) of patient-led melanoma surveillance. This qualitative study was nested within a pilot RCT conducted at dermatologist and skin specialist GP-led melanoma clinics in New South Wales, Australia. We conducted semistructured interviews with 8 of the total 11 clinicians who were involved in the trial, including 4 dermatologists (3 provided teledermatology, 2 were treating clinicians), 1 surgical oncologist, and 3 GPs with qualifications in skin cancer screening (the remaining 3 GPs declined an interview). Thematic analysis was used to analyze the data with reference to the concepts of "medical overuse" and "high-value care." Clinicians identified several potential benefits, including increased access to dermatology services, earlier detection of melanomas, reassurance for patients between scheduled visits, and a reduction in unnecessary clinic visits. However, they also identified some potential concerns regarding the use of the technology and remote monitoring that could result in diagnostic uncertainty. These included poor image quality, difficulty making assessments from a 2D digital image (even if good quality), insufficient clinical history provided, and concern that suspicious lesions may have been missed by the patient. Clinicians thought that uncertainty arising from these concerns, together with perceived potential medicolegal consequences from missing a diagnosis, might lead to increases in unnecessary clinic visits and procedures. Strategies suggested for achieving high-value care included managing clinical uncertainty to decrease the potential for medical overuse and ensuring optimal placement of patient-led teledermoscopy within existing clinical care pathways to increase the potential for benefits. Clinicians were enthusiastic about the potential and experienced benefits of mobile teledermoscopy; however, managing clinical uncertainty will be necessary to achieve these benefits in clinical care outside of trial contexts and minimize potential harms from medical overuse. Australian and New Zealand Clinical Trials Registry ACTRN12616001716459; https://anzctr.org.au/Trial/Registration/TrialReview.aspx?id=371865.


Pain and haemorrhage are the most common reasons for emergency department use and hospital admission in adults following ambulatory surgery: results of a population-based cohort study.
Advances in healthcare delivery have allowed for the increase in the number of ambulatory surgery procedures performed in Canada. Despite these advances, patients return to hospital following discharge. However, the reason for unplanned healthcare use after ambulatory surgery in Canada is not well understood. To examine unplanned healthcare use, specifically emergency department visit and hospital admissions, in the 3 days after ambulatory surgery in Ontario, Canada. This population-based retrospective cohort study was conducted using de-identified administrative databases. Participants were residents in the province of Ontario, Canada; 18 years and older; and underwent common ambulatory surgical procedures between 2014 and 2018. The outcomes included emergency department (ED) visit and hospital admission. Incidence rates were calculated for the total cohort, for each patient characteristic and for surgical category. The odds ratios and 95% confidence intervals were calculated for each outcome using bivariate and multivariate logistic regression. 484,670 adults underwent select common surgical procedures during the study period. Patients had healthcare use in the first 3 days after surgery, with 14,950 (3.1%) ED visits and 14,236 (2.9%) admissions. The incidence of ED use was highest after tonsillectomy (8.1%), cholecystectomy (4.2%) and appendectomy (4.0%). Incidence of admissions was highest after appendectomy (21%). Acute pain (19.7%) and haemorrhage (14.2%) were the most frequent reasons for an ED visit and "convalescence following surgery" (49.2%) followed by acute pain (6.2%) and haemorrhage (4.5%) were the main reasons for admission. These findings can assist clinicians in identifying and intervening with patients at risk of healthcare use after ambulatory surgery. Pain management strategies that can be tailored to the patient, and earlier follow-up for some patients may be required. In addition, administrative decision-makers could use the results to estimate the impact of specific ambulatory procedures on hospital resources for planning and allocation of resources.


Liver transplantation for hepatocellular carcinoma.
Orthotopic liver transplantation (OLT) is the best available option for early hepatocellular carcinoma (HCC), although its application is limited by stringent selection criteria, costs, and deceased donor graft shortage, particularly in Asia, where living donor liver transplant (LDLT) has been developed. This article reviews the present standards for patient selection represented by size-and-number criteria with particular references to Milan Criteria and novel prediction models based on results achieved in patients exceeding those limits, with consideration of the expanded indication represented by the UCSF Criteria. The expected outcomes after deceased donor liver transplant (DDLT) or LDLT are favorable if predetermined selection criteria are applied. However, selection bias, difference in waiting time, and ischemia-regeneration injuries of the graft among DDLT vs LDLT may influence long-term results. In the article, the differences between East and West in first-line treatments for HCC (resection vs transplantation), indications, and ethics for the donor, are summarized as well as possible novel predictors of tumor biology (especially DNA mutation and fractional allelic loss, FAI) to be considered for better outcome prediction. Liver transplantation remains the most promising product of modern surgery and represents a cornerstone in the management of patients with HCC.


[Peripheral nerve blocks in paediatric anaesthesia].
Most children undergoing surgery can benefit from regional anaesthetic techniques, either as the sole anaesthetic regimen or, as usual in paediatric practice, in combination with general anaesthesia. The use of peripheral nerve blocks (PNBs) in paediatric anaesthesia is an effective way to decrease the side-effects and complications associated with central blocks. In spite of their many advantages, including easy performance end efficacy, peripheral nerve blocks are still underused. This article discusses a general approach to PNBs in children and provides data concerning the practice of this regional technique in different age groups. Data from 1,650 procedures were prospectively collected during the period from March 1, 2007 to February 29, 2008. The type of PNB, if any, as well as the patient age were noted. Our patients were divided into four groups: 0-3 years, 4-7 years, 8-12 years and 13-18 years. During the investigated period, PNBs as a sole technique or in anaesthetized children were performed in 7.45% of cases. Ilioinguinal/iliohypogastric nerve block and penile block were the most common (70% of all PNBs) distributed mainly among the children between 4-7 years of age (p < 0.05). In older children, extremity PNBs predominate in regard to other types of blocks. PNBs are most frequently performed under general anaesthesia (85%), so the perineural approach requires a safe technique to avoid nerve damage. The observed differences in PNB usage seem to be related to patient age and correlate with common pathology and also with technical availability of PNB performance.


The influence of different metal-chelate conjugates of pentixafor on the CXCR4 affinity.
The overexpression of the chemokine receptor 4 (CXCR4) in different epithelial, mesenchymal, and hematopoietic cancers makes CXCR4 an attractive diagnostic and therapeutic target. However, targeting the CXCR4 receptor with small cyclic pentapeptide-based radiopharmaceuticals remains challenging because minor structural modifications within the ligand-linker-chelate structure often significantly affect the receptor affinity. Based on the excellent in vivo properties of CXCR4-directed pentapeptide [(68)Ga]pentixafor (cyclo(-D-Tyr-N-Me-D-Orn(AMB-DOTA)-L-Arg-L-2-Nal-Gly-)), this study aims to broaden the spectrum of applicable (radio)metal-labeled pentixafor analogs. Cyclic pentapeptides, based on the pentixafor scaffold, were synthesized by a combined solid- and solution-phase peptide synthesis. The CXCR4 receptor affinities of the cold reference compounds were determined in competitive binding assays using CXCR4-expressing Jurkat T - cell leukemia cells and [(125)I]FC131 as the radioligand. Metalated pentixafor derivatives with cyclic and acyclic chelators were synthesized by solid-phase peptide synthesis and evaluated in vitro. The resulting CXCR4 affinities (IC50) were highly dependent on the chelator and metal used. Two pentapeptides, Ga-NOTA and Bi-DOTA conjugates, offer an improved affinity compared to [(68)Ga]pentixafor. Based on the pentapeptide [(68)Ga]pentixafor, a broad range of metal-labeled analogs were investigated. The affinities of the new compounds were found to be strongly dependent on both the chelator and the metal used. Bi-labeled pentixafor showed high receptor affinity and seems to be a promising ligand for further preclinical evaluation and future α-emitter-based endoradiotherapy.


Pioglitazone hydrochloride stimulates insulin secretion in HIT-T 15 cells by inducing Ca2+ influx.
Pioglitazone hydrochloride (AD-4833), one of the thiazolidinedione analogs, is a new anti-diabetic agent which improves peripheral insulin resistance in diabetic patients. We determined the direct effect of AD-4833 on insulin secretion in HIT-T 15 cells. The effects of AD-4833 (10(-7) M to 10(-5) M) on insulin secretion were examined in 3 and 7 mM glucose-containing F-12 K media. The addition of 10(-5) M AD-4833 significantly increased insulin secretion in both media, but its stimulatory effect was more potent in the medium containing 7 mM glucose. The addition of 10(-5) M AD-4833 caused an immediate, significant increase in cytosolic free Ca2+ concentration ([Ca2+]i). Nifedipine at all concentrations from 10 to 1000 nM significantly attenuated insulin secretion by 10(-5) M AD-4833. In addition, 10(-5) M AD-4833 failed to stimulate insulin secretion in the CA(2+)-free Kreb's-Ringer bicarbonate buffer. These data indicated that AD-4833 stimulates in vitro insulin secretion in HIT-T 15 cells, perhaps by inducing Ca2+ influx.


Design and Synthesis of Novel α-Methylchalcone Derivatives, Anti-Cervical Cancer Activity, and Reversal of Drug Resistance in HeLa/DDP Cells.
In this study, a collection of newly developed α-methylchalcone derivatives were synthesized and assessed for their inhibitory potential against human cervical cancer cell lines (HeLa, SiHa, and C33A) as well as normal human cervical epithelial cells (H8). Notably, compound 3k exhibited substantial inhibitory effects on both HeLa and HeLa/DDP cells while demonstrating lower toxicity toward H8 cells. Furthermore, the compound 3k was found to induce apoptosis in both HeLa and HeLa/DDP cells while also inhibiting the G2/M phase, resulting in a decrease in the invasion and migration capabilities of these cells. When administered alongside cisplatin, 3k demonstrated a significant reduction in the resistance of HeLa/DDP cells to cisplatin, as evidenced by a decrease in the resistance index (RI) value from 7.90 to 2.10. Initial investigations into the underlying mechanism revealed that 3k did not impact the expression of P-gp but instead facilitated the accumulation of rhodamine 123 in HeLa/DDP cells. The results obtained from CADD docking analysis demonstrated that 3k exhibits stable binding to microtubule proteins and P-gp targets, forming hydrogen bonding interaction forces. Immunofluorescence analysis further revealed that 3k effectively decreased the fluorescence intensity of α and β microtubules in HeLa and HeLa/DDP cells, resulting in disruptions in cell morphology, reduction in cell numbers, nucleus coagulation, and cell rupture. Additionally, Western blot analysis indicated that 3k significantly reduced the levels of polymerized α and β microtubule proteins in both HeLa and HeLa/DDP cell lines while concurrently increasing the expression of dissociated α and β microtubule proteins. The aforementioned findings indicate a potential correlation between the inhibitory effects of 3k on HeLa and HeLa/DDP cells and its ability to inhibit tubulin and P-gp.


Effect of gamma-radiation on fowl sperm function in vitro and in vivo.
Doses of up to 300 Gy of ionizing radiation had little effect on fowl sperm morphology, ATP content and motility when measured in vitro. Fertility of eggs from hens inseminated with spermatozoa receiving 50 Gy, in terms of post-oviducal development, was less than 4%. However, 35% appeared 'fertile' by macroscopic examination of the germinal disc of unincubated eggs. These contained few embryonic cells, although the vitelline membrane contained many trapped spermatozoa. After doses of 100 Gy or more, inseminated spermatozoa were not found in the vitelline membrane and no fertile or apparently fertile eggs were produced; nor did such spermatozoa enter the utero-vaginal sperm-host glands. Genetic transformation using fowl spermatozoa irradiated with doses in excess of 100 Gy appears to be an unlikely prospect.


Urine metabolic phenotyping in children with nocturnal enuresis and comorbid neurobehavioral disorders.
Nocturnal enuresis (NE) is a common problem among 10% school-aged children. The etiologies underlying childhood NE is complex and not fully understood nowadays. Nevertheless, increasing evidence suggests a potential link between neurobehavioral disorders and enuresis in children. In this study, we aimed to explore novel metabolomic insights into the pathophysiology of NE and also, its association with pediatric psychiatric problems. Urine collected from 41 bedwetting children and 27 healthy control children was analyzed by using 1H-nuclear magnetic resonance spectroscopy from August 2017 to December 2018. At regular follow-up, there were 14 children with refractory NE having a diagnosis of attention deficient hyperactivity disorder (ADHD) or anxiety. Eventually, we identified eight significantly differential urinary metabolites and particularly increased urinary excretion of betaine, creatine and guanidinoacetate linked to glycine, serine and threonine metabolism were associated with a comorbidity of neurobehavioral disorders in refractory bedwetting children. Notably, based on physiological functions of betaine acting as a renal osmolyte and methyl group donor, we speculated its potential role in modulation of renal and/or central circadian clock systems, becoming a useful urinary metabolic marker in diagnosis of treatment-resistant NE in children affected by these two disorders.


Diauxic Inhibition: Jacques Monod's Ignored Work.
Diauxie is at the origin of research that led Jacques Monod (1910-1976), François Jacob (1920-2013), and André Lwoff (1902-1994) to win the Nobel Prize in Physiology or Medicine in 1965 for their description of the first genetic regulatory model. Diauxie is a term coined by Jacques Monod in 1941 in his doctoral dissertation that refers to microbial growth in two phases. In this article, we first examine Monod's thesis to demonstrate how and why Monod interpreted diauxie as a phenomenon of enzyme inhibition or suppression of adaptive enzymes. We also briefly investigate prior enzyme suppression studies, before Monod's work, which indicate that he is the first person to observe diauxic growth. Second, we analyse Monod's post-thesis publications throughout his scientific career, revealing that diauxic inhibition was a significant part of Monod's scientific activities and greatly fascinated Monod until the end of his life. Paradoxically, Monod's work and interest on diauxic inhibition are still neglected in historical recounts, focused mostly on Monod's enzymatic adaptation studies. Indeed, we uncovered a statement by Monod's colleague, Lwoff, who transformed a quotation from Monod by replacing the word phenomenon with enzymatic adaptation, which we believe has influenced historians. Finally, we offer hypotheses to explain why Lwoff altered Monod's statement.


A new approach in analyzing the accident severity of pedestrian crashes using structural equation modeling.
According to official statistics in Iran, there were 17000 fatalities in road traffic crashes in 2018 that 25% of all crash fatalities belong to pedestrians. In most of the researches related to pedestrians' safety, one aspect of the traffic crash (e.g. the injury or crash severity) is almost considered for the investigation. In order to perform a complete study of the crash, accident size can be utilized which involves different aspects of the crash. Accident size is described in terms of the number of fatalities and injured individuals and the number of damaged and involved vehicles in a crash. According to the fact that accident size has multiple indicators and it is not measured directly, traditional methodologies cannot be applied. So, in the present study the effective factors on the accident size of pedestrian crashes are investigated through structural equation modeling. For the purpose of this study, 3718 pedestrian-involved crash data occurred in Isfahan province is used for the modeling. The independent variables are weather conditions, road surface conditions, time, horizontal and vertical alignments, road type and location, driver's gender and age, vehicle type, pedestrian's age, gender and clothing color. The results indicated that highways, the pedestrians' invisibility, female and old-aged pedestrians, heavy vehicles, old-aged and female drivers are related to the increase of the accident size in pedestrian crashes. These results denote that the mentioned variables are associated with the higher number of injuries, fatalities, the higher number of involved and damaged vehicles in a crash. Present study shows the importance of considering safety improvement measures in highways, educating the people in the society about the traffic safety, the separation of pedestrian and motor vehicle traffic flow and considering the old people in policies and programs for mitigating the accident size.


Corded Ware cultural complexity uncovered using genomic and isotopic analysis from south-eastern Poland.
During the Final Eneolithic the Corded Ware Complex (CWC) emerges, chiefly identified by its specific burial rites. This complex spanned most of central Europe and exhibits demographic and cultural associations to the Yamnaya culture. To study the genetic structure and kin relations in CWC communities, we sequenced the genomes of 19 individuals located in the heartland of the CWC complex region, south-eastern Poland. Whole genome sequence and strontium isotope data allowed us to investigate genetic ancestry, admixture, kinship and mobility. The analysis showed a unique pattern, not detected in other parts of Poland; maternally the individuals are linked to earlier Neolithic lineages, whereas on the paternal side a Steppe ancestry is clearly visible. We identified three cases of kinship. Of these two were between individuals buried in double graves. Interestingly, we identified kinship between a local and a non-local individual thus discovering a novel, previously unknown burial custom.


Protection against Vibrio vulnificus infection by active and passive immunization with the C-terminal region of the RtxA1/MARTXVv protein.
Vibrio vulnificus is a foodborne pathogen that is prevalent in coastal waters worldwide. Infection with V. vulnificus causes septicemia with fatality rates exceeding 50% even with aggressive antibiotic therapy. Several vaccine studies to prevent V. vulnificus infection have been performed but have had limited success. In this study, we identified the C-terminal region (amino acids 3491 to 4701) of the V. vulnificus multifunctional autoprocessing RTX (MARTXVv or RtxA1) protein, RtxA1-C, as a promising antigen that induces protective immune responses against V. vulnificus. Vaccination of mice with recombinant RtxA1-C protein with adjuvant elicited a robust antibody response and a dramatic reduction in blood bacterial load in mice infected intraperitoneally. Vaccination resulted in significant protection against lethal challenge with V. vulnificus. Furthermore, intraperitoneal passive immunization with serum raised against the recombinant RtxA1-C protein demonstrated marked efficacy in both prophylaxis and therapy. These results suggest that active and passive immunization against the C-terminal region of the RtxA1 protein may be an effective approach in the prevention and therapy of V. vulnificus infections.


Immunodiagnosis of aspergillosis.
Aspergillus species are ubiquitous fungi and have been implicated as the causative agents of a variety of lung disorders in humans. These disorders include allergic, saprophytic, and systemic manifestations. The allergic disorders mainly affect atopic persons, and invasive or systemic diseases affect immunosuppressed individuals. Immunodiagnosis can help the practitioner diagnose these diseases. Demonstration of circulating antibodies is a useful criterion, but the lack of dependable and standardized antigens is a limiting factor in the diagnosis of most Aspergillus-induced diseases. Despite this limitation, however, immunodiffusion and enzyme-linked immunosorbent assays have been widely used for the detection of antibodies in the sera of patients with aspergillosis. Similarly, crude and semipurified antigens are being used to demonstrate skin hypersensitivity in patients, and several methods have been useful in the detection of antigenemia in patients with invasive aspergillosis. With a growing number of reports on the incidence of aspergillosis and an increase in the number of immunosuppressed individuals in the population, more rapid methods and more reliable reagents for immunodiagnosis are needed. With recent attempts at obtaining reliable reagents for through hybridoma technology and molecular biological techniques, substantial progress toward efficient immunodiagnosis may be achieved.


Sensitization in early age to food allergens in children with atopic dermatitis.
Clinical and laboratory evidence increasingly supports the notion that food allergy plays a role in the pathogenesis of atopic dermatitis (AD). However, the prevalence of clinically significant food hypersensitivity among children with AD remains an unanswered question. To prospectively determine the prevalence of IgE-mediated food hypersensitivity among patients referred to a dermatology department for evaluation of AD, and to analyze the clinical relevance of these sensitizations in AD. We studied 44 infants of both sexes, aged less than 12 months old, who attended the dermatology department with symptoms of AD. Compliance with Hanifin-Rajka criteria was confirmed and the severity of AD was evaluated using the SCORAD index. IgE-mediated sensitization to cow's milk, alpha-lactalbumin, beta-lactoglobulin, casein, egg-white, egg-albumin, ovomucoid and foods introduced into the diet was studied using the skin prick test (SPT) and measurement of specific serum IgE (sIgE) by CAP System fluorescein-enzyme immunoassay. Cow's milk, as well as suspected foods from the clinical history or those with a positive SPT and/or sIgE, were withdrawn from the diet to evaluate improvement in AD, and an open controlled challenge test was carried out. Of the 44 patients studied, sensitization to foods was detected in 27 (61 %). No changes were observed in AD during the elimination diet or when the eliminated foods were subsequently reintroduced into the diet. The results of open controlled food challenges were positive in 12 patients (27 %). A high prevalence of food sensitization was found in infants with AD. The most frequent sensitization observed was to egg, although with little clinical relevance since this food had not been introduced into the diet. In the sample studied, the clinical relevance of the observed food hypersensitivities was confirmed in relation to AD. Further studies are required to confirm these results.


Food and Drug Administration, American Academy of Ophthalmology, American Academy of Optometry, American Association for Pediatric Ophthalmology and Strabismus, American Optometric Association, American Society of Cataract and Refractive Surgery, and Contact Lens Association of Ophthalmologists Co-Sponsored Workshop: Controlling the Progression of Myopia: Contact Lenses and Future Medical Devices.
The prevalence of myopia is high and increasing. Approximately 5 billion people around the world are expected to be myopic by the year 2050. Methods to slow the progression of myopia and therefore potentially decrease the associated sight-threatening complications have been the subject of a number of investigations. A workshop, sponsored by the United States Food and Drug Administration (FDA) Center for Devices and Radiological Health, American Academy of Ophthalmology, American Academy of Optometry, American Association for Pediatric Ophthalmology and Strabismus, American Optometric Association, American Society of Cataract and Refractive Surgery, and Contact Lens Association of Ophthalmologists, Inc, convened myopia experts from around the world to discuss principles to consider in the design of clinical trials investigating the effectiveness and safety of myopia control devices. Experts discussed parameters such as study endpoints, duration, enrollment criteria, patient-reported outcomes, recruitment, and retention. The discussions among the experts, FDA, and audience members should help to facilitate the development and evaluation of reasonably safe and effective myopia control devices.


Inflammation and collagenase production in rats with adjuvant arthritis reduced with 13-cis-retinoic acid.
Oral administration of 13-cis-retinoic acid (40 or 160 milligrams per kilogram of body weight daily) significantly reduced the inflammation associated with developing and established adjuvant arthritis, an experimentally induced arthritis in rats that resembles human rheumatoid arthritis. The amount of collagenase secreted in tissue culture by adherent cells isolated from the inflamed joints of adjuvant rats treated with 13-cis-retinoic acid also decreased as compared to the amount secreted by cells from vehicle-treated adjuvant rats. Collagenase is important in the joint destruction accompanying rheumatoid arthritis. The successful use of retinoids in the treatment of this proliferative but nonmalignant disorder demonstrates a new application of these compounds.


Relationship of glycemic control to total diabetes-related costs for managed care health plan members with type 2 diabetes.
Glycosylated hemoglobin (A1c) is a well-established measure of glycemic control, and evidence suggests that maintaining an acceptable A1c level may be associated with lower treatment costs in adults with diabetes. Understanding the impact on total treatment costs of staying at the target A1c level is of great importance to managed care organizations. The goal of this study was to determine whether type 2 diabetes patients at or below the target A1c level of 7% had lower diabetes-related costs compared with patients above an A1c level of 7%. This study was a retrospective database analysis using eligibility data, medical and pharmacy claims data, and laboratory data from a large U.S. health care organization. Patients were included in the study if they had 2 or more claims for type 2 diabetes (International Classification of Diseases, Ninth Revision, Clinical Modification [ICD-9-CM] codes 250.x0 or 250.x2) and at least 1 A1c value (first such date defined as the index date) during the 12-month period from January 1, 2002, through December 31, 2002. Patients with 2 or more medical claims for type 1 diabetes (ICD-9-CM codes 250.x1 or 250.x3) were excluded from the study. Study patients were divided into 2 groups, those at the target A1c level (7%) and those at the above-target A1c level (>7%), and were followed for a period of 1 year after their index date. Demographic, clinical, and cost variables were extracted from the administrative database. Multiple linear regression analysis was used to compare treatment costs between patients at the target A1c level and patients above target level. A total of 3,121 patients (46.0%) were identified as being at the target A1c level, and 3,659 patients (54%) were identified as being above the target A1c level during the study period. After controlling for confounding factors, the predicted total diabetes-related cost for the above-target group during the 1-year follow-up period was 1,540 US dollars per patient, 32% higher than the total diabetes-related cost (1,171 US dollars) for the at-target group (P <0.001). Results of this analysis suggest that managed care members with type 2 diabetes who stayed continuously at the target A1c level of 7% or less over a 1-year follow-up period incurred lower diabetes-related costs compared with managed care members with type 2 diabetes who were continuously over the target A1c level of 7%.


Antibacterial mechanism of gold nanoparticles on Streptococcus pneumoniae.
Streptococcus pneumoniae is a causal agent of otitis media, pneumonia, meningitis and severe cases of septicemia. This human pathogen infects elderly people and children with a high mortality rate of approximately one million deaths per year worldwide. Antibiotic-resistance of S. pneumoniae strains is an increasingly serious health problem; therefore, new therapies capable of combating pneumococcal infections are indispensable. The application of gold nanoparticles has emerged as an option in the control of bacterial infections; however, the mechanism responsible for bacterial cell lysis remains unclear. Specifically, it has been observed that gold nanoparticles are capable of crossing different structures of the S. pneumoniae cells, reaching the cytosol where inclusion bodies of gold nanoparticles are noticed. In this work, a novel process for the separation of such inclusion bodies that allowed the analysis of the biomolecules such as carbohydrates, lipids and proteins associated with the gold nanoparticles was developed. Then, it was possible to separate and identify proteins associated with the gold nanoparticles, which were suggested as possible candidates that facilitate the interaction and entry of gold nanoparticles into S. pneumoniae cells.


Mathematical modelling of pseudorabies virus (syn. Aujeszky's disease virus) outbreaks aids eradication programmes: a review.
Pseudorabies virus will be eradicated from the Netherlands if a typical infectious pig (Rind) infects, on average, less than one other pig. In this review, we used a stochastic SIR model to estimate Rind using data from the field and from experiments. Rind in sow herds was estimated to be significantly less than 1 and in rearing and finishing pigs Rind was higher than 1. However, if Rind is higher than 1, PRV can still be eradicated if one infectious herd infects less than one other herd during the period that the herd is infectious(Rherd <1). Some future developments in Dutch pig husbandry (e.g. group-housing of sows) and possible risks after halting vaccination are also quantitatively evaluated.


Charge-induced phase separation in lipid membranes.
Phase separation in lipid bilayers that include negatively charged lipids is examined experimentally. We observed phase-separated structures and determined the membrane miscibility temperatures in several binary and ternary lipid mixtures of unsaturated neutral lipid, dioleoylphosphatidylcholine (DOPC), saturated neutral lipid, dipalmitoylphosphatidylcholine (DPPC), unsaturated charged lipid, dioleoylphosphatidylglycerol (DOPG((-))), saturated charged lipid, dipalmitoylphosphatidylglycerol (DPPG((-))), and cholesterol. In binary mixtures of saturated and unsaturated charged lipids, the combination of the charged head with the saturation of the hydrocarbon tail is a dominant factor in the stability of membrane phase separation. DPPG((-)) enhances phase separation, while DOPG((-)) suppresses it. Furthermore, the addition of DPPG((-)) to a binary mixture of DPPC/cholesterol induces phase separation between DPPG((-))-rich and cholesterol-rich phases. This indicates that cholesterol localization depends strongly on the electric charge on the hydrophilic head group rather than on the ordering of the hydrocarbon tails. Finally, when DPPG((-)) was added to a neutral ternary system of DOPC/DPPC/cholesterol (a conventional model of membrane rafts), a three-phase coexistence was produced. We conclude by discussing some qualitative features of the phase behaviour in charged membranes using a free energy approach.


Comparison of cutting and pencil-point spinal needle in spinal anesthesia regarding postdural puncture headache: A meta-analysis.
Postdural puncture headache (PDPH), mainly resulting from the loss of cerebral spinal fluid (CSF), is a well-known iatrogenic complication of spinal anesthesia and diagnostic lumbar puncture. Spinal needles have been modified to minimize complications. Modifiable risk factors of PDPH mainly included needle size and needle shape. However, whether the incidence of PDPH is significantly different between cutting-point and pencil-point needles was controversial. Then we did a meta-analysis to assess the incidence of PDPH of cutting spinal needle and pencil-point spinal needle. We included all randomly designed trials, assessing the clinical outcomes in patients given elective spinal anesthesia or diagnostic lumbar puncture with either cutting or pencil-point spinal needle as eligible studies. All selected studies and the risk of bias of them were assessed by 2 investigators. Clinical outcomes including success rates, frequency of PDPH, reported severe PDPH, and the use of epidural blood patch (EBP) were recorded as primary results. Results were evaluated using risk ratio (RR) with 95% confidence interval (CI) for dichotomous variables. Rev Man software (version 5.3) was used to analyze all appropriate data. Twenty-five randomized controlled trials (RCTs) were included in our study. The analysis result revealed that pencil-point spinal needle would result in lower rate of PDPH (RR 2.50; 95% CI [1.96, 3.19]; P < 0.00001) and severe PDPH (RR 3.27; 95% CI [2.15, 4.96]; P < 0.00001). Furthermore, EBP was less used in pencil-point spine needle group (RR 3.69; 95% CI [1.96, 6.95]; P < 0.0001). Current evidences suggest that pencil-point spinal needle was significantly superior compared with cutting spinal needle regarding the frequency of PDPH, PDPH severity, and the use of EBP. In view of this, we recommend the use of pencil-point spinal needle in spinal anesthesia and lumbar puncture.


Tobacco purchasing in Australia during regular tax increases: findings from the International Tobacco Control Policy Evaluation Project.
We examined Australian tobacco purchasing trends, the average self-reported price paid within each purchase type and the association between type of tobacco product purchased and participant characteristics, including quit intentions, between 2007 and 2020. We analysed data collected from adults who smoked factory-made and/or roll-your-own (RYO) cigarettes in nine waves (2007-2020) of the International Tobacco Control Policy Evaluation Project Australia Survey (nsample=5452, nobservations=11 534). The main outcome measures were type of tobacco products purchased: RYO, carton, pack or pouch size and brand segment. Logistic regression, fit using generalised estimating equations, was estimated the association between the outcome and participant characteristics. The reported price-minimising purchasing patterns increased from 2007 to 2020: any RYO (23.8-43.9%), large-sized pack (2007: 24.0% to 2016: 34.3%); shifting from large-sized to small-sized packs (2020: 37.7%), and economy brand (2007: 37.2% to 2020: 59.3%); shifting from large (2007: 55.8%) to small economy packs (2014: 15.3% to 2020: 48.1%). Individuals with a lower income, a higher nicotine dependence level and no quit intention were more likely to purchase RYO and large-sized packs. RYO, large-sized packs and products with a low upfront cost (eg, small RYO pouches and small-sized economy brand packs) may appeal to people on low incomes. Australia's diverse tobacco pack and pouch sizes allow the tobacco industry to influence tobacco purchases. Standardising pack and pouch sizes may reduce some price-related marketing and especially benefit people who have a low income, are highly addicted and have no quit intention.


A role of cis-dichlorodiammineplatinum(II) in squamous cell lung cancer.
Forty-one patients with advanced squamous cell lung cancer and no prior chemotherapy were entered in a prospectively randomized trial comparing dianhydrogalactitol plus Adriamycin (DA) versus DA plus cis-dichlorodiammineplatinum(II) (DAP). The DAP regimen was superior to the DA regimen in regression rate (53% versus 27%), median regression duration (255 versus 122 days), median time to tumor progression (approximately 175 versus 58 days), and median survival time (185 versus 126 days). Patients who were greater than 60 years old responded particularly well to the DAP regimen and accounted for most of the survival advantage. Nausea, vomiting, and myelosuppression were more frequent and severe with the DAP regimen. This study seems to indicate a role of cis-dichlorodiammineplatinum(II) in patients with advanced squamous cell lung cancer. The particular advantage noted for older patients needs further evaluation.


Plasma miR-126 is a potential biomarker for early prediction of type 2 diabetes mellitus in susceptible individuals.
Type 2 diabetes mellitus (T2DM) is a major public health problem in China. Diagnostic markers are urgently needed to identify individuals at risk of developing T2DM and encourage them to adapt to a healthier life style. Circulating miRNAs present important sources of noninvasive biomarkers of various diseases. Recently, a novel plasma microRNA signature was identified in T2DM. Here, we evaluated the T2DM-related miRNA signature in plasma of three study groups: normal (fasting glucose (FG), 4.8-5.2 mmol/L), T2DM-susceptible (FG, 6.1-6.9 mmol/L), and T2DM individuals (FG, ≥ 7.0 mmol/L) and tested the feasibility of using circulating miRNAs to identify individuals at risk of developing T2DM. Among the 5 miRNAs included in the signature, miR-29b and miR-28-3p are not detectable. miR-15a and miR-223 have comparable expression levels among three groups. Notably, miR-126 is the only miRNA that showed significantly reduced expression in susceptible individuals and T2DM patients compared to normal individuals, suggesting that miR-126 in circulation may serve as a potential biomarker for early identification of susceptible individuals to T2DM.


The Fundamental Characteristics of a Translational Scientist.
Translational science is defined as the field of investigation focused on understanding the scientific and operational principles underlying each step of the translational process. Further development of the field is advanced by describing the key desirable characteristics of individuals who seek to uncover these principles to increase the efficiency and efficacy of translation. The members of Translation Together, a newly launched international collaborative effort to advance translational innovation, present here a consensus representation of the fundamental characteristics of a translational scientist. We invite all stakeholders to contribute in the ongoing efforts to develop the field and educate the next generation of translational scientists.


Association between habitual coffee consumption and skeletal muscle mass in middle-aged and older Japanese people.
Coffee consumption has been suggested, in animal studies, to inhibit the progression of sarcopenia, possibly through its anti-inflammatory effects; however, few studies have been carried out in humans. We aimed to examine whether coffee consumption was related to indicators of sarcopenia in a Japanese population, and whether the association was mediated by reduced inflammation. This study was a cross-sectional design. Participants were community residents (n = 6369) aged 45-74 years. We measured skeletal muscle mass index (SMI; kg/m2 ) by a bioelectrical impedance method, and grip strength with a Smedley-type dynamometer. Habitual coffee consumption was assessed by a self-administered questionnaire. Serum high-sensitivity C-reactive protein was measured as an inflammatory marker. The association between habitual coffee consumption and SMI or grip strength was analyzed with a linear regression model adjusted for covariates. A significant positive association was found between coffee consumption and SMI (men: β = 0.023; Ptrend = 0.004, women: β = 0.011; Ptrend = 0.012). Further adjustment for high-sensitivity C-reactive protein did not materially alter the results (men: β = 0.023; Ptrend = 0.005, women: β = 0.009; Ptrend = 0.024). The relationship between coffee consumption and grip strength did not reach statistical significance; however, a positive trend was observed (men: β = 0.208; Ptrend = 0.085, women: β = 0.092; Ptrend = 0.167). We found that coffee consumption was positively associated with SMI independently of inflammation in middle-aged and older Japanese people. Reduced inflammation by coffee does not seem to be an important mediator, and further investigations are required to explore the mechanisms of this association. Geriatr Gerontol Int 2021; 21: 950-958.


Does ozonation of drinking water reduce the risk of bladder cancer?
Epidemiologic studies of drinking water disinfection byproducts have focused primarily on the carcinogenic potential of chlorination byproducts. Because drinking water has been ozonated in France for decades, we were able to assess the carcinogenic risk of the disinfection byproducts generated by both ozonation and chlorination. We used data from a case-control study of bladder cancer conducted between 1985 and 1987 in 7 French hospitals. We compared 281 cases and 272 controls for whom we could reconstruct at least 70% of the residential exposure to drinking water contaminants over a 30-year period. When we took potential confounders and exposure to chlorination byproducts into account, the risk of bladder cancer decreased as duration of exposure to ozonated water increased (OR = 0.60 [95% CI = 0.3-1.3] for 1-9 years; OR = 0.31 [0.1-0.7] for 10 years or more). Simultaneously, the risk of bladder cancer increased with duration of exposure to chlorinated surface water and with the estimated trihalomethane content of the water. Our data suggest that ozonation reduces the risk associated with the chlorination of surface water and that ozonation alone could have an independent beneficial effect on bladder cancer risk. Our results are consistent with experimental evidence that ozonation in combination with chlorination decreases the concentration of trihalomethane in treated water and eliminates some of the mutagenicity of raw water.


Microtube embedded hydrogel bioprinting for vascularization of tissue-engineered scaffolds.
Vascular tissue engineering has been considered promising as one of the alternatives for viable artificial tissues and organs. Macro- and microscale hollow tubes fabricated with various techniques have been widely studied to mimic blood vessels. To date, the fabrication of biomimetic capillary vessels with sizes ranging from 1 to 10 µm is still challenging. In this paper, core-sheath microtubes were electrospun to mimic capillary vessels and were embedded in carboxymethyl cellulose/sodium alginate hydrogel for bioprinting. The results showed improved printing fidelity and promoted cell attachment. The tube concentration and tube length both had significant influences on filament size and merging area. Printed groups with higher microtube concentration showed higher microtube density, with filament/nozzle size ratio, and printed/designed grid area ratio closer to 100%. In the in vitro experiments, microtubes were not only compatible with human umbilical vein endothelial cells but also provided microtopographical cues to promote cell proliferation and morphogenesis in three-dimensional space. In summary, the microtubes fabricated by our groups have the potential for the bioprinting of vascularized soft tissue scaffolds.


Afferent facilitation induced by iontophoretic application of acidic amino acids in the ampullary electroreceptors of Plotosus.
In an attempt to identify the afferent transmitter in acousticolateralis receptors, effects of some acidic amino acids were examined in ampullary electroreceptors of the marine catfish Plotosus anguillaris, which have only afferent innervation. The ampulla (sensory epithelium) was first hyperpolarized in situ to suppress receptor-cell activity and release of transmitter, and hence resting afferent discharges completely. In the absence of transmitter, amino acids were applied to the ampulla by iontophoresis through three-barreled electrodes. Afferent impulses were recorded from the nerve trunk by a suction electrode, and single-unit responses were analyzed by their instantaneous frequency (F) records. L-Glutamate (Glu) induces single-unit responses postsynaptically at discrete spots over the ampulla. The F records showed graded and slow time courses, and F peaks increased sigmoidally against Glu doses with Hill coefficients of 2-3. Response maxima reached 120-210 Hz in doses of one log unit over the threshold. Glu response showed little desensitization. At the Glu-sensitive spots, kainate (KA) and quisqualate (QA) induced more persistent excitation and L-homocysteate (HCA) weaker excitation than did Glu, with similar latency. L-Aspartate (Asp) induced small slow responses with long latency in only one third of the tested Glu spots. D-Glutamate (D-Glu), D-aspartate (D-Asp), L-cysteate (CA), L-cysteine sulfinate (CSA), and N-methyl-DL-aspartate generally induced no response. Some aspartate analogues, L- and D-Asp, CA, and CSA induced persistent potentiation of Glu responses. Apparent affinity increase and change in response shape during potentiation indicated suppression of Glu uptake. However, apparent slope change in the log dose-response curves suggested that additional mechanisms were involved. In contrast, QA, KA, Glu, and HCA induced the excitation only. Comparable topical application of the transmitter was simulated by stimulating a few receptor cells at Glu spots to induce focal responses. Some aspects of the focal and Glu responses were compared. Pharmacological and neurochemical evidence so far available on this material is discussed with respect to the Glu hypothesis. Glu, even though a potent agonist in terms of afferent excitation, seems to act on extrasynaptic receptor sites in the nonmyelinated nerve terminals.


[Tuberculosis in Norwegian children--diagnostic challenges].
Each year, 20-25 Norwegian children below the age of 18 are diagnosed with tuberculosis in Norway. As a demonstration of various difficulties in the work-up and diagnosis of tuberculosis, we present eight infected children aged 15 months to 10 years. Children often contract the infection from adults and may develop serious manifestations including miliary tuberculosis and meningitis. The symptoms are often not specific and tuberculosis may be mistaken for other diseases. Delay and inappropriate diagnostics may have deleterious consequences. The main message is to start treatment upon clinical suspicion of tuberculosis. It is mandatory to sample the necessary biological material for microbiological tests before starting treatment.


Social behaviour in bees influences the abundance of Sodalis (Enterobacteriaceae) symbionts.
Social interactions can facilitate transmission of microbes between individuals, reducing variation in gut communities within social groups. Thus, the evolution of social behaviours and symbiont community composition have the potential to be tightly linked. We explored this connection by characterizing the diversity of bacteria associated with both eusocial and solitary bee species within the behaviourally variable family Halictidae using 16S amplicon sequencing. Contrary to expectations, we found few differences in bacterial abundance or variation between social forms; most halictid species appear to share similar gut bacterial communities. However, several strains of Sodalis, a genus described as a symbiont in a variety of insects but yet to be characterized in bees, differ in abundance between eusocial and solitary bees. Phylogenetic reconstructions based on whole-genome alignments indicate that Sodalis has independently colonized halictids at least three times. These strains appear to be mutually exclusive within individual bees, although they are not host-species-specific and no signatures of vertical transmission were observed, suggesting that Sodalis strains compete for access to hosts. The symbiosis between halictids and Sodalis therefore appears to be in its early stages.


Fetal and childhood growth patterns associated with bone mass in school-age children: the Generation R Study.
Low birth weight is associated with lower bone accrual in children and peak bone mass in adults. We assessed how different patterns of longitudinal fetal and early childhood growth influence bone properties at school age. In 5431 children participating in a population-based prospective cohort study, we measured fetal growth by ultrasound at 20 and 30 weeks gestation, and childhood growth at birth, 1, 2, 3, and 4 years of age. We analyzed these growth measurements in relation to total body (less head) BMD measured by DXA at age 6. We used conditional growth modeling; a technique which takes into account correlation between repeatedly measured growth measures. Our results showed that estimated fetal weight gain, femur length growth between 20 and 30 weeks of gestation, femur length growth between 30 weeks and birth, as well as all height and weight growth measurements from birth to 4 years of age were all positively associated with BMC, bone area (BA), and BMD (all p < 0.01). Fetal femur length growth between 30 weeks and birth was positively associated with BMC and BA (both p < 0.001), but not with BMD. Overall, childhood growth measurements exerted a larger influence on bone measures than fetal growth measures. The strongest effect estimate was observed during the first year of life. Children born small (<10th percentile) for gestational age (SGA) had lower BMC and BA, but not BMD, than children born appropriate for gestational age (AGA), whereas children born large (>90th percentile) for gestational age (LGA) had higher BMC and BA (all p < 0.001). These differences were no longer present in children showing subsequent accelerated and decelerated infant growth, respectively. We conclude that both fetal and childhood growth patterns are associated with bone mineral accrual, showing the strongest effect estimates in infancy. Compensatory infant growth counteracts the adverse consequences of fetal growth restriction on bone development.