Manganese: a review of occupational and environmental toxicology.
The following topics are dealt with in this brief review concerning the hygienic and toxicologic aspects of manganese: Occurrence, production, uses; experimental pathology; clinical toxicology; occupational and environmental hygiene aspects of human exposure. The preventive measures outlined: reduction of dust concentration to comply with the established MAC value, in extreme situation even with the use of personal protection; preventive pre-employment examinations to exclude from work all individuals with liver and kidney lesions, blood dyscrasia, organic CNS lesions and progressive lung diseases; elaboration of suitable exposure tests, especially the method of hair analysis, to ensure the biological monitoring of environmental exposure to manganese.


A Clinicopathological and Immunofluorescence Study of Intraepidermal Immunobullous Diseases.
Autoimmune blistering diseases (AIBDs) are characterized by autoantibodies directed against antigens of skin. Direct immunofluorescence (DIF) study helps in confirming the diagnosis where histopathology alone is noncontributory. This study aimed to evaluate the clinicopathological and DIF features of intraepidermal AIBD and to assess their relative diagnostic significance. It was an institution-based observational study. A total of 34 patients were studied over a period of 1½ years in the Department of Pathology in collaboration with the Department of Dermatology of a tertiary care hospital. The clinical, pathological, and DIF features were evaluated and documented. Data were analyzed by statistical tests using GraphPad InStat. Pemphigus vulgaris (PV) was the predominant type with 18 (53%) cases followed by 15 (44%) cases of pemphigus foliaceus (PF) and a single case of pemphigus erythematosus (PE). The age of the patients ranged from 17 to 85 years. Overall, there was a female preponderance in the study group. The most common presenting feature was pruritus (58.82%). Tzanck smear showed the presence of acantholytic cells in thirty (88.24%) patients. Characteristic histopathological features were present in all the cases of PV and PF except one case of PF which was found to be a case of PE. DIF study showed intraepidermal deposition of intercellular immunoglobulin G (IgG) and C3 both in PV and PF. The case of PE showed epidermal "antinuclear antibody" staining with IgG. Immunofluorescence study may be used as an additional tool for confirmation of diagnosis where histopathology alone is inconclusive.


Consensus statement on safety measures for pressurized intraperitoneal aerosol chemotherapy.
Pressurized intraperitoneal aerosol chemotherapy (PIPAC) is a promising treatment for peritoneal cancer that entails, however, potential risks for the caregivers in the operating room (OR). This study aimed to reach a consensus within the PIPAC community on a comprehensive safety protocol. Active PIPAC centers were invited to participate in a two-round Delphi process on 43 predefined items: concise summaries of the existing evidence were presented together with questions formulated using the population, intervention, comparator, and outcome framework. According to the Grading of Recommendations Assessment, Development, and Evaluation, the strength of recommendation was voted by panelists, accepting a consensus threshold of ≥50% of the agreement for any of the four grading options, or ≥70% in either direction. Forty-seven out of 66 invited panelists answered both rounds (response rate 76%). The consensus was reached for 41 out of 43 items (95.3%). Strong and weak recommendations were issued for 30 and 10 items, respectively. A positive consensual recommendation was issued to activate laminar airflow without specific strength, neither strong nor weak. No consensus was reached for systematic glove change for caregivers with a high risk of exposure and filtering facepiece mask class 3 for caregivers with low risk of exposure. A high degree of consensus was reached for a comprehensive safety protocol for PIPAC, adapted to the risk of exposure for the different caregivers in the OR. This consensus can serve as a basis for education and help reach a high degree of adherence in daily practice.


Relationships between intergroup contact and prejudice among minority and majority status groups.
Considerable research has shown that greater intergroup contact corresponds with lower intergroup prejudice, yet little is known regarding how the relationships between contact and prejudice may vary for members of minority and majority status groups. The present research examined differences in contact-prejudice relationships among members of minority and majority status groups, using data from a larger meta-analytic study of the effects of intergroup contact. Results indicate that the relationships between contact and prejudice tend to be weaker among members of minority status groups than among members of majority status groups. Moreover, establishing Allport's (1954) proposed conditions for optimal intergroup contact significantly predicts stronger contact-prejudice relationships among members of majority status groups, but not among members of minority status groups. Implications of these findings for future research on contact between minority and majority status groups are discussed.


Association of Biochemically Verified Post-Diagnosis Smoking and Nonmuscle-Invasive Bladder Cancer Recurrence Risk.
Our goal was to determine the association between biochemically verified post-diagnosis smoking exposure and nonmuscle-invasive bladder cancer (NMIBC) recurrence risk. We conducted a prospective study of 354 NMIBC patients with a smoking history undergoing care between 2015 and 2018. Patients contributed at least 2 biospecimens during followup which were tested for cotinine to determine biochemically verified post-diagnosis smoking exposure (yes/no). Our primary endpoint was time to first recurrence after study start date. We examined whether post-diagnosis smoking exposure was associated with recurrence risk in multivariable Cox proportional hazards models that accounted for demographics, clinicopathological variables, time since diagnosis and pack-years. Patients were predominantly White, male and had a median age of 68 years. Most patients had Ta disease (62%) and tumors of high grade (68%). Intravesical bacillus Calmette-Guérin was given to 63% of the cohort. Patients were followed for a median of 3.6 years since study start. Post-diagnosis smoking exposure was detected in 22% of patients, and 38.7% (137) of patients experienced a recurrence during followup. In multivariable models, only bacillus Calmette-Guérin treatment and prior recurrence rate were significantly associated with recurrence. There was no association between post-diagnosis smoking exposure and recurrence risk (HR: 0.73, 95% CI: 0.45-1.20). In a cohort of patients with predominantly high risk NMIBC, post-diagnosis smoking exposure was not associated with NMIBC recurrence. However, smoking cessation support remains a critical component of cancer care given that the benefits of quitting extend far beyond NMIBC recurrence.


Role of nitric oxide-dependent pathways in ethanol-induced anxiolytic effects in rats.
Nitric oxide (NO) is a neuromodulator and an intercellular and retrograde messenger that mediates several functions in the central nervous system. The effects of ethanol (EtOH) on neuronal NO-dependent pathways have been the focus of recent research. Most studies have concentrated on the actions of chronic EtOH exposure. In this study, we examined the role of NO-dependent pathways in the acute actions of EtOH. We used the elevated plus-maze test to study the role of NO-dependent pathways in the behavior of rats treated with acute EtOH. We tested the effects of 7-nitroindazole, a reversible competitive inhibitor of nitric oxide synthase. We also studied the effects of the cyclic guanylate monophosphate (cGMP) analog, 8-Bromoguanosine cyclic 3',5'-monophosphate sodium salt, and the NO donors S-nitroso-N-acetylpenicillamine or sodium nitroprusside. When injected by either intraperitoneal (6 mg/kg) or intrahippocampal (20 nmol) routes, 7-nitroindazole increased the percentage of open arm entries and time spent in open arms for rats injected with EtOH (1.0 g/kg, ip). This dose of EtOH did not produce an anxiolytic effect when administered alone. Additional experiments were performed with a dose of 1.2 g/kg of EtOH (ip), which produced an anxiolytic effect. Intrahippocampal administration of the cGMP analog, 8-Bromoguanosine cyclic 3',5'-monophosphate sodium salt (40 nmol), or the NO donors S-nitroso-N-acetylpenicillamine (20 or 40 nmol) or sodium nitroprusside (20 or 40 nmol) blocked the anxiolytic effect of this dose of EtOH. These results indicate that inhibition of NO-dependent pathways enhances, whereas stimulation of these pathways decreases, the efficacy of EtOH to produce anxiolytic effects in rats. We postulate that NO-dependent increases in guanylate cyclase activity and cGMP levels oppose the anxiolytic effects produced by acute EtOH administration.


Fulminant Hepatic Failure Secondary to Herpes Hepatitis in a Patient With Myasthenia Crisis: An Elusive Diagnosis.
Herpes hepatitis is a rare cause of fulminant hepatic failure contributing to less than 1% of all cases. It is most often seen in persons who are immunosuppressed and in pregnant women. The presentation is usually non-specific and rapidly progressive, thus making antemortem diagnosis of this condition rare. We present a patient who was on chronic immunosuppressive therapy for systemic lupus erythematosus and subsequently developed multi-organ failure with anicteric transaminitis as a result of disseminated herpes infection. The diagnosis was only made post-mortem. A confounding factor in this case was the fact that the patient underwent plasmapheresis, which skewed the interpretation of liver function tests in the setting of acute liver failure.


Serum beta-hydroxybutyrate measurement in patients with uncontrolled diabetes mellitus.
To assess whether routinely measuring serum beta-hydroxybutyrate (beta-OHB) concentration might help judge the severity of or assist in treating patients with suspected ketoacidosis. Serum beta-OHB level was estimated by a standard enzymatic method in 64 episodes in adults admitted to a municipal hospital. Of the 85 specimens analyzed, 60 were taken before treatment from a nearly consecutive group of diabetic patients with ketosis, 21 were follow-up specimens, and 4 were from nondiabetic patients with ketosis. In the 85 specimens, the correlation between serum carbon dioxide and beta-OHB levels was -0.69, and that between anion gap and beta-OHB level was 0.75. For just the initial specimens, the respective correlation coefficients were -0.60 and 0.52. The correlations between serum beta-OHB and carbon dioxide levels and the anion gap were close, but not sufficiently so for the beta-OHB measurements to be routinely useful alone to assess the severity of the ketoacidosis. Full laboratory assessment of the severity and characteristics of ketoacidosis also requires knowledge of serum carbon dioxide level, anion gap, often blood pH, and ideally serum acetoacetate and lactate concentrations as well as serum beta-OHB concentration.


Gamma-aminobutyric acid amides of nortriptyline and fluoxetine display improved pain suppressing activity.
The GABA amides of the antidepressants nortriptyline and fluoxetine, 1 and 2, were compared to their respective parent compounds in rodent models of pain. The amides significantly reduced early nociceptive and late inflammatory responses compared to nortriptyline or fluoxetine, where 1 exhibited overall better efficacy than 2. Amide 1 was most efficacious in lowering cytokine secretion, edema and hyperalgesia induced by formalin and lambda-carrageenan, respectively. Thus, 1 is a promising candidate for the treatment of pain.


Dystroglycan is a dual receptor for agrin and laminin-2 in Schwann cell membrane.
We have shown previously that alpha-dystroglycan with a molecular mass of 120 kDa is a Schwann cell receptor of laminin-2, the endoneurial isoform of laminin comprised of the alpha2, beta1, and gamma1 chains. In this paper, we show that Schwann cell alpha-dystroglycan is also a receptor of agrin, an acetylcholine receptor-aggregating molecule having partial homology to laminin alpha chains in the C terminus. Immunochemical analysis demonstrates that the peripheral nerve isoform of agrin is a 400-kDa component of the endoneurial basal lamina and is co-localized with alpha-dystroglycan surrounding the outermost layer of myelin sheath of peripheral nerve fibers. Blot overlay analysis demonstrates that both endogenous peripheral nerve agrin and laminin-2 bind to Schwann cell alpha-dystroglycan. Recombinant C-terminal fragment of the peripheral nerve isoform of agrin also binds to Schwann cell alpha-dystroglycan, confirming that the binding site for Schwann cell alpha-dystroglycan resides in the C terminus of agrin molecule. Furthermore, the binding of recombinant agrin C-terminal fragment to Schwann cell alpha-dystroglycan competes with that of laminin-2. All together, these results indicate that alpha-dystroglycan is a dual receptor for agrin and laminin-2 in the Schwann cell membrane.


[Pollution Characteristics of Water-soluble Ions in PM2.5 During the Lantern Festival of 2021 in Zibo City].
Based on the online ion data, we have analyzed the cause of a PM2.5 pollution episode, which happened during the Lantern Festival in Zibo in 2021. The pollution characteristics of water-soluble ion components were analyzed, the formation mechanism of secondary inorganic ions (SNA) was discussed, and the changes in the liquid water content (LWC) and pH value of particulate matter before and after pollution were comparatively analyzed. The results showed that the pollution period before the Lantern Festival (T1) and the pollution period at night (T2) ρ(WSIIs) during the Lantern Festival were 46.83 μg·m-3 and 71.18 μg·m-3, respectively, which were 2.3 times and 3.6 times that of the cleaning period, respectively. Among them, the growth multiple of SNA during the T1 period was greater[ρ(NO3-) was 2.9 times, ρ(SO42-) was 2.8 times, and ρ(NH4+) was 2.4 times] than the growth multiple of PM2.5 (2.1 times), which showed that the increase in SNA concentration during the T1 period was the main reason for the increase in PM2.5 concentration. Furthermore, the Cl-, K+, and Mg2+ concentrations, which were 4.0, 14.8, and 16.5 times that of the cleaning period, respectively, increased significantly during the T2 period, indicating that the fireworks and firecrackers caused the rapid increase in the PM2.5 concentration during the T2 period. The LWC during the pollution period was 49.37 μg·m-3, which was 2.9 times that of the cleaning period. LWC was mainly affected by RH and NH4+ during the T1 period and was also affected by Mg2+ during the T2 period. The average pH during the pollution period in Zibo was 4.79±1.54, which was 0.14 lower than that during the cleaning period. The pH during the T1 period was affected by the combined effects of SO42- and NH4+, which made it decrease 0.53 compared to that during the cleaning period. The pH value during the T2 period may be affected by the K+, Cl-, and Mg2+ emitted from the fireworks and firecrackers, causing the pH to increase 0.65 compared to that during the cleaning period. The formation mechanism showed that SO42- was mainly generated by heterogeneous hydrolysis during the pollution episode, whereas NO3- was mainly generated by homogeneous reactions. On the whole, during the pollution episode, the increase in PM2.5 concentration before the Lantern Festival was mainly caused by the increase in SNA concentration, and the increase the night of the Lantern Festival was mainly caused by setting off fireworks and firecrackers.


[Croatian-Glagolithic marginalia medica].
A large number of Croatian Glagolitic manuscripts was investigated with the aim to identify those which are out of the main content - as certain marginalia medica croatico-glagolitica. Such a model was a specific and valuable source for research of medical history and a possible addition to historical factography as well. The analyzed marginalia were recipes, prayers, short chronicales and signa agoniae. Some recipes reflect the transfer of folk experience as well as of official medical knowledge through centuries. The text entitled Zlamenĵa po kih se poznava da nemoĉnik ima brzo umreti (=signa agoniae) is the unique such composition preserved in Croatian Glagolitic writing. It shows the influence of various medical systems and trace of Hippocratism in Glagolitic heritage.


Superatoms: Electronic and Geometric Effects on Reactivity.
The relative role of electronic and geometric effects on the stability of clusters has been a contentious topic for quite some time, with the focus on electronic structure generally gaining the upper hand. In this Account, we hope to demonstrate that both electronic shell filling and geometric shell filling are necessary concepts for an intuitive understanding of the reactivity of metal clusters. This work will focus on the reactivity of aluminum based clusters, although these concepts may be applied to clusters of different metals and ligand protected clusters. First we highlight the importance of electronic shell closure in the stability of metallic clusters. Quantum confinement in small compact metal clusters results in the bunching of quantum states that are reminiscent of the electronic shells in atoms. Clusters with closed electronic shells and large HOMO-LUMO (highest occupied molecular orbital-lowest unoccupied molecular orbital) gaps have enhanced stability and reduced reactivity with O2 due to the need for the cluster to accommodate the spin of molecular oxygen during activation of the molecule. To intuitively understand the reactivity of clusters with protic species such as water and methanol, geometric effects are needed. Clusters with unsymmetrical structures and defects usually result in uneven charge distribution over the surface of the cluster, forming active sites. To reduce reactivity, these sites must be quenched. These concepts can also be applied to ligand protected clusters. Clusters with ligands that are balanced across the cluster are less reactive, while clusters with unbalanced ligands can result in induced active sites. Adatoms on the surface of a cluster that are bound to a ligand result in an activated adatom that reacts readily with protic species, offering a mechanism by which the defects will be etched off returning the cluster to a closed geometric shell. The goal of this Account is to argue that both geometric and electronic shell filling concepts serve as valuable organizational principles that explain a wide variety of phenomena in the reactivity of clusters. These concepts help to explain the fundamental interactions that allow for specific clusters to be described as superatoms. Superatoms are clusters that exhibit a well-defined valence. A superatom cluster's properties may be intuitively understood and predicted based on the energy gained when the cluster obtains its optimal electronic and geometric structure. This concept has been found to be a unifying principle among a wide variety of metal clusters ranging from free aluminum clusters to ligand protected noble metal clusters and even metal-chalcogenide ligand protected clusters. Thus, the importance of electronic and geometric shell closing concepts supports the superatom concept, because the properties of certain clusters with well-defined valence are controlled by the stability that is enhanced when they retain their closed electronic and geometric shells.


Induction of macrophage antiprotozoal activity by monocyte chemotactic and activating factor.
To determine if monocyte chemotactic and activating factor (MCAF) induces intracellular antimicrobial activity, human monocyte-derived macrophages were treated with MCAF and challenged with Toxoplasma gondii and Leishmania donovani. Pretreatment with MCAF induced macrophages to inhibit protozoal replication by approximately 50%. These findings suggest a potential host defense role for MCAF in the inflammatory response to intracellular pathogens.


[Medical prescriptions in haemodialysis patients: critical analysis].
The aim of the study is to analyse haemodialysis patients' prescriptions accordance with legal registrations in 64 chronic haemodialysis patients during a one-month follow-up period. Patients are taking 9.6+/-3 different drugs. A mean of 2.4+/-1.3 prescriptions per patient disagree with the recommended drug use. The directions for use in this specific population are defective in 89%. Moreover, at least one drug-drug interaction is found in 78% of our patients. Only 8% of the studied hemodialysis patients benefit from prescriptions in agreement with the legal recordings, as a consequence of the lack of drug studies in this particular population. Therefore, clinicians have to rely on their own experience to establish their prescriptions, which can involve their responsibility in case of litigation. Furthermore, the frequency of drug-drug interactions in these polymedicated patients implies that a close pharmaceutical follow-up should be implemented.


Androgen receptor interacts with telomeric proteins in prostate cancer cells.
The telomeric complex, shelterin, plays a critical role in protecting chromosome ends from erosion, and disruption of these complexes can lead to chromosomal instability culminating in cell death or malignant transformation. We reported previously that dominant-negative mutants of one of the telomeric proteins called TIN2 cause death of androgen receptor (AR)-negative but not AR-positive prostate cancer cells, raising the question of a possible role of AR in the structural stability of telomeric complexes. Consistent with this possibility, in the present study, we observed that the AR antagonist Casodex (bicalutamide) disrupted telomeric complexes in AR-positive LNCaP cells but not in AR-negative PC-3 cells. Immunofluorescent studies revealed colocalization of TIN2 and AR. Reciprocal immunoprecipitation studies showed association of AR with telomeric proteins. Furthermore, telomeric proteins were overexpressed in prostate cancer cells compared with normal prostate epithelial cells, and sucrose density gradient analysis showed co-sedimentation of AR with telomeric proteins in a shelterin-like mega complex. Together, these observations suggest an allosteric role of AR in telomere complex stability in prostate cancer cells and suggest that AR-antagonist Casodex-mediated cell death may be due to telomere complex disruption.


Restorative health: lessening the impact of previous abuse and violence in the lives of vulnerable girls.
"Restorative health" is the idea that those who have been denied access to the social determinants of health, particularly as young children, should have the right to restoration of healthy functioning. In interviewing a group of vulnerable young women, ranging in age from 15 to 22, we discovered how they experience the link between health and justice in their lives. Participants were living on the street, in extreme poverty, or both. Traumatic early childhood events continued to affect their ability to function healthy. We conclude and suggest that certain rights-based principles need to ground the development of interventions with this group.


Child and adolescent psychiatry: toward the twenty-first century.
The history of child and adolescent psychiatry is reviewed, from its beginning in the Hull House movement in Chicago a century ago through subsequent social and scientific transformations. The field's relationships to academic medicine, to progress in the basic sciences, and to child advocacy are described. Current challenges to the field, including the gaps in and barriers to care, the emergence of for-profit enterprises, and the lack of consensus regarding health care regulation, are discussed in terms of the evolving social mission of the profession. Priorities for the next century are identified.


Probing ternary complex equilibria of crown ether ligands by time-resolved fluorescence spectroscopy.
Ternary complex formation with solvent molecules and other adventitious ligands may compromise the performance of metal-ion-selective fluorescent probes. As Ca(II) can accommodate more than 6 donors in the first coordination sphere, commonly used crown ether ligands are prone to ternary complex formation with this cation. The steric strain imposed by auxiliary ligands, however, may result in an ensemble of rapidly equilibrating coordination species with varying degrees of interaction between the cation and the specific donor atoms mediating the fluorescence response, thus diminishing the change in fluorescence properties upon Ca(II) binding. To explore the influence of ligand architecture on these equilibria, we tethered two structurally distinct aza-15-crown-5 ligands to pyrazoline fluorophores as reporters. Due to ultrafast photoinduced electron-transfer (PET) quenching of the fluorophore by the ligand moiety, the fluorescence decay profile directly reflects the species composition in the ground state. By adjusting the PET driving force through electronic tuning of the pyrazoline fluorophores, we were able to differentiate between species with only subtle variations in PET donor abilities. Concluding from a global analysis of the corresponding fluorescence decay profiles, the coordination species composition was indeed strongly dependent on the ligand architecture. Altogether, the combination of time-resolved fluorescence spectroscopy with selective tuning of the PET driving force represents an effective analytical tool to study dynamic coordination equilibria and thus to optimize ligand architectures for the design of high-contrast cation-responsive fluorescence switches.


Vitamin D levels and the risk of prostate cancer and prostate cancer mortality.
Vitamin D has a role in bone turnover and potentially bone-metastatic spread of prostate cancer (PCa). The aim of this observational study was to address the association between levels of serum vitamin D, diagnosis of PCa and subsequent mortality in men who underwent a biopsy of the prostate. All men who underwent prostatic biopsy in the Danish PCa Registry (DaPCaR) and who had a serum vitamin D measurement during the period 2004 to 2010 (n = 4,065) were identified. Men were categorized by clinical cut-offs based on seasonally adjusted serum vitamin D levels in <25 (deficient), 25-50 (insufficient), 50-75 (sufficient) and >75 nmol/L (high) serum vitamin D. Logistic regression model for association between vitamin D and risk of PCa diagnosis and multivariate survival analyses were applied. No association between serum vitamin D and risk of PCa was found. Overall survival was lowest for serum vitamin D deficiency and a significantly higher PCa specific mortality (HR: 2.37, 95%CI: 1.45-3.90, p < .001) and other cause mortality (HR: 2.08, 95%CI: 1.33-3.24, p = .001) was found for PCa patients with serum vitamin D deficiency compared to serum vitamin D sufficiency. No association was found between serum vitamin D categories and risk of PCa in men who underwent biopsy of the prostate. Men with PCa and serum vitamin D deficiency had a higher overall and PCa specific mortality compared to men with a sufficient level of serum vitamin D.


Surgical intervention for maternal ovarian torsion in pregnancy.
Maternal ovarian torsion in pregnancy is a rare complication. This study was conducted to review the clinical manifestations, and to compare the outcome between laparoscopy and laparotomy in women undergoing surgery for ovarian torsion (OT) during pregnancy. Using the International Classification of Disease, Ninth Revision, and Clinical Modification, we reviewed the clinical records of patients with OT during pregnancy between 1997 and 2008 at a university hospital. Twenty pregnant women were identified with surgically proven OT, 10 in the first trimester, eight in the second, and two in the third. Thirteen (65%) cases were suspected before operation to be adnexal torsion. The most common symptom and sign were pelvic pain (95%) and an adnexal or pelvic mass (95%), followed by nausea and vomiting (65%), elevated white blood cell count >12×10(9)/L (45%), and fever (10%). Most patients in the first trimester (75%) and a minority in the second and third trimesters (37.5%) received management via laparoscopy. Patients undergoing laparoscopy treatment had smaller ovarian masses and a shorter postoperative hospital stay than those receiving laparotomy. None of these patients had significant complications during or after surgery. However, the outcomes of pregnancy varied: 12 (60%) term deliveries, three (15%) preterm deliveries at over 31 gestational weeks, one missed abortion and four elective abortions in the first trimester. The diagnosis of OT during pregnancy is often missed due to nonspecific clinical features and uncommon objective findings. Detorsion only or detorsion plus ancillary procedures via laparoscopy is recommended to treat pregnant women suffering from OT, owing to the advantages of a shorter hospital stay and favorable surgical and pregnancy outcomes.


A comparison of psychosocial and physical functioning in patients with motor neurone disease and metastatic cancer.
Although their disease processes and treatments are different, patients with motor neurone disease (MND) and those with late-stage cancer share a common situation--one in which the quality of life, rather than a cure, becomes the focus of care. We report here a comparison of 126 patients with MND and 125 with metastatic cancer on a range of physical and psychosocial measures. Compared to cancer patients, MND patients were younger, had greater social contacts, but were more physically impaired. Cancer patients had more pain and were on more medication (opioids, steroids, and analgesics). Although the Beck depression scores were similar in both groups, MND patients had significantly higher scores for demoralization, hopelessness, and suicidal ideation. Cancer patients, on the other hand, scored significantly higher on anhedonia. We suggest this difference in the quality of depression represents a difference in illness experience of the two groups and has relevance for the ways we treat depression in the medically ill.


Encoding of prospective tasks in the human prefrontal cortex under varying task loads.
Successful realization of planned actions requires the brain to encode intentions over delays. Previous research has indicated that several regions in the rostral or anterior prefrontal cortex (PFC) encode delayed intentions. However, different processes may encode the same future task depending on task load during the delay. This difference may depend on the computational resources available when the delay is occupied with an ongoing task and when it is task-free. Here we directly investigated and compared the representation of delayed intentions in the human brain in the presence and absence of ongoing task load during the delay. We acquired fMRI data in combination with an event-based prospective memory design where human subjects remembered to perform the same future tasks over occupied and task-free delays. We used time-resolved multivoxel pattern classification and found that: (1) rostrolateral PFC (BA 46) encoded the delayed intention during both delay types; (2) rostromedial PFC (BA 10) encoded the intentions during occupied delays; whereas (3) a variety of more posterior regions, including the anterior cingulate cortex (BA 24), the supplementary motor area (BA 6), and the precuneus, encoded intentions during task-free delays. Overall, the medial PFC encoded delayed intentions more rostrally in the presence of an ongoing delay task and more caudally in its absence. Thus, rostromedial PFC may play a specialized role in the encoding of prospective memory that depends on higher computational demands (e.g., given higher task load during the delay). In contrast, the rostrolateral PFC is a more general area that encodes future intentions regardless of task load.


Targeting angiogenesis-dependent calcified neoplasms using combined polymer therapeutics.
There is an immense clinical need for novel therapeutics for the treatment of angiogenesis-dependent calcified neoplasms such as osteosarcomas and bone metastases. We developed a new therapeutic strategy to target bone metastases and calcified neoplasms using combined polymer-bound angiogenesis inhibitors. Using an advanced "living polymerization" technique, the reversible addition-fragmentation chain transfer (RAFT), we conjugated the aminobisphosphonate alendronate (ALN), and the potent anti-angiogenic agent TNP-470 with N-(2-hydroxypropyl)methacrylamide (HPMA) copolymer through a Glycine-Glycine-Proline-Norleucine linker, cleaved by cathepsin K, a cysteine protease overexpressed at resorption sites in bone tissues. In this approach, dual targeting is achieved. Passive accumulation is possible due to the increase in molecular weight following polymer conjugation of the drugs, thus extravasating from the tumor leaky vessels and not from normal healthy vessels. Active targeting to the calcified tissues is achieved by ALN's affinity to bone mineral. The anti-angiogenic and antitumor potency of HPMA copolymer-ALN-TNP-470 conjugate was evaluated both in vitro and in vivo. We show that free and conjugated ALN-TNP-470 have synergistic anti-angiogenic and antitumor activity by inhibiting proliferation, migration and capillary-like tube formation of endothelial and human osteosarcoma cells in vitro. Evaluation of anti-angiogenic, antitumor activity and body distribution of HPMA copolymer-ALN-TNP-470 conjugate was performed on severe combined immunodeficiency (SCID) male mice inoculated with mCherry-labeled MG-63-Ras human osteosarcoma and by modified Miles permeability assay. Our targeted bi-specific conjugate reduced VEGF-induced vascular hyperpermeability by 92% and remarkably inhibited osteosarcoma growth in mice by 96%. This is the first report to describe a new concept of a narrowly-dispersed combined polymer therapeutic designed to target both tumor and endothelial compartments of bone metastases and calcified neoplasms at a single administration. This new approach of co-delivery of two synergistic drugs may have clinical utility as a potential therapy for angiogenesis-dependent cancers such as osteosarcoma and bone metastases.


Adverse experiences in early childhood and their longitudinal impact on later behavioral problems of children living in poverty.
Adverse childhood experiences (ACEs) are an identified risk factor for the social and emotional development of children. What is less known is the long-term effects of ACEs when poverty and ACEs coincide. Using longitudinal cohort-panel data, we examined whether exposure to ACEs by the age of three among poor children would longitudinally result in behavioral problems at ages three, five, nine, and 15, after controlling for mothers' socioeconomic status and their children's characteristics. We used a subsample of 2750 children and their parents living in urban poverty from the Fragile Families and Child Wellbeing study. Logistic regression modeling was used to obtain adjusted odds ratios of ACE categories predicting behavioral problems after accounting for family socioeconomic position. Our findings indicate that experiencing ACEs in early childhood was significantly associated with later behavioral outcomes from childhood to adolescence. Exposure to multiple ACEs before the age of three was significantly associated with the top-risk behavior group at age five; the odd ratios were 2.0 (CI = 1.3-3.1) and 2.9 (CI = 1.8-4.6) for two ACEs and three or more ACEs, respectively. At both ages nine and 15, children experiencing two or more ACEs had 1.9 to 3.2 times higher odds to demonstrate more the top 10th percentile of behavioral problems. Among covariates, mothers' race and education, and children's gender and temperament were identified as significant factors to determine behavior problems. The findings support policies and programs for families with children who have experienced economic disadvantages and early childhood adversity.


Domain organization of the yeast histone chaperone FACT: the conserved N-terminal domain of FACT subunit Spt16 mediates recovery from replication stress.
The abundant nuclear complex termed FACT affects several DNA transactions in a chromatin context, including transcription, replication, and repair. Earlier studies of yeast FACT, which indicated the apparent dispensability of conserved sequences at the N terminus of the FACT subunit Cdc68/Spt16, prompted genetic and biochemical studies reported here that suggest the domain organization for Spt16 and the other FACT subunit Pob3, the yeast homolog of the metazoan SSRP1 protein. Our findings suggest that each FACT subunit is a multidomain protein, and that FACT integrity depends on Pob3 interactions with the Spt16 Mid domain. The conserved Spt16 N-terminal domain (NTD) is shown to be without essential function during normal growth, but becomes important under conditions of replication stress. Genetic interactions suggest that some functions carried out by the Spt16 NTD may be partially redundant within FACT.


Human cytomegalovirus structural proteins: immune reaction against pp150 synthetic peptides.
In the present study, several peptides of the major structural antigen (pp150) of human cytomegalovirus (CMV) have been chemically synthesized and tested by a modified slot blotting procedure for their ability to bind CMV-specific immunoglobulin G (IgG) and IgM present in human sera. The sequences of the peptides were deduced on the basis of either (i) their presence in a fusion protein already known to be frequently recognized by human antibody or (ii) their high content of hydrophilic amino acids as deduced from the published nucleotide sequence. An important IgM-binding epitope was found to be located in the last 38 amino acids at the carboxy terminus of the molecule. This region reacts with anti-CMV IgM present in the great majority (83.3%) of IgM-positive human sera, and adsorption experiments have shown that IgM titers to the entire pp150 decrease 25 to 50% in most sera previously absorbed with this region. The overall results obtained endorse the continued synthesis of other sequences in order to define a group of peptides sensitive and specific enough to replace the virus and infected cells as an antigenic substrate in the serological evaluation of anti-CMV antibody.


[Tissue factor pathway inhibitors].
Activation of the extrinsic pathway of blood coagulation plays the key role in the process of blood coagulation. The hemostasis is influenced by the activity of procoagulant factors and its inhibitors. In this work we summarize existing data on tissue factor pathway inhibitors: TFPI and TFPI-2. Despite structural similarity between them, they exhibit many differences in synthesis, distribution and mechanism of action. TFPI inhibits the activity of factor Xa and the complex of tissue factor and factor VIIa (TF/VIIa). Conversely, TFPI-2 does not inhibits factor Xa, but it is a strong inhibitor of factor XIa, plasma kallikrein, plasmin and trypsin.


Two adjacent AP-1-like binding sites form the electrophile-responsive element of the murine glutathione S-transferase Ya subunit gene.
An electrophile-responsive element (EpRE) in the 5' flanking region of the mouse glutathione S-transferase Ya subunit gene was recently found to be responsible for the induction of gene expression by xenobiotics that contain or acquire by metabolism an electrophilic center. We now find that this EpRE is composed of two adjacent 9-base-pair motifs related in sequence to the AP-1 binding site, a transcriptional enhancer originally identified as the phorbol 12-myristate 13-acetate (PMA) response element and known to be regulated by the binding of protein products of c-jun and c-fos genes. Synthetic oligonucleotides representing each of the AP-1-like binding sites of the EpRE and the AP-1 site consensus sequence were prepared and assayed for their enhancer activity and inducibility by tert-butylhydroquinone, beta-naphthoflavone, and PMA. Single AP-1-like sequences showed a lower enhancer activity than an AP-1 consensus sequence and no inducibility. Two adjacent AP-1-like sites were found to act synergistically and to confer inducibility beyond that observed for a single AP-1 consensus sequence. Examination of the PMA-responsive region of a number of genes shows the presence of adjacent AP-1-like sites and indicates that the structure of the EpRE found in the Ya gene may occur more generally and may be important in regulating the magnitude of the electrophilic response. The present study demonstrates the binding and transactivation of the EpRE by Jun and Fos and indicates that the AP-1 site is part of the EpRE. The induction by PMA or tert-butylhydroquinone appears to be independent of protein kinase C activity since it is not affected by inhibitors of this enzyme.


Coinfection of Cotton Plants with Watermelon Mosaic Virus and a Novel Polerovirus in China.
Cotton is the most important fiber crop worldwide. To determine the presence of viruses in cotton plants showing leaf roll and vein yellowing symptoms in Henan Province of China, a small RNA-based deep sequencing approach was performed. Analysis of the de novo-assembled contigs followed by reverse transcription PCR allowed the reconstruction of watermelon mosaic virus and an unknown virus. The genome of the unknown virus was determined to be 5870 nucleotides in length, and has a genomic organization with characteristic features of previously reported poleroviruses. Sequence analysis revealed that the virus was closely related to, but significantly different from, cotton leafroll dwarf virus, a polerovirus of the family Solemoviridae. This virus had less than 90% amino acid sequence identity in the products of both ORF0 and ORF1. According to the polerovirus species demarcation criteria set by the International Committee on Taxonomy of Viruses, this virus should be assigned to a new polerovirus species, for which we propose the name "cotton leaf roll virus".


Cholinesterases from plant tissue: v. Cholinesterase is not pectin esterase.
Several properties of the cholinesterase from Phaseolus aureus Roxb. and of pectin (methyl) esterases from both Phaseolus aureus and Lycopersicon esculentum (L.) Mill. are contrasted. Cholinesterase activity is inhibited by all of the concentrations of NaCl tested, from 0.05 m to 0.9 m, a property which differs sharply from published data pertaining to pectin esterase. Although crude preparations of cholinesterase contain pectin esterase activity, further purification by gel filtration of the cholinesterase results in a nearly complete elimination of the pectin esterase activity. The activity of neither the pectin esterase from Lycopersicon esculentum nor that from Phaseolus aureus is affected by 25 mum neostigmine, a potent inhibitor of the cholinesterase activity extracted from Phaseolus aureus.


Binocular non-stereoscopic cues can deceive clinical tests of stereopsis.
Stereoscopic vision plays a critical role in visual perception; however, it is difficult to assess. In clinical settings, stereoacuity is assessed with clinical stereotests. Observers can use monocular cues to deceive some of the most common stereotests, such as the Titmus test. The Randot test has been found free of monocular cues, and here we confirm that result by testing observers under monocular viewing. However, there is a common misconception that only monocular cues can be used to deceive stereotests. Here we demonstrate that binocular non-stereoscopic cues can also be used to pass the Randot, by testing participants with the test rotated, a condition that abolishes stereopsis, and comparing the performance to a monocular viewing condition. We also assessed the Random Dot Butterfly test and discovered considerable amounts of non-stereoscopic cues, including binocular cues in the Circles that can be used to deceive the test. Participants with amblyopia had more difficulty using non-stereoscopic cues than neurotypical observers. We gathered normal-viewing Randot stereoacuities for 110 participants (90 neurotypical and 20 with amblyopia) and compared them to psychophysical stereoacuities (our gold standard). The Randot test showed low positive normalized predictive values for detecting stereoblindness. It could perfectly detect stereo-impairment but with a low sensitivity.


Rapid and low-cost method to prove the nature of no documented tachycardia in children and teenagers without pre-excitation syndrome.
Aims Symptoms in children are often difficult to interpret. The purpose of this study was to report the results of transoesophageal electrophysiological study (EPS) performed in children complaining of sudden onset tachycardia with normal non-invasive studies. Methods and results Eighty-two children and teenagers (mean age 15 +/- 3 years) presented with suspected but no documented paroxysmal supraventricular tachycardia (SVT). ECG was normal. Non-invasive studies were negative; 23 children had syncope with tachycardias. They underwent transoesophageal EPS in our out-patient clinic. The mean duration of transoesophageal EPS was 11 +/- 5 min. Electrophysiological study was negative in 25 children. AV nodal re-entrant tachycardia could be induced in 37 children, 11 of them associated with syncope. Wolff-Parkinson-White syndrome (WPW) was diagnosed in five children in which atrioventricular re-entrant tachycardia was inducible. Atrioventricular re-entrant tachycardia due to a concealed AP was induced in 14 children. Verapamil-sensitive ventricular tachycardia was induced in one patient. Factors associated with tachycardia inducibility were an older age (15.5 +/- 2 vs. 14 +/- 4 years) (P < 0.05) and the absence of syncope (81 vs. 52%) (P < 0.05). During a mean follow-up of 3 +/- 1 year, no patient with negative EPS developed documented tachycardia. In 17 children with inducible SVT, radiofrequency ablation of the re-entrant circuit was subsequently performed. Conclusion Transoesophageal EPS is a fast method for proving the nature of paroxysmal tachycardia in children and teenagers presenting with normal ECG and for demonstrating WPW syndrome not visible on standard ECG. The negative predictive value of transoesophageal EPS for the diagnosis of SVT was 100%.


Physician-assisted suicide: the dangers of legalization.
The authors examine physician-assisted suicide in the light of what is known about suicide and terminal illness, exploring the potential for abuse if legalization occurs. The elderly, those frightened by illness, and the depressed of all ages would be potential victims. The authors discuss the cases that have received public attention as illustrative of these abuses.


Pigment epithelium-derived factor: a multimodal tumor inhibitor.
Pigment epithelium-derived factor (PEDF), a noninhibitory member of the serine protease inhibitor (serpin) family, is a well-known potent endogenous inhibitor of angiogenesis. It has been known for years to be aberrantly expressed in ocular disorders, but in recent years, down-regulation has been shown to be prevalent in a range of cancers as well. This review describes the trimodal anticancer activities of this interesting protein: antiangiogenesis, apoptosis-mediated tumor suppression, and tumor cell differentiation. The key to successful antitumor therapy with this protein is the ability to synthesize the recombinant form of the protein (or its active shortened forms) and deliver at therapeutic doses or alternatively to use gene transfer technology to prolong the effect in vivo. Although there is a substantial amount of work carried out at the preclinical stage with this protein, more groundwork has to be done before PEDF is tested against cancer in clinical trials.


[Twenty years of Spanish psychological research in Psicothema (1989-2008)].
Twenty years of Spanish psychological research in Psicothema (1989-2008). Scientific collaboration and gender equality are two spheres of great interest for the organizations responsible for instigating scientific policies. We have identified the research groups and women's contribution to one of the most outstanding Spanish psychology journals. We have selected papers published during the 1989-2008 period in Psicothema. A bibliometric and network analysis was carried out using the Pajek Software tool. We have analysed 1,718 papers, written by 2,423 authors, 53.45% men and 47.54% women. We have identified 86 research groups made up of 293 researchers. We have observed a high increase of productivity and collaboration in the 1999-2008 period, as well as the consolidation, growth, and evolution of many incipient groups identified in the 1989-1998 period, benefiting from the increased interest and prestige of the journal. Despite the increase in the number of women, rising from 35.71% in 1989 to 48.48 in 2008, we have observed an imbalance with regard to productivity and the order of signatures, where women are relegated to intermediate positions.


[Clinical evaluation of 99mTc-HMPAO labeled leukocyte imaging in ulcerative colitis].
Inflammatory imaging using 99mTc-HMPAO-labeled mixed leukocytes was assessed for use in treating 11 cases diagnosed as ulcerative colitis: 10 cases with total colitis and 1 with left-sided colitis. They consisted of 8 patients with relapse-remitting type and 3 with chronic continuous type. Radionuclide abdominal images were obtained at 1 hr, 4 hr and 24 hr after intravenous injection of 200 MBq prepared 99mTc leukocytes. Obvious colonic activity noted at 4 hr served as the basis for positive comparative criterion in the present study. The diagnostic efficacy of radionuclide imaging was compared with endoscopic findings (based on Matts' classification) and the clinical manifestations as reference. The sensitivity and specificity of this imaging were 83.3% and 85.7%, respectively, these values being consistent with endoscopic findings and clinical manifestations at sites of disease activity. All of positive images changed to negative after treatment by leukocyte apheresis or glucocorticoid. Based on these results, 99mTc leukocyte imaging can be used to accurately evaluate severity and treatment response in ulcerative colitis. Leukocytes may be closely related to the pathogenesis of ulcerative colitis.


Keratoacanthoma Centrifugum Marginatum with Spontaneous Regression and Its Possible Differential Diagnosis.
Keratoacanthoma centrifugum marignatum (KCM) is a rare variant of keratoacanthoma, which is characterized by the dense infiltration of inflammatory cells throughout the dermis, especially around the keratinocytic islands. Therefore, it is sometimes difficult to differentiate between KCM and cutaneous T-cell lymphomas. In this report, we describe a case of KCM with spontaneous regression that showed dense infiltration of CD3+CD8+ T cells. Our present case suggested the importance of investigating tumor-infiltrating lymphocytes to avoid the misdiagnosis of KCM as cutaneous T-cell lymphoma.


Adaptive SPECT imaging with crossed-slit apertures.
Preclinical single-photon emission computed tomography (SPECT) is an essential tool for studying the progression, response to treatment, and physiological changes in small animal models of human disease. The wide range of imaging applications is often limited by the static design of many preclinical SPECT systems. We have developed a prototype imaging system that replaces the standard static pinhole aperture with two sets of movable, keel-edged copper-tungsten blades configured as crossed (skewed) slits. These apertures can be positioned independently between the object and detector, producing a continuum of imaging configurations in which the axial and transaxial magnifications are not constrained to be equal. We incorporated a megapixel silicon double-sided strip detector to permit ultrahigh-resolution imaging. We describe the configuration of the adjustable slit aperture imaging system and discuss its application toward adaptive imaging, and reconstruction techniques using an accurate imaging forward model, a novel geometric calibration technique, and a GPU-based ultra-high-resolution reconstruction code.


Nervous system sports-related injuries.
With the evolution of sports medicine as a subspecialty comes a redirection of attention to the medical and athletic communities to the prompt recognition and timely treatment of many potentially serious conditions. Nervous system complications of sports injuries have gone largely unaddressed in the literature. It is the goal of this communication to review the wide range of nervous system injuries due to sports activities, discuss the mechanisms of such injuries, and focus attention on their prevention and management. The role of the neurologist and neurosurgeon as a sports medicine team member will be discussed, as will their roles in planning and development of programs and equipment at the secondary school, collegiate, and professional levels.


Mate Choice Drives Evolutionary Stability in a Hybrid Complex.
Previous studies have shown that assortative mating acts as a driver of speciation by countering hybridization between two populations of the same species (pre-zygotic isolation) or through mate choice among the hybrids (hybrid speciation). In both speciation types, assortative mating promotes speciation over a transient hybridization stage. We studied mate choice in a hybrid vertebrate complex, the allopolyploid fish Squalius alburnoides. This complex is composed by several genomotypes connected by an intricate reproductive dynamics. We developed a model that predicts the hybrid complex can persist when females exhibit particular mate choice patterns. Our model is able to reproduce the diversity of population dynamic outcomes found in nature, namely the dominance of the triploids and the dominance of the tetraploids, depending on female mate choice patterns and frequency of the parental species. Experimental mate choice trials showed that females exhibit the preferences predicted by the model. Thus, despite the known role of assortative mating in driving speciation, our findings suggest that certain mate choice patterns can instead hinder speciation and support the persistence of hybrids over time without speciation or extinction.


Standardization versus customization of glucose reporting.
Bergenstal et al. (Diabetes Technol Ther 2013;15:198-211) described an important approach toward standardization of reporting and analysis of continuous glucose monitoring and self-monitoring of blood glucose (SMBG) data. The ambulatory glucose profile (AGP), a composite display of glucose by time of day that superimposes data from multiple days, is perhaps the most informative and useful of the many graphical approaches to display glucose data. However, the AGP has limitations; some variations are desirable and useful. Synchronization with respect to meals, traditionally used in glucose profiles for SMBG data, can improve characterization of postprandial glucose excursions. Several other types of graphical display are available, and recently developed ones can augment the information provided by the AGP. There is a need to standardize the parameters describing glycemic variability and cross-validate the available computer programs that calculate glycemic variability. Clinical decision support software can identify and prioritize clinical problems, make recommendations for modifications of therapy, and explain its justification for those recommendations. The goal of standardization is challenging in view of the diversity of clinical situations and of computing and display platforms and software. Standardization is desirable but must be done in a manner that permits flexibility and fosters innovation.


Hybridocytochemical detection of mRNA for calcitonin, CGRP, NPY and somatostatin in thyroid parafollicular (C) cells in three rodent species.
The present study was aimed at hybridocytochemical (HCC) detection and interspecies comparison of mRNA for calcitonin (CT), calcitonin gene-related peptide (CGRP), neuropeptide Y (NPY) and somatostatin (SS) in thyroid C cells of two rodent families of wild Microtidae: pine voles and common voles and also of laboratory Muridae, Wistar rats. Studies were performed on adult males. The HCC method in situ and immunomax technique were used to detect mRNA. DNA oligonucleotide probes labeled with digoxigenin were used in the HCC method. The obtained results were compared to the results of immunocytochemical (ICC) examinations, where rabbit or mouse antibodies against human CT, SS, NPY and rat CGRP, as well as chromogranin A were performed. In the present study, HCC reaction has demonstrated the presence of mRNA for CT and CGRP in all thyroid C cells in all the species examined. However, mRNA for NPY and SS was observed in very few C cells in rat and in many more C cells in the two species of wild rodents. The distribution of the positive cells corresponded with that of ICC detected cells.


The FTO rs9939609 Variant Is Associated with Cardiometabolic Disease Risk and Dietary Energy Intakes in Children with Mental Health Disorders.
Second-generation antipsychotics (SGAs) are used to treat children for mental health disorders but in some children they cause cardiometabolic complications including weight gain and type 2 diabetes. Genetic variants can place a child at risk of developing these metabolic complications. The fat mass and obesity-associated (FTO) rs9939609 A allele has been associated with obesity and dietary energy intakes in healthy children but its relation to metabolic complications in SGA-treated children is not known. This study investigated the association of the FTO rs9939609 variant and SGA treatment with cardiometabolic complications and dietary intakes in children with mental health disorders. A cross-sectional population of children (≤18 y; n = 506) with mental health disorders that were SGA-treated (n = 197) and SGA-naïve (n = 309) were recruited through the Department of Psychiatry at BC Children's Hospital. Dietary intakes were estimated using 3-d food records in a subset of children (n = 73). Genotype frequencies were not different between SGA-treated (TT genotype 42.6%, TA genotype 38.6%, AA genotype 18.8%) and SGA-naïve (TT 41.1%, TA 39.5%, AA 19.4%) children. Children with the A allele had lower BMI z-sores compared with the TT genotype (0.84 ± 1.19 compared with 1.19 ± 1.36; P = 0.005, adjusted for ethnicity). We observed an interaction between FTO genotype and SGA status on fasting glucose (P = 0.036). SGA-naïve children with the A allele had higher fasting glucose than those with the TT genotype (4.96 ± 0.35 compared with 4.81 ± 0.35 mmol/L; P = 0.001), in adjusted models (age, sex, ethnicity, and BMI z-score). This was not observed in SGA-treated children. Children with the A allele had higher daily total energy intakes compared with the TT genotype (1994 ± 619 compared with 1814 ± 484 kcal/d; P = 0.048), in adjusted models (age, sex, ethnicity, and BMI z-score); no effect of SGA-treatment was observed. Our findings suggest the A allele of the FTO rs9939609 variant is associated with higher BMI in children with mental health disorders, but only in those not treated with SGAs.


Efficient paclitaxel production using protoplasts isolated from cultured cells of Taxus cuspidata.
Efficient isolation of protoplasts from Taxus cuspidata cultured cells, localization of paclitaxel in the cultured cells, and efficient production of paclitaxel by protoplasts were studied. Hemicellulase, potassium citric acid solution, and degassing treatments were effective in increasing the yield of protoplasts isolated from T. cuspidata cultured cells. Protoplasts yields (3.2 - 6.4 x 10(6) number/g-fresh weight cells) were achieved by combining the various treatments with specific culture and cell phases. It was found that about 30% and 35% of paclitaxel in the cells was located in cell walls and/or between the cell wall and cell membrane (CW) of suspension cells in the growth phase and in the stationary phase, respectively. About 30% and 43% of paclitaxel in the cells was located in CW of the cells grown in solid culture in growth phase and in the stationary phase, respectively. In comparison with cell suspension culture, protoplasts in a static culture and the protoplasts immobilized in agarose gel in shaking culture resulted to about 6 times increase in the extracellular paclitaxel accumulation.


Effect of Vacancy Defects on the Vibration Frequency of Graphene Nanoribbons.
Graphene is a type of two-dimensional material with special properties and complex mechanical behavior. In the process of growth or processing, graphene inevitably has various defects, which greatly influence the mechanical properties of graphene. In this paper, the mechanical properties of ideal monolayer graphene nanoribbons and monolayer graphene nanoribbons with vacancy defects were simulated using the molecular dynamics method. The effect of different defect concentrations and defect positions on the vibration frequency of nanoribbons was investigated, respectively. The results show that the vacancy defect decreases the vibration frequency of the graphene nanoribbon. The vacancy concentration and vacancy position have a certain effect on the vibration frequency of graphene nanoribbons. The vibration frequency not only decreases significantly with the increase of nanoribbon length but also with the increase of vacancy concentration. As the vacancy concentration is constant, the vacancy position has a certain effect on the vibration frequency of graphene nanoribbons. For nanoribbons with similar dispersed vacancy, the trend of vibration frequency variation is similar.


Fake news and fake research: Why meta-research matters more than ever.
Research is in a crisis of credibility, and this is to the peril of all paediatricians. Billions of dollars are being wasted each year because research is not planned, badly conducted or poorly reported, and this is on a background of rapidly reducing research budgets. How can paediatricians, families and patients make informed treatment choices if the evidence base is absent or not trustworthy? This article discusses why meta-research now matters more than ever, how it can help solve this crisis of credibility and how this should lead to more efficient and effective clinical care. The field of meta-research or research-on-research is the ultimate big picture approach to identifying and solving issues of bias, error, misconduct and waste in research. Meta-researchers value authenticity over aesthetics and quality over quantity. The utility of meta-research does not rely on accusations or critical assessments of individual research, but through highlighting where and how the scientific method and research standards across all fields can be improved. Meta-researchers study, analyse and critique the research pathway, focusing on elements such as methods (how to conduct), evaluation (how to test), reporting (how to communicate), reproducibility (how to verify) and incentives (how to reward). In the current climate it is now more critical than ever that we make use of meta-research and prioritise high-quality high-impact research, ultimately leading to improved patient outcomes.


Physiological dose of estrogen regulates extrathymic T cells in female mice.
Bilateral ovariectomy was performed in female C57BL/6 mice to investigate whether extrathymic T cells in the liver are regulated by physiological doses of estrogen or progesterone. Extrathymic T cells were identified using a two-color immunofluorescence test for CD3 and interleukin-2 receptor beta-chain (IL-2R beta) antigens. The T cells were identified as intermediate CD3+ cells with a high expression of IL-2R beta. It was further demonstrated that the proportion and number of intermediate CD3+ cells decreased in response to ovariectomy, an effect that was countered by estrogen administration. These results suggest that the activation of extrathymic T cells may be regulated by physiological levels of estrogen.


Determination of zinc in erythrocytes, granulocytes and serum by flame atomic absorption spectrophotometry.
Zinc was determined in granulocytes, erythrocytes and serum by flame atomic absorption spectrophotometry. Using a Teflon sampling cup, the sample volume for a single granulocyte zinc determination was reduced to 100 microliter. Zinc in serum and erythrocytes was measured by aspirating the sample at a rate of 5 ml/min. The total volume of blood used was 23-28 ml. The intra-assay coefficient of variation of the method for granulocytes was 0.01 and of the method for serum and erythrocytes 0.018. The corresponding inter-assay coefficient of variation was 0.037 and 0.03, respectively. Recovery of zinc determined by the method of aspirating the small volume (100 microliter) was 103% and by the method of aspirating 5 ml/min 97%. The coefficient of correlation between the two methods was r = 0.9968.


Calreticulin mutant mice develop essential thrombocythemia that is ameliorated by the JAK inhibitor ruxolitinib.
Mutations of calreticulin (CALR) are detected in 25-30% of patients with essential thrombocythemia (ET) or primary myelofibrosis and cause frameshifts that result in proteins with a novel C-terminal. We demonstrate that CALR mutations activated signal transducer and activator of transcription 5 (STAT5) in 293T cells in the presence of thrombopoietin receptor (MPL). Human megakaryocytic CMK11-5 cells and erythroleukemic F-36P-MPL cells with knocked-in CALR mutations showed increased growth and acquisition of cytokine-independent growth, respectively, accompanied by STAT5 phosphorylation. Transgenic mice expressing a human CALR mutation with a 52 bp deletion (CALRdel52-transgenic mice (TG)) developed ET, with an increase in platelet count, but not hemoglobin level or white blood cell count, in association with an increase in bone marrow (BM) mature megakaryocytes. CALRdel52 BM cells did not drive away wild-type (WT) BM cells in in vivo competitive serial transplantation assays, suggesting that the self-renewal capacity of CALRdel52 hematopoietic stem cells (HSCs) was comparable to that of WT HSCs. Therapy with the Janus kinase (JAK) inhibitor ruxolitinib ameliorated the thrombocytosis in TG mice and attenuated the increase in number of BM megakaryocytes and HSCs. Taken together, our study provides a model showing that the C-terminal of mutant CALR activated JAK-STAT signaling specifically downstream of MPL and may have a central role in CALR-induced myeloproliferative neoplasms.


Educational progress, behaviour, and motor skills at 10 years in early treated congenital hypothyroidism.
To assess educational attainments, behaviour, and motor skills at 10 years of age in a group of children with congenital hypothyroidism identified by neonatal screening. 59 children with congenital hypothyroidism born in 1978-81, 31 cases with pretreatment thyroxine (T4) values of 40 nmol/l or below (group I) and 28 less severe cases with T4 values over 40 nmol/l (group II), together with 59 classroom control children matched for age, sex, social class, and main language spoken at home. The Neale analysis of reading ability; the child health and education study written test of mathematics; Rutter behaviour questionnaires for parents and teachers; the Oseretsky test of motor proficiency (short form). On all measures the congenital hypothyroidism children in group I had less satisfactory scores for educational attainments, behaviour, and motor skills than those in group II and controls. For reading the differences were small and did not reach statistical significance, but the deficits in mathematics and total motor skills were statistically significant (p < 0.01). There were more striking and statistically significant differences in behaviour scores, particularly with respect to attentional difficulties. Although less striking, these were also apparent in the group II children with mild hypothyroidism. At the age of 10 years severe congenital hypothyroidism is associated with some mild impairment in educational and motor attainments. Behaviour problems are also common, even in some children with less severe congenital hypothyroidism.


Questioning the specificity and sensitivity of ELISA for bullous pemphigoid diagnosis.
The reported sensitivity and specificity of enzyme-linked immunosorbent assay (ELISA) for bullous pemphigoid (BP) diagnosis is approximately 87% and 98%, respectively. These statistics suggest that ELISA is a reliable diagnostic test; therefore, the use of ELISA for BP diagnosis has increased. We report the case of a man who was diagnosed with BP and was treated for 3 years based on a positive ELISA for IgG against BP180. After reevaluation, his revised diagnosis was not consistent with BP based on clinical presentation, histopathology, and direct immunofluorescence (DIF). Reviewing reports of ELISA for BP diagnosis in the literature revealed several issues including dissimilar diagnostic procedures and patient populations, multiple reports of positive ELISA in patients without BP, and lack of explanation for these false-positives. This case report and review of the literature is a cautionary tale regarding the use of ELISA as an independently reliable test for BP diagnosis.


Seasonal prevalence of Anopheles stephensi larvae and existence of two forms of the species in an urban garden in Calcutta City.
A study on the occurrence and seasonal prevalence of Anopheles stephensi (principal vector of malaria in Calcutta) larvae in an urban garden in central Calcutta revealed their presence in collections of fresh and/or rain water in various kinds of temporary and permanent breeding habitats. Breeding sites in shady areas were preferred in comparison to those receiving direct sunlight. The density of An. stephensi larvae was highest in earthen and lowest in wooden pots throughout the study period, as supported by ANOVA and Wilcoxon's signed rank test. During monsoons the larval density was uniformly high in all four, i.e. earthen, stone, cement and wooden kinds of pots. The variation in the density of An. stephensi larvae due to the effect of season and nature of pots was highly significant. Co-existence of An. stephensi larvae with Aedes aegypti, Ae. albopictus and Cx. quinquefasciatus was also observed at times in certain water collections. An. stephensi population in the urban garden in central Calcutta was found to comprise of two forms the type and variety mysorensis.


Movement Evoked Pain and Mechanical Hyperalgesia after Intramuscular Injection of Nerve Growth Factor: A Model of Sustained Elbow Pain.
Lateral epicondylalgia presents as lateral elbow pain provoked by upper limb tasks. An experimental model of elbow pain provoked by movement/muscle contraction and maintained over several days is required to better understand the mechanisms underlying sustained elbow pain. This study investigated the time course and pain location induced by nerve growth factor (NGF) injection into a wrist extensor muscle, and whether movement and muscle contraction/stretch provoked pain. On Day 0 twenty-six painfree volunteers were injected with NGF (N = 13) or isotonic saline (randomized) into the extensor carpi radialis brevis (ECRB) muscle of the dominant arm. On Day 2 pain was induced in all participants by hypertonic saline injection into ECRB. A Likert scale and patient-rated tennis elbow evaluation (PRTEE) was used to assess pain and functional limitation (Days 0-10). Pain intensity during contraction/stretch of ECRB, and pressure pain thresholds (PPTs) were recorded before and after injections on Days 0 and 2, and Days 4 and 10. Compared with isotonic saline, NGF evoked: i) greater Likert pain ratings from 12 hours post-injection until Day 6, ii) greater PRTEE scores on Days 2 and 4, iii) greater pain during ECRB contraction/stretch on Day 2, and iv) lower PPTs on Day 4. This article presents a novel experimental human pain model suitable to study the sustained effects of lateral elbow pain on sensorimotor function and to probe the mechanisms underlying persistent musculoskeletal pain.


Suppression of mitochondrial complex I influences cell metastatic properties.
Despite the fact that mitochondrial dysfunction has an important role in tumorigenesis and metastasis, the underlying mechanism remains to be elucidated. Mitochondrial Complex I (NADH:ubiquinone oxidoreductase) is the first and the largest protein complex of the mitochondrial electron-transport chain (ETC),which has an essential role in maintaining mitochondrial function and integrity. In this study, we separately knocked down two subunits of mitochondrial complex I, GRIM-19 or NDUFS3, and investigated their effects on metastatic behaviors and explored the possible mechanisms. Our data showed that stable down-modulation of GRIM-19 or NDUFS3 decreased complex I activity and reactive oxygen species (ROS) production; led to enhanced cell adhesion, migration, invasion, and spheroid formation; and influenced the expressions of extracellular matrix (ECM) molecules and its related proteins. We also observed that the expressions of GRIM-19, NDUFS3, and ECM elements were correlated with invasive capabilities of breast cancer cell lines. These results suggest that inhibition of complex I affects metastatic properties of cancer cells, and mitochondrial ROS might play a crucial role in these processes by regulating ECM.


Anti-glomerular basement membrane (Goodpasture) disease exacerbated by invasive pulmonary aspergillosis.
Prior reports have associated the unusual presentations of anti-glomerular basement membrane (anti-GBM) disease and pulmonary aspergillosis in an immunocompetent host. We present the case of a previously healthy 24-year-old man who presented with symptoms of hemoptysis, cough, and dyspnea. His environmental risk factors included tobacco use (smoking), hydrocarbon exposure, and an unidentified mold present in his home. Laboratory evaluation revealed positive serum anti-GBM autoantibodies and positive detection of galactomannan in the bronchoalveolar lavage fluid. The diagnosis of anti-GBM disease was confirmed with a kidney biopsy. Management included therapy with voriconazole for aspergillosis and prednisone, plasmapheresis, and eventually cyclophosphamide for anti-GBM disease.


[Misleading presentation of rhombencephalitis due to Listeria monocytogenes].
Rhombencephalitis due to Listeria monocytogenes may present in a variety of forms. We describe the case of a 76 years old male presenting with an acute symmetrical palsy of fifth, seventh, ninth-tenth cranial nerves, ophthalmoparesia and miosis, without evident signs of long tract lesion, mimicking a peripheral nervous system or neuromuscular junction disease. The results of the blood cultures and the magnetic resonance imaging gave the diagnosis of rhombencephalitis due to Listeria. Rhombencephalitis due to Listeria monocytogenes may present as a symmetric palsy of cranial nerves, therefore it can mimic a cranial nerves multineuritis syndrome.


Invasive papillary carcinomas of the extrahepatic bile ducts: a clinicopathologic and immunohistochemical study of 13 cases.
Carcinomas of the extrahepatic bile ducts are uncommon neoplasms that are morphologically heterogeneous and associated with a poor prognosis. We have previously shown that the noninvasive and minimally invasive papillary carcinomas of the extrahepatic bile ducts behave as in situ carcinomas and are associated with a better prognosis. We reviewed the clinical records of 13 patients with invasive papillary carcinomas of the extrahepatic bile ducts and analyzed the microscopic features and selected immunohistochemical reactivity (p53, Mib-1, and Dpc4) that might correlate with patient survival. In addition, we present the updated SEER (Surveillance, Epidemiology, and End Results) data of the National Cancer Institute for the invasive extrahepatic bile duct carcinomas compiled from 1975 to 1998. The 13 patients with papillary carcinoma had a male to female ratio of 1:1, and their ages ranged from 33 to 89 years. Painless jaundice and abdominal pain were the most common complaints. Five tumors were located in the distal portion, one in the mid portion, and six in the proximal portion of the common bile duct. One papillary carcinoma arose in the right hepatic duct. The Whipple procedure was performed in six patients, common bile duct resection in six, and right hepatic lobectomy in one. The cell phenotype of the papillary carcinomas was biliary in nine and intestinal in three. One tumor had both biliary and intestinal phenotypes. Four tumors dedifferentiated (two to undifferentiated small cell carcinomas, one to small [oat] cell carcinoma, and one to giant cell carcinoma). Two papillary carcinomas extended into the pancreas and three into the liver. Only one patient had lymph node metastases at presentation. Follow-up was available in 10 patients. Six patients died of disease from 2 weeks to 2 years and 1 month after surgery. Four patients are alive with no evidence of disease from 4 months to 8 years and 8 months after surgery. Of 174 invasive papillary carcinomas compiled by the SEER program, 71 were confined to the ductal wall, and 61 had regional lymph node metastases. Papillary carcinomas confined to the ductal wall have better 10-year relative survival rates than adenocarcinomas limited to the wall (21% versus 12%). Likewise papillary carcinomas with lymph node metastasis have better prognosis than adenocarcinoma with nodal metastases (10-y survival rate of 12% versus 5%). Currently, the histologic type and the stage of the disease are the most important prognostic factors in these papillary carcinomas. Separation of invasive and noninvasive or minimally invasive papillary carcinoma is critical in estimating the patient outcome. Our findings suggest that there is no correlation between p53, Ki-67, and Dpc4 expression in these tumors and survival of the patients.


New approaches in predicting and diagnosing preeclampsia: Congo Red Dot Paper Test (Review).
Preeclampsia (PE), a complication of pregnancy that is characterized by de novo hypertension and proteinuria, remains a leading cause of morbidity and mortality during pregnancy, influencing 2.5-7% of singleton and 7-21% of twin pregnancies. At present, diagnosis is based on traditional but unreliable and nonspecific clinical markers, and treatment of PE is suboptimal, with minimal effect on maternal and fetal mortality and morbidity. With the hope of developing an affordable and simple procedure for PE prediction for developing countries, a previous study examined the use of Congo red staining of misfolded and damaged proteins in the urine of women with PE. This feature has diagnostic and prognostic potential since it precedes the onset of clinical manifestations and correlates with disease severity. The test is inexpensive, popular within the medical staff, easy to use, and identifies women with PE in only 3 min. Obstetrical providers benefit from the Congo Red Dot Paper Test analysis, since a negative result promotes lesser waiting times in triage, prevents unneeded admissions, and diminishes the health costs associated per case.


High glucose-ROS conditions enhance the progression in cholangiocarcinoma via upregulation of MAN2A2 and CHD8.
Diabetes is a major risk factor in the development and progression of several cancers including cholangiocarcinoma (CCA). However, the molecular mechanism by which hyperglycemia potentiates progression of CCA is not clearly understood. Here, we showed that a high glucose condition significantly increased reactive oxygen species (ROS) production and promoted aggressive phenotypes of CCA cells, including proliferation and migration activities. Mannosidase alpha class 2a member 2 (MAN2A2), was upregulated at both mRNA and protein levels in a high glucose- and ROS-dependent manner. In addition, cell proliferation and migration were significantly reduced by MAN2A2 knockdown. Based on our proteome and in silico analyses, we further found that chromodomain helicase DNA-binding protein 8 (CHD8) was induced by ROS signaling and regulated MAN2A2 expression. Overexpression of CHD8 increased MAN2A2 expression, while CHD8 knockdown dramatically reduced proliferation and migration as well as MAN2A2 expression in CCA cells. Moreover, both MAN2A2 and CHD8 were highly expressed with positive correlation in CCA tumor tissues. Collectively, these data suggested that high glucose conditions promote CCA progression through ROS-mediated upregulation of MAN2A2 and CHD8. Thus, glucose metabolism is a promising therapeutic target to control tumor progression in patients with CCA and diabetes.


Dual matrilineal geographic distribution of Korean type 2 diabetes mellitus-associated -11,377 G adiponectin allele.
The present study was performed to identify the susceptible single nucleotide polymorphisms (SNPs) for the prediction of Korean type 2 diabetes mellitus (T2DM) and to clarify the matrilineal origin of Korean T2DM‑specific SNPs. Fourteen SNPs from the adiponectin (ADIPOQ), hepatocyte nuclear factor 4α, phosphoenolpyruvate carboxykinase 1 and glucokinase genes in the Korean population were analyzed. Only one SNP, ‑11,377 C/G on the ADIPOQ gene, was finally determined to be responsible for the incidence of Korean T2DM (P=0.028). The G‑T‑T‑A haplotype at positions ‑11,377, +45, +276 and +349 on the ADIPOQ gene was also associated with a high incidence of Korean T2DM (P=0.023). In addition, the susceptibility of Korean individuals to T2DM appears to be affected by their matrilineal origin. Of note, the group of Southern origin, consisting of mitochondrial DNA macrohaplogroups F and R, was predisposed to T2DM, whereas the group of Northern origin, consisting of haplogroups A and Y, was resistant to T2DM. This implied that the differential genetics between the two groups, which were formed from the initial peopling of the proto‑Korean population via Southern and Northern routes to the present time, may explain their differing susceptibility to T2DM. In conclusion, from Southern Asia Northward, a matrilineal origin of Korean individuals appears to be responsible for the prevalence of Korean T2DM caused by the ‑11,377 G allele.


Quasi-steady carbon plasma source for neutral beam injector.
Carbon plasma is successfully sustained during 1000 s without any carrier gas in the bucket type ionization chamber with cusp magnetic field. Every several seconds, seed plasmas having ∼3 ms duration time are injected into the ionization chamber by a shunting arch plasma gun. The weakly ionized carbon plasma ejected from the shunting arch is also ionized by 2.45 GHz microwave at the electron cyclotron resonance surface and the plasma can be sustained even in the interval of gun discharges. Control of the gun discharge interval allows to keep high pressure and to sustain the plasma for long duration.


MEROPS: the database of proteolytic enzymes, their substrates and inhibitors.
Peptidases, their substrates and inhibitors are of great relevance to biology, medicine and biotechnology. The MEROPS database (http://merops.sanger.ac.uk) aims to fulfill the need for an integrated source of information about these. The database has hierarchical classifications in which homologous sets of peptidases and protein inhibitors are grouped into protein species, which are grouped into families, which are in turn grouped into clans. Recent developments include the following. A community annotation project has been instigated in which acknowledged experts are invited to contribute summaries for peptidases. Software has been written to provide an Internet-based data entry form. Contributors are acknowledged on the relevant web page. A new display showing the intron/exon structures of eukaryote peptidase genes and the phasing of the junctions has been implemented. It is now possible to filter the list of peptidases from a completely sequenced bacterial genome for a particular strain of the organism. The MEROPS filing pipeline has been altered to circumvent the restrictions imposed on non-interactive blastp searches, and a HMMER search using specially generated alignments to maximize the distribution of organisms returned in the search results has been added.


Liver Abnormalities in Systemic Lupus Erythematosus: A Prospective Observational Study.
Objectives In this study, we aimed to examine and analyze liver abnormalities among patients with systemic lupus erythematosus (SLE), including both newly diagnosed patients and those being followed up, as well as the prevalence of lupus hepatitis. Methods This was a prospective observational study. Clinical data, liver function tests (LFTs), and the findings from the ultrasonography of the abdomen among the patients were prospectively recorded and evaluated. Results Overall, 28 of the total 135 (20.7%) patients had liver abnormalities, including biochemical and those detected via ultrasonography. Ten patients had transaminitis, defined as aspartate aminotransferase (AST) or alanine aminotransferase (ALT) levels >2 times the upper limit of normal (ULN). Nine patients had elevated alkaline phosphatase (ALP) or gamma-glutamyl transferase (GGT) of >2 times ULN. In three patients, transaminitis was due to anti-tubercular therapy (ATT)-induced hepatitis; in seven (5.2%), no specific cause for transaminitis could be identified, and hence they were classified as cases of lupus hepatitis. On comparing clinical features between patients with (n=7) and without lupus hepatitis (n=128), the condition was more prevalent in newly diagnosed SLE patients compared to those who had been on follow-up [six (85.7%) vs. 30 (23.6%), p=0.002]. All seven patients with lupus hepatitis had complete resolution of the transaminitis on follow-ups. However, one patient who had received ATT (isoniazid, rifampicin, ethambutol, and pyrazinamide) died. Ultrasonography showed fatty liver in seven patients and chronic liver disease in one patient. Conclusion In this study, transaminitis due to lupus hepatitis was seen in newly diagnosed lupus patients and was not associated with disease activity. Before diagnosing lupus hepatitis, drug-induced liver disease has to be ruled out, and if persistent LFT abnormalities are present, further workup is suggested to rule out overlap with primary biliary cirrhosis and/or autoimmune hepatitis.


Granular common acute lymphoblastic leukaemia in adults: a morphological study.
2 cases of acute lymphoblastic leukaemia characterised by the presence of cytoplasmic inclusions morphologically similar to azurophil granules are described. Azurophil granulation of blasts is one of the cardinal features which differentiate acute myeloid from acute lymphoblastic leukaemia. Although such granulation of lymphoblasts has caused diagnostic confusion in the past, we found that the granules could be distinguished from myeloid azurophil granules both morphologically and by their characteristic cytochemical staining reactions. They were negative for peroxidase/sudan black and chloroacetate esterase, but gave coarse scattered granular positivity for both acid phosphatase and alpha-naphthyl acetate esterase. Both the electron microscopic appearance of the granules and their cytochemical staining reactions suggested that they were lysosomes. Granular ALL does not appear to be associated with any morphological subtype or karyotype; but is strongly associated with the common ALL phenotype. Its prognostic significance remains uncertain. It occurs more frequently than the small number of previous reports might suggest and, given the potential for misdiagnosis, should be more widely recognised.


Activation of DegP chaperone-protease via formation of large cage-like oligomers upon binding to substrate proteins.
Cells use molecular chaperones and proteases to implement the essential quality control mechanism of proteins. The DegP (HtrA) protein, essential for the survival of Escherichia coli cells at elevated temperatures with homologues found in almost all organisms uniquely has both functions. Here we report a mechanism for DegP to activate both functions via formation of large cage-like 12- and 24-mers after binding to substrate proteins. Cryo-electron microscopic and biochemical studies revealed that both oligomers are consistently assembled by blocks of DegP trimers, via pairwise PDZ1-PDZ2 interactions between neighboring trimers. Such interactions simultaneously eliminate the inhibitory effects of the PDZ2 domain. Additionally, both DegP oligomers were also observed in extracts of E. coli cells, strongly implicating their physiological importance.


An integrated digital PCR system with high universality and low cost for nucleic acid detection.
Digital PCR is the most advanced PCR technology. However, due to the high price of the digital PCR analysis instrument, this powerful nucleic acid detection technology is still difficult to be popularized in the general biochemistry laboratory. Moreover, one of the biggest disadvantages of commercial digital PCR systems is the poor versatility of reagents: each instrument can only be used for a few customized kits. Herein, we built a low-cost digital PCR system. The system only relies on low-cost traditional flat-panel PCR equipment to provide temperature conditions for commercial dPCR chips, and the self-made fluorescence detection system is designed and optically optimized to meet a wide range of reagent requirements. More importantly, our system not only has a low cost (<8000 US dollars) but also has a much higher universality for nucleic acid detection reagents than the traditional commercial digital PCR system. In this study, several samples were tested. The genes used in the experiment were plasmids containing UPE-1a fragment, TP53 reference DNA, hepatitis B virus DNA, leukemia sample, SARS-COV-2 DNA, and SARS-COV-2 RNA. Under the condition that DNA can be amplified normally, the function of the dPCR system can be realized with simpler and low-price equipment. Some DNA cannot be detected by using the commercial dPCR system because of the special formula when it is configured as the reaction solution, but these DNA fluorescence signals can be clearly detected by our system, and the concentration can be calculated. Our system is more applicable than the commercial dPCR system to form a new dPCR system that is smaller and more widely applicable than commercially available machinery.


Fabry disease and G6PD in three family members with priapism: is the nitric oxide pathway to blame?
Fabry disease is an X-linked multisystem disorder due to alpha galactosidase A deficiency leading to glycosphingolipid accumulation with a predilection for the vascular endothelium and affecting the cardiovascular, renal, and neurologic systems. To report a familial cluster of priapism in three males from a family with Fabry disease and glucose-6-phosphate dehydrogenase (G6PD) deficiency and discuss possible mechanisms. Patient charts, Fabry registry, and literature review. Priapism has been reported in 6 males among the 1,558 males of the Fabry registry. Eight additional case reports of priapism in patients with Fabry disease and two reports of patients with G6PD were collected from the literature. Derangement in the nitric oxide (NO) pathway, which can occur in both Fabry disease and G6PD, is suggested as a hypothesis for the priapism in our patients. It is suggested that priapism should be included in the list of clinical symptoms of Fabry patients and that Fabry disease should be added to the differential diagnosis of priapism. Furthermore, the association of G6PD and Fabry disease with priapism emphasizes the need for further study to explore the role of NO metabolism in the etiology of Fabry disease manifestations.


Coronary heart disease in China.
Coronary heart disease (CHD) is the second leading cause of cardiovascular death in the Chinese population. It accounts for 22% of cardiovascular deaths in urban areas and 13% in rural areas. Although mortality from CHD in China is relatively low compared with Western levels, the burden of CHD has been increasing. This is partly because of a worsening profile of risk factors, such as an increased prevalence of hypertension, hyperlipidaemia, overweight/obesity, diabetes, etc and partly because of an increase in the aged population. Large-scale, randomised controlled trials on thrombolytic, blood-pressure-lowering, antiplatelet and blood-cholesterol-lowering treatment as well as cardiac intervention have been conducted for Chinese patients with myocardial infarction. The studies provide important information for the prevention and management of chronic CHD and acute myocardial infarction in the Chinese population.


Sustainable and Continuous Synthesis of Enantiopure l-Amino Acids by Using a Versatile Immobilised Multienzyme System.
The enzymatic synthesis of α-amino acids is a sustainable and efficient alternative to chemical processes, through which achieving enantiopure products is difficult. To more address this synthesis efficiently, a hierarchical architecture that irreversibly co-immobilises an amino acid dehydrogenase with polyethyleneimine on porous agarose beads has been designed and fabricated. The cationic polymer acts as an irreversible anchoring layer for the formate dehydrogenase. In this architecture, the two enzymes and polymer colocalise across the whole microstructure of the porous carrier. This multifunctional heterogeneous biocatalyst was kinetically characterised and applied to the enantioselective synthesis of a variety of canonical and noncanonical α-amino acids in both discontinuous (batch) and continuous modes. The co-immobilised bienzymatic system conserves more than 50 % of its initial effectiveness after five batch cycles and 8 days of continuous operation. Additionally, the environmental impact of this process has been semiquantitatively calculated and compared with the state of the art.


A comparison of traditional vs. Canadian tailored prophylaxis dosing of prophylactic factor infusions in children with haemophilia A and B in a single hemophilia treatment center.
Prophylactic infusion of clotting factor concentrates is a developing standard of care for individuals with haemophilia. The ideal schedule and techniques of prophylactic infusions remain incompletely defined. Our aim was to determine the optimal techniques and schedules for factor prophylaxis in paediatric patients. A retrospective electronic medical record review of all children treated with prophylactic factor infusions in a single Haemophilia Treatment Center was conducted. Comparison of traditional vs. Canadian dosing regimens and primary vs. secondary prophylaxis was made. Failure of prophylaxis was defined as the first serious bleed. A total of 58 children were identified for review. Five cases were excluded (four due to high titre inhibitors and one due to repeated non-compliance), thus there were 53 total cases: 46 with severe haemophilia, 2 with moderate haemophilia, 5 with mild haemophilia, 44 with haemophilia A and 9 with haemophilia B; 32 Traditional dosing and 21 Canadian dosing regimens. Patients on primary prophylaxis had a decreased failure rate (25%) compared to children treated with secondary prophylaxis (67%) regardless of technique of prophylaxis. When compared to a 'Traditional' factor prophylaxis schedule, the 'Canadian' tailored prophylaxis protocol was comparable with the exception of a decreased use of implanted venous devices in the 'Canadian' group. Ongoing bleeding (primarily joint bleeds) occurs with all prophylactic regimens. The lowest incidence of treatment failure was noted in children who began primary prophylaxis at a young age and before initial joint bleeds. Primary prophylaxis is superior to secondary prophylaxis regardless of dosing regimen. Traditional and Canadian dosing regimens were equivalent in outcome when measured over several years of follow-up.


The mechanism of ORFH79 suppression with the artificial restorer fertility gene Mt-GRP162.
The restoration fertility complex (RFC) was previously identified in Honglian (HL)-cytoplasmic male sterility (CMS) rice (Oryza sativa), and glycine-rich protein 162 (GRP162) is responsible for binding to the CMS-associated transcript atp6-orfH79. Here, we engineered a recombinant GRP162 containing the mitochondrial transit peptide, termed Mt-GRP162, as an artificial restorer of fertility (Rf) gene. Mt-GRP162 was confirmed to bind to CMS-associated RNA and to localize to the mitochondria. The transgenic plants showed restored fertility with partially functional pollen. We found that the expression of ORFH79 decreased in transgenic plants, while the expression of atp6-orfH79 was not changed. These findings indicate that Mt-GRP162 restores fertility by suppressing the expression of the cytotoxic protein ORFH79 at the post-transcriptional level rather than via the cleavage of atp6-orfH79 in the presence of RFC. These findings contribute to our understanding of the mechanisms of restoration through diverse pathways.


Approach of self-adaptive illumination for optical imaging systems.
An optical imaging signal is vulnerable to undesired features such as ambient light illumination and partial specular reflection from the target; the success of extracting target features from images depends largely on appropriate design of illumination. This paper presents an approach for self-adaptive illumination for optical imaging systems. The proposed illumination system projects a reference image to a target surface as an initial structured illumination, and then adjusts the projected image automatically to compensate the negative influences of undesired features. After this self-adaptive control process, undesired features would appear mostly invisible in the captured images. The signal-to-noise ratio would be improved dramatically well before subsequent image processing. In the validation experiments, several images with uniform brightness were offered as reference images; the captured images could achieve high brightness uniformity, even when the target surface was uneven or was illuminated by ambient light. In a further experiment of selective vessel illumination on a human palm, simulated vessel regions were selectively illuminated. Undesired features, like palm prints, almost disappeared in the images captured.