In-vitro antioxidant,in-vivo anti-inflammatory,and acute toxicity study of Indonesian propolis capsule from Tetragonula sapiens

Propolis is widely used as traditional medicine since ancient times. It was necessary to conduct the pre-clinical study because of its relevant curative properties. This study aimed to investigate in-vitro antioxidant, standardize quality parameters, study acute toxicity, and determine in-vivo anti-inflammatory. Three spectrophotometric methods were used to determine antioxidant activity. The standardization includes physical, chemical, and microbiological evaluation. Furthermore, an acute toxicity test was conducted using 20 female Sprague Dawley (SD) strain rats divided into 4 groups with different dose of propolis. The in vivo anti-inflammatory test was carried out using the carrageenan induction method on rats' soles. A total of 36 female SD rats were classified into 6 groups as follows, Group normal, negative control, diclofenac sodium, and three propolis groups (72; 144; and 288 mg/kg BW). The results demonstrated the IC₅₀ values of the DPPH and ABTS scavenging activity 9.694 ppm and 2.213 ppm, respectively. The FRAP reducing power was 189.05 mg AaE/g. The physical appearance of propolis capsule was vegicaps as white – white, size 0, with light brown granule. Moreover, the content weight was 418.88 mg with a disintegration time of 7 min 53 s, while the water, flavonoid, and polyphenol contents were 9.07%, 1.59%, and 98.0821 mg GAE/g respectively. The content of heavy metal and microbial contamination were not detected. The acute toxicity results showed LD₅₀ ≥ 5 g/kg BW, no toxicity symptoms, and no abnormalities in all rats. The anti-inflammatory inhibition percentage for groups III, IV, V, and VI was 11.86%, 6.53%, 7.81%, and 6.63% respectively, while the anti-inflammatory drugs effectiveness percentage compared to positive controls were 55.00%, 65.83%, and 55.83% respectively. Based on these results, it can be concluded that propolis capsules fulfilled the standardization requirements, and it is likely to be non-toxic, and effective as antioxidant and anti-inflammatory.     

Structure-function relationships and molecular genetics of the 3β-hydroxysteroid dehydrogenase gene family

The isoenzymes of the 3β-hydroxysteroid dehydrogenase/5-ene-4-ene-isomerase (3β-HSD) gene family catalyse the transformation of all 5-ene-3β-hydroxysteroids into the corresponding 4-ene-3-keto-steroids and are responsible for the interconversion of 3β-hydroxy- and 3-keto-5-androstane steroids. The two human 3β-HSD genes and the three related pseudogenes are located on the chromosome 1p13.1 region, close to the centromeric marker D1Z5. The 3β-HSD isoenzymes prefer NAD⁺ to NADP⁺ as cofactor with the exception of the rat liver type III and mouse kidney type IV, which both prefer NADPH as cofactor for their specific 3-ketosteroid reductase activity due to the presence of Tyr³⁶ in the rat type III and of Phe³⁶ in mouse type IV enzymes instead of Asp³⁶ found in other 3β-HSD isoenzymes. The rat types I and IV, bovine and guinea pig 3β-HSD proteins possess an intrinsic 17β-HSD activity psecific to 5-androstane 17β-ol steroids, thus suggesting that such “secondary” activity is specifically responsible for controlling the bioavailability of the active androgen DHT. To elucidate the molecular basis of classical form of 3β-HSD deficiency, the structures of the types I and II 3β-HSD genes in 12 male pseudohermaphrodite 3β-HSD deficient patients as well as in four female patients were analyzed. The 14 different point mutations characterized were all detected in the type II 3β-HSD gene, which is the gene predominantly expressed in the adrenals and gonads, while no mutation was detected in the type I 3β-HSD gene predominantly expressed in the placenta and peripheral tissues. The mutant type II 3β-HSD enzymes carrying mutations detected in patients affected by the salt-losing form exhibit no detectable activity in intact transfected cells, at the exception of L108W and P186L proteins, which have some residual activity (1%). Mutations found in nonsalt-loser patients have some residual activity ranging from 1 to 10% compared to the wild-type enzyme. Characterization of mutant proteins provides unique information on the structure-function relationships of the 3β-HSD superfamily.     

Pharmacological cyclin-dependent kinase inhibitors inhibit replication of wild-type and drug-resistant strains of herpes simplex virus and human immunodeficiency virus type 1 by targeting cellular,not viral,proteins

Pharmacological cyclin-dependent kinase (cdk) inhibitors (PCIs) block replication of several viruses, including herpes simplex virus type 1 (HSV-1) and human immunodeficiency virus type 1 (HIV-1). Yet, these antiviral effects could result from inhibition of either cellular cdks or viral enzymes. For example, in addition to cellular cdks, PCIs could inhibit any of the herpesvirus-encoded kinases, DNA replication proteins, or proteins involved in nucleotide metabolism. To address this issue, we asked whether purine-derived PCIs (P-PCIs) inhibit HSV and HIV-1 replication by targeting cellular or viral proteins. P-PCIs inhibited replication of HSV-1 and -2 and HIV-1, which require cellular cdks to replicate, but not vaccinia virus or lymphocytic choriomeningitis virus, which are not known to require cdks to replicate. P-PCIs also inhibited strains of HSV-1 and HIV-1 that are resistant to conventional antiviral drugs, which target viral proteins. In addition, the anti-HSV effects of P-PCIs and a conventional antiherpesvirus drug, acyclovir, were additive, demonstrating that the two drugs act by distinct mechanisms. Lastly, the spectrum of proteins that bound to P-PCIs in extracts of mock- and HSV-infected cells was the same. Based on these observations, we conclude that P-PCIs inhibit virus replication by targeting cellular, not viral, proteins.     

An automatic diagnostic system of coronary artery lesions in Kawasaki disease using intravascular optical coherence tomography imaging

Intravascular optical coherence tomography (IV‐OCT) is a light‐based imaging modality with high resolution, which employs near‐infrared light to provide tomographic intracoronary images. Morbidity caused by coronary heart disease is a substantial cause of acute coronary syndrome and sudden cardiac death. The most common intracoronay complications caused by coronary artery disease are intimal hyperplasia, calcification, fibrosis, neovascularization and macrophage accumulation, which require efficient prevention strategies. OCT can provide discriminative information of the intracoronary tissues, which can be used to train a robust fully automatic tissue characterization model based on deep learning. In this study, we aimed to design a diagnostic model of coronary artery lesions. Particularly, we trained a random forest using convolutional neural network features to distinguish between normal and diseased arterial wall structure. Then, based on the arterial wall structure, fully convolutional network is designed to extract the tissue layers in normal cases, and pathological tissues regardless of lesion type in pathological cases. Then, the type of the lesions can be characterized with high precision using our previous model. The results demonstrate the robustness of the model with the approximate overall accuracy up to 90%.      

Phylogenetic diversity of Bradyrhizobium strains nodulating Calicotome spinosa in the Northeast of Algeria

Fifty-two slow-growing strains were isolated from root nodules of Calicotome spinosa grown in the Northeast of Algeria and grouped in 24 rep-PCR clusters. One representative strain for each profile was further phylogenetically characterized. The nearly complete 16S rRNA gene sequence indicated that all strains were affiliated to Bradyrhizobium. Multi-Locus Sequence Analysis (MLSA) of the atpD, glnII and recA genes and of the 16S-23S rRNA internal transcribed spacer (ITS) showed that these strains formed four divergent clusters: one close to Bradyrhizobium canariense and Bradyrhizobium lupini and three others separate from all the described species, representing three putative new Bradyrhizobium species. A phylogenetic analysis based on the nodC gene sequence affiliated the strains to either of the two symbiovars, genistearum or retamae.     

Von Hippel-Lindau (VHL) disease with pheochromocytoma in the Black Forest region of Germany: evidence for a founder effect

We identified a germline missense mutation at nucleotide 505 (T to C) of the VHL tumor suppressor gene in 14, apparently unrelated, VHL type 2A families from the Black Forest region of Germany. This mutation was previously identified in two VHL 2A families living in Pennsylvania (USA). All affected individuals in the 16 families shared the same VHL haplotype indicating a founder effect. This missense mutation at codon 169 (Tyr to His) would probably cause an alteration in the structure of the putative VHL protein. The association of this distinct mutation with the pheochromocytoma phenotype in VHL may help to elucidate the genetic mechanism of carcinogenesis in this multi tumor cancer syndrome.     

Effects of intraspecific larval competition on the development of the African buffalo fly Haematobia thirouxi potans

The effects of larval crowding on development, survival and size of the African buffalo fly, Haematobia thirouxi potans (Bezzi), were examined in small (10 or 20 g) and 1 000 g masses of cattle dung in the laboratory at 25°C. Dung was infested with fly eggs at densities which ranged from 0.05 to 32.0 eggs per g dung; in most cases 90–100% of eggs hatched. The duration of larval developmentin 10 g dung masses was extended by larval crowding at densities above 0.8 eggs per g dung and the development period with 32 eggs per g dung (11 days) was double that observed in uncrowded dung masses. Both size and survival decreased with increasing larval density, but stunting was observed at lower densities than those which reduced survival and so size was the more sensitive index of the adequacy of the larval environment. The effects of larval crowding became evident at lower larval densities in the 1 000 g dung masses than in the 10 g dung masses. There was a minimum size below which third instar larvae failed to pupariate. The probability of survival was related to pupal size and a high proportion of the smaller individuals failed to eclose.

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Effets de la competition intraspécifique sur le développement des larves, la taille des pupes et la survie d'Haematobia thirouxi potans

Résumé Les effets du surpeuplement larvaire sur le développement, la survie et la taille d'Haematobia thirouxi potans, ont été examinés sur de petites quantités de bouse (10 et 20 g) et de plus importantes (1 000 g), au laboratoire à 25°C. Les bouses étaient contaminées avec des oeufs de la mouche à des densités de 0,05 à 32 oeufs par gramme de bouse. Dans la plupart des cas le taux d'éclosion était de 90–100%. La durée du dévelopement larvaire dans 10 g de bouse a été prolongée pour les densités supérieures à 0,8 oeuf per g, et la durée développement pour 32 oeufs per g (11 jours) était le double de celle obtenue grand il n'y avait pas surpeuplement. La taille et la survie diminuent toutes les deux avec la densité larvaire, mais la miniaturisation était observée à des densités où la survie n'était pas modifiée; ainsi la taille est un indice plus sensible de l'adéquation de l'habitat larvaire. Les effets du surpeuplement larvaires deviennent clairs à des densités plus faibles avec 1 000 g de la bouse qu'avec 10 g. Il y a eu une taille minimale endessous de laquelle les larves étaient incapables de former une pupe. La probabilité d'éclosion était liée à la taille de la pupe, et une plus forte proportion de petits individus ne sont pas parvenu à éclore.

     

Molecular Nature of Spontaneous Mutations in Mouse Lactate Dehydrogenase-a Processed Pseudogenes

The presence of at least ten mouse LDH-A pseudogenes was demonstrated in the genomic blot analysis, and four different processed pseudogenes have thus far been isolated and characterized. In this report, the nucleotide sequences to two different mouse lactate dehydrogenase-A processed pseudogenes, M11 and M14, were determined and compared with the protein-coding sequences of the mouse and rat LDH-A functional genes. In the pseudogene M11, the sequence of 64 nucleotides from codon no. 257 to 278 was tandemly duplicated. In the pseudogene M14, the sequence of 22 nucleotides from codon no. 68 to 75 was replaced by an inserted repetitive sequence of 242 nucleotides homologous to a mouse truncated R element. The pattern of nucleotide substitutions accumulated in mouse LDH-A pseudogenes M11 and M14, as well as that of pseudogene M10 identified previously, was analyzed, and the substitution frequencies of the C or G at the CG dinucleotide were found to be high.     

Von Hippel-Lindau (VHL) disease: distinct phenotypes suggest more than one mutant allele at the VHL locus

Summary As part of an attempt to locate the von Hippel-Lindau locus (VHL) on chromosome 3, we evaluated 41 families with von Hippel-Lindau disease from the United States and Canada. One large family was identified whose disease phenotype was distinct from typical VHL. The most common disease manifestation was pheochromocytoma occuring in 57% (27/47) of affected family members. Few (4/47) affected family members had symptomatic spinal or cerebellar hemangioblastomas; no affected family member had renal cell carcinoma (0/47) or pancreatic cysts (0/24). Previously, genetic analysis demonstrated that the disease manifestations in this family were linked to RAF1 and D3S18, markers shown to be linked to typical VHL. These results suggest that there are mutant alleles at the VHL locus associated with distinct tissue specificities.