Potential cross-species transmission of highly pathogenic avian influenza H5 subtype (HPAI H5) viruses to humans calls for the development of H5-specific and universal influenza vaccines.
In recent years, highly pathogenic avian influenza H5 subtype (HPAI H5) viruses have been prevalent around the world in both avian and mammalian species, causing serious economic losses to farmers. HPAI H5 infections of zoonotic origin also pose a threat to human health. Upon evaluating the global distribution of HPAI H5 viruses from 2019 to 2022, we found that the dominant strain of HPAI H5 rapidly changed from H5N8 to H5N1. A comparison of HA sequences from human- and avian-derived HPAI H5 viruses indicated high homology within the same subtype of viruses. Moreover, amino acid residues 137A, 192I, and 193R in the receptor-binding domain of HA1 were the key mutation sites for human infection in the current HPAI H5 subtype viruses. The recent rapid transmission of H5N1 HPAI in minks may result in the further evolution of the virus in mammals, thereby causing cross-species transmission to humans in the near future. This potential cross-species transmission calls for the development of an H5-specific influenza vaccine, as well as a universal influenza vaccine able to provide protection against a broad range of influenza strains.


Characterization of the protective effects of L-nitroarginine methyl ester (L-NAME) against the toxicity of sulphur mustard in vitro.
The protective effects of L-nitroarginine methyl ester (L-NAME) against the toxicity of sulphur mustard (HD) was characterized in primary cultures of chick embryo forebrain neurons. These effects were not associated with the known nitric oxide synthase-inhibiting characteristics of this compound. No protection was evident in immature (1 day old) cultures. However, L-NAME pre-treatment resulted in a concentration-dependent protection against the toxicity of HD in mature (5 days and older) cultures, with maximal protection reaching greater than 220% at 5 mM L-NAME in terms of increasing the LC50 of control HD-treated cultures. Maximal protective effects were also achieved when L-NAME treatment was delayed up to 3 h post-HD exposure. These effects were reduced by 5 h and absent when the L-NAME was added to the cultures 8 h after HD exposure. Protection against the toxicity of HD was dependent on the continued presence of L-NAME in the medium and was persistent up to 48 h after HD exposure. This compound is one of the most effective drugs yet identified in protecting against the toxicity of HD and is the only one that exerts its effects therapeutically.


Variation in access to sugar-sweetened beverages in vending machines across rural, town and urban high schools.
The 2010 Dietary Guidelines for Americans include reducing consumption of sugar-sweetened beverages. Among the many possible routes of access for youth, school vending machines provide ready availability of sugar-sweetened beverages. The purpose of this study was to determine variation in high school student access to sugar-sweetened beverages through vending machines by geographic location - urban, town or rural - and to offer an approach for analysing school vending machine content. Cross-sectional observational study. Between October 2007 and May 2008, trained coders recorded beverage vending machine content and machine-front advertising in 113 machines across 26 schools in New Hampshire and Vermont, USA. Compared with town schools, urban schools were significantly less likely to offer sugar-sweetened beverages (P = 0.002). Rural schools also offered more sugar-sweetened beverages than urban schools, but this difference was not significant. Advertisements for sugar-sweetened beverages were highly prevalent in town schools. High school students have ready access to sugar-sweetened beverages through their school vending machines. Town schools offer the highest risk of exposure; school vending machines located in towns offer up to twice as much access to sugar-sweetened beverages in both content and advertising compared with urban locations. Variation by geographic region suggests that healthier environments are possible and some schools can lead as inspirational role models.


A magnetic hydrogel for the efficient retrieval of kidney stone fragments during ureteroscopy.
Only 60-75% of conventional kidney stone surgeries achieve complete stone-free status. Up to 30% of patients with residual fragments <2 mm in size experience subsequent stone-related complications. Here we demonstrate a stone retrieval technology in which fragments are rendered magnetizable with a magnetic hydrogel so that they can be easily retrieved with a simple magnetic tool. The magnetic hydrogel facilitates robust in vitro capture of stone fragments of clinically relevant sizes and compositions. The hydrogel components exhibit no cytotoxicity in cell culture and only superficial effects on ex vivo human urothelium and in vivo mouse bladders. Furthermore, the hydrogel demonstrates antimicrobial activity against common uropathogens on par with that of common antibiotics. By enabling the efficient retrieval of kidney stone fragments, our method can lead to improved stone-free rates and patient outcomes.


Effects of prolactin on central dopaminergic neurons.
The activities of various dopaminergic neurons in the brain have been estimated biochemically by quantifying the rate of synthesis or turnover of dopamine in brain regions containing the terminals of these neurons. In addition, the activity of tuberoinfundibular dopaminergic neurons have been estimated from the concentration of dopamine in pituitary stalk blood. Manipulations which increase concentrations of prolactin in cerebrospinal fluid or serum (systemic or intracerebroventricular injections of prolactin; injections of dopaminergic antagonists or estrogens; implantation of pituitaries under the renal capsule; prolactin secreting tumors) increase the activity of tuberoinfundibular dopaminergic neurons. The activities of other dopaminergic neuronal systems are not affected by prolactin.


Predictors of social services supervision of babies of mothers with mental illness after admission to a psychiatric mother and baby unit.
There is insufficient information on the predictors of parenting difficulties in mothers with severe mental illness. Using data from mother and baby units in the UK we aimed to examine the social and clinical characteristics of mothers whose babies were supervised by social services on discharge. A case-control study was carried out using data from mother and baby units and facilities entered onto the Marce database. Of 1197 mothers, 23% were discharged with their babies under some form of social services supervision. Factors independently associated with an increased risk of supervision included social class (OR 3.16, 95% CI 1.99-5.03), single marital status (OR 2.10, 95% CI 1.38-3.20), behavioural disturbance (OR 1.69, 95% CI 1.08-2.65) and psychiatric illness in the partner (OR 2.67, 95% CI 1.59-4.49). The diagnostic groups independently associated with the highest risk of having a supervised baby were schizophrenia (OR 5.16, 95% CI 2.61-10.21) and personality disorder (OR 9.29, 95% CI 3.46-24.91). Mothers with schizophrenia are at particularly high risk of having their baby supervised by social services. Preventative interventions should be targeted at socio-economic difficulties, early detection of psychiatric disorders postpartum and treatment of perinatal mental illness in the context of the whole family.


Perinatal tuberculosis associated hemophagocytic lymphohistiocytosis.
Hemophagocytic lymphohistiocytosis [HLH] is a reactive disorder characterized by generalised non-malignant histiocytic proliferation with prominent hemophagocytosis. It may be either primary [familial (FLH)] or secondary [infection or malignancy associated]. Organisms incriminated for infection associated hemophagocytic syndrome (IAHS) include viruses, bacteria, spirochetes, fungi and parasites. Reports of IAHS associated with tuberculosis in neonates are rare. The authors report a case of perinatal tuberculosis presenting as hemophagocytic lymphohistiocytosis.


Distribution of mesophilic Aeromonas species in raw and ready-to-eat fish and meat products in Switzerland.
A total of 829 poultry, meat, shellfish and fish products commonly consumed in Switzerland were qualitatively and quantitatively examined for the presence of mesophilic Aeromonas spp. Overall, aeromonads occurred in 24.1% of the samples. Raw food products were frequently contaminated (e.g. 94.1% in minced meat), with colony counts up to 6.0 x 10(6)/g. Some ready-to-eat products had a relatively high percentage of positive samples as well, such as cooked ham in slices (38.2%), mortadella (12.9%), smoked cooked sausage (15.6%), hot and cold smoked fish (10.9-14.3%) and gravad salmon (10.5%). Colony counts, however, were somewhat lower (up to 1.7 x 10(3)/g). The high contamination rate of cooked or hot smoked foods suggests recontamination after cooking or smoking, e.g., at the slicing and packaging stage. 61.2% of the identified strains were Aeromonas hydrophila, followed by 22.5% Aeromonas caviae and 16.3% Aeromonas sobria.


[Benign positional vertigo, an armchair for diagnosis and for treatment: description and significance].
The benign positional vertigo is a very frequent pathology. It requires to establish the diagnosis, to fixe the head in some positions to get various nystagmus which are observed directly or by video-nystagmoscopy or analyzed by video-nystagmography. To describe the diagnostic and therapeutical interests of a special armchair, now available, whose characteristics are to be able to swivel around two axes of vertical and horizontal rotations, the patient being completely interdependent of the armchair. It thus makes it possible to place the patient's head in position wished with a high degree of accuracy and facility. After a short description of the characteristics of armchairs classically used, the authors describe in detail this new armchair Its geometrical characteristics are reported and its capacities, like its mode of use: fixing of the patient with armchair then mobilization of the unit "armchair-patient" and setting in position of the head at the point of the space desired by the ENT. The authors report the main advantages of this armchair. It allows a great accuracy of movement given to the semicircular canals, a possible mobilization with an amplitude until there ever reached, a significant reduction of the proprioceptive entries of patient, a perfect safety of the examination, a possible mobilization of obese or arthritic patients and, finally, a very increased comfort for the patient himself. Finally future technological developments possible are brought back. Thanks to the use of the armchair of mechanical assistance, the diagnosis and treatment of benign positional vertigo appear more certain, more precise and more practical at the same time for the medical doctor and the patients. A multicentric study is in process to show its interest in this pathology.


Association of amnioinfusion volume at the time of surgery for twin-twin transfusion syndrome and latency to delivery.
To evaluate the impact of amnioinfusion and other peri-operative factors on pregnancy outcomes in the setting of Twin-twin transfusion syndrome (TTTS) treated via fetoscopic laser photocoagulation (FLP). Retrospective study of TTTS treated via FLP from 2010 to 2019. Pregnancies were grouped by amnioinfusion volume during FLP (<1 L vs. ≥1 L). The primary outcome was latency from surgery to delivery. An amnioinfusion statistic (AIstat) was created for each surgery based on the volume of fluid infused and removed and the preoperative deepest vertical pocket. Regression analysis was planned to assess the association of AIstat with latency. Patients with amnioinfusion of ≥1 L at the time of FLP had decreased latency from surgery to delivery (61 ± 29.4 vs. 73 ± 28.8 days with amnioinfusion <1 L, p < 0.001) and increased preterm prelabor rupture of membranes (PPROM) <34 weeks (44.7% vs. 33.5%, p = 0.042). Amnioinfusion ≥1 L was associated with an increased risk of delivery <32 weeks (aRR 2.6, 95% CI 1.5-4.5), 30 weeks (aRR 2.4, 95% CI 1.5-3.8), and 28 weeks (aRR 1.9, 95% CI 1.1-2.3). Cox-proportional regression revealed that AIstat was inversely associated with latency (HR 1.1, 95% CI 1.1-1.2). Amnioinfusion ≥1 L during FLP was associated with decreased latency after surgery and increased PPROM <34 weeks.


Removal of common bile duct stones by the combination of percutaneous balloon dilatation and extracorporeal shock-wave lithotripsy.
Endoscopic papillotomy may not be possible in patients who have previously undergone gastric surgery. We describe the successful treatment of such a patient with ultrasound-guided, percutaneous transhepatic biliary drainage, followed by balloon dilatation of the sphincter of Oddi and extracorporeal shock-wave lithotripsy. Combinations of new treatment forms may be necessary and effective when single forms of treatment fail.


Determination of Histidine Protonation States in Proteins by Fast Magic Angle Spinning NMR.
Histidine residues play important structural and functional roles in proteins, such as serving as metal-binding ligands, mediating enzyme catalysis, and modulating proton channel activity. Many of these activities are modulated by the ionization state of the imidazole ring. Here we present a fast MAS NMR approach for the determination of protonation and tautomeric states of His at frequencies of 40-62 kHz. The experiments combine 1H detection with selective magnetization inversion techniques and transferred echo double resonance (TEDOR)-based filters, in 2D heteronuclear correlation experiments. We illustrate this approach using microcrystalline assemblies of HIV-1 CACTD-SP1 protein.


Upper lid orbicularis oculi muscle strip and sequential brow suspension with autologous fascia lata is beneficial for selected patients with essential blepharospasm.
Severe cases of blepharospasm resistant to botulinum toxin represent a challenging clinical problem. Over the last 10 years, we have adopted a staged surgical management of these cases with an initial upper lid orbicularis myectomy (combined with myectomy of procerus and corrugator supercilius as appropriate) and then 4-6 months later a brow suspension with autologous fascia lata. The aim of this study was to assess the outcome of this staged surgical approach. A questionnaire was sent to all patients who had undergone the procedure and the clinical records reviewed. Fourteen patients had undergone the procedure of which 13 were alive. They were sent a questionnaire and 10 of them responded. All had both procedures. Eight of the 10 reported great benefit from the surgery, one some benefit, and one was worse off. All patients still required botulinum toxin injections after the surgery. Majority, but not all, of the patients in our series greatly benefitted from this staged surgical approach.


Excess Free Fructose Beverages and Allergy in Children and Adolescents: Results From NHANES 2005-2006.
Our purpose was to investigate the relationship between intake of excess free fructose beverages and allergy among children and adolescents. We analyzed data of 860 children (aged 6 to 12 years) and 1,142 adolescents (aged 13 to 19 years) from the National Health and Nutrition Examination Survey 2005-2006. Logistic regression analyses were performed to determine the associations between consumption of excess free fructose beverages and allergic symptoms or allergic sensitization. The pattern of findings was not entirely consistent, but some analyses supported the hypothesis of an association between intake of excess free fructose beverages and allergy. After controlling for the potential confounders, children who consumed nondiet fruit drinks at least 5 times per week had a nearly 2.5-fold greater odds of allergic sensitization than did those who consumed such drinks only 1 to 3 times per month (OR = 2.446; 95% CI, 1.583-3.780). Adolescents who consumed excess free fructose beverages at least 5 times per week or 1 to 4 times per week had about fivefold greater odds of presence of allergic symptoms than did those who seldom consumed these beverages (OR = 5.164; 95% CI, 1.866-14.297 and OR = 4.112; 95% CI, 1.857-9.107, respectively). Adolescents who consumed apple juice at least 5 times per week had a twofold greater odds of presence of allergic sensitization than did the seldom consumers (OR = 2.215; 95% CI, 1.178-4.164). These findings provide some support for the hypothesis of a link between intake of excess free fructose beverages and allergic symptoms or allergic sensitization in children and adolescents.


Seasonal variation of lupus nephritis in a cohort of Egyptian patients.
Systemic lupus erythematosus is an autoimmune multisystem disease; renal affection is one of its most common manifestations. The effect of environmental factors on lupus nephritis flares is not fully understood. This is a retrospective study that included 200 patients with lupus nephritis flares. All patients had confirmed diagnosis of lupus nephritis on histopathological examination. Lupus nephritis flares were defined by either (1) nephritic flare: defined as increased proteinuria or serum creatinine concentration; abnormal urinary sediment or a reduction in creatinine clearance, or (2) proteinuria flare defined as persistent increase in proteinuria > 0.5-1.0 g/day after achieving complete remission; doubling to > 1 g/day after achieving partial remission. The time of renal flare (month of the year) was recorded to determine the effect of seasonal variation on lupus nephritis flares. The median age for the patients was 33 years (IQR = 13); 92% of patients were females. The median duration of lupus was 7 years (IQR = 6). The median serum creatinine was 1.4 mg/dl, median serum urea level was 32, and median UPCR was 2.4 gm/dl. The highest incidence of flares occurred in June (14%) and July (12.5%) (p = 0.003). Seasonal pattern of LN flare was observed in our study in Egyptian cohort of patients, with most flares observed during meteorological summertime. Larger studies are needed to confirm this seasonal pattern. Key Points • Flares of lupus nephritis are common in patients with systemic lupus erythromatosus. • A seasonal pattern of flares of lupus nephritis was observed in our study. This seasonal pattern has been observed by previous studies in variable ethnicities and variable climatic circumstances.


Evidence for endothelin-1-mediated vasoconstriction in severe chronic heart failure.
Heart failure is commonly associated with high plasma concentrations of endothelin-1, a powerful vasoconstrictor produced by endothelium. The role of endogenously released endothelin-1 in the maintenance of vascular tone in chronic heart failure was assessed by acute administration of an endothelin receptor antagonist, bosentan. 24 patients with chronic heart failure received randomly and double blind two intravenous infusions of either placebo or bosentan (100 mg followed after 60 min by 200 mg). Systemic haemodynamics and plasma endothelin-1 and big-endothelin-1 concentrations were determined before and repeatedly during the 120 min observation period. Baseline endothelin-1 and big-endothelin-1 concentrations, which were above the normal range in all patients, correlated directly with the extent of pulmonary hypertension, with left and right heart filling pressures, and with pulmonary vascular resistance and inversely with cardiac index. Compared with placebo, bosentan reduced mean arterial pressure by 7.7% (95% CI 7.1-9.7), pulmonary artery pressure by 13.7% (10.5-16.9), right atrial pressure by 18.2% (12.0-24.4), and pulmonary artery wedged pressure by 8.6% (5.3-12.0); it increased cardiac index by 13.6% (9.1-18.2), decreased systemic vascular resistance by 16.5% (13.2-19.8), and decreased pulmonary vascular resistance by 33.2% (22.4-44.0). Heart rate did not change. Plasma endothelin-1 concentrations rose more than twofold from baseline in bosentan recipients while big-endothelin-1 concentrations were unchanged. These findings indicate that, in patients with chronic heart failure who have high circulatory endothelin-1 concentrations, this peptide contributes to maintenance of vascular tone. The acute haemodynamic effects of bosentan suggest that chronic endothelin antagonism could be beneficial in such patients.


Seizure disorder in the men of a Veterans Administration nursing home.
This study examined the descriptive epidemiology of seizure disorder in 129 male residents of a Veterans Administration Nursing Home. Eighty-seven of the residents were institutionalized because of nonpsychiatric disorders (60 for chronic neurologic diseases, and 27 for other medical conditions). Forty-two were institutionalized because of a chronic psychosis (39 for schizophrenia, three for affective disorders). We determined for each resident an extensive clinical data base of 54 items including measures of hematologic, nutritional, metabolic and endocrine status, as well as continuing medications. In the nonpsychiatric group, 16 of the 87 men had a seizure disorder. In the psychiatric group, this proportion was only three of 42. The prevalence of epilepsy in the nonpsychiatric group was 20-40 times greater than in the aged-matched general population of men. In the nonpsychiatric group, the onset of seizures followed the onset of organic brain disease. Forty-five percent of seizure disorders occurred in men who had experienced a cerebrovascular accident, and 23% in men with other types of chronic brain disease. The seizures of the nonpsychiatric men had been observed to be generalized clonic-tonic in 45%, and partial complex in 22%. Ninety-four percent of the nonpsychiatric men with epilepsy received anticonvulsants, and none had experienced more than one seizure during the preceding year. Univariate statistical analysis of the 54 item data base showed that the occurrence of seizure disorder correlated inversely with age, blood urea nitrogen, serum creatinine and serum bilirubin, and directly with plasma testosterone, hemoglobin, use of anticonvulsants, and use of psychotherapeutic agents.(ABSTRACT TRUNCATED AT 250 WORDS)


Nerve growth to tooth buds after homotopic or heterotopic autotransplantation.
Feline permanent incisor tooth buds (bell stage) were autotransplanted to mandibular alveolar sockets (homotopic site) or to the submandibular subcutis or the leg (heterotopic sites). This was done in 34 kittens aged 1-2 months. After survival times of 3-8 months the animals were fixed by glutaraldehyde perfusion. A total of 56 mineralized teeth, which had developed at the recipient sites, were removed, demineralized and processed for light microscopic (LM) general evaluation. Fourty-four teeth, which were judged to be grossly normal in the LM, were selected for electron microscopic (EM) analysis with respect to the occurrence of pulpal nerve fibres. The highest proportion of normal teeth (16 of 16) was obtained from the alveolar site, followed by the submandibular (11 of 14) and hindlimb (17 of 26) sites. Most of the grossly normal grafts possessed pulpal axons (37 of 44). The alveolar grafts were all innervated and exhibited a largely normal appearance qualitatively and in terms of percentage of myelinated fibres. The proportion of innervated pulps was lower among the heterotopic mandibular (10 of 11) and hindlimb (11 of 17) grafts. In addition, signs of nerve fibre degeneration appeared more frequently at the heterotopic sites. On the basis of these findings, and in view of the results of other workers, we conclude that tooth germs are attractive targets for all divisions of the trigeminal nerve and for cutaneous nerves outside the trigeminal system. However, the morphological picture tends to become increasingly abnormal with increasing distance from the normal locus.


Lenvatinib in patients with unresectable hepatocellular carcinoma who do not meet the REFLECT trial eligibility criteria.
This study aimed to determine the efficacy and safety of lenvatinib for patients with unresectable hepatocellular carcinoma (HCC) who did not meet REFLECT eligibility criteria (phase 3 clinical trial). In this multicenter retrospective study, patients with unresectable HCC treated with lenvatinib between 2018 and 2019 and had adequate clinical data were included. Objective response rate, progression-free-survival (PFS) and safety were evaluated according to meeting or not meeting the REFLECT eligibility criteria and according to the criteria of the REFLECT trial. Of the 105 patients included, 61% (64 of 105) did not meet the REFLECT eligibility criteria. Safety and median PFS of lenvatinib were similar between the patients who did and those who did not meet the criteria. Among the patients who did not meet the criteria, 28, 27, 14, six, seven and five had a history of tyrosine kinase inhibitor (TKI) treatment, Child-Pugh score B, HCC in ≥50% of the liver, reduced platelet count, bile duct invasion and main portal vein invasion, respectively. The efficacy and safety of lenvatinib for patients with or without Child-Pugh-score B or HCC in ≥50% of the liver were similar. Although treatment outcome was not significantly different, patients with TKI treatment history tended to have longer median PFS, whereas those with main portal vein invasion tended to have shorter median PFS. Lenvatinib was effective for patients who did not meet the REFLECT inclusion criteria. However, the treatment outcome may vary according to several factors, such as a history of TKI treatment and tumor invasion.


Associations of cord blood metabolites with perinatal characteristics, newborn anthropometry, and cord blood hormones in project viva.
Metabolomics has emerged as a powerful tool to characterize biomarkers and elucidate physiological processes underlying adverse health outcomes. Little is known of these relationships during gestation and infancy, which are critical period for development of metabolic disease risk. To identify cord blood metabolite patterns associated with birth size; and to investigate relations of the birth size-associated metabolite patterns, and a branched chain amino acid (BCAA) metabolite pattern with a range of newborn and perinatal characteristics. Using untargeted mass-spectrometry, we quantified metabolites in cord blood of 126 mother-child pairs. After excluding 103 xenobiotics, we used principal components analysis (PCA) to consolidate the remaining 606 metabolites into principal components ("factors"). Next, we identified factors associated with gestational age-and sex-standardized birthweight z-score (BW/GA) and examined associations of the BW/GA-associated pattern(s) and the BCAA pattern with cord blood insulin, leptin, adiponectin, insulin-like growth factor (IGF)-1, IGF-2, and IGF binding protein 3 (IGFBP-3) using multivariable linear regression. Finally, we examined associations of maternal/perinatal characteristics with the cord blood metabolite patterns. Mean BW/GA z-score was 0.27±0.98 units. About half of the infants were male (52.4%) and white (57.1%). Of the 6 factors identified from PCA, one was associated with higher BW/GA: Factor 5, which comprised metabolites involved in energy production (malate, succinate, fumarate) and nucleotide turnover (inosine 5-monophosphate, adenosine 5-monophosphate, cytidine 5-monophosphate) pathways. In multivariable analysis, Factor 5 was related to higher cord blood leptin (1.64 [95% CI: 0.42, 2.87] ng/mL) and IGF-1 even after adjusting for IGFBP-3 (3.35 [0.25, 6.44] ng/mL). The BCAA pattern was associated with higher BW/GA (0.20 [0.03, 0.36] z-scores) and IGFBP-3 (106.5 [44.7, 168.2] ng/mL). No maternal characteristics were associated with either metabolite pattern; however, infants born via Cesarean delivery exhibited a higher score for Factor 5, and gestation length was inversely associated with the BCAA pattern. Metabolites in energy production and DNA/RNA turnover pathways in cord blood are associated with larger size at birth, and higher leptin and IGF-1. Similarly, the BCAA pattern was associated with larger birth size and IGFBP-3.


TREM2-independent oligodendrocyte, astrocyte, and T cell responses to tau and amyloid pathology in mouse models of Alzheimer disease.
Non-neuronal responses in neurodegenerative disease have received increasing attention as important contributors to disease pathogenesis and progression. Here we utilize single-cell RNA sequencing to broadly profile 13 cell types in three different mouse models of Alzheimer disease (AD), capturing the effects of tau-only, amyloid-only, or combined tau-amyloid pathology. We highlight microglia, oligodendrocyte, astrocyte, and T cell responses and compare them across these models. Notably, we identify two distinct transcriptional states for oligodendrocytes emerging differentially across disease models, and we determine their spatial distribution. Furthermore, we explore the impact of Trem2 deletion in the context of combined pathology. Trem2 knockout mice exhibit severely blunted microglial responses to combined tau and amyloid pathology, but responses from non-microglial cell types (oligodendrocytes, astrocytes, and T cells) are relatively unchanged. These results delineate core transcriptional states that are engaged in response to AD pathology, and how they are influenced by a key AD risk gene, Trem2.


A review of post-operative feeding in patients undergoing resection and reconstruction for oral malignancy and presentation of a pre-operative scoring system.
Percutaneous endoscopic gastrostomy (PEG) and nasogastric tubes (NGT) are routine after resection and reconstruction of oral cancer. The selection of the most appropriate method of feeding can be challenging, as both methods carry morbidity. This makes correct selection paramount. The objectives of this retrospective review were to identify the benefits and complications of feeding with PEG and NGT in patients with oral malignancy. We retrospectively reviewed 144 patients who had undergone oral cancer resection and reconstruction, to compare PEG and NGT feeding and to identify the key factors that aid selection of the most appropriate feeding method. We used these factors to develop the Key to Appropriate Replacement Enteral Nutrition (KAREN) scoring system. One hundred and twenty of the 144 patients were managed with PEG, and of these, 9 used it for less than 28 days. The mean (range) duration of use was 13 (5-63) days, and 1.9 (1-5) tubes/patient were used. The KAREN scoring system assigned the correct method of feeding in 92% of cases. The scoring system requires prospective validation but could provide clinicians with a tool to assist in a sometimes difficult decision.


Avulsion of flexor profundus, secondary to enchondroma of the distal phalanx.
Avulsion injury of the profundus tendon secondary to enchondroma of the distal phalanx is extremely rare. A case is described in which the enchondroma cavity was filled with an iliac bone graft and the avulsed fragment attached to the bone graft and the distal phalanx with a bone screw. Full function of the digit was regained.


Identification of two amino acids within E2 important for the pathogenicity of chimeric classical swine fever virus.
Our previous study demonstrated that a chimeric classical swine fever virus (CSFV) vSM/CE2 containing the E2 gene of the vaccine C-strain on the genetic background of the virulent CSFV strain Shimen (vSM) was attenuated in swine but reversed to virulence after serial passages in PK15 cells. To investigate the molecular basis of the pathogenicity, the genome of the 11th passage vSM/CE2 variant (vSM/CE2-p11) was sequenced, and two amino acid mutations, T745I and M979K, within E2 of vSM/CE2-p11 were observed. Based on reverse genetic manipulation of the chimeric cDNA clone pSM/CE2, the mutated viruses vSM/CE2/T745I, vSMCE2/M979K and vSM/CE2/T745I;M979K were rescued. The data from infection of pigs demonstrated that the M979K amino acid substitution was responsible for pathogenicity. Studies in vitro indicated that T745I and M979K increased infectious virus production and replication. Our results indicated that two residues located at sites 745 and 979 within E2 play a key role in determining the replication in vitro and pathogenicity in vivo of chimeric CSFV vSM/CE2.


Microarray analysis of gene expression differences in microglia after exposure to graphene quantum dots.
Graphene quantum dots (GQDs) have been broadly applied in biomedicine in recent years. So far, researches have reported that GQDs might contribute to the injury of the central nervous system (CNS), yet the latent toxicological mechanism is not clear. This study aims to investigate the underlying biological mechanism responsible for the neurotoxicity of nitrogen-doped GQDs (N-GQDs) and amino-functionalized GQDs (A-GQDs) by use of genome-wide transcription microarray. The findings showed that 174 and 1341 genes were altered significantly in the BV2 cells treated by 25 μg/mL N-GQDs and 100 μg/mL N-GQDs compared with the control, respectively. As for the BV2 cells exposed to 100 μg/mL A-GQDs, 1396 diversely expressed genes were detected. By comparing the 100 μg/mL N-GQDs exposed group with 100 μg/mL A-GQDs exposed group, the expression of 256 genes was extensively altered, including 58 upregulated genes and 198 downregulated genes. From Gene Ontology (GO) analysis, the altered genes were mainly enriched in functions of ion channel activation and cellular processes. Based on the KEGG pathway and signal-net analysis, the toxicity of GQDs in BV2 cells was closely related to calcium signaling pathway, cell cycle and endocytosis. And the pathways that the shared mRNAs involved all served as the crucial roles in the neurotoxicity of GQDs despite the chemical functionalization (N-GQDs or A-GQDs). In addition, the consequences from qRT-PCR, Western blot, intracellular calcium level measurements and comet assay further confirmed that calcium dyshomeostasis, DNA damage and cell cycle arrest were the key factors responsible for the GQDs-induced neurotoxicity through affecting several classical signaling pathways. In conclusion, our research will supply essential data for further studies on mechanisms of GQDs-induced neurotoxicity by use of genome-wide screening.


A new predictive tool for the early risk assessment of gestational diabetes mellitus.
The Italian National Institute of Health has recently introduced a selective screening based on the risk profile of pregnant women, which while recommending against screening of women at low risk (LR) for GDM, it recommends an early test for women at high risk (HR) for GDM. Herein, we assessed the accuracy and cost-effectiveness of this screening and developed a new index that improves these requirements. We retrospectively enrolled 3974 pregnant women. GDM was diagnosed with a 2h 75-g OGTT at 16-18 weeks (early test) or 24-28 weeks of gestation, according to the IADPSG guidelines. 55.6% of HR women had GDM, although only 38.4% underwent early screening. Among 2654 women at medium risk, 20.9% had GDM; paradoxically, among 770 LR women, that would not have been screened, 26.6% received a GDM diagnosis. Based on these unsatisfactory results, we elaborated the Capula's index, that reduced both screening tests (p<0.001) and potentially undetected GDM cases (p<0.001), and corrected the paradoxical prevalence estimates of GDM obtained with the current Italian guidelines. Also, Capula's index improved correlation of GDM risk profile with obstetric and neonatal adverse events. Capula's index improves accuracy of selective screening for GDM.


Academic-Community Partnership to Develop a Patient-Centered Breast Cancer Risk Reduction Program for Latina Primary Care Patients.
This collaborative study sought to address Latina breast cancer (BC) disparities by increasing health literacy (HL) in a community health center situated on the US-Mexico border region of San Diego County. An academic-community partnership conducted formative research to develop a culturally tailored promotora-based intervention with 109 individuals. The Spanish language program, entitled Nuestra Cocina: Mesa Buena, Vida Sana (Our Kitchen: Good Table, Healthy Life), included six sessions targeting HL, women's health, BC risk reduction, and patient-provider communication; sessions include cooking demonstrations of recipes with cancer-risk-reducing ingredients. A pilot study with 47 community health center Latina patients was conducted to examine the program's acceptability, feasibility, and ability to impact knowledge and skills. Pre- and post-analyses demonstrated that participants improved their self-reported cancer screening, BC knowledge, daily fruit and vegetable intake, and ability to read a nutrition label (p < 0.05). Results of the pilot study demonstrate the importance of utilizing patient-centered culturally appropriate noninvasive means to educate and empower Latina patients.


Biomaterials derived from hard palate mucosa for tissue engineering and regenerative medicine.
Autologous materials have superior biosafety and are widely used in clinical practice. Due to its excellent trauma-healing ability, the hard palate mucosa (HPM) has become a hot spot for autologous donor area research. Multiple studies have conducted an in-depth analysis of the healing ability of the HPM at the cellular and molecular levels. In addition, the HPM has good maneuverability as a donor area for soft tissue grafts, and researchers have isolated various specific mesenchymal stem cells (MSCs) from HPM. Free soft tissue grafts obtained from the HPM have been widely used in the clinic and have played an essential role in dentistry, eyelid reconstruction, and the repair of other specific soft tissue defects. This article reviews the advantages of HPM as a donor area and its related mechanisms, classes of HPM-derived biomaterials, the current status of clinical applications, challenges, and future development directions.


Stereoscan studies of rediae, cercariae, cysts, excysted metacercariae and migratory stages of Fasciola hepatica.
The external surface of the redial body of Fasciola hepatica is provided with microvillus-like projections or short lamellae, and short cilium-like structures are common anteriorly. The anterior part of the cercarial body possesses a pattern of regularly arranged small depressions each containing a spine. Both long and short cilium-like structures occur anteriorly. The tail is spineless and provided with dorsolateral folds. The outer cyst wall is formed by granules secreted from the tegument all over the body apart from the ventral sucker. Most granules transform into fibrillae which form the thick outer spongy layer. The precursor of the inner cyst wall is at the beginning closely attached to the metacercarial surface, but later the membrane-like cyst wall extends, and when fully formed the metacercaria lies free in the flattened circular inner cyst. The ventral plug is formed by the ventral sucker. The tegument of newly excysted metacercariae is provided with simple pointed spines, but later during migration in the mouse the spines become flattened and multipointed. Very young migratory stages may be attached with host cells.


Requirement of caveolae microdomains in extracellular signal-regulated kinase and focal adhesion kinase activation induced by endothelin-1 in primary astrocytes.
Endothelin-1 (ET-1) mitogenic activity in astrocytes is mediated by the activation of the extracellular signal-regulated kinase (ERK) pathway together with the Rho-dependent activation of the focal adhesion kinase (FAK) pathway. To clarify the mechanisms responsible for the coordinate activation of both pathways in the ET-1 signal propagation, the involvement of caveolae microdomains, suggested to play a role in signal transduction, was evaluated. In this study, it is reported that caveolae of primary astrocytes are enriched in endothelin receptor (ETB-R). Furthermore, signaling molecules such as the adaptor proteins Shc and Grb2, and the small G protein Rho, also reside within these microdomains. Selective disassembly of caveolae by filipin III impairs the ET-1-induced tyrosine phosphorylation of proteins including ERK and FAK. In agreement with these observations, astrocytes pretreated with filipin III also failed to form stress fibers and focal adhesions and did not undergo the associated morphological changes in response to ET-1. This study reveals that structural integrity of caveolae is necessary for the adhesion-dependent mitogenic signals induced by ET-1 in astrocytes, through compartmentation of ETB-R with the upstream signaling molecules of the ERK and FAK pathways.


Possible effects of repeated exposure to ibuprofen and acetaminophen on the intestinal immune response in young infants.
There has been an exponential increase in the frequency of immune deviations in young children. Consequently, research investigating environmental causes for this increase has become a Public Health priority. We have summarized the experimental observations and epidemiological data that could link repeated acetaminophen and ibuprofen exposure in early infancy to this increase. Recent observations on the maturational immunity of the intestinal sub-mucosal lamina propria underscore indeed the importance of prostaglandins (PGE2s). PGE2 appearing at this sub-mucosal level is a product of arachidonic acid metabolism mediated by type-2 cyclooxygenase (COX-2) situated on the membrane of many immune cells. Moreover, it seems that acetaminophen - like ibuprofen - also carries a non-selective inhibitory action on peripheral COXs, besides its central action. This inhibitory action of acetaminophen on COX2 only relates to physiological, low arachidonic acid concentrations. This explains the difference in anti-inflammatory effects. The impact of repeated inhibition of mucosal PGE2 synthesis due to COX-inhibitor exposure on maturational immunity has been demonstrated in animal experiments. Repeatedly exposed young animals do not develop tolerance to food antigens and exhibit autoimmune deviations. Several recent epidemiological studies have also reported on the magnitude of acetaminophen and ibuprofen exposure in children and the increase in immune deviations, it is important to better understand the potential negative impact of repeated inhibitions of prostaglandin synthesis by COX2s during infancy. Since acetaminophen and ibuprofen are commonly administered analgesics and antipyretics, a well-designed prospective strategy for pharmacovigilance and -epidemiology of COX-inhibitor exposure in infancy is urgently needed.


Effect of High-Volume Injectate in Lumbar Transforaminal Epidural Steroid Injections: A Randomized, Active Control Trial.
There have been many studies proving the effectiveness of lumbar transforaminal epidural steroid injections (TFESIs) for the treatment of radicular pain. Dexamethasone has been suggested as an alternative to particulate steroids. However, no controlled trials have investigated the effect of different injected volumes for a same dose of dexamethasone. To compare the effects of a high-volume injectate with those of a low-volume injectate using the same dose of dexamethasone for 2 groups in lumbar TFESI. A prospective, randomized, active control trial. The outpatient clinic of a single academic medical center. A total of 66 patients were randomized to receive lumbar transforaminal epidural dexamethasone injections with either a low-volume injectate (3 mL, N = 30) or a high-volume injectate (8 mL, N = 32). The primary outcome measures for this study were the incidence of the patients achieving meaningful pain relief and a reduction on the Visual Analogue Scale (VAS, range 0-100) at 4 weeks after the procedure. The definition of "meaningful pain relief" was ≥ 50% from baseline. The secondary outcomes included the Roland-Morris Disability Questionnaire (RMDQ, range 0-24) score and adverse effects. The outcomes were assessed 4 weeks after the procedure. Four weeks after the procedure, in the DL8 group, the incidence of achieving meaningful pain relief was higher compared with DL3 group (19, 59.4% vs. 9, 30%, P = 0.024). Both groups demonstrated a significant improvement in their VAS and RMDQ scores (P < 0.05). The VAS of the high-volume injectate group (DL8) was significantly lower than that of the low-volume injectate group (DL3) (33.3 ± 25 vs. 46.3 ± 25, P = 0.036). There was no significant difference in the RMDQ score between the 2 groups. We enrolled a small number of patients and did not check the long-term outcomes. Injectate at a volume of 8 mL was more effective than injectate at a volume of 3 mL for radicular pain in a lumbar transforaminal steroid injection, although both of the injectates contained the same dose of dexamethasone.


Fatty acid composition of normal and malignant cells and cytotoxicity of stearic, oleic and sterculic acids in vitro.
The aim of this study was to investigate the hypothesis that saturated fatty acids are differentially cytotoxic to cancer cells. Three studies were undertaken to: (1) measure the toxicities of stearic and oleic acids to normal and malignant cells in vitro, (2) assess if there is any relationship between toxicity and relative fatty acid composition and (3) determine whether the relative fatty acid composition of a cancer cell line could be modified by sterculic acid, an inhibitor of delta-9-desaturase. Stearic (18:0) and oleic (18:1) acids inhibited the colony-forming abilities of five human cancer cell lines and two non-neoplastic cell lines in a dose-dependent fashion. The concentration of oleic acid required to reduce colony formation ability by 50% was 2.5-6.0-fold greater than that of stearic acid. Addition of sterculic acid to a cancer cell line resulted in steady-state levels of stearic acid and increasing percentage of oleic acid.


(-)-Epigallocatechin gallate inhibits hepatitis C virus (HCV) viral protein NS5B.
In this study, we elucidated a small molecule inhibitor on viral protein NS5B identified through a high-throughput screening strategy using optical nanoparticle-based RNA oligonucleotide. We have previously shown that quantum dots (QDs)-RNA oligonucleotide can specifically recognize the HCV viral proteins. We have also demonstrated that conjugated QDs-RNA oligonucleotide can specifically and sensitively interact with designed biochips [1,2]. Among the flavonoids examined, (-)-epigallocatechin gallate (EGCG) demonstrated a remarkable inhibition activity on HCV viral protein, NS5B. (-)-Epigallocatechin gallate, at 0.005 μg mL(-1) or more, concentration-dependently attenuated the binding affinity on a designed biochip as evidenced by QDs-RNA oligonucleotide. At a concentration of 0.1 μg mL(-1), (-)-epigallocatechin gallate showed a 50% inhibition activity on QDs-RNA oligonucleotide biochip assay. We screened a small molecule inhibitor on the viral protein, NS5B, identified through a high-throughput screening strategy using on-chip optical nanoparticle-based RNA oligonucleotide on chip. In this designed strategy, the convenient and efficient screening and development of an on-chip viral protein inhibitor using a QDs-RNA oligonucleotide assay is achievable with high sensitivity and simplicity. In addition, this platform is expected to be applicable toward the inhibitor screening of other types of diseases.


Anti-poliovirus activity of protease inhibitor AG-7404, and assessment of in vitro activity in combination with antiviral capsid inhibitor compounds.
The National Research Council has recommended that at least one, preferably two, polio antiviral drugs be developed as a supplement to the tools currently available for control of polio outbreaks post-eradication. The primary application of such drugs is expected to be the resolution of chronic poliovirus excretion in persons with primary immunodeficiency disorders. We have assessed the in vitro activity of AG-7404 (also known as "compound 1"), an inhibitor of picornaviral 3C protease, against a large panel of programmatically important poliovirus strains and its activity in combination with two poliovirus capsid inhibitors, V-073 and BTA798. AG-7404 was active against all viruses in this panel, with EC50 values ranging from 0.080 to 0.674 μM. Similarly, BTA798 was active against all viruses in this panel, with EC50 values ranging from 0.003 to 0.591μM. By comparison, values for V-073 were 0.003-0.126 μM. BTA798 was active against V-073-resistant variants with an alanine to valine change in VP3 at position 24. However, BTA798 was inactive against the V-073-resistant strains with amino acid substitutions at VP1 amino acids 194 (equivalent to 192 in type 3) and 236. As expected from its different mechanism of action, AG-7404 was fully active against all V-073-resistant variants, with EC₅₀ values ranging from 0.218 to 0.819 μM, compared to values of 0.202-0.407 μM for the V-073-susceptible parental strains. In vitro drug combination experiments demonstrated synergy between AG-7404 and either V-073 or BTA798, whereas the combination of the two capsid inhibitors acted additively.


High-resolution structures of a heterochiral coiled coil.
Interactions between polypeptide chains containing amino acid residues with opposite absolute configurations have long been a source of interest and speculation, but there is very little structural information for such heterochiral associations. The need to address this lacuna has grown in recent years because of increasing interest in the use of peptides generated from d amino acids (d peptides) as specific ligands for natural proteins, e.g., to inhibit deleterious protein-protein interactions. Coiled-coil interactions, between or among α-helices, represent the most common tertiary and quaternary packing motif in proteins. Heterochiral coiled-coil interactions were predicted over 50 years ago by Crick, and limited experimental data obtained in solution suggest that such interactions can indeed occur. To address the dearth of atomic-level structural characterization of heterochiral helix pairings, we report two independent crystal structures that elucidate coiled-coil packing between l- and d-peptide helices. Both structures resulted from racemic crystallization of a peptide corresponding to the transmembrane segment of the influenza M2 protein. Networks of canonical knobs-into-holes side-chain packing interactions are observed at each helical interface. However, the underlying patterns for these heterochiral coiled coils seem to deviate from the heptad sequence repeat that is characteristic of most homochiral analogs, with an apparent preference for a hendecad repeat pattern.


A comparison of gene repair strategies in cell culture using a lacZ reporter system.
Synthetic oligonucleotides and DNA fragments of less than 1 kilobase (kb) have been shown to cause site-specific genetic alterations in mammalian cells in culture and in vivo. We have used a lacZ reporter gene system to compare the efficiency of episomal and chromosomal gene repair in human embryonic kidney epithelial cells (HEK293), Chinese Hamster Ovary fibroblasts (CHOK1), human bronchial epithelial cells (16HBE), and mouse embryonic stem (ES) cells. The lacZ gene contains a G to A nucleotide change, (Glu to Lys mutation) that abrogates beta-galactosidase activity. We compared the efficiency of different gene repair methods to correct this mutation and restore beta-galactosidase activity. We evaluated PCR-generated double-stranded DNA fragments of 0.52-1.9 kb, single-stranded DNA oligonucleotides of 20, 35, or 80 bases containing internal phosphorothioate links, and a 68 base RNA:DNA oligonucleotide. All of the oligonucleotides and DNA fragments showed some gene repair ability with an episomal plasmid. Short DNA fragments of 0.52 kb or greater gave the highest frequencies of episomal gene repair while single-stranded DNA oligonucleotides gave the highest frequency of chromosomal repair. In the context of a chromosomal target, antisense DNA oligonucleotides gave 5-fold higher frequencies of gene repair than their sense counterparts. The RNA:DNA chimeric oligonucleotide gave little or no gene repair on either a chromosomal or episomal target.


Adjuvant treatment using bleomycin in squamous cell carcinoma of penis: study of 19 cases.
Nineteen patients with squamous cell carcinoma of the penis had surgical treatment or a combination of surgery and radiation. They also had chemotherapy using bleomycin as a single agent or in combination with vincristine and methotrexate. The total doses of bleomycin varied between 89 mg/m2 and 212 mg/m2. The best results were obtained in well differentiated carcinomas.


An unusual combination of a bilateral aberrant suprascapular artery with neurovascular structures variants.
The report describes a bilateral suprascapular artery (SPSA) of atypical origin in coexistence with neurovascular aberrant structures. The variants were identified in a 91-year-old formalin-embalmed male cadaver, derived from a body donation program after a signed informed consent. The left-sided SPSA emanated from the 1st part of the axillary artery, coursed between the brachial plexus lateral and medial cords, accompanied by the suprascapular nerve, and passed below the superior transverse scapular ligament. Ipsilateral coexisted variants were the lateral thoracic artery multiplication, the subscapular trunk formation, and the musculocutaneous nerve duplication. In the right supraclavicular area, a SPSA duplication was identified. The main artery emanated from the thyrocervical trunk in common with the transverse cervical artery and the accessory SPSA emanated from the dorsal scapular artery. Both SPSAs coursed over the superior transverse scapular ligament, while the suprascapular nerve ran below the ligament. The current study reported a bilateral aberrant SPSA, originating from the AA 1st part (left side) and from the dorsal scapular artery (right side), which coexisted with adjacent neurovascular structures' variants. The left SPSA atypically coursed below the superior transverse scapular ligament. Such an unusual combination of variations, present bilaterally in the current study, may be challenging for radiologists and surgeons.


Non-Rhizobial Endophytes Associated with Nodules of Vigna radiata L. and Their Combined Activity with Rhizobium sp.
Root nodules of legume plants are devoted for hosting endophytic symbiotic bacteria that fix atmospheric nitrogen but recently proved as a niche for various non-rhizobial endophytes (NRE) also. In the present investigation, one rhizobial and two NRE were isolated and characterized as Rhizobium sp. AAU B3, Bacillus sp. AAU B6 and Bacillus sp. AAU B12. These isolates were studied for in vitro biocontrol activity against two pathogenic fungi. NRE isolates exhibited antifungal activity against root rot causing Macrophomina phaseolina (ITCC-6749) isolated from Vigna radiata and wilt causing pathogen Fusarium udum Butler isolated from Cajanus cajan in liquid as well as on solid medium. Furthermore, their antagonism was increased markedly when combined with Rhizobium sp. Moreover, Bacillus sp. AAU B6 showed amplification of the zwittermicin A gene (~ 950 bp) which is evident for the production of antibiotics. All three isolates showed HCN production in vitro also, Bacillus sp. AAU B12 exhibited amplification of its gene hcnC. Pathogenic fungal hyphae became thin, transparent, and bent as well as fungi lost their normal growth and branching patterns when exposed to volatile compounds produced by NRE. All the 3 isolates produced siderophores, however siderophore production was increased considerably when all three strains are mixed together. Furthermore, all the three isolates produced cell wall degrading enzymes (chitinase, protease, and cellulase) but lipolytic activity was exhibited only by Rhizobium sp. AAU B3. When NRE inoculated in combination of Rhizobium; overcomes the disease severity against M. phaseolina under pot study. Thus, from present study it is concluded that co-inoculation of NRE and Rhizobium sp. can be exploited as biocontrol bio-agents against M. phaseolina in green gram at field levels.


The elderly nutrition program: an effective national framework for preventive nutrition interventions.
To guide national policy, Congress mandated the 1992 research evaluation of the Elderly Nutrition Program (ENP), the nation's oldest framework for providing community- and home-based preventive nutrition and health-related services to older persons. This article summarizes key findings on the program's influence on nutritional health, the targeting and costs of its nutrition services, and the study's policy implications. The research included a nationally representative sample of ambulatory and homebound ENP participants And a matched sample of nonparticipants drawn from the US Health Care Financing Administration's Medicare beneficiary listings. Interviews conducted in respondents' homes considered demographic and health characteristics and assessed anthropometry and physical functioning, nutrient intake and socialization patterns, and utilization of ENP program services (participants only). Administrative and service delivery data were gathered from all levels of the ENP infrastructure. The ENP program currently provides congregate and home-delivered meals and other nutrition- and health-related services to about 7% of the older population overall, including an estimated 20% of the nation's poor elders. Compared with nonparticipants, ambulatory and homebound ENP participants are better nourished (4% to 31% higher mean daily nutrient intakes; P<.001) and achieve higher levels of socialization (17% higher average monthly social contacts; P<.001). Federal spending on ENP nutrition services is efficiently leveraged by funding from other public and private sources, allowing ENP to more than double the nutrition services it provides to program participants. ENP is a well-targeted, effective, and efficient federal program available to dietetics and other professionals for providing elderly persons with community-based and home-delivered nutrition and related services. The ENP infrastructure offers a potential model for preventive nutrition intervention programs in ambulatory and homebound at-risk older populations.


Improved visual discrimination learning in mice with partial 5-HT2B gene deletion.
The neurotransmitter serotonin (5-hydroxytryptamine, 5-HT) has been linked to multiple aspects of cognition. For example, in rodents, discrimination and reversal learning are altered by experimentally induced changes in brain serotonin levels, and reduced expression of the 5-HT2B receptor subtype in mice and humans is associated with decreased serotonergic tone and increased behavioral impulsivity. Serotonin modulates cognitive flexibility as well as fear and anxiety, but the specific contributions of 5-HT2B receptors to these behaviors is unknown. The current study assessed mice with partial Htr2b deletion for performance on a touchscreen-based pairwise visual discrimination and reversal learning task followed by a test of cued fear learning. Male Htr2b heterozygous mice (+/-) and littermate controls (+/+) were trained to discriminate between two visual stimuli presented on a touch-sensitive screen, one which predicted delivery of a 14-mg food pellet and the other which was not rewarded. Once discrimination performance criterion was attained, the stimulus-reward contingencies were reversed. Htr2b +/- mice were faster to reach discrimination criterion than +/+ controls, and made fewer errors. Htr2b +/- mice were also slower to make responses and collect rewards. Conversely, measures of reversal learning were not different between genotypes. Pavlovian cued fear conditioning was also normal in Htr2b +/-mice. These data demonstrate a selective improvement in touchscreen-based discrimination learning in mice with partial deletion of the 5-HT2B receptor, and provide further insight into the role of the 5-HT2B receptor in cognition.


The neglected role of the electrocardiogram in the diagnostic work-up of hypertensive patients: a study in clinical practice.
The electrocardiogram (ECG) is listed among the routine diagnostic examinations for hypertensive patients according to international guidelines. The aim of this survey was to investigate whether an ECG is routinely prescribed before an echocardiographic examination by general practitioners for the evaluation of subclinical cardiac damage in uncomplicated hypertensive patients in a suburban area of Italy. A total of 60 consecutive hypertensive patients (45% men, mean age 60 ± 14 years) referred to a single out-patient cardiology service by their general practitioners for the assessment of cardiac organ damage by echocardiography were enrolled in the study. Patients' demographic data and medical history were collected at the echocardiographic laboratory on a questionnaire administered by the attending physician. Less than 40% of the study sample had undergone an ECG during the 12-month period preceding the echocardiographic examination. Notably, only 60% of newly diagnosed hypertensive patents had an ECG done after the discovery of their hypertensive status. These findings show that an ECG is not routinely prescribed as a first-line examination in the assessment of cardiac organ damage related to hypertension by a large fraction of general practitioners in current clinical practice in a northern area of Italy. This contrasts with available evidence that the combined assessment of left ventricular hypertrophy by ECG and echocardiogram provides the best evaluation of cardiac damage and stratification of cardiovascular risk in hypertension.


A new surgical approach to the third ventricle with interruption of the striothalamic vein.
In the removal of tumours that develop within the third ventricle, most approaches are not entirely satisfactory. Therefore, a new approach has been devised: transfrontal exposure of the anterior portion of the frontal horn; coagulation and section of the striothalamic vein in order to open up the roof of the third ventricle; use of a blunt spatula introduced in the foramen of Monro and pushed backwards under the choroïd plexus. This approach has been used in ten cases. Postoperative mortality has been nil; the surgical approach has not apparently been responsible for any sequelae. It is simple and gives a good view of the third ventricle.


6-Nitro-2,3-bis-(thio-phen-2-yl)quinoxaline.
The title compound, C16H9N3O2S2, was synthesized via a condensation reaction in refluxing acetic acid. One thienyl ring is nearly coplanar with the quinoxaline unit [dihedral angle = 3.29 (9)°], the other makes an angle of 83.96 (4)°.


Discrepancy in plasma melarsoprol concentrations between HPLC and bioassay methods in patients with T. gambiense sleeping sickness indicates that melarsoprol is metabolized.
With the use of a specific high-performance liquid chromatography (HPLC) method and a bioassay which determines trypanocidal activity, concentrations of melarsoprol were assessed in plasma, urine and cerebrospinal fluid (CSF) from 8 patients with late-stage Trypanosoma gambiense sleeping sickness. The aim was to unravel to what extent the bioassay codetermines biologically active metabolites of melarsoprol. Subjects were given one dose of melarsoprol i.v. per day for 4 days (1.2, 2.4, 3.0-3.6, 3.0-3.6 mg per kg b.w., respectively). Plasma samples were obtained before the first melarsoprol injection, immediately after and at 1 h, 24 h and 5 days after the 4th injection. Urine was collected before melarsoprol therapy and at 24 h after the 4th injection. CSF samples were taken once before treatment and at 24 h after the 4th injection. HPLC analyses showed that plasma concentrations immediately after the 4th injection varied from 2200 to 15,900 nmol/l; dropping to 0-1800 nmol/l at 1 h; and to undetectable levels at 24 h. In urine small amounts of melarsoprol were recovered. Melarsoprol could not be detected in CSF by HPLC. Immediately after injection, bioassay analyses showed plasma concentrations of the same magnitude as HPLC assays but at 1 h they were 4-65-fold higher than the levels assessed by HPLC. Even 24 h and 5 days after the 4th injection there was significant but decreasing activity. Urine levels were 40-260-fold higher than the measured HPLC concentrations, whereas there was low but detectable activity in CSF. Results indicate that melarsoprol is rapidly eliminated from plasma. The significant trypanocidal activity determined by bioassay and simultaneous low or undetectable levels of melarsoprol assayed by HPLC indicate that the compound is transformed into metabolites with parasiticidal activity.


Bio-inspired adhesive and self-healing hydrogels as flexible strain sensors for monitoring human activities.
Nowadays, hydrogels have been intensively explored due to their promising potential application in bioelectronics. However, the lack of self-healing performance limits the lifespan and stability of materials. Here, a stretchable, self-healing and conductive hydrogel is prepared via physical crosslinking including hydrogen bonding, hydrophobic association and complexation effect, endowing the hydrogels with stretchability of 1150%, tensile strength of 112 kPa, flexibility and puncture resistance. Besides, the hydrogels possess extraordinary conductive property with a conductivity of 0.0720 S cm-1 and exhibit stable changes of resistance signals with a gauge factor of 2.506. Meanwhile, the hydrogels also exhibit adhesive performance due to the presence of catechol groups, and the maximum peeling adhesive strength could reach 23 N m-1. It is envisioned that the highly stretchable, self-healable, adhesive and conductive hydrogels would widen the application as the flexible strain sensors for soft robots, artificial limb, movement-monitoring equipment.


Salicylamide inhibitors of influenza virus fusion.
Structural variation of the quinolizidine heterocycle of the influenza fusion inhibitor BMY-27709 was examined by several topological dissections in order to illuminate the critical features of the ring system. This exercise resulted in the identification of a series of synthetically more accessible decahydroquinolines that retained the structural elements of BMY-27709 important for antiviral activity. The 2-methyl-cis-decahydroquinoline 6f was the most potent influenza inhibitor identified that demonstrated an EC50 of 90 ng/mL in a plaque reduction assay.


Dynamic models of BOLD contrast.
This personal recollection looks at the evolution of ideas about the dynamics of the blood oxygenation level dependent (BOLD) signal, with an emphasis on the balloon model. From the first detection of the BOLD response it has been clear that the signal exhibits interesting dynamics, such as a pronounced and long-lasting post-stimulus undershoot. The BOLD response, reflecting a change in local deoxyhemoglobin, is a combination of a hemodynamic response, related to changes in blood flow and venous blood volume, and a metabolic response related to oxygen metabolism. Modeling is potentially a way to understand the complex path from changes in neural activity to the BOLD signal. In the early days of fMRI it was hoped that the hemodynamic/metabolic response could be modeled in a unitary way, with blood flow, oxygen metabolism, and venous blood volume-the physiological factors that affect local deoxyhemoglobin-all tightly linked. The balloon model was an attempt to do this, based on the physiological ideas of limited oxygen delivery at baseline and a slow recovery of venous blood volume after the stimulus (the balloon effect), and this simple model of the physiology worked well to simulate the BOLD response. However, subsequent experiments suggest a more complicated picture of the underlying physiology, with blood flow and oxygen metabolism driven in parallel, possibly by different aspects of neural activity. In addition, it is still not clear whether the post-stimulus undershoot is a hemodynamic or a metabolic phenomenon, although the original venous balloon effect is unlikely to be the full explanation, and a flow undershoot is likely to be important. Although our understanding of the physics of the BOLD response is now reasonably solid, our understanding of the underlying physiological relationships is still relatively poor, and this is the primary hurdle for future models of BOLD dynamics.


[Study of the cutaneous scar after external dacryocystorhinostomy].
To objectively analyze the cutaneous scar after external dacryocystorhinostomy and to compare the cosmetic result of a nasal incision and an inferior eyelid incision. All patients consecutively operated between January 2007 and December 2009 were notified of an examination. The cutaneous scar was photographed and de visu analyzed by the two authors who gave marks : 20, no visible scar ; 10, visible scar but cosmetically considered not detrimental ; 0, visible scar and considered unesthetic. Eighteen of the 60 patients notified came for examination. One hundred percent of ten patients operated via an inferior eyelid incision were graded 20 versus 25% of eight patients operated via a nasal incision. Of these patients, 62.5% were graded 10. Only one patient was marked 0 (nasal incision). The inferior eyelid incision seemed to give much better cosmetic results than the nasal incision.


A single Bayesian estimator for iohexol clearance estimation in ICU, liver failure and renal transplant patients.
Iohexol clearance has been proposed to estimate the glomerular filtration rate (GFR). A population pharmacokinetics (popPK) model was developed from heterogeneous patients. A Bayesian estimator (MAP-BE) based on a limited sampling strategy (LSS) was derived and evaluated in external patients. Full pharmacokinetic data (7-12 samples) from 172 patients receiving iohexol for measurement of their GFR (unstable and stable ICU patients, liver failure patients and kidney transplant patients) were split into development (n = 136) and validation (n = 36) datasets. A PopPK model was developed in Monolix and was used to develop MAP-BE based on LSS. Its performance for GFR estimation was evaluated in the validation set. A two-compartment model with first-order elimination best described the data. The final model included the type of patients on volume of distribution (Vd), clearance and intercompartmental constants, serum creatinine on clearance and body weight on Vd. The best LSS included samples at 0.1-1-9 h exhibiting a relative mean prediction error (MPE) (RMSE) = -3.7% (14.3%) and better performance than the Bröchner-Mortensen formula (-3.0%/17%). Split by type of patients, the highest interindividual variability and imprecision was observed in unstable ICU patients (MPE (RMSE) = 3.7% (18.8%)) while the best performances were obtained for renal transplant patients (MPE (RMSE) = 1.0% (5.8%)). All LSS that included samples before 9 hours for the third sample were associated with an increased imprecision. A single MAP-BE of iohexol based on a three-sample LSS for four heterogeneous populations was developed and allowed accurate estimation of GFR in kidney transplant patients, slightly biased in stable ICU patients and slightly imprecise in unstable ICU patients.


Exploratory study of how Cognitive Multisensory Rehabilitation restores parietal operculum connectivity and improves upper limb movements in chronic stroke.
Cognitive Multisensory Rehabilitation (CMR) is a promising therapy for upper limb recovery in stroke, but the brain mechanisms are unknown. We previously demonstrated that the parietal operculum (parts OP1/OP4) is activated with CMR exercises. In this exploratory study, we assessed the baseline difference between OP1/OP4 functional connectivity (FC) at rest in stroke versus healthy adults to then explore whether CMR affects OP1/OP4 connectivity and sensorimotor recovery after stroke. We recruited 8 adults with chronic stroke and left hemiplegia/paresis and 22 healthy adults. Resting-state FC with the OP1/OP4 region-of-interest in the affected hemisphere was analysed before and after 6 weeks of CMR. We evaluated sensorimotor function and activities of daily life pre- and post-CMR, and at 1-year post-CMR. At baseline, we found decreased FC between the right OP1/OP4 and 34 areas distributed across all lobes in stroke versus healthy adults. After CMR, only four areas had decreased FC compared to healthy adults. Compared to baseline (pre-CMR), participants improved on motor function (MESUPES arm p = 0.02; MESUPES hand p = 0.03; MESUPES total score p = 0.006); on stereognosis (p = 0.03); and on the Frenchay Activities Index (p = 0.03) at post-CMR and at 1-year follow-up. These results suggest enhanced sensorimotor recovery post-stroke after CMR. Our results justify larger-scale studies.


An electron microscopic examination for the presence of acid groups in the organic matrix of mineralization nodules in foetal bone.
Several biochemical and histochemical investigations of the role of glycosaminoglycans (GAG) in bone and cartilage mineralization have been performed, but the exact relationship between GAG and mineralization is not clear. We performed an electron microscopic histochemical study of the composition of organic bone matrix at sites of beginning mineralization in mouse foetal radii, directing special attention to the presence and location of GAG. To demonstrate these substances, use was made of positive colloidal thorium dioxide and ruthenium red. The results demonstrate the presence of material containing acid groups in the young mineralization nodules. It is concluded that part of the demonstrated acid groups might be sulphate groups of glycosaminoglycans.


Molecular fingerprinting of Salmonella enterica subsp. enterica Typhimurium and Salmonella enterica subsp. enterica derby isolated from tropical seafood in South India.
Salmonella enterica subsp. enterica Typhimurium and Salmonella enterica subsp. enterica Derby strains isolated from different seafood were genotyped by PCR-ribotyping and ERIC-PCR assays. This study has ascertained the genetic relatedness among serovars prevalent in tropical seafood. PCR-ribotyping exhibited genetic variation in both Salmonella serovars, and ribotype profile (II) was most predominant, which was observed in 10/18 of Salmonella enterica subsp. enterica Typhimurium and 7/17 Salmonella enterica subsp. enterica Derby isolates. Cluster analysis of ERIC-PCR for Salmonella enterica subsp. enterica Typhimurium strains exhibited nine different banding patterns and four strains showed >95% genetic homology within the cluster pairs. ERIC-PCR produced more genetic variations in Salmonella enterica subsp. enterica Typhimurium; nevertheless, both methods were found to be comparable for Salmonella enterica subsp. enterica Derby isolates. Discrimination index of PCR-ribotyping for Salmonella enterica subsp. enterica Typhimurium isolates was obtained at 0.674 and index value 0.714 was observed for Salmonella enterica subsp. enterica Derby strains. Molecular fingerprinting investigation highlighted the hypothesis of diverse routes of Salmonella contamination in seafood as multiple clones of Salmonella enterica subsp. enterica Typhimurium and Salmonella enterica subsp. enterica Derby were detected in same or different seafood throughout the study period.


Aztreonam: the first monobactam.
There are several areas in which the use of aztreonam seems logical. Infections caused by organisms sensitive to aztreonam that are known to be multiresistant to other agents can be treated directly with aztreonam in single, directed therapy, thus making the use of more toxic agents unnecessary. In types of infection in which both gram positive and gram negative bacteria are present, aztreonam can replace the usual aminoglycoside component of the therapeutic regimen. In settings of mixed infections suspected of being caused by drug-resistant strains of Enterobacteriaceae and/or P. aeruginosa, aztreonam can be combined with an agent active against gram positive organisms or with one active against anaerobes. Aztreonam has proven to be effective, safe therapy for serious and life-threatening infections caused by multiresistant aerobic gram negative bacteria. It should be used in combination with drugs that inhibit gram positive species if the etiology of the infection is not known, particularly in the immunocompromised, neutropenic patient. Doses of 1 g every 8 to 12 hours will be adequate for treatment of infections caused by most Enterobacteriaceae. Whether 2 g doses every 8 hours would be preferred for treatment of systemic Pseudomonas infections remains to be determined. Urinary infections caused by gram negative bacteria can be treated with 500 mg administered IM once or twice daily. The dosage of aztreonam should be adjusted in patients with renal failure. Clearly, aztreonam is a useful addition to the antimicrobial agents available to the physician.


Regularized logistic regression for obstructive sleep apnea screening during wakefulness using daytime tracheal breathing sounds and anthropometric information.
Obstructive sleep apnea (OSA) is a prevalent health problem. Developing a technology for quick OSA screening is momentous. In this study, we used regularized logistic regression to predict the OSA severity level of 199 individuals (116 males) with apnea/hypopnea index (AHI) ≥ 15 (moderate/severe OSA) and AHI < 5 (non-OSA) using their tracheal breathing sounds (TBS) recorded during daytime, while they were awake. The participants were guided to breathe through their nose, and then through their mouth at their deep breathing rate. The least absolute shrinkage and selection operator (LASSO) feature selection approach was used to select the discriminative features from the power spectra of the TBS and the anthropometric information. Using a five-fold cross-validation procedure, five different training sets and their corresponding blind-testing sets were formed. The average blind-testing classification accuracy over the five different folds was found to be 79.3% ± 6.1 with the sensitivity (specificity) of 82.2% ± 7.2% (75.8% ± 9.9%). The accuracy for the entire dataset was found to be 81.1% with sensitivity (specificity) of 84.4% (77.0%). The feature selection and classification procedures were intelligible and fast. The selected features were physiologically meaningful. Overall, the results show that TBS analysis can be used as a quick and reliable prediction of the presence and severity of OSA during wakefulness without a sleep study. Graphical abstract Wakefulness screening of obstructive sleep apnea using tracheal breathing sounds and anthropometric information by means of regularized logistic regression with the least absolute shrinkage and selection operator approach for feature selection and classification.


Forest dynamics and its driving forces of sub-tropical forest in South China.
Tree mortality and recruitment are key factors influencing forest dynamics, but the driving mechanisms of these processes remain unclear. To better understand these driving mechanisms, we studied forest dynamics over a 5-year period in a 20-ha sub-tropical forest in the Dinghushan Nature Reserve, South China. The goal was to identify determinants of tree mortality/recruitment at the local scale using neighborhood analyses on some locally dominant tree species. Results show that the study plot was more dynamic than some temperate and tropical forests in a comparison to large, long-term forest dynamics plots. Over the 5-year period, mortality rates ranged from 1.67 to 12.33% per year while recruitment rates ranged from 0 to 20.26% per year. Tree size had the most consistent effect on mortality across species. Recruitment into the ≥1-cm size class consistently occurred where local con-specific density was high. This suggests that recruitment may be limited by seed dispersal. Hetero-specific individuals also influenced recruitment significantly for some species. Canopy species had low recruitment into the ≥1-cm size class over the 5-year period. In conclusion, tree mortality and recruitment for sixteen species in this plot was likely limited by seed dispersal and density-dependence.


Clinical benefits of PD-1/PD-L1 inhibitors in advanced hepatocellular carcinoma: a systematic review and meta-analysis.
Significant improvement of objective response rate and overall survival period has been achieved in several types of solid tumors by treatment with PD-1/PD-L1 inhibitors, which shed some light on hepatocellular carcinoma (HCC). Currently, a number of clinical trials concerning the application of checkpoint inhibitors in HCC are ongoing, some of which have shown favorable expectations. Hereby, we conducted a meta-analysis of existing studies to reveal the efficacy and safety of checkpoint inhibitors in advanced HCC. Medline, Embase, Cochrane Library, and Web of Science were searched from inception to January 31, 2020. The clinical trials reporting the efficacy of PD-1/PD-L1 inhibitors in advanced HCC patients were eligible. Overall results of complete response (CR), partial response (PR), stable disease (SD), progression of disease (PD), objective response rate (ORR), disease control rate (DCR), overall survival (OS), progression-free survival (PFS) and rate of adverse events (AE) with their 95% confidence intervals (95%CI) were calculated as the primary focus of the meta-analysis. Subgroup analyses were conducted primarily according to the categories of PD-1 inhibitor or PD-L1 inhibitor and combination therapy or monotherapy. In addition, pooled results of PD-1/PD-L1 monoclonal antibodies (mAb) combining with anti-VEGF agents were calculated separately. A total of 20 studies with 1232 patients were included. The overall CR, PR and SD rate were 0.01 (95% CI 0.01-0.03), 0.17 (95% CI 0.14-0.22) and 0.39 (95% CI 0.34-0.43), respectively. The overall ORR and DCR were 0.20 (95% CI 0.16-0.24) and 0.60 (95% CI 0.54-0.67), respectively. The overall PFS and OS were 3.58 months (95% CI 2.65-4.50) and 12.24 months (95% CI 10.48-14.00), respectively. For patients treated with PD-1/PD-L1 mAb combing with anti-VEGF agent, ORR was 29% (95% CI 0.15-0.43) and DCR was 77% (95% CI 0.70-0.84). For all included studies, the overall rate of AE was 0.63 (95% CI 0.45-0.78) and serious adverse events (SAE) was 0.11 (95% CI 0.06-0.22). PD-1/PD-L1 inhibitors showed favorable outcomes concerning response rates and survival periods in advanced HCC. Updated results from high-quality clinical trials are expected to validate these findings.


Serum cystatin C was a marker of poststroke fatigue in hypertensive intracerebral hemorrhage.
The relationship between poststroke fatigue (PSF) and serum Cystatin C (Cys-C) levels in hypertensive intracerebral hemorrhage (HICH) patients has not been determined. In this study, we investigated the association between serum Cys-C levels and PSF in HICH patients. A total of 125 patients with HICH were enrolled. Fatigue assessment was performed 6 months after HICH onset. The presence of PSF was defined as Fatigue Severity Scale (FSS) of 4 or more. Serum Cys-C levels were measured within 24 hr after admission. The correlation between FSS score and Cys-C level was analyzed by Spearman's correlation. Receiver operating characteristic (ROC) curves for PSF were calculated using Cys-C values. Of enrolled 125 patients in the study, 36.0% who developed PSF were divided to the PSF group, which had higher Cys-C levels compared with the no-PSF group. There was significant positive correlation between FSS score and serum Cys-C level. Receiver operating characteristic curves for PSF revealed an area under the curve of 0.86 for Cys-C. High admission Cys-C (>0.75mg/L) yielded specificity of 93.7%, positive predictive value of 87.5%, and negative predictive value of 88.2%. In multivariate analysis, Cys-C increased by 1 mg/dl (0.1 mg/L), and the risk of PSF in patients increased by 2.55 times (odds ratio = 2.55, 95% CI: 1.65-3.95, p < .001). High Cys-C levels have predictive value for PSF and can be used as one screening indicator for PSF occurrence.


Hyperinsulinemia or Insulin Resistance: What Impacts the Progression of Alzheimer's Disease?
Type 2 diabetes mellitus (T2D), which is often accompanied by hyperinsulinemia and insulin resistance, is associated with an increased risk for developing mild cognitive impairment and Alzheimer's disease (AD); however, the underlying mechanisms for this association are still unclear. Recent findings have shown that hyperinsulinemia and insulin resistance can coexist or be independent events. This makes it imperative to determine the contribution of these individual conditions in impacting AD. This literature review highlights the recent developments of hyperinsulinemia and insulin resistance involvement in the progression and pathogenesis of AD.


Ketamine inhibits calcium elevation and hydroxyl radical and nitric oxide production in lipopolysaccharide-stimulated NR8383 alveolar macrophages.
Macrophages play a critical role in mediating inflammatory processes; activated macrophages respond to endotoxin by releasing pro-inflammatory cytokines including tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6. Ketamine, a widely used anesthetic agent, has unequivocally anti-inflammatory effects in vivo and in vitro. However, the detailed mechanisms for the anti-inflammatory effects of ketamine in microglia have not been elucidated yet. This study aimed to evaluate the effects of ketamine on lipopolysaccharide (LPS)-induced nitric oxide (NO), hydroxyl radical (·OH) production, and intracellular calcium accumulation in macrophages. Macrophages were pretreated with ketamine at the concentrations of 10, 100, and 1,000 μM 1 h before LPS stimulation. The production of NO and ·OH in the culture supernatant of macrophages was assayed by Griess Reagent Kit. LPS enhanced NO and ·OH production and provoked a significant intracellular calcium elevation. With the concentrations higher than 100 μM, ketamine inhibited LPS-induced NO and ·OH accumulation and intracellular calcium elevation. However, a low concentration of ketamine (10 μM) did not exert anti-inflammatory effects. These results suggest that intracellular calcium elevation is, at least, partially involved in the inhibitory effect of ketamine.


The NAD salvage pathway in mesenchymal cells is indispensable for skeletal development in mice.
NAD is an essential co-factor for cellular energy metabolism and multiple other processes. Systemic NAD+ deficiency has been implicated in skeletal deformities during development in both humans and mice. NAD levels are maintained by multiple synthetic pathways but which ones are important in bone forming cells is unknown. Here, we generate mice with deletion of Nicotinamide Phosphoribosyltransferase (Nampt), a critical enzyme in the NAD salvage pathway, in all mesenchymal lineage cells of the limbs. At birth, NamptΔPrx1 exhibit dramatic limb shortening due to death of growth plate chondrocytes. Administration of the NAD precursor nicotinamide riboside during pregnancy prevents the majority of in utero defects. Depletion of NAD post-birth also promotes chondrocyte death, preventing further endochondral ossification and joint development. In contrast, osteoblast formation still occurs in knockout mice, in line with distinctly different microenvironments and reliance on redox reactions between chondrocytes and osteoblasts. These findings define a critical role for cell-autonomous NAD homeostasis during endochondral bone formation.


Production of teicoplanin by valine analogue-resistant mutant strains of Actinoplanes teichomyceticus.
Teicoplanin is a glycopeptide antibiotic produced by Actinoplanes teichomyceticus. A strain improvement to increase the productivity of the major component, teicoplanin A2-2, was carried out. As the fatty moiety of teicoplanin A2-2 is derived from L-valine, L-valine analogue (valine hydroxamate)-resistant mutants were derived. One of the mutants, 98-1-227, overproduced valine and produced a higher titer of total teicoplanin with higher A2-2 content. In a pilot fermentor (7 m3), the total productivity of teicoplanin was 1,800 units/ml and the A2-2 content was 58%.


Trans-stromal epithelial cyst after multiple lamellar keratoplasties - Case report and review of literature.
Single Observational Case Report. A 67-year-old male of Persian descent had a complex systemic and ocular history prior to a right penetrating keratoplasty (PK) reported here. The clinical diagnoses leading to the PK included Cogan's syndrome, chronic uveitis, secondary glaucoma, and corneal stromal scarring, presumed secondary to a corneal ulcer diagnosed on the second visit to our clinic. The specimen described here had been in place for 11 months and 17 days after the third failed Descemet stripping endothelial keratoplasty (DSEK). Visual acuities had ranged from 20/100 to 20/400 in both eyes. Visual acuity in the right eye just before surgery was 20/400. Intraocular pressures were 22 mmHg in both eyes with functioning Ahmed glaucoma shunts. The stromal cyst was not suspected preoperatively and no clinical imaging was performed. Histopathology including serial sections and immunohistochemistry. Histologic study demonstrated a trans-stromal corneal epithelial cyst without goblet cells that extended through a 500 μm gap in the donor tissue surface and edge. This large stromal cyst was an unusual complication of serial posterior lamellar keratoplasties and we postulate that multiple prior posterior lamellar grafts may have been a risk factor for this complication. Anterior segment imaging with either anterior segment optical coherence tomography or high-resolution ultrasound would likely have detected this stromal cyst.


Genetic studies on metallothionein synthesis in the mouse: the induction of metallothionein by cadmium in inbred strains.
Differences in the Cd-induced levels of liver metallothionein in inbred mouse strains are described. All strains examined show substantial increases in metallothionein production after the administration of 25 mumol CdCl2/kg, but the maximal levels achieved after 48 hr differed such that CBA/Ca greater than DBA/2 greater than C57BL/10 = C3H/He greater than BALB/c. These differences were generally not paralleled by differences in the amount of Cd2+ bound to metallothionein or in the total tissue content of Cd2+. Incorporation of [35S]cysteine into metallothionein suggest that differential synthesis affecting both isoproteins (MT1 and MT2) equally is largely responsible for the strain difference between C57BL/10 and DBA/2, and interstrain crosses between these two strains reveal codominant inheritance of Cd2+-induced metallothionein level and indicate that a single gene, either linked or identical to the cdm gene on chromosome 3, is involved.


[Prevention of postoperative pancreatic complications following duodenopancreatectomy using somatostatin].
Somatostatin was prophylactically administered to 10 patients who had surgery for pancreatic diseases. The postoperative course uncomplicated with no increase in serum amylase levels. Bile induced acute pancreatitis in six beagles was prophylactically treated with somatostatin. The results demonstrate that SRIF not only inhibits basal but also pancreatic-induced blood amylase and enzyme activities in the dog. SRIF-treated animals were in good general condition compared with untreated controls, macroscopic and histologic aspects of the pancreas were significantly improved.


The effects of Cu2+ and Pb2+ on the solution structure of calf thymus DNA: DNA condensation and denaturation studied by Fourier transform ir difference spectroscopy.
The interaction of calf thymus DNA with Cu2+ and Pb2+ was studied in aqueous solution at pH 6.5 with metal/DNA (P) (P = phosphate) molar ratios (r) 1/80, 1/40, 1/20, 1/10, 1/4, 1/2, and 1, using Fourier Transform ir (FTIR) spectroscopy. Correlations between the ir spectral changes, metal ion binding mode, DNA condensation, and denaturation, as well as conformational features, were established. Spectroscopic evidence has shown that at low metal/DNA (P) molar ratios 1/80 and 1/40, copper and lead ions bind mainly to the PO2- of the backbone, resulting in increased base-stacking interaction and duplex stability. The major copper ion base binding via G-C base pairs begins at r > 1/40, while the lead ion base binding occurs at r > 1/20 with the A-T base pairs. The denaturation of DNA begins at r = 1/10 and continues up to r = 1/2 in the presence of copper ions, whereas a partial destabilization of the helical structure was observed for the lead ion at high metal ion concentration (r = 1/2). Metal-DNA binding also results in DNA condensation. No major departure from the B-family structure was observed, upon DNA interaction with these metal ions.


Selenium, selenoprotein P, and oxidative stress levels in SARS-CoV-2 patients during illness and recovery.
This study aimed to assess tendency of oxidative stress in COVID-19 patients depending on severity. The study was conducted with 80 post-COVID-19 disease patients and 40 acutely ill patients. Content of selenium in blood plasma was detected by a fluorimetric method with di-amino-naphthalene using acidic hydrolysis. Selenoprotein P, malondialdehyde and 4-hydroxynonenal and their metabolite adducts were evaluated by spectrophotometric methods using commercial assay kits. Obtained results showed that selenium content in blood for post-COVID-19 disease patients was of a similar lower norm for Latvian inhabitants. Selenium and seleno-protein P contents for acute patients were significantly decreased compared with post-COVID-19 disease patients. In conclusion, COVID-19 involves induction of antioxidant systems-in case of severe disease, patients have significantly low concentration of selenium, seleno-protein P and higher level of oxidative stress, which, in turn, confirms the more intense formation of free radicals in the body.


Mechanism of inactivation of thyroid peroxidase by thioureylene drugs.
We have investigated the mechanism by which the thioureylene drugs, 1-methyl-2-mercaptoimidazole (MMI) and 6-n-propylthiouracil (PTU), inactivate thyroid peroxidase (TPO). Our results indicate that inactivation of TPO by MMI and PTU involves a reaction between the drugs and the oxidized heme group produced by interaction between TPO and H2O2. This conclusion is supported by the following observations. First, addition of a low concentration of H2O2 to a solution of TPO shifted lambda max of the Soret band from 411 to 420 nm, reflecting the formation of an oxidized form of TPO (TPOox). Addition of MMI or PTU to TPOox produced a Soret spectrum that was significantly different from the spectrum of native TPO or TPOox, whereas addition of MMI or PTU to native TPO produced no significant change in the heme spectrum. Second, studies with radiolabeled MMI and PTU combined with simultaneous assays of enzyme activity (guaiacol assay) showed that firm binding of the drugs to TPO and inactivation of the enzyme occurred on addition of the drugs to TPOox. However, neither binding nor inactivation occurred on addition of the drugs to native TPO. Third, the presence of a low concentration of iodide prevented the shift in the Soret spectrum, the binding of labeled drug, and the loss of enzyme activity associated with the addition of thioureylene drugs to TPO + H2O2. Under these conditions we assume that the enzyme was present as TPO X Iox, a form in which the heme is present in the same reduced state as in native TPO. This would explain the protective action of iodide on the inactivation of TPOox by MMI and PTU.


Two-dimensional 1H NMR of two chemically modified analogs of the basic pancreatic trypsin inhibitor. Sequence-specific resonance assignments and sequence location of conformation changes relative to the native protein.
Two-dimensional nuclear magnetic resonance was used to obtain sequence specific assignments for the 1H NMR spectra of two chemically modified analogs of the basic pancreatic trypsin inhibitor. In one analog the disulfide bond 14-38 was cleaved, in the second derivative the N-terminus was transaminated. From measurements of the chemical shifts and determination of the sequence locations of slowly exchanging backbone amide protons it was found that conformational differences between the native inhibitor and the chemical modifications occur exclusively near the modification sites and that the internal hydrogen bonds are nearly fully preserved. Intriguing conformation differences with respect to the native protein are that for five residues in the transaminated inhibitor and for one residue in the reduced inhibitor multiple local conformers are indicated, and that the four internal water molecules observed in the crystal structure of the native inhibitor appear not to be preserved after reduction of the disulfide bond 14-38.


Purification to homogeneity of the major "4S" PAH binding protein from "non responsive" DBA/2N mouse liver by affinity chromatography.
DBA/2N is a genetically non responsive inbred strain of mice in which administration of polycyclic aromatic hydrocarbons (PAHs) does not induce microsomal monooxygenase activity. DBA/2N mouse liver cytosol contains a polycyclic aromatic hydrocarbon-binding protein that sediments, in a sucrose gradient, at 4S ("4S" PAH-BP). Its binding kinetic and physicochemical properties indicate that this protein is practically indistinguishable from the "4S" PAH-BP identified and characterized in liver cytosol of rats and other PAH responsive rodents including C57 B1/6J mice. "4S" PAH-BP was purified to homogeneity from DBA/2N mouse liver by ammonium sulfate fractionation of the cytosol, followed by Sephadex G-200 chromatography and, finally, affinity chromatography using 1-aminopyrene-Sepharose 6B. This procedure yielded about 50 micrograms of protein from 50-60 g of mouse liver, with a recovery of 18%. "4S" PAH-BP as a complex with 3H-(benzo-a-pyrene) was more than 99% pure. A single band was seen on polyacrylamide gel electrophoresis under non denaturing conditions. H-BaP comigrated with the protein band. 3H-BaP bound to the protein was displaced by PAHs with a specificity identical to that obtained using crude cytosol. On electrophoresis in SDS gels, the purified protein migrated as a single protein band with an apparent molecular weight of 40,000.