Axonal transport of neuropeptides in the cervical vagus nerve of the rat.
Accumulations of the neuropeptides substance P (SP), somatostatin (ST), and vasoactive intestinal polypeptide (VIP) proximal to a crush in the cervical vagus nerve of the rat have been measured using sensitive radioimmunoassays. Each of the peptides was rapidly transport towards the peripheral terminals of vagal afferent fibres, with average rates of flow ranging from 0.8 to 2.7 mm h-1. In the rabbit vagus nerve, SP was transported with an average rate of 4 mm h-1, which is more than double the rate for this peptide in the rat. Double crush experiments in rabbit vagus nerves indicated that the rapidly transported proportion of the total content of SP in the nerve free was about 34%. From this, the rate of transport of SP in the rapidly transported pool in the rabbit vagus nerve can be calculated to be 12 mm h-1 (280 mm day-1). Since such double crush experiments were not possible in the rat, it is not clear whether the different average rates of transport of SP in the rat and the rabbit reflect real differences in the rate of rapid transport in the two species. In common with rapid axonal transport of other neurotransmitters, the transport of SP and ST in the rat vagus nerve was blocked by colchicine, a drug that disrupts microtubules.


A novel sunflower broomrape race with unusual virulence potentially caused by a mutation.
The sunflower broomrape (Orobanche cumana Wallr.) gene pools of the Guadalquivir Valley and Cuenca province in Spain had predominantly race-F virulence. A new race G was observed recently in the Guadalquivir Valley potentially due to the genetic recombination of the avirulence genes of both gene pools. In this research, we have studied populations with atypical virulence from Cuenca. These populations parasitize on DEB2 sunflower line, resistant to all race-G populations evaluated. Ten populations collected in Cuenca province were evaluated with sunflower differential lines and genotyped with 67 SNP markers. Although genetic recombination with individuals of the Guadalquivir Valley gene pool has been observed in most populations, recombination of avirulence genes was discarded as the cause of the new virulence because the population with the highest degree of attack on DEB2 showed no introgression from an external gene pool. Accordingly, a point mutation is proposed as the putative cause of the new virulence. The present study provided a detailed characterization of each population, including the accurate classification of the individuals belonging to each of the classical Spanish gene pools, F1 hybrids, and those that evolved from hybridization between both gene pools. This information is essential to understand how sunflower broomrape populations are evolving in Spain, which in turn may be helpful to understand the dynamics of sunflower broomrape populations in other areas of the world and use this information to develop durable strategies for resistance breeding.


Plasma carcinoembryonic antigen concentrations and immunohistochemical patterns of epithelial marker antigens in patients with large bowel carcinoma.
Carcinoembryonic antigen (CEA), secretory component (SC), and epithelial IgA were traced by paired immunofluorescence staining in 102 large bowel carcinomas from 99 patients. The immunohistochemical results were evaluated semiquantitatively in relation to histological tumour grade, clinicopathological stage, and preoperative plasma CEA concentration. CEA expression was significantly increased (p less than 0.05) in the following order: histologically normal colon mucosa, transitional mucosa adjacent to tumours, neoplastic epithelium; the reverse was true for the expression of SC and epithelial IgA (p less than 0.01). CEA was significantly more abundant in the moderately and poorly differentiated tumors than in the well differentiated ones (p less than 0.05), whereas the latter showed better expression of SC (p less than 0.05) and epithelial IgA (p approximately 0.06). In the transitional mucosa, CEA staining tended to be inversely related to histological tumour grade, whereas SC and epithelial IgA were significantly better seen in this zone when the adjacent tumour was well differentiated than when it was moderately or poorly differentiated (p less than 0.01). Furthermore, the expression of SC and epithelial IgA in the transitional mucosa decreased with increasing invasiveness of the tumours, whereas the opposite relation was indicated for CEA expression. Plasma CEA concentrations were not clearly correlated with histological levels than the localised well differentiated tumours tended to be associated with lower levels than the localised moderately differentiated ones (p approximately 0.06). Moreover, the latter variety was associated with lower plasma CEA concentrations than disseminated tumours of comparable differentiation (p less than 0.01).


Highly diastereoselective synthesis of nucleoside adducts from the carcinogenic benzo[a]pyrene diol epoxide and a computational analysis.
A diastereoselective synthesis of the nucleoside adducts corresponding to a cis ring-opening of the carcinogen (+/-)-7 beta, 8 alpha-dihydroxy-9 alpha,10 alpha-epoxy-7,8,9,10-tetrahydrobenzo[a]pyrene (BaP DE-2) by 2'-deoxyadenosine and 2'-deoxyguanosine is described. The key intermediate (+/-)-10alpha-amino-7beta,8alpha,9alpha-trihydroxy-7,8,9,10-tetrahydrobenzo[a]pyrene was synthesized by a highly diastereoselective dihydroxylation wherein phenylboronic acid was a water surrogate. The resulting boronate ester was converted to a tetraol derivative in which two of the four hydroxyl groups (trans 7, 8) were protected as benzoate esters while the remaining two (cis 9, 10) were free. The cis glycol entity was then subjected to a reaction with 1-chlorocarbonyl-1-methylethylacetate to yield an intermediate chloro monoacetoxy dibenzoate. Displacement of the halide with azide, complete cleavage of the esters, and catalytic reduction of the azide yielded the requisite amino triol. Fluoride displacement from appropriately protected nucleoside derivatives, 6-fluoropurine 2'-deoxyribonucleoside and 2-fluoro-2'-deoxyinosine, by the amino triol then yielded diastereomeric pairs of diol epoxide-adducted 2'-deoxyadenosine (dA) and 2'-deoxyguanosine (dG) nucleosides. Small aliquots of these adducts were separated for characterization purposes. The present approach provides the first diastereoselective synthesis of the cis adducts of BaP DE-2 with 2'-deoxyguanosine as well as the first synthesis of both dA and dG adducts from a common intermediate. An informative analysis of the 1H NMR spectra of the cis adducts synthesized and comparisons to the trans adducts are reported. To gain insight into the diastereoselectivity in the key dihydroxylation step, a computational analysis, including molecular mechanics (MMFF94) and semiempirical AM1 geometry optimizations, yielded results that are in fairly good agreement with the experimental observations.


Isolated carpal scaphoid dislocation.
Isolated carpal scaphoid dislocations are rare. Because of this, treatment strategies can be conflicting and vague. The current authors present a case of isolated scaphoid dislocation that was treated initially with closed reduction and percutaneous pinning. Failure of this index treatment necessitated open reduction and internal fixation providing an adequate short-term outcome. Current treatment strategies now advocate ligament reconstruction as a first-line treatment to restore normal anatomy and preserve function.


Coenzyme Q10 stability in pediatric liquid oral dosage formulations.
OBJECTIVE, To study the stability of Coenzyme Q10 (CoQ10), used as therapeutic agent in treatment of cardiovascular and mitochondrial diseases, in three liquid formulations: two soybean oil solutions (50 mg/ml), one of them with the addition of vitamin E and an O/W emulsion (20 mg/ml) for pediatric use. Furthermore, optimize and validate a stabilityindicating HPLC method for the analysis of CoQ10 in the studied formulations. METHOD, All samples were stored at 25°C. CoQ10 content of each formulation was analyzed in duplicate using fast microbore high performance liquid chromatography (Micro HPLC) at 0, 3, 6, 15, 30, 60 and 110 days. RESULTS, All formulations stayed stable at 25°C during the 110 days of the study. However, the oil solutions presented greater content variations through all the study period. CONCLUSIONS, The CoQ10 emulsion can be stored for at least 110 days at 25 °C and it has proven to be safer when narrow dose adjustment is required. The proposed analytical method was suitable for the study of stability of different formulations meeting the validation parameters according to international guidelines.


Stability of Doxycycline Absorbed on Root Canal Dentin After Obturation with Gutta-Percha/AH26 and Resilon/RealSeal at Different Time Intervals.
Eradication of microorganisms present in the root canal system is paramount for the successful outcome of root canal therapy. The purpose of this study was to compare the of doxycycline absorbed from MTAD into root canal dentin after obturation with gutta-percha/AH26 and Resilon/RealSeal at different time intervals. Fifty-one extracted human teeth were instrumented. Thirty samples were obturated with either gutta-percha/AH26 or Resilon/self-etch RealSeal after final irrigation with MTAD. Fifteen samples were kept unobturated (positive control); six samples were obturated with either gutta-percha/AH26 or Resilon/self-etch RealSeal without MTAD irrigation (negative control). After aging for 1, 3 or 6 weeks, dentin debri were collected, the Doxycycline compound was extracted and its amount was quantified using high performance liquid chromatography. The statistical significance of the change in Doxycycline concentrations was tested with two-way ANOVA. The mean concentration of Doxycycline in dentin for one, three and six-week guttapercha/AH26 samples was 1.8±0.36, 1.22±0.22 and 0.67±0.11 respectively, whereas these concentrations in Resilon/RealSeal samples were 1.60±0.26, 0.80±0.14 and 0.59±0.01 respectively. Regarding the positive control group, these concentrations were 2.09±0.11, 1.54±0.12 and 0.72±0.07 respectively for 1, 3 and 6-week intervals. No Doxycycline was detected in negative control groups. The Doxycycline concentrations showed a significant difference forobturating materials (p=0.008). These concentrations were higher in the gutta-percha/AH26 samples than Resilon/RealSeal samples in each time interval. The remaining amount of Doxycycline bonded to dentin was higher when root canals were obturated with gutta-percha/AH26 compared to Resilon/RealSeal. The stability of Doxycycline showed a time dependent decrease.


Risk of infection according to the gamma globulin level in the 100 days following allogeneic stem cell transplantations.
Immunoglobulin replacement therapy is recommended in case of severe hypogammaglobulinemia after allogeneic hematopoietic stem cell transplantation (allo-HSCT). However, the supposed increased risk of infection in case of hypogammaglobulinemia has not been confirmed in allo-HSCT. In this study, we assessed the relationship between the gamma globulin level and the risk of infection during the 100 days following the allo-HSCT. We gathered the weekly laboratory tests from day 7 to day 100 of 76 allograft patients, giving a total of 1 044 tests. 130 infections were documented clinically, by imaging, or microbiologically. Average gamma globulin levels between D-7 and D100 did not differ between patients with or without infection (642 ± 232 and 671 ± 246 mg/dL, respectively, P = .65). Gamma globulin level <400 mg/dl was not associated with the occurrence of infection between the test studied and the next one (aOR 1.33 [0.84-2.15], P = .24). The gamma globulin level was not predictive of bacterial or fungal infections (AUC 0.54 [95%CI: 0.47-0.61]) nor of viral reactivations (AUC 0.51 [95%CI: 0.43-0.60]). This confirmed that the humoral deficiency is a minor part of the immune deficiency in the 100 days post-transplant. This questions the relevance of the indications of immunoglobulin substitution during this period.


Expression of cystatin C in the female reproductive tract and its effect on human sperm capacitation.
Cystatin C (CST3), a cysteine protease inhibitor in seminal plasma, is expressed in animal uteri. However, its expression in the human female reproductive tract and its effect on human sperm capacitation are unclear. The cellular localization of CST3 was observed using immunohistochemistry. The binding of CST3 to sperm was examined using immunocytochemistry. Sperm motility parameters were analyzed using computer-assisted sperm analysis. Sperm capacitation was evaluated by analyzing cholesterol content, protein tyrosine phosphorylation levels, and the acrosome reaction. Immunohistochemical staining demonstrated that CST3 is prominently expressed in the female reproductive tract, including the epithelial lining and cervix and endometrium fluids, particularly at times near ovulation. It can bind to human sperm on the post-acrosomal head region and the mid and principal piece of the tail. CST3 enhances sperm motility and inhibits the signal initiating sperm capacitation, i.e., efflux of cholesterol from the sperm plasma membrane and a late sperm capacitation event, i.e., the increase in the sperm protein tyrosine phosphorylation. The suppressive trend on sperm acrosome reaction further supports CST3's ability to inhibit sperm capacitation. These findings suggest that cervical CST3 may prevent precocious capacitation and acrosome reaction, thus preserving sperm fertilizing ability before it reaches the fallopian tube. Additionally, CST3 may help sperm enter the upper reproductive tract by enhancing sperm motility.


Complete rupture of unscarred uterus with delayed symptoms: Case report and possible mechanism.
Uterine ruptures typically occur suddenly, have obvious symptoms, and may require urgent treatment. We experienced a case of complete rupture of an unscarred uterus that was undetected for 9 days. We report the clinical course and possible factors that led to it. Nine days after delivery, complete rupture of the posterior uterine wall was diagnosed by magnetic resonance imaging. The colon and ovaries were tightly adhered to the posterior uterine body. When this adhesive lesion was detached, 7 cm horizontal defects of the muscular layer were revealed. The ruptured wound was repaired. Magnetic resonance imaging 1 year postoperatively showed no thinning of the repaired lesion and suggested successful reperfusion. Mild clinical course despite complete uterine rupture was presumably because of adhesion of the ovaries and intestines to the rupture site.


Results of coronary bypass grafting in treatment of left main stenosis.
Study comparing the results of coronary artery bypass grafting (CABG) with and without cardiopulmonary bypass (CPB) in the treatment of left main stenosis (LMS) has not yet been made in Bosnia and Herzegovina. The main aim of this study was to compare result of CABG performed on 176 patients, in Cardiovacular clinic of University Clinical Center in Tuzla from May 1999 to January 2005, by these two methods in LMS group of patients in the early and late postoperative period. The study was divided in two parts. In the first part, early postoperative period (30 days after the surgery) has been analysed, which encompasses results of CABG in 92 patients revascularisied without CPB (OPCAB) method (Group A), and 84 patients with LMS revascularisied with CPB method (ONCAB, CCAB) (Group B). In the second part, late postoperative period (one year after surgery) has been analysed. Patients from both groups were contacted and interviewed. Total number (276 versus 278), same as average number of grafts per patients (3.0 +/- 1.45 versus 3.31 +/- 0.86 p = 0.096), was insignificantly higher in group B. Perioperative and postoperative results revealed significant differences between two groups in reduced mechanical ventilation time (2.9 versus 7.3 hours, p = 0.039), less blood transfusion requirement (200.3 versus 419.9 ml, p = 0.035) and postoperative length of stay (7.4 versus 8.3 days). Inotrop support requirement was significantly higher in group B during the surgery (14.1% versus 29.8%, p = 0.019) and postoperative period longer than 12 hours (7.6 versus 22.6%; p = 0.009). Significant difference for mortality was noticed in early postoperative period in group A (0.0 versus 5.95%, p = 0.023). There was no significant difference for any of additional procedures (7.9 versus 7.8, p = 0.802), for major complications (8.0 versus 9.1%; p = 0.985) or for mortality (7.9 versus 5.2%; p = 0.692). Patients' survival after CABG for period of 12, 24, 36 and 48 months after surgery was not statistically significant (97.0 versus 96.1%, p = 0.802; 95.5 versus 96.1%, p = 0.857; 93.2 versus 94.8%, p = 0.913; 92.0 versus 94.8%, p = 0.692). This study revealed that OPCAB is effective and safe method for treatment of LMS, and there are certain advantages in comparison to classical method in short-term follow up, with no difference in long-term results.


Acute Appendicitis Caused by an Echinococcal Brood Capsule Unmasks an Asymptomatic Hepatic Hydatid Cyst.
Hydatidosis is an endemic helminthic disease in the cattle-grazing regions of Asia. It is usually caused by the cestode Echinococcus granulosus. Internal organs, particularly the liver and lungs, are predominantly affected, but the appendix is only rarely involved with the formation of characteristic hydatid cyst. We present a unique case of appendiceal hydatidosis in an 18-year-old woman with acute appendicitis. Her preoperative abdominal ultrasound revealed an asymptomatic hydatid cyst in the liver. An echinococcal brood capsule was detected postoperatively within the appendicular lumen under microscope.


Reverse transcriptase associated with avian sarcoma-leukosis viruses. I. Comparison of intra-virion content of multiple enzyme forms.
The RNA-dependent DNA polymerase (the reverse transcriptase) was solubilized from three related strains of avian sarcoma virus (ASV B77, ASV tsLA334, and ASV QV2) as well as avian myeloblastosis virus (AMV) and a chicken endogenous virus (RAV-O), by a combination of non-ionic detergent treatment and CsCl step-gradient centrifugation, and was subsequently separated into individual enzyme forms by poly(C)-agarose column chromatography. The newly developed two-step method allowed us to purify the three molecular forms (alpha-, alpha beta- and beta-form) of highly active enzyme rapidly and quantitatively from all the five virus strains examined. The molar ratio of the three enzyme forms differed among the virus strains: For the three sarcoma viruses, the major species was the alpha beta-form enzyme, the putative holoenzyme, and the alpha- and beta-form enzymes were less than a few percent and 15-25%, respectively, while the alpha-form enzyme content was higher for the two leukosis viruses than for the three sarcoma viruses. Both the total DNA polymerase activity and the content of the two enzyme subunits in purified virions of the three sarcoma virus was in the following order: ASV tsLA334 greater than ASV B77 greater than ASV QV2, which paralleled the virus yield at a permissive temperature in roller bottle cultures of chick embryo fibroblasts. No alteration was found in the thermolability of DNA polymerases between tsLA334, which carries ts mutations affecting both virus growth and cell-transformation, and other viruses.


Conductance of ion channels and nanopores with charged walls: a toy model.
We consider ion transport through protein ion channels in lipid membranes and water-filled nanopores in silicon films. It is known that, due to the large ratio of dielectric constants of water and the surrounding material, an ion placed inside the channel faces a large electrostatic self-energy barrier. The barrier leads to an exponentially large resistance of the channel. We study reduction of the electrostatic barrier by immobile charges located on the internal walls of the channel. We show that the barrier practically vanishes already at relatively small concentration of wall charges.


Ethical dilemmas in organ donation and transplantation.
Since the first successful organ transplantation in 1953, we have seen an explosive development in transplantation surgery, particularly during the 1980s. With it followed an abundance of legal controversies and ethical dilemmas. Optimal use of viable organs necessitated precise definition of brain death in heart-beating cadavers with artificially maintained ventilation and circulation. Viable organs must remain well perfused to be suitable for procurement and transplantation into carefully selected recipients on an equal-opportunity basis. Due consideration must be given to both medical and social indications. At present, homografts dominate the field of organ transplantation; however, because of the shortage of human organs, both artificial organs (especially hearts) and xenografts are expected to become increasingly common in the near future. No doubt, the use of such modern technology will introduce additional ethical problems.


The relationship of single words to two-word sentences.
The relationship between the single-word utterances and 2-word sentences of 12 children was examined as part of a longitudinal study of language development. Children whose single-word utterances usually named objects produced sentences which named many objects. Children whose single words were frequently interjections produced sentences which expressed desire for an object. 2-word sentences seem to have some of their structural and functional roots in the single-word phase.


Getting the message across: does the use of drama aid education in palliative care?
Drama is a promising means of delivering educational messages in palliative care. Research studies have found drama to be an effective means of delivering educational messages in other domains of learning, such as teaching health education to children and adults and engaging the general public in health policy development. This paper discusses the potential of drama for palliative care education and provides an example of the use of a drama to deliver messages about death and dying at a conference on palliative care. The paper suggests a theoretical framework for how future drama productions could be developed to educate the community, health professionals and students about palliative care.


Determination of ascorbic acid and carotenoids in food commodities by liquid chromatography with mass spectrometry detection.
Two methods, one to determine ascorbic acid and one to determine lycopene and beta-carotene, in vegetables and fruits by liquid chromatography coupled with mass spectrometry (LC-MS) have been established. The chromatographic separation of the studied compounds and their MS parameters were optimized to improve selectivity and sensitivity. In both methods, separation was carried out with two coupled columns, first a C(18) and then a dC(18), using as mobile phase 70% methanol (0.005% acetic acid) and 30% acetic acid 0.05% for ascorbic acid determination and a mixture of methanol, tetrahydrofuran, and acetonitrile (60:30:10 v/v/v) for carotenoid analysis in isocratic mode. The molecular ion was selected for the quantification in selective ion monitoring (SIM) mode. Ascorbic acid was detected with electrospray ionization probe (ESI) in negative mode, while chemical ionization atmospheric pressure (APCI) in positive mode was used for the target carotenoids. The methodology for ascorbic acid analysis is based on an extraction with polytron using methanol and a mixture of methaphosphoric acid and acetic acid. Extraction of the carotenoids was carried out with tetrahydrofuran/methanol (1:1) (v/v). The proposed methods were applied, after their corresponding validations, to the analysis of four varieties of tomatoes, tomato in tin enriched and dried tomato, and to the analysis of mango and kiwi fruits, to compare the content in these compounds. Moreover, the influence of the process of freezing and the effect that the manipulation/preservation has in the content of ascorbic acid in tomato have also been studied.


Superconducting and charge density wave transition in single crystalline LaPt2Si2.
We present results of our comprehensive studies on single crystalline LaPt2Si2. Pronounced anomaly in electrical resistivity and heat capacity confirms the bulk nature of superconductivity (SC) and charge density wave (CDW) transition in the single crystals. While the charge density wave transition temperature is lower, the superconducting transition temperature is higher in single crystal compared to the polycrystalline sample. This result confirms the competing nature of CDW and SC. Another important finding is the anomalous temperature dependence of upper critical field H C2(T). We also report the anisotropy in the transport and magnetic measurements of the single crystal.


A comparison of care consumer and care provider perspectives on the quality of life of persons with persistent and severe psychiatric disabilities.
Agreement on care consumer quality of life (QOL) in different life domains between persons with a psychiatric disability and their care providers was examined. The former persons tended to report higher QOL than their care providers and care providers and care consumers evinced limited agreement on actual care consumers' QOL and on the substance of care consumer QOL Care providers' confidence in their judgment of care consumer QOL, but not duration of care provider-care consumer contact, was associated with more care provider-care consumer agreement. Thus, ways of increasing care providers' understanding of care consumers' perceptions of problems and goals should be investigated.


Upholding the Tripartite Mission in Times of Crisis: Purpose and Perseverance in the COVID-19 Pandemic.
Academic otolaryngology has historically comprised a tripartite mission of research, education, and clinical care. This mission is greatly strained by the COVID-19 pandemic. Research laboratories are fallow, surgical cases are deferred, and clerkships are canceled. Otolaryngologists are adapting to new circumstances ranging from virtual patient care to urgent procedures in protective gear to deployment to provisional field hospitals. Amid these operational challenges, the specialty is demonstrating extraordinary resilience and grit, discovering new ways to serve learners, colleagues, and communities. Statements from leaders in otolaryngology reveal selfless acts and purposefulness. Necessity has spurred innovation in education, science, and novel models of care. Paragons strike notes of hope, inspiring us to persevere and serve. This commentary explores the present challenges and offers a vision for upholding the academic mission.


Two ways to inactivate the Ki-67 protein-Fragmentation by nanoparticles, crosslinking with fluorescent dyes.
Light can manipulate molecular biological processes with high spatial and temporal precision and optical manipulation has become increasingly popular during the last years. In combination with absorbing dyes or gold nanoparticles light is a valuable tool for cell and protein inactivation with high precision. Here we show distinct differences in the underlying mechanisms whether gold nanoparticles or fluorescent dyes are used for the inactivation of the Ki-67 protein. The proliferation-associated protein Ki-67 was addressed by the antibody MIB-1. In vitro studies showed a fragmentation of the Ki-67 protein after laser irradiation of 15 nm gold nanoparticle antibody conjugates with nanosecond pulsed laser, while continuous wave (cw) irradiation of fluorescein isothiocyanate (FITC)- and Alexa 488-labeled antibodies led to specific crosslinking of Ki-67. The irradiation energy for the gold nanoparticles was above cavitation bubble formation threshold. We observed a fragmentation of the target protein and also of the gold particles. The understanding of the underlying inactivation mechanisms is important for the application and further development of these two techniques, which can harness nanotechnology to introduce molecular selectivity to biological systems.


One-step production of α-ketoglutaric acid from glutamic acid with an engineered L-amino acid deaminase from Proteus mirabilis.
Currently, α-ketoglutaric acid (α-KG) is industrially produced by multi-step chemical synthesis, which can cause heavy environmental pollution. Here we reported a simple one-step approach for the production of α-KG by transforming l-glutamic acid with an engineered l-amino acid deaminase (l-AAD) from Proteus mirabilis. First, to facilitate the purification of membrane-bound l-AAD, one N-terminal transmembrane region (from 21 to 87th nucleotide) was removed from l-AAD to block the binding of l-AAD with membrane, and the relatively low-usage codons were replaced by high-usage codons in Escherichia coli to improve the expression level. However, inclusion bodies formed when expressing the ΔN-LAAD in E. coli BL 21, and then the soluble and active ΔN-LAAD was obtained by the solubilization and renaturation of ΔN-LAAD. Furthermore, the biochemical properties of the refolded ΔN-LAAD were characterized and compared with those of full-length l-AAD. Finally, the ΔN-LAAD was used to synthesize α-KG and the maximal formation rate of α-KG reached 12.6% (w/w) at 6h under the following conditions: 12g/L l-glutamic acid, 0.1g/L ΔN-LAAD, 5mM MgCl2, temperature 45°C and pH 8.0. Compared with the multi-step chemical synthesis, the transformation approach has less environmental pollution and has a great potential for α-KG production.


Human immunodeficiency virus: The potential of medicinal plants as antiretroviral therapy.
Human immunodeficiency virus (HIV) infection is fast becoming widespread in the world with 37.7 million people living with HIV in 2020. Antiretroviral therapy involving chemical drugs has declined acquired immunodeficiency syndrome (AIDS)-related mortality and improved the life quality of AIDS/HIV sufferers. However, the emergence of drug resistance and side effects are the main obstacles for the long-term use of these chemicals as antiretroviral therapy. Recently, a lot of emphasis is being put on finding naturally occurring drug candidates that show activity against HIV and can be potentially used as antiretroviral therapy. In this study, different medicinal plants, Pistacia khinjuk, Teucrium stocksianum, Uncaria tomentosa, Pistacia integerrima, Trigonella gharuensis, and Artocarpus lakoocha, were explored for their anti-HIV potential. Syncytium and p24 assays were performed to determine antiviral activity, while the MTT assay was used to determine cytotoxicity. Results showed that extracts from all six plants inhibited HIV replication in vitro. Also, extracts from Pistacia khinjuk, Teucrium stocksianum, Uncaria tomentosa, and Pistacia integerrima showed low cytotoxicity with a 50% cytotoxicity concentration value of >200 μM. Results of this study indicate that there is potential in these natural extracts to become candidate drugs to be used as complementary and alternative medicine for HIV infection.


The new conundrum: do estrogens have any cardiovascular benefits?
Clearly, a new era has begun, with increasing numbers of the scientific/medical community asking whether estrogens have any cardiovascular benefits. Doubts have arisen from two randomized prospective trials. The Heart and Estrogen/progestin Replacement Study (women who were generally beyond 65 years of age with preexisting coronary heart disease) found no benefit in reducing coronary events by a combination of estrogens and a progestin. Later, the Estrogen Replacement Atherosclerosis Trial reported that no benefit could be shown for either conjugated equine estrogens only or the combined therapy group for women with preexisting coronary artery stenosis. There are lessons to be taken from monkey models about the new conundrum. Estrogens have beneficial effects in the early stages of atherogenesis, but have little or no beneficial effects in the final stages of plaque complications, instability and coronary heart disease events. Using the monkey model, we have addressed the question of when "primary prevention" should begin. We examined the effect of contraceptive steroid treatment of stressed animals at high risk to progressing atherosclerosis due to their estrogen deficiency and subsequently examined the effect of estrogen replacement therapy or no estrogen replacement therapy following surgical menopause. The most robust inhibition of atherosclerosis progression was found in those animals given contraceptive steroids premenopausally and subsequently estrogen replacement postmenopausally. The notion of starting contraceptive therapy during the perimenopausal period to be followed immediately with estrogen replacement postmenopausally is likely to be the most favorable approach to the inhibition of atherosclerosis progression.


Genetic and epigenetic analysis of SSAT gene dysregulation in suicidal behavior.
It has recently been proposed that the SSAT gene plays a role in the predisposition to suicidal behavior. SSAT expression was found to be down-regulated in the brain of suicide completers. In addition, a single nucleotide polymorphism (SNP) rs6526342 was associated both with variation in SSAT expression and with suicidal behavior. In this study, we aimed to characterize the relationship between SSAT dysregulation and suicide behavior. To this end, we measured SSAT expression levels in the ventral prefrontal cortex (VPFC) of suicide completers (n = 20) and controls (n = 20) and found them to be significantly down-regulated in suicide victims (P = 0.007). To identify the basis of the regulation of SSAT expression, we performed an association analysis of 309 SNPs with SSAT transcript levels in 53 lymphoblastoid cell lines from the CEPH collection. We then examined the methylation status of the SSAT promoter region in males and females suicide completers and control subjects whose SSAT brain expression had been measured. We found no evidence to support a role for SNPs in controlling the level of SSAT expression. SSAT promoter methylation levels were not different between suicide completers and controls and did not correlate with SSAT expression levels. In addition, we found no indication of a genetic association between suicidal behavior and SNPs located within the SSAT gene. Our study provides new results which show that dysregulation of SSAT expression does play a role in suicide behavior. However, our data do not support any association between rs6526342 and variation in SSAT expression or suicidal behavior.


Part 1: The Theoretical Basis for Recovery-Oriented Management of Substance Use Disorders in Primary Care.
There is strong evidence in the literature that screening and brief counseling interventions are effective in detecting alcohol problems and decreasing alcohol consumption among patients in primary care settings but somewhat weaker evidence regarding screening and brief intervention for drug problems. In 2014, two published studies made news and caused concern among proponents of substance Screening, Brief Intervention and Referral to Treatment (SBIRT) programs, when their authors concluded that brief interventions were not effective for decreasing drug use among primary care patients identified through screening, and advised that widespread adoption of screening and brief intervention for drug use was not warranted.An evaluation of the theoretical foundations for evidence-based SBIRT services was conducted to produce possible explanations for why traditional SBIRT works well for individuals with unhealthy alcohol use but not as well for those with more serious substance use disorders, including drug use and alcohol/drug dependence. Smith and Liehr's evaluation framework for middle-range theory was utilized to analyze the Chronic Care Model, which was featured prominently in early SBIRT literature, and the newer Recovery Management model, which provides a philosophical framework for organizing modern addictions services and quality-of-life enhancements (Part 1 of this two-part series).Programs are more likely to succeed if guided by theory, and examination of relevant components of theory-based interventions can be useful in developing practical strategies for meeting program objectives. A new, theory-based, recovery-oriented framework for primary care SBIRT is introduced in Part 2 ("SBIRT+RM(C): A Proposed Model for Recovery-Oriented Primary Care").


Removal of bacteriophages MS2 and phiX174 during transport in a sandy anoxic aquifer.
The objectives of our study were to determine (i) removal of bacteriophage MS2 and phiX174, as surrogates for human pathogenic viruses, in an anoxic aquifer and (ii) the safe length of the microbial protection zone in anoxic aquifers. 3.5 Log units of MS2 were removed by adsorption and inactivation during 63 days residence time, which was 1.4 log units lower than removal of phiX174. These removal rates were considerably lower than previously reported for MS2 and phiX174 in oxic aquifers and consequently longer protection zones around anoxic aquifers might be needed. Therefore, the observed log removal of MS2 was used in a risk assessment approach to determine the required safe length of the microbial protection zone. In case of a leaking sewer in the vicinity of a well in an anoxic aquifer, the risk assessment demonstrated that a microbial protection zone of 110 m may be needed to meet a risk of infection of 10(-4) persons per year. This length can be two to three times larger than the length of the protection zone currently used in a number of countries.


Tuberculosis of the breast.
Tuberculosis of the breast is a rare disease with non specific manifestations. It should be included in the differential diagnosis of breast lesions in immune compromised patients especially in tuberculosis endemic areas of the world. We report a case of a 31 year old HIV positive lady who presented with an ulcerated mass on the left breast of 3 months' duration. Incision biopsy of the lesion confirmed tuberculosis of the breast. Treatment with standard anti tuberculous drugs resulted In complete resolution. A high index of suspicion is required to make a diagnosis of breast tuberculosis. The disease can be treated conservatively with standard anti tuberculous drugs while surgery is reserved for rare cases.


Novel object recognition memory: measurement issues and effects of MDMA self-administration following short inter-trial intervals.
The present study was undertaken to examine effects of self-administered MDMA on novel object exploration (NOR) memory. Self-administration was conducted during daily 2 h tests that continued until a total of 165 mg/kg was self-administered (range = 13-41 days for individual rats). Control rats were placed in the self-administration boxes during daily sessions but did not receive any drug. One or 10 weeks following the last self-administration session, memory was assessed using a standard NOR task. When exploration time was used as the dependent measure for the control rats, there was no consistent pattern of change as a function of inter-trial interval (ITI) and exploration times failed to reveal decay in the function relating exploration to ITI. When number of approaches was examined as a function of ITI, however, there was a preference for the novel object following the short ITIs (1-15 min) and the function relating preference to ITI decayed with longer ITIs. When tested 7 days following the last self-administration session, rats that self-administered MDMA failed to demonstrate NOR even following the shortest ITI of 1 min. The data support the idea that MDMA self-administration produces cognitive deficits and are consistent with the idea that attentional processes become disrupted. There was, however, recovery of NOR memory when rats were tested following an extended drug-free period of 70 days. Thus, the deficits are transient and recovery was apparent.


Psychological adjustment to hand injuries: nursing management.
Traumatic hand injuries can be devastating experiences both physically and psychologically. The plastic surgery nurse who can identify the various responses that routinely occur with hand injured patients is better prepared to normalize their experiences and implement the necessary interventions toward overall rehabilitation. This article will provide an overview of specific characteristics and emotional reactions following hand injuries, factors that influence adjustment, and nursing management that promotes optimal recovery.


Analysis of nasal and labial deformities in cleft lip, alveolus and palate patients by a new rating scale: preliminary report.
In this study the nasal deformities in patients with cleft lip, alveolus and palate (CLAP) were analysed and the relevant role of the perinasal-perioral muscular balance, and the inborn dislocation of the alar cartilages is presented. 50 CLAP patients were analysed in whom 29 primary cheiloplasties, 12 lip revisions and 9 rhinoplasties were performed. The lip repair was done by a modification of Millard's technique, the nose by either a closed or open-sky rhinoplasty. The severity of the cleft appearance was evaluated pre- and postoperatively, according to a pre-agreed visual rating scale. There were 4 degrees of severity of the deformity preoperatively (mild, moderate, severe and very severe), and postoperatively 5 categories of outcome (excellent, very good, good, satisfactory and poor) depending on the scores obtained by summing up the points corresponding to different types of deformity. This scale is closely related to the American Cleft Palate classification of clefts. 17 excellent, 4 very good, 2 good, 5 satisfactory and 1 poor result were obtained in the group of primary cheiloplasty. Eight excellent, 4 very good results were obtained by the lip revisions. Seven excellent and 2 satisfactory results were obtained following rhinoplasty. During the primary lip repair, it is important to correct the abnormal position of ala nasi, the nasal floor and the base of the columella. When correct insertion of m. transversus nasi to the nasal spine is achieved and a good repair of m. orbicularis oris, symmetry of the alae and normal growth of lip and columella was obtained even in most severe bilateral cases. In cases of diastasis of the orbicularis and transversus nasi muscles, in combination with other soft tissue deformities or scars, a secondary musculo-periosteal revision is recommended. The defect of the soft tissue triangle of the nose is best corrected via an open rhinoplasty.


Ocular Manifestations of Inflammatory Bowel Disease.
Inflammatory bowel disease (IBD) is a chronic disease connected to the immune system, with a predilection for the gastrointestinal tract. However, a large proportion of the patients have extraintestinal manifestations (EIM), and the ocular system is affected in some patients. The clinical presentation of ocular pathology is broad, ranging from asymptomatic cases to blindness, leading to high morbidity. Ocular complications can be primary and, in general, are associated with episodes of acute flare-ups that subside with immunological management of the digestive disease. Secondary complications arise from the primary ones and as adverse effects of immunological treatment. In addition, on many occasions, the ocular manifestation may appear prior to the presentation of digestive symptoms. The presence of a multidisciplinary team that includes a gastroenterologist and an ophthalmologist is important in order to achieve early diagnosis of ocular complications, thereby preventing, treating, and avoiding unfavorable and irreversible long-term ocular sequelae.


A novel TUBG1 mutation with neurodevelopmental disorder caused by malformations of cortical development.
Neurodevelopmental disorder caused by malformations of cortical development is a rare neurological disease. Heterozygous missense variants in the TUBG1 gene lead to malformations of human cortical development, which further result in intellectual disability, developmental retardation, and epilepsy. To the best of our knowledge, only thirteen patients and a total of nine pathogenic TUBG1 variants have been described in the published literature. This study reports the case details and genetic data analysis of a girl (aged 8 years, 9 months) with developmental delay, psychomotor regression, epilepsy, and left external ear deformity. A novel TUBG1 mutation was identified by whole-exome sequencing and Sanger sequencing, confirming that this mutation may be the cause of the neurodevelopmental disorders. This case report characterizes the phenotypic spectrum, molecular genetic findings, and functional consequences of novel pathogenic TUBG1 variants in neurodevelopmental disorders caused by cortical development malformations.


Recovery of alveolar bone by the guided bone regeneration technique.
Bone defects associated with the removal of endosteal implants often create challenging problems for clinicians. The guided bone regeneration (GBR) technique has been recently used in the treatment of such defects, and promising results have been obtained. This report describes the use of Gore-Tex tissue augmentation material (GTAM) membranes after the removal of Bioceram screw implants in two patients who wore the implants for 10 years and seven years, respectively. After removal, the residual bone defects were treated by the GBR technique. Bone regeneration was confirmed in the defects around the newly placed implants, and decreased bone mass was avoided in both patients. In case 1, a free gingival graft was used to increase the area of attached gingiva around the implants, since the status of peri-implant soft tissue is also crucial to the outcome of therapy. This technique was very effective in maintaining an adequate width of attached gingiva.


Safety and efficacy of the HIV-1 attachment inhibitor prodrug BMS-663068 in treatment-experienced individuals: 24 week results of AI438011, a phase 2b, randomised controlled trial.
BMS-663068 is an oral prodrug of BMS-626529, an attachment inhibitor that binds to HIV-1 gp120, blocking viral attachment to host CD4 cells. AI438011 is an ongoing trial investigating the efficacy, safety, and dose-response of BMS-663068 in treatment-experienced, HIV-1-infected patients. Herein we present the results of the primary analysis. AI438011 is a phase 2b, randomised, active-controlled trial, at 53 hospitals and outpatient clinics across ten countries in North and South America, Europe, and Africa. Individuals with an HIV-1 RNA viral load of at least 1000 copies per mL and a BMS-626529 half-maximum inhibitory concentration lower than 100 nmol/L were randomly assigned (1:1:1:1:1) to receive either BMS-663068 at 400 mg twice daily, 800 mg twice daily, 600 mg once daily, or 1200 mg once daily or ritonavir-boosted atazanavir (300 mg of atazanavir and 100 mg of ritonavir once daily), each with 400 mg of raltegravir twice daily and 300 mg of tenofovir disoproxil fumarate once daily as a backbone. The sponsor, participants, and investigators were masked for BMS-663068 dose but not for allocation. Primary endpoints were the proportion of patients with an HIV-1 RNA viral load less than 50 copies per mL (response rate) at week 24 and the frequency of serious adverse events and adverse events leading to discontinuation, up to the week 24 analysis. The primary analyses included all patients who received at least one dose of study drug (modified intention-to-treat population). This study is registered at ClinicalTrials.gov, NCT01384734. Between July 26, 2011, and July 16, 2012, 581 participants were assessed for eligibility. Of these, 254 patients were randomly assigned to receive either BMS-663068 (n=52 for the 400 mg twice daily group, n=50 for the 800 mg twice daily group, n=51 for the 600 mg once daily group, and n=50 for the 1200 mg once daily group) or ritonavir-boosted atazanavir (n=51). 200 patients received at least one dose of BMS-663068, and 51 patients received at least one dose of ritonavir-boosted atazanavir. At week 24, 40 (80%) of 50 patients in the BMS-663068 400 mg twice daily group, 34 (69%) of 49 patients in the 800 mg twice daily group, 39 (76%) of 51 patients in the 600 mg once daily group, and 36 (72%) of 50 patients in the 1200 mg once daily group had an HIV-1 RNA viral load less than 50 copies per mL, compared with 38 (75%) of 51 patients in the ritonavir-boosted atazanavir group. Serious adverse events were noted in 13 (7%) of 200 patients in the BMS-663068 groups and five (10%) of the 51 patients in the ritonavir-boosted atazanavir group. Four (2%) of the 200 patients in the BMS-663068 groups and two (4%) of the 51 patients in the ritonavir-boosted atazanavir group discontinued because of adverse events. No serious adverse events or adverse events leading to discontinuation were BMS-663068-related. Grade 2-4 adverse events related to study drug(s) occurred in 17 (9%) of 200 patients across the BMS-663068 groups and 14 (27%) of 51 patients in the ritonavir-boosted atazanavir group. For the BMS-663068 groups these events were mostly single instances with no dose relation and for the ritonavir-boosted atazanavir group these were mostly gastrointestinal or hepatobiliary disorders associated with hyperbilirubinaemia. In a comparison with ritonavir-boosted atazanavir, efficacy and safety of BMS-663068 up to the week 24 analysis support continued development of BMS-663068, which is being assessed in a phase 3 trial in heavily treatment-experienced individuals. Bristol-Myers Squibb.


Development and Validation of a Multiclass Method for Analysis of Veterinary Drug Residues in Milk Using Ultrahigh Performance Liquid Chromatography Electrospray Ionization Quadrupole Orbitrap Mass Spectrometry.
This paper presents the development and validation of a multiclass method for the analysis of veterinary drug residues in milk using ultrahigh performance liquid chromatography electrospray ionization quadrupole Orbitrap mass spectrometry (UHPLC/ESI Q-Orbitrap). The 12 classes of veterinary drugs (a total of 125) included in this study were endectocides, fluoroquinolones, ionophores, macrolides, nitroimidazole, NSAIDs, β-lactams, penicillins, phenicols, sulfonamides, tetracyclines, and aminoglycosides. Veterinary drug residues in milk were extracted using a modified salting-out supported liquid extraction (SOSLE) method, which entailed the precipitation of milk proteins using an extraction buffer (oxalic acid and EDTA, pH 3) and acetonitrile, a salting-out acetonitrile/water phase separation using ammonium sulfate, and solid-phase extraction (SPE) using polymeric reversed-phase sorbent cartridges. The final extracts were concentrated and reconstituted into a buffer solution and analyzed using UHPLC/ESI Q-Orbitrap mass spectrometry. The developed method was validated using a nested experimental design to evaluate the method performance characteristics, such as overall recovery, intermediate precision, and measurement uncertainty. The method was able to quantify or screen up to 105 veterinary drugs from 11 different classes, except aminoglycosides. The limits of quantification were as low as 1.0 μg/kg, with an analytical range from 1.0 to 100.0 μg/kg in milk.


Heterogeneous catalytic degradation of phenolic substrates: catalysts activity.
This review article explored the catalytic degradation of phenol and some phenols derivates by means of advanced oxidation processes (AOPs). Among them, only the heterogeneous catalyzed processes based on catalytic wet peroxide oxidation, catalytic ozonation and catalytic wet oxidation were reviewed. Also selected recent examples about heterogeneous photocatalytic AOPs will be presented. In details, the present review contains: (i) data concerning catalytic wet peroxide oxidation of phenolic compounds over metal-exchanged zeolites, hydrotalcites, metal-exchanged clays and resins. (ii) Use of cobalt-based catalysts, hydrotalcite-like compounds, active carbons in the catalytic ozonation process. (iii) Activity of transition metal oxides, active carbons and supported noble metals catalysts in the catalytic wet oxidation of phenol and acetic acid. The most relevant results in terms of catalytic activity for each class of catalysts were reported.


Sub-wavelength grating mode transformers in silicon slab waveguides.
We report on several new types of sub-wavelength grating (SWG) gradient index structures for efficient mode coupling in high index contrast slab waveguides. Using a SWG, an adiabatic transition is achieved at the interface between silicon-on-insulator waveguides of different geometries. The SWG transition region minimizes both fundamental mode mismatch loss and coupling to higher order modes. By creating the gradient effective index region in the direction of propagation, we demonstrate that efficient vertical mode transformation can be achieved between slab waveguides of different core thickness. The structures which we propose can be fabricated by a single etch step. Using 3D finite-difference time-domain simulations we study the loss, polarization dependence and the higher order mode excitation for two types (triangular and triangular-transverse) of SWG transition regions between silicon-on-insulator slab waveguides of different core thicknesses. We demonstrate two solutions to reduce the polarization dependent loss of these structures. Finally, we propose an implementation of SWG structures to reduce loss and higher order mode excitation between a slab waveguide and a phase array of an array waveguide grating (AWG). Compared to a conventional AWG, the loss is reduced from -1.4 dB to < -0.2 dB at the slab-array interface.


Effect of various oxygen free radical scavengers in preventing tissue injury caused by Escherichia coli in pyelonephritic mice.
Reactive oxygen species have been found to be responsible for the tissue injury caused in experimental pyelonephritis in mice. The extent of lipid peroxidation (as assayed by malondialdehyde formation) was found to be increased significantly (p less than .001) in the infected group as compared to the normal mice. Superoxide dismutase and catalase (oxygen free radical scavengers) showed a significant decrease (p less than .001) in the extent of lipid peroxidation even in the presence of infection. Dimethyl sulfoxide, a hydroxyl ion scavenger, was however found to be effective only at 4 and 7 days postinfection (p less than .001). Allopurinol, an inhibitor of xanthine oxidase, did not significantly (p greater than .05) inhibit the formation of lipid peroxides, even upto 7 days postinfection. There was a significant decrease (p less than .05) in the activities of renal brush border membrane enzymes used as markers of renal tissue damage (i.e. alkaline phosphatase, leucine amino-peptidase and gamma-glutamyl transpeptidase) in the infected group as compared to the normal group. In the presence of superoxide dismutase, dimethylsulfoxide and catalase except allopurinol, the activities of all the enzymes but maltase were found to be increased significantly (p less than .05) as compared to the infected group. There was a significant increase (p less than .01) in the bacterial count in the presence of superoxide dismutase and DMSO in infected mice as compared to the infected control mice. However, no significant difference was observed in the catalase and allopurinol treated groups.


Parameter estimation and prediction for coronavirus disease outbreak 2019 (COVID-19) in Algeria.
The wave of the coronavirus disease outbreak in 2019 (COVID-19) has spread all over the world. In Algeria, the first case of COVID-19 was reported on 25 February, 2020, and the number of confirmed cases of it has increased day after day. To overcome this difficult period and a catastrophic scenario, a model-based prediction of the possible epidemic peak and size of COVID-19 in Algeria is required. We are concerned with a classical epidemic model of susceptible, exposed, infected and removed (SEIR) population dynamics. By using the method of least squares and the best fit curve that minimizes the sum of squared residuals, we estimate the epidemic parameter and the basic reproduction number ℜ 0. Moreover, we discuss the effect of intervention in a certain period by numerical simulation. We find that ℜ 0 = 4.1, which implies that the epidemic in Algeria could occur in a strong way. Moreover, we obtain the following epidemiological insights: the intervention has a positive effect on the time delay of the epidemic peak; the epidemic size is almost the same for a short intervention; a large epidemic can occur even if the intervention is long and sufficiently effective. Algeria must implement the strict measures as shown in this study, which could be similar to the one that China has finally adopted.


A Journey through the Cytoskeleton with Protein Kinase CK2.
Substrate pleiotropicity, a very acidic phosphorylation consensus sequence, and an apparent uncontrolled activity, are the main features of CK2, a Ser/Thr protein kinase that is required for a plethora of cell functions. Not surprisingly, CK2 appears to affect cytoskeletal structures and correlated functions such as cell shape, mechanical integrity, cell movement and division. This review outlines our current knowledge of how CK2 regulates cytoskeletal structures, and discusses involved pathways and molecular mechanisms.


Silica-induced alveolar cell tumors in rats.
Min-U-Sil5, a form of alpha quartz, has been shown to induce peripheral lung tumors in rats exposed to the dust by inhalation. The animals were exposed to a nominal particle concentration of 12.4 mg/m3 for 8 hr/day, 4 days/week, for 2 years. The induced tumors were large and peripheral, and, when examined by electron microscopy, were found to be composed predominantly of alveolar type II cells. These cells were found in papillary, acinar, and solid forms of the tumors and were characterized by lamellar inclusion bodies. This is in contrast to the mouse, in which the papillary form was associated with Clara cells and the acinar form was linked with the type II cell. In this study, the Clara cell was a minor component of the tumor mass. No clear risk is established in man linking silica exposure to increased lung tumor rates.


[Examination of the differential hematocrit and the plasma viscosity under the influence of ionizing radiation].
Systematic investigations into the impact of haemorheological parameters through ionizing rays are not available in literature. The main factors influencing the flow capacity of the blood were determined in vitro by using a set of special methods. Even by using high dosages of irradiation (600 R) only those changes in the flow capacity of the blood could be identified which lay in or near the dispersion area of the methods. These investigations carried out in vitro only take into account the immediate effect of the blood and not rheological changes caused by the intact organism after irradiation.


High-throughput optofluidic particle profiling with morphological and chemical specificity.
We present a method for high-throughput optofluidic particle analysis that provides both the morphological and chemical profiles of individual particles in a large heterogeneous population. This method is based on an integration of a time-stretch optical microscope with a submicrometer spatial resolution of 780 nm and a three-color fluorescence analyzer on top of an inertial-focusing microfluidic device. The integrated system can perform image- and fluorescence-based screening of particles with a high throughput of 10,000 particles/s, exceeding previously demonstrated imaging particle analyzers in terms of specificity without sacrificing throughput.


Bacterial Signatures of "Red-Operculum" Disease in the Gut of Crucian Carp (Carassius auratus).
Fish gut microbiota play important roles in fish immunity, nutrition, and the adaptation to environmental changes. To date, few studies have focused on the interactions among environmental factors, fish diseases, and gut microbiota compositions. We compared the gut bacterial communities of healthy crucian carps (Carassius auratus) with those of individuals affected by "red-operculum" disease and corresponding water and sediment microbiota in four fish farm ponds. Distinct gut bacterial communities were observed in healthy and diseased fish. The bacterial communities of diseased fish were less diverse and stable than those of healthy individuals. The differences in bacterial community compositions between diseased and healthy fish were explained by the changes in the relative abundances of some specific bacterial OTUs, which belonged to the genera such as Vibrio, Aeromonas, and Shewanella, and they were prevalent in diseased fish, but rare or even absent in environmental samples. Water temperature and ammonia concentration were the two most important environmental factors that impacted gut microbiota in diseased fish. These results highlighted the surge of some potential pathogens as bacterial signatures that were associated with "red-operculum" disease in crucian carps.


Muscle cell and motor protein function in patients with a IIa myosin missense mutation (Glu-706 to Lys).
The pathogenic events leading to the progressive muscle weakness in patients with a E706K mutation in the head of the myosin heavy chain (MyHC) IIa were analyzed at the muscle cell and motor protein levels. Contractile properties were measured in single muscle fiber segments using the skinned fiber preparation and a single muscle fiber in vitro motility assay. A dramatic impairment in the function of the IIa MyHC isoform was observed at the motor protein level. At the single muscle fiber level, on the other hand, a general decrease was observed in the number of preparations where the specific criteria for acceptance were fulfilled irrespective of MyHC isoform expression. Our results provide evidence that the pathogenesis of the MyHC IIa E706K myopathy involves defective function of the mutated myosin as well as alterations in the structural integrity of all muscle cells irrespective of MyHC isoform expression.


CYP1B1 Activates Wnt/β-Catenin Signaling through Suppression of Herc5-Mediated ISGylation for Protein Degradation on β-Catenin in HeLa Cells.
Cytochrome P450 1B1 (CYP1B1) acts as a hydroxylase for estrogen and activates potential carcinogens. Moreover, its expression in tumor tissues is much higher than that in normal tissues. Despite this association between CYP1B1 and cancer, the detailed molecular mechanism of CYP1B1 on cancer progression in HeLa cells remains unknown. Previous reports indicated that the mRNA expression level of Herc5, an E3 ligase for ISGylation, is promoted by CYP1B1 suppression using specific small interfering RNA, and that ISGylation may be involved in ubiquitination related to β-catenin degradation. With this background, we investigated the relationships among CYP1B1, Herc5, and β-catenin. RT-PCR and western blot analyses showed that CYP1B1 overexpression induced and CYP1B1 inhibition reduced, respectively, the expression of Wnt/β-catenin signaling target genes including β-catenin and cyclin D1. Moreover, HeLa cells were treated with the CYP1B1 inducer 7,12-dimethylbenz[α]anthracene (DMBA) or the CYP1B1 specific inhibitor, tetramethoxystilbene (TMS) and consequently DMBA increased and TMS decreased β-catenin and cyclin D1 expression, respectively. To determine the correlation between CYP1B1 expression and ISGylation, the expression of ISG15, a ubiquitin-like protein, was detected following CYP1B1 regulation, which revealed that CYP1B1 may inhibit ISGylation through suppression of ISG15 expression. In addition, the mRNA and protein expression levels of Herc5 were strongly suppressed by CYP1B1. Finally, an immunoprecipitation assay revealed a direct physical interaction between Herc5 and β-catenin in HeLa cells. In conclusion, these data suggest that CYP1B1 may activate Wnt/β-catenin signaling through stabilization of β-catenin protein from Herc5-mediated ISGylation for proteosomal degradation.


Butorphanol and pentazocine in patients with severe postoperative pain.
Either butorphanol tartrate (1, 2, or 4 mg) or pentazocine (30 or 60 mg) was given intramuscularly (5 dose groups) to 262 patients, who were scheduled for major operations, if they had severe pain after full awakening in the recovery room, postoperatively under double-blind conditions. There were at least 50 patients in each of the five dosage groups. Pain intensity and relief were scored numerically for each treated patient at 1/2 1, 2, 3 and 4 hr, while under direct surveillance, and the patients were seen again for follow-ups during the first 24 hr postoperatively. Appreciable pain relief developed within 30 min at all dose levels, with a peak analgesic effect apparent at about 1 hr. Satisfactory relief persisted for 4 hr in the majority of patients in each group. With the lowest dose of butorphanol tartrate (1 mg), which gave good pain relief for only 2 to 4 hr, approximately 60% of the patients had to be remedicated within 4 hr. Vital signs were not appreciably affected, but the patients who received the high dose of analgesics (butorphanol tartrate 4 mg or pentazocine lactate 60 mg) were often quite drowsy. The incidence of other side effects was negligible. The relative potency assay showed that butorphanol was approximately 20 times as potent as pentazocine for up to 4 hr.


Poly[diaqua-(μ(4)-benzene-1,2,4,5-tetra-carboxyl-ato)[μ(2)-1,4-bis-(3-pyridyl-meth-yl)piperazine]dizinc(II)].
In the title compound, [Zn(2)(C(10)H(2)O(8))(C(16)H(20)N(4))(H(2)O)(2)](n), the Zn(II) atom is in a distorted tetra-hedral environment, being coordinated by one N atom from a 1,4-bis-(3-pyridyl-meth-yl)piperazine (3-bpmp) ligand, two O atoms from two carboxyl-ate groups of the pyromellitate anion and one water mol-ecule. The distortion of the tetrahedral coordination may be ascribed to the hydrogen bonds between the carboxyl-ate groups and the adjacent water mol-ecules. Each Zn(II) atom links to three organic ligands and each pyromellitate ligand coordinates to four Zn(II) atoms, forming a (3,4)-connected infinite three-dimensional framework. O-H⋯N inter-actions also occur.


Prevalence and Determinants of Unmet Social Needs Among Rural and Urban Veterans.
To examine the prevalence and determinants of nine unmet social needs among rural compared with urban Veterans. Retrospective study using survey data collected in 2020 merged with Veterans Health Administration (VA) administrative data. For each unmet need, separate logistic regression modes were run predicting the odds of rural compared with urban Veterans endorsing the need adjusting for sociodemographic characteristics and comorbidities. 2,801 Veterans responded to the survey (53.7% response rate). Veterans experienced high rates of need (e.g., 22% reported food insecurity). Unmet need prevalence varied minimally between rural and urban Veterans and where they did, rural Veterans were less likely to endorse the need (e.g., loneliness). For many unmet needs, Black compared with White Veterans were at higher risk. Regional unmet need disparities were also observed. As VA considers expanding unmet need interventions, tailoring interventions to the sub-populations most at risk may be warranted.


Transesophageal echocardiography in an emergency setting. A district general hospital experience.
Transesophageal echocardiography is an extremely useful technique for the study of various cardiovascular pathologies. In the particular setting of emergency, it is of great value for prompt diagnosis and appropriate therapy. It was our aim to evaluate, in our hospital, the benefits obtained by the use of transesophageal echocardiography in an emergency setting. We retrospectively studied patients who underwent transesophageal echocardiography (TEE) in an emergency setting, from June 1997 to December 2002, evaluating demographic characteristics, indication to perform TEE, benefit obtained (diagnosis or exclusion of initial diagnosis), and technique-related complications. There were 97 transesophageal echocardiograms performed in an emergency setting in the period under consideration, accounting for 19.3% of the total number of exams. Fifty-two patients (53.6%) were male, mean age 63.9 +/- 12.7. Nineteen patients (19.6%) were on assisted ventilation. The indications to perform TEE were: possible massive or submassive pulmonary thromboembolism in 32 patients (33.0%); suspected aortic dissection in 19 (19.6%); shock with inconclusive transthoracic echocardiogram in ten (10.3%); possible endocarditis in eight (8.2%); possible prosthetic valve dysfunction in seven (7.2%); intracardiac mass in six (6.2%); search for cardiac source of embolism in five (5.2%); possible mechanical complication of acute myocardial infarction in four (4.1%); pre-electrical cardioversion study in four patients with atrial fibrillation (4.1%); and suspected congenital heart disease in two (2.1%). TEE examination yielded additional information and helped in the therapeutic decision in 88 patients (90.7%), leading to a diagnosis in 49 (50.6%), which was different from the initial diagnostic hypothesis in four, and exclusion of the suspected diagnosis in 39 (40.1%). There was only one minor complication (1.0%) and no TEE-related mortality. We concluded that transesophageal echocardiography is an extremely useful and safe cardiovascular diagnostic technique in an emergency setting in a district general hospital, enabling a diagnosis to be reached or excluded in almost all patients, which is essential for implementing appropriate therapy.


Silkworms suppress the release of green leaf volatiles by mulberry leaves with an enzyme from their spinnerets.
In response to herbivory, plants emit a blend of volatile organic compounds that includes green leaf volatiles (GLVs) and terpenoids. These volatiles are known to attract natural enemies of herbivores and are therefore considered to function as an indirect defense. Selection should favor herbivores that are able to suppress these volatile emissions, and thereby make themselves less conspicuous to natural enemies. We tested this possibility for silkworms, which were observed to leave secretions from their spinnerets while feeding on mulberry leaves. When we ablated the spinnerets of silkworms, no secretions were observed. Leaves infested by intact silkworms released smaller amounts of GLVs than leaves infested by ablated silkworms, indicating that the spinneret secretion suppressed GLV production. This difference in GLV emissions was also reflected in the behavioral response of Zenillia dolosa (Tachinidae), a parasitoid fly of silkworms. The flies laid fewer eggs when exposed to the volatiles from intact silkworm-infested leaves than when exposed to the volatiles from ablated silkworm-infested leaves. We identified a novel enzyme in the secretion from the spinneret that is responsible for the GLV suppression. The enzyme converted 13(S)-hydroperoxy-(9Z,11E,15Z)-octadecatrienoic acid, an intermediate in the biosynthetic pathway of GLVs, into its keto-derivative in a stereospecific manner. Taken together, this study shows that silkworms are able to feed on mulberry in a stealthy manner by suppressing GLV production with an enzyme in secretions of their spinnerets, which might be a countermeasure against induced indirect defense by mulberry plants.


High Expression of MORC2 is Associated with Poor Clinical Outcomes and Immune Infiltrates in Colon Adenocarcinoma.
Microrchidia 2 (MORC2) is a universally expressed molecule that has recently been identified as a chromatin modulator and elevated in many malignancies. However, its prognostic value and immunological role of MORC2 in colon adenocarcinoma (COAD) have never been illustrated. The clinical parameters and MORC2 expression datasets of COAD patients were obtained from The Cancer Genome Atlas (TCGA). Cancer and adjacent tissue specimens from surgically resected COAD patients were collected, and quantitative real-time PCR was used to detect MORC2 expression. Differentially expressed genes related to MORC2 were discovered and used for functional enrichment analysis. The diagnostic and prognostic values of MORC2 in COAD were conducted using receiver operating characteristics (ROC), Kaplan-Meier survival curve analysis, PrognoScan, Gene Expression Profiling Interactive Analysis (GEPIA) public databases and nomograms. Eventually, the association of MORC2 with tumor microenvironment was analyzed by using TIMER and GSVA package of R (v3.6.3). MORC2 expression was upregulated in COAD tissues, and the RT-qPCR results further verified the reliability of our differential analysis at the transcriptional level. Additionally, higher expression of MORC2 was correlated to a poor prognosis for COAD patients. MORC2 was an independent prognostic factor for COAD and could be a diagnostic factor for early COAD. Furthermore, MORC2 expression was positively correlated with immune cells such as NK cells, TFH cells and so on. The findings demonstrated that overexpression of MORC2 was correlated with worse prognosis and immune infiltrates of COAD. MORC2 can serve as a reliable diagnostic and prognostic biomarker and a target of immunotherapy for COAD patients.


Redundant sensory information does not enhance sequence learning in the serial reaction time task.
In daily life we encounter multiple sources of sensory information at any given moment. Unknown is whether such sensory redundancy in some way affects implicit learning of a sequence of events. In the current paper we explored this issue in a serial reaction time task. Our results indicate that redundant sensory information does not enhance sequence learning when all sensory information is presented at the same location (responding to the position and/or color of the stimuli; Experiment 1), even when the distinct sensory sources provide more or less similar baseline response latencies (responding to the shape and/or color of the stimuli; Experiment 2). These findings support the claim that sequence learning does not (necessarily) benefit from sensory redundancy. Moreover, transfer was observed between various sets of stimuli, indicating that learning was predominantly response-based.


Chamomile Flower, Myrrh, and Coffee Charcoal, Components of a Traditional Herbal Medicinal Product, Diminish Proinflammatory Activation in Human Macrophages.
A traditional herbal medicinal product, containing myrrh, chamomile flower, and coffee charcoal, has been used in Germany for the relief of gastrointestinal complaints for decades. Clinical studies suggest its use in the maintenance therapy of inflammatory bowel disease. However, the pharmacological mechanisms underlying the clinical effects are not yet fully understood.The present study aims to elucidate immunopharmacological activities of myrrh, chamomile flower, and coffee charcoal by studying the influence of each plant extract on gene expression and protein release of activated human macrophages.The plant extracts effect on gene and protein expression of activated human monocyte-derived macrophages was investigated by microarray gene expression analysis and assessment of the release of pro- and anti-inflammatory mediators (TNFα, chemokine CXCL13, and interleukin-10) using an ELISA test system.The extracts of myrrh, chamomile flower, and coffee charcoal influenced gene expression of activated human macrophages within the cytokine/chemokine signaling pathway. Particularly, chemokine gene expression was suppressed. Subsequently, the production of CXCL13 and, to a minor extent, cytokine TNFα was inhibited by all herbal extracts. Chamomile flower and coffee charcoal extracts enhanced interleukin-10 release from activated macrophages. The observed effects on protein release were comparable to the effect of budesonide, which decreased TNFα and CXCL13 and enhanced interleukin-10 release.The components of the herbal medicinal product influence the activity of activated human macrophages on both gene and protein level. The induced alterations within chemokine/cytokine signaling could contribute to a positive effect on the immunological homeostasis, which is disturbed in patients with chronic intestinal inflammation.


Hip structural analysis, trabecular bone score, and bone mineral density in post-menopausal women with type-2 diabetes mellitus: a multi-center cross-sectional study in the south of Iran.
This study aimed to evaluate bone mineral density (BMD), trabecular microarchitecture, and proximal hip geometry in diabetic postmenopausal women, where BMD alone cannot reflect bone strength adequately. We found significantly lower trabecular bone score and BMD at the distal radius and total forearm in diabetic subjects compared to controls. The limitations resulting from the exclusive assessment of bone mineral density (BMD) in people with diabetes can lead to underestimation of microarchitectural and geometric changes, both of which play an essential role in the fracture risk. Therefore, we aimed to evaluate BMD, trabecular bone score (TBS), and hip structural analysis (HSA) in diabetic type-2 post-menopausal women and compare them with healthy postmenopausal subjects. BMD was assessed at the lumbar spine, femoral sites, distal radius, and total forearm using dual-energy X-ray absorptiometry (DXA); TBS was measured based on DXA images using the software at the same region of interest as the BMD measurements; geometric assessment at the proximal femur was performed by the HSA program. A total of 348 ambulatory type-2 diabetic postmenopausal women and 539 healthy postmenopausal women were enrolled. TBS and BMD at the distal radius and total forearm were significantly (P value < 0.05) lower in cases compared to controls after age and body mass index (BMI) adjustment. In addition, degraded bone microarchitecture was significantly (P value < 0.05) more prevalent in diabetic subjects than in non-diabetic controls after adjusting for age and BMI. A number of geometric indices of the proximal hip were significantly lower in the controls than in those with diabetes (P-value < 0.05). This study may highlight the utility of the TBS and BMD at the distal radius and total forearm in subjects with type-2 diabetes mellitus, where the BMD at central sites may not adequately predict fracture risk.


Prevention of contrast-induced nephropathy in vascular surgery patients.
Presently, only hydration and N-acetylcysteine have been shown to be effective in decreasing the incidence of radiographic contrast-induced nephropathy. We investigated the role of N-acetylcysteine and various hydration protocols in vascular surgery patients undergoing angiography. A single-center, randomized, placebo-controlled trial was conducted in patients with stable, preexisting renal dysfunction undergoing elective, outpatient angiography. Patients were randomized to outpatient oral hydration and N-acetylcysteine, inpatient hydration plus N-acetylcysteine, or our standard therapy of inpatient intravenous hydration alone. Two of twenty-eight (7%) patients who received outpatient oral hydration and N-acetylcysteine developed contrast-induced nephropathy, while two of 25 (8%) who recieved inpatient hydration plus N-acetylcysteine developed contrast-induced nephropathy and two of 25 (8%) who received standard therapy of inpatient intravenous hydration alone developed contrast-induced nephropathy. There was no statistical difference in incidence of contrast-induced nephropathy between the groups. No statistically significant independent risk factors were identified among the patients who developed contrast-induced nephropathy. N-Acetylcysteine did not confer additional benefit to patients treated with inpatient intravenous hydration. Outpatient oral hydration plus N-acetylcysteine was as effective at preventing contrast-induced nephropathy as inpatient therapies and avoided costly hospital admission.


Dietary modulation of rat colonic cAMP-dependent protein kinase activity.
Malignant transformation of cells is associated with enhanced proliferation and alterations in cAMP-dependent protein kinase (PKA) activity. To investigate the role of PKA in normal colonic cell proliferation, PKA was characterized in rat colonic mucosa. In addition, rats were fed diets containing different fats (corn oil, fish oil) and fibers (pectin, cellulose, fiber free) to elicit varying levels of colonic cell proliferation in order to study this signaling system under normal physiologic conditions. Overall, PKA activities were higher in cytosolic compared to membrane fractions. PKA type II (PKA II) isozyme contributed 89 +/- 1% and 96 +/- 1% of total PKA activity in cytosolic and membrane fractions, respectively. Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis revealed the presence of mRNA for both the alpha and beta isoforms of the regulatory subunits of PKA II. PKA activities were 21-33% higher in distal membrane and total distal fractions in rats fed a cellulose/corn oil diet compared to animals consuming the other fiber/fat diets. These effects were seen only in the distal colon, where the number of cells per crypt column was elevated only in animals fed the cellulose/corn oil diet relative to other diets. Diet-induced mitogenic responses did not involve significant changes in the relative activity of PKA I and II isozymes. These data demonstrate that dietary effects on PKA activity in the distal colon may be related to changes in cell differentiation as indicated by the number of cells per crypt column.


The structural and functional implications of linked SNARE motifs in SNAP25.
We investigated the functional and structural implications of SNAP25 having two SNARE motifs (SN1 and SN2). A membrane-bound, intramolecular FRET probe was constructed to report on the folding of N-terminal SN1 and C-terminal SN2 in living cells. Membrane-bound constructs containing either or both SNARE motifs were also singly labeled with donor or acceptor fluorophores. Interaction of probes with other SNAREs was monitored by the formation of SDS-resistant complexes and by changes in FRET measured in vitro using spectroscopy and in the plasma membrane of living cells using TIRF microscopy. The probes formed the predicted SDS-resistant SNARE complexes. FRET measurements revealed that syntaxin induced a close association of the N-termini of SN1 and SN2. This association required that the SNARE motifs reside in the same molecule. Unexpectedly, the syntaxin-induced FRET was prevented by VAMP. Both full-length SNAP25 constructs and the combination of its separated, membrane-bound constituent chains supported secretion in permeabilized chromaffin cells that had been allowed to rundown. However, only full-length SNAP25 constructs enabled robust secretion from intact cells or permeabilized cells before rundown. The experiments suggest that the bidentate structure permits specific conformations in complexes with syntaxin and VAMP and facilitates the function of SN1 and SN2 in exocytosis.


Using Planar Phi29 pRNA Three-Way Junction to Control Size and Shape of RNA Nanoparticles for Biodistribution Profiling in Mice.
RNA is rapidly emerging as a versatile building block for nanoparticle assembly due to its simplicity in base pairing, while exhibiting diversity in function such as enzymatic activity similar to some proteins. Recent advances in RNA nanotechnology have generated significant interests in applying RNA nanoparticles for various applications in nanotechnology and nanomedicine. In particular, assessing the effect of size and shape on cell entry and intracellular trafficking as well as in vivo biodistribution of nanoparticles is challenging due to the lack of nanoparticles rich in structure while varying in size and shape. RNA nanotechnology exemplified by the packaging RNA (pRNA) of bacteriophage phi29 DNA packaging motor has provided a different prospect in nanoparticle designs. Of note, there is a robust three-way junction (3WJ) motif in pRNA which can serve as an adaptable scaffold to construct thermodynamically stable 2D planar and 3D globular RNA architectures with tunable shapes and sizes, and harboring various targeting, therapeutic, and imaging modules. This chapter focuses on the methods for constructing pRNA-3WJ based nanoparticles with controllable sizes and shapes, and assessment of their biodistribution profiles in cancer mouse models after systemic injection and ocular mouse models following subconjunctival injection.


Excitatory amino acid receptors in glial progenitor cells: molecular and functional properties.
We have analyzed the molecular and biophysical properties of glutamate-gated channels in cells of the oligodendrocyte lineage, using both the CG-4 primary cell line (Louis et al: J. Neurosci. Res. 31:193-204, 1992a) and oligodendrocyte progenitors purified from the rat cerebral cortex. CG-4 progenitor cells, as well as primary progenitors, were stained with a specific anti-GABA antibody. In whole-cell patch-clamp recordings, rapid perfusion of the agonists L-glutamate, kainate, and AMPA produced rapidly desensitizing currents in CG-4 cells. NMDA was ineffective. Both rapidly desensitizing and steady-state components of responses to kainate were inhibited by the kainate/AMPA receptor antagonist CNQX. Northern blot analysis of total mRNA isolated from CG-4 cells revealed co-expression of both AMPA- and kainate-preferring glutamate receptor subunits. The activation of glutamate receptors in CG-4 cells caused a rapid and transient elevation of mRNAs for the immediate early gene NGFI-A.


Plasma Lipidomic Signature of Rectal Adenocarcinoma Reveals Potential Biomarkers.
Rectal adenocarcinoma (RAC) is a common malignant tumor of the digestive tract and survival is highly dependent upon stage of disease at diagnosis. Lipidomic strategy can be used to identify potential biomarkers for establishing early diagnosis or therapeutic programs for RAC. To evaluate the lipoperoxidation biomarkers and lipidomic signature in the plasma of patients with RAC (n = 23) and healthy controls (n = 18). Lipoperoxidation was evaluated based on malondialdehyde (MDA) and F2-isoprostane levels and the lipidomic profile obtained by gas chromatography and high resolution mass spectrometry (ESI-q-TOF) associated with a multivariate statistical technique. The most abundant ions identified in the RAC patients were those of protonated phosphatidylcholine and phosphatidylethanolamine. It was found that a lisophosphatidylcholine (LPC) plasmalogen containing palmitoleic acid [LPC (P-16:1)], with highest variable importance projection score, showed a tendency to be lower in the cancer patients. A reduction of n - 3 polyunsaturated fatty acids was observed in the plasma of these patients. MDA levels were higher in patients with advanced cancer (stages III/IV) than in the early stages groups and the healthy group (p < 0.05). No differences in F2-isoprostane levels were observed among these groups. This study shows that the reduction in plasma levels of LPC plasmalogens associated with an increase in MDA levels may indicate increased oxidative stress in these patients and identify the metabolite LPC (P-16:1) as a putatively novel lipid signature for RAC.


Evaluation of developmental toxicity of coniine to rats and rabbits.
Conium maculatum (poison hemlock, CM) is teratogenic in several domestic species, presumably due to its piperidine alkaloids, including coniine, which has been verified to be teratogenic in cattle. Coniine/CM teratogenicity culminates in production of arthrogryposis. The purpose of this study was to evaluate coniine-induced teratogenicity in two laboratory animal species, Sprague-Dawley rats and New Zealand white rabbits. Pregnant rats were given coniine (25 mg/kg body weight) by oral gavage at 8-hour intervals on gestation days 16-18. Pregnant rabbits were given coniine (40 mg/kg body weight) by oral gavage at 8-hour intervals on gestation days 20-24. Rats were killed on day 19 and rabbits on day 29. Fetuses were immediately removed, weighed, and examined for external abnormalities. Alternate fetuses were either stained for skeletal examinations with alizarin red-S or fixed in Bouin's solution for visceral examination. Symptoms of maternal intoxication due to coniine administration were observed in both the rat and the rabbit, and higher doses were uniformly lethal. Rabbits treated with coniine appeared to lose more weight and eat less than controls, but there was no statistically significant difference between groups. Fetal weights were significantly lower in coniine-exposed rat and rabbit fetuses indicating fetotoxicity. The only statistically significant treatment-related visceral or skeletal malformation was a reduction of cranial ossification of rabbit fetuses, probably related to maternal toxicity. Coniine-exposed rabbit litters tended to be affected by arthrogryposis (no bony deformities noted on skeletal exam) more than controls (2/6 vs. 0/9).


Resistance of SARS-CoV-2 variants to neutralization by antibodies induced in convalescent patients with COVID-19.
Administration of convalescent plasma or neutralizing monoclonal antibodies (mAbs) is a potent therapeutic option for coronavirus disease 2019 (COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. However, SARS-CoV-2 variants with mutations in the spike protein have emerged in many countries. To evaluate the efficacy of neutralizing antibodies induced in convalescent patients against emerging variants, we isolate anti-spike mAbs from two convalescent COVID-19 patients infected with prototypic SARS-CoV-2 by single-cell sorting of immunoglobulin-G-positive (IgG+) memory B cells. Anti-spike antibody induction is robust in these patients, and five mAbs have potent neutralizing activities. The efficacy of most neutralizing mAbs and convalescent plasma samples is maintained against B.1.1.7 and mink cluster 5 variants but is significantly decreased against variants B.1.351 from South Africa and P.1 from Brazil. However, mAbs with a high affinity for the receptor-binding domain remain effective against these neutralization-resistant variants. Rapid spread of these variants significantly impacts antibody-based therapies and vaccine strategies against SARS-CoV-2.


Hearing Outcomes in Stickler Syndrome: Variation Due to COL2A1 and COL11A1.
Stickler syndrome (SS) is a heterogeneous inherited connective tissue disorder, often due to a mutation in COL2A1 or COL11A1. Mutations in these genes cause collagen abnormalities affecting ocular, auditory, orofacial, and skeletal systems, including hearing loss, micrognathia, and cleft palate. Understanding the variability of hearing phenotypes based on genetic mutation has a significant impact on treatment and long-term care. A retrospective chart review of pediatric patients with a confirmed diagnosis of SS between January 2003 and December 2018 at a tertiary pediatric hospital was performed. Patients were excluded if they did not have genetic evaluation, craniofacial/ear, nose, and throat evaluation, and/or audiologic testing. Charts were reviewed for the following information: age, race, sex, SS diagnosis, genetic variant of SS, and audiological testing data. There were 29 confirmed patients with SS who met criteria, 16 with type I (COL2A1) and 13 with type II (COL11A1). Of the 13 patients with type II, 12 (92%) demonstrated hearing loss, ranging in severity from mild to severe. In type I, 25% of patients had mild or resolved hearing loss. Results suggest that patients with type II SS are more likely to have congenital hearing loss than type I. Data also suggest that the COL11A1 mutation shows consistently more severe hearing loss than the COL2A1 mutation.


African swine fever in North Sumatra and West Java provinces in 2019 and 2020, Indonesia.
African swine fever (ASF) is a highly lethal and contagious viral haemorrhagic disease of domestic and wild pigs, caused by the ASF virus (ASFV). After entering China in 2018, the disease has continued to spread through Asia. In September 2019, a team from the Indonesian Research Center for Veterinary Science, Bogor, investigated outbreaks in backyard pigs in the Dairi and Humbang Hasundutan districts of North Sumatra province. In January 2020, three pigs purchased from a pig seller in Bogor District, West Java province were also tested. Real-time PCR results confirmed ASFV DNA in sixteen out of twenty-nine samples, with nine positive samples from North Sumatra and seven from West Java. Four partial or full-length genes (i.e. p72, p54, pB602L and CD2v) and a 356-bp fragment between the I73R and I329L genes were sequenced from representative samples. Phylogenetic analysis established that the ASFV in the samples from both North Sumatra and West Java were identical, indicating a common source of infection, and that they belonged to the p72 genotype II and serogroup 8. The sequences from the Indonesian ASFVs were also identical to other genotype II ASFV from domestic pigs in Vietnam, China and Russia.


Factors affecting separation and detection of bile acids by liquid chromatography coupled with mass spectrometry in negative mode.
Bile acids (BAs) are cholesterol metabolites with important biological functions. They undergo extensive host-gut microbial co-metabolisms during the enterohepatic circulation, creating a vast structural diversity and resulting in great challenges to separate and detect them. Based on the bioanalytical reports in the past decade, this work developed three chromatographic gradient methods to separate a total of 48 BA standards on an ethylene-bridged hybrid (BEH) C18 column and high-strength silica (HSS) T3 column and accordingly unraveled the factors affecting the separation and detection of them by liquid chromatography coupled with mass spectrometry (LC-MS). It was shown that both the acidity and ammonium levels in mobile phases reduced the electrospray ionization (ESI) of BAs as anions of [M-H]-, especially for those unconjugated ones without 12-hydroxylation. It was also found that the retention of taurine conjugates on the BEH C18 column was sensitive to the strength of formic acid and ammonium in mobile phases. By using the volatile buffers with an equivalent ammonium level as mobile phases, we comprehensively demonstrated the effects of the elution pH value on the retention behaviors of BAs on both the BEH C18 column and HSS T3 column. Based on the retention data acquired on a C18 column, we presented the ionization constants (pK a) of various BAs with the widest coverage beyond those of previous reports. When we made attempts to establish the structure-retention relationships (SRRs) of BAs, the lack of discriminative structural descriptors for BA stereoisomers emerged as the bottleneck problem. The methods and results presented in this work are especially useful for the development of reliable, sensitive, high-throughput, and robust LC-MS bioanalytical protocols for the quantitative metabolomic studies. Graphical Abstract Nonlinear curve fitting of capacity factors and elution pH value for the separation of common unconjugated bile acids.


Case-control study of idiopathic pulmonary fibrosis and environmental exposures.
The authors conducted a matched case-control study of interstitial lung disease (ILD) using a self-administered questionnaire. All cases in the practice of two pulmonary specialists were identified. Cases were excluded if they were thought to have occupational exposures as a contributor, hypersensitivity pneumonitis or sarcoidosis, or some other well-recognized cause. Three controls were matched by sex and age (one year) as identified through orthopedic surgeons at the same institutions. Two rounds of a questionnaire were mailed; 17 cases (37.7%) and 94 controls (32.4%) responded to the questionnaire, although many of the case addresses were no longer valid. Individuals with ILD were 16.0 times as likely to report mold exposure and 11.1 times as likely to report silica as were controls in their workplace. Odds ratios associating ILD with moisture indicators in the home were in the same range as previously published associations between such indicators and wheezing. ILD may have environmental and occupational causes that warrant more systematic exploration.


MMP-21 is expressed by macrophages and fibroblasts in vivo and in culture.
Matrix metalloproteinase (MMP)-21 and MMP-26 (matrilysin-2) are two recently cloned epithelial metalloproteases. Here we examined their expression in various benign skin disorders, in which macrophages and fibroblasts have been implicated as well as in cultures of these cells. Expression of MMP-21 was detected by immunohistochemistry in a subset of macrophages of granulomatous skin lesions and in fibroblasts in dermatofibromas. MMP-21 mRNA was found in THP-1, U937, HEL 299 and Hs68 cells. Furthermore, MMP-21 protein was detected by immunohistochemistry in cultures of the same cell lines. In culture MMP-21 was upregulated by phorbol myristate acetate in THP-1 cells and by retinoic acid (RA) in U937 cells, and downregulated by transforming growth factor beta 1 (TGF-beta1) in HEL 299 as assessed by Taqman quantitative polymerase chain reaction (PCR). Expression of MMP-26 was detected by immunohistochemistry in granulomatous skin diseases and actinic elastosis. MMP-26 at both mRNA and protein levels was only found in HEL 299 cells. In culture it was downregulated by TGF-beta1, RA and IL-1beta as assessed by Taqman quantitative PCR. Our results suggest these two novel MMPs are not only associated with cancer but may be important in connective tissue remodelling and pathobiology of various benign skin disorders.