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The discovery of a novel 2-aminotetrahydropyridine class of BACE1 inhibitors is described. Their pKaand lipophilicity were modulated by a pending sulfonyl group, while good permeability and brain penetration were achieved via intramolecular hydrogen bonding. BACE1 selectivity over BACE2 was achieved in the S3 pocket by a novel bicyclic ring system. An optimization addressing reactive metabolite formation, cardiovascular safety, and CNS toxicity is described, leading to the clinical candidate JNJ-67569762 (12), which gave robust dose-dependent BACE1-mediated amyloid β lowering without showing BACE2-dependent hair depigmentation in preclinical models. We show that 12has a favorable projected human dose and PK and hence presented us with an opportunity to test a highly selective BACE1 inhibitor in humans. However, 12was found to have a QT effect upon repeat dosing in dogs and its development was halted in favor of other selective leads, which will be reported in the future.

Multiple programmed death-ligand 1 (PD-L1) immunohistochemistry assays performed using different antibodies including DAKO 22C3, DAKO 28-8, and Ventana SP142 PD-L1—predictive markers for response to various immune checkpoint inhibitors in NSCLC—have been approved in several countries. The differences in multiple PD-L1 immunohistochemistry assay results in predicting the therapeutic response to combined chemoimmunotherapy in patients with NSCLC remain unclear.

Accumulation of amyloid β peptides (Aβ) is thought to be one of the causal factors of Alzheimer’s disease (AD). The aspartyl protease β-site amyloid precursor protein cleaving enzyme 1 (BACE1) is the rate-limiting protease for Aβ production, and therefore, BACE1 inhibition is a promising therapeutic approach for the treatment of AD. Starting with a dihydro-1,3-thiazine-based lead, Compound J, we discovered atabecestat 1(JNJ-54861911) as a centrally efficacious BACE1 inhibitor that was advanced into the EARLY Phase 2b/3 clinical trial for the treatment of preclinical AD patients. Compound 1demonstrated robust and dose-dependent Aβ reduction and showed sufficient safety margins in preclinical models. The potential of reactive metabolite formation was evaluated in a covalent binding study to assess its irreversible binding to human hepatocytes. Unfortunately, the EARLY trial was discontinued due to significant elevation of liver enzymes, and subsequent analysis of the clinical outcomes showed dose-related cognitive worsening.

Lung adenocarcinoma with negative TTF-1 expression is believed to be a poor prognostic factor for certain systemic treatments. Nevertheless, the impact of TTF-1 expression on combined chemoimmunotherapy remains unclear. We aimed to investigate the relationship between tumor TTF-1 expression and the efficacy of combined chemoimmunotherapy in patients with advanced lung adenocarcinoma.

In recent years, programmed cell death-ligand 1 (PD-L1) inhibitor plus platinum-etoposide chemotherapy was found to have favorable clinical outcomes in patients with extensive-stage SCLC (ES-SCLC). The usefulness of early tumor shrinkage (ETS) has been reported in various types of cancers. Nevertheless, there have been few reports evaluating ETS in ES-SCLC. Therefore, this study aimed to evaluate the role of ETS in the clinical outcomes of patients with ES-SCLC receiving chemoimmunotherapy.

EGFR tyrosine kinase inhibitors are standard therapeutic agents for patients with advanced NSCLC harboring EGFR mutations. Nevertheless, some patients exhibit primary resistance to EGFR tyrosine kinase inhibitors in the first-line treatment setting. AXL, a member of the TYRO3, AXL, and MERTK family of receptor tyrosine kinases, is involved in primary resistance to EGFR tyrosine kinase inhibitors in EGFR-mutated NSCLC.

l-ribose isomerase (L-RI) from Cellulomonas parahominisMB426 can convert l-psicose and d-tagatose to l-allose and d-talose, respectively. Partially purified recombinant L-RI from Escherichia coliJM109 was immobilized on DIAION HPA25L resin and then utilized to produce l-allose and d-talose. Conversion reaction was performed with the reaction mixture containing 10% l-psicose or d-tagatose and immobilized L-RI at 40 °C. At equilibrium state, the yield of l-allose and d-talose was 35.0% and 13.0%, respectively. Immobilized enzyme could convert l-psicose to l-allose without remarkable decrease in the enzyme activity over 7 times use and d-tagatose to d-talose over 37 times use. After separation and concentration, the mixture solution of l-allose and d-talose was concentrated up to 70% and crystallized by keeping at 4 °C. l-Allose and d-talose crystals were collected from the syrup by filtration. The final yield was 23.0% l-allose and 7.30% d-talose that were obtained from l-psicose and d-tagatose, respectively.

The use of immune checkpoint inhibitors (ICIs) with chemotherapy has increased the survival of patients with advanced NSCLC. Nevertheless, the efficacy of ICI treatment for NSCLC with EGFRmutations is limited. Previous studies have not evaluated the efficacy of ICI treatment after osimertinib treatment in real-world settings.

To date, the efficacy and safety of programmed death-ligand 1 (PD-L1) inhibitor plus platinum-etoposide chemotherapy for patients with extensive-stage SCLC (ES-SCLC), with real-world evidence, stratified on the basis of age and performance status (PS), have not been fully investigated. The aim of this study was to evaluate the efficacy and safety of PD-L1 inhibitor plus platinum-etoposide chemotherapy in patients with ES-SCLC.

6-Deoxy-l-glucose, 6-deoxy-l-altrose, and 6-deoxy-l-allose were produced from l-rhamnose with an immobilized enzyme that was partially purified (IE) and an immobilized Escherichia colirecombinant treated with toluene (TT). 6-Deoxy-l-psicose was produced from l-rhamnose by a combination of l-rhamnose isomerase (TT-PsLRhI) and d-tagatose 3-epimerase (TT-PcDTE). The purified 6-deoxy-l-psicose was isomerized to 6-deoxy-l-altrose and 6-deoxy-l-allose with l-arabinose isomerase (TT-EaLAI) and l-ribose isomerase (TT-AcLRI), respectively, and then was epimerized to l-rhamnulose with immobilized d-tagatose 3-epimerase (IE-PcDTE). Following purification, l-rhamnulose was converted to 6-deoxy-l-glucose with d-arabinose isomerase (TT-BpDAI). The equilibrium ratios of 6-deoxy-l-psicose:6-deoxy-l-altrose, 6-deoxy-l-psicose:6-deoxy-l-allose, and l-rhamnulose:6-deoxy-l-glucose were 60:40, 40:60, and 27:73, respectively. The production yields of 6-deoxy-l-glucose, 6-deoxy-l-altrose, and 6-deoxy-l-allose from l-rhamnose were 5.4, 14.6, and 25.1%, respectively. These results indicate that the aldose isomerases used in this study acted on 6-deoxy aldohexoses.

The L-rhamnose isomerase gene (rhi) of Mesorhizobium lotiwas cloned and expressed in Escherichia coli, and then characterized. The enzyme exhibited activity with respect to various aldoses, including D-allose and L-talose. Application of it in L-talose production from galactitol was achieved by a two-step reaction, indicating that it can be utilized in the large-scale production of L-talose.

The xylitol dehydrogenase gene (xdh) of Bacillus palliduswas cloned and overexpressed in Escherichia coliusing pQE60 vector, for the first time. The open reading frame of 759 bp encoded a 253 amino acid protein with a calculated molecular mass of 27,333 Da. The recombinant xylitol dehydrogenase (XDH) was purified to homogeneity by three-step column chromatography, producing a single SDS–PAGE band of 28 kDa apparent molecular mass. The enzyme exhibited maximal activity at 55 °C in glycine-NaOH buffer pH 11.0, with 66% of initial enzyme activity retained after incubation at 40 °C for 1 h. In further application of the recombinant bacterium to L-xylulose production from xylitol (initial concentration 5%) using a resting cell reaction, 35% L-xylulose was produced within 24 h. This result indicates that this recombinant XDH is applicable in the large-scale production of L-xylulose.

D-Psicose, a new alternative sweetener, was produced from allitol by microbial oxidation of the newly isolated strain Enterobacter aerogenesIK7. Cells grown in tryptic soy broth medium (TSB) supplemented with D-mannitol at 37 °C were found to have the best oxidation potential. The cells, owing to broad substrate specificity, oxidized various polyols (tetritol, pentitol, and hexitol) to corresponding rare ketoses. By a resting cell reaction, 10% of allitol was completely transformed to the product D-psicose, which thus becomes economically feasible for the mass production of D-psicose. Finally, the product was crystallized and confirmed to be D-psicose by analytical methods.

The L-arabinose metabolic gene cluster, araA, araB, araD, araG, araHand araR, encoding L-arabinose isomerase (L-AI) and its accessory proteins was cloned from Mycobacterium smegmatisSMDU and sequenced. The deduced amino acid sequence of araAdisplayed highest identity with that of Bacillus subtilis(52%). These six genes comprised the L-arabinose operon, and its genetic arrangement was similar to that of B. subtilis. The L-AI gene (araA), encoding a 501 amino acid protein with a calculated molecular mass of 54,888 Da, was expressed in Escherichia coli. The productivity and overall enzymatic properties of the recombinant L-AI were almost same as the authentic L-AI from M. smegmatis. Although the recombinant L-AI showed high substrate specificity, as did L-AI from other organisms, this enzyme catalyzed not only isomerization of L-arabinose-L-ribulose and D-galactose-D-tagatose but also isomerization of L-altrose-L-psicose and L-erythrulose-L-threose. In combination with L-AI from M. smegmatis, L-threose and L-altrose can be produced from cheap and abundant erythritol and D-fructose respectively, indicating that this enzyme has great potential for biological application in rare sugar production. Transcription analysis using various sugars revealed that this enzyme was significantly induced not only by L-arabinose and D-galactose but also by L-ribose, galactitol, L-ribulose, and L-talitol. This different result of transcription mediated by sugars from that of E. colisuggests that the transcriptional regulation of araAfrom M. smegmatisagainst sugar is loose compared with that from E. coli, and that it depends on the hydroxyl configuration at C2, C3 and C4 positions of sugars.

Several 1-C-alkyl-2,3,4,6-tetra-O-benzyl-α-D-glucopyranosides were stereoselectively synthesized in good yields by the reaction of the 1-C-alkyl-2,3,4,6-tetra-O-benzyl-?-D-glucopyranosyl acetates with alcohols in the presence of 5 mol% Sc(OTf)3. The nature of the alkyl groups at the anomeric centers of the glycosyl donors had almost no influence on the reactivity and stereoselectivity of this glycosidation.

The gene encoding xylanase F3 (xynF3) was isolated from a genomic library of Aspergillus oryzaeKBN616, used for making shoyu koji.The structural part of xynF3was found to be 1468 bp. The nucleotide sequence of cDNA amplified by RT-PCR showed that the open reading frame of xynF3was interrupted by ten short introns and encoded 323 amino acids. Direct N-terminal amino acid sequencing showed that the precursor of XynF3 had a signal peptide of 22 amino acids. The predicted amino acid sequence of XynF3 has strong similarity to other family 10 xylanases from fungi. The xynF3gene was successfully overexpressed in A. oryzaeand the XynF3 was purified. The molecular mass of XynF3 estimated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis was 32,000. This was almost the same as the molecular mass of 32,437 calculated from the deduced amino acid sequence. The purified XynF3 showed an optimum activity at pH 5.0 and 58°C. It had a Km of 6.5 mg/ml and a Vmaxof 435 μmol•min−1•mg−1when birch wood xylan was used as a substrate. Expression of the xynF3gene was analyzed using an Eschelichia coliβ-glucuronidase gene as a reporter. The result indicated that xynF3is expressed in the medium containing wheat bran as a carbon source.

The gene encoding xylanase G2 (xynG2) was isolated from a genomic library of Aspergillus oryzaeKBN616, used for making shoyu koji. The structural part of xynG2was found to be 767 bp. The nucleotide sequence of cDNA amplified by RT-PCR showed that the open reading frame of xynG2was interrupted by a single intron which was 71 bp in size and encoded 232 amino acids. Direct N-terminal amino acid sequencing showed that the precursor of XynG2 had a signal peptide of 44 amino acids. The predicted amino acid sequence of XynG2 has strong similarity to other family 11 xylanases from fungi. The xynG2gene was successfully overexpressed in A. oryzaeand the overpexpressed XynG2 was purified. The molecular weight of XynG2 estimated on sodium dodecyl sulfate-polyacrylamide gel electrophoresis was 21,000. This was almost the same as the molecular weight of 20,047 calculated from the deduced amino acid sequence. The purified XynG2 showed an optimum activity at pH 6.0 and 58°C. It had a Km of 5.1 mg/ml and a Vmax of 123 μmol/min/mg when birch wood xylan was used as a substrate.

• A patient with secondary (sporadic type) hypokalemic periodic paralysis with relative hypertension had reninism with a high concentration of plasma angiotensin I (ANG-I) but no hyperaldosteronism or high angiotensin II value. Angiotensin-converting enzyme (ACE) activity was usually normal. Results of other hormonal analyses were also normal. However, the glomerular filtration rate and filtration fraction of the kidneys were greatly elevated. Despite severe hypokalemia, the patient's potassium clearance was high. No evidence of distinct hyperplasia of the juxtaglomerular cells was obtained. These results suggest that decreased affinity of ACE to the substrate ANG-I (so-called ACE dysfunction syndrome) produced the reninism and high concentration of plasma ANG-I, and that the latter induced an increase in the glomerular filtration rate of the kidneys with sequential occurrence of secondary hypokalemic periodic paralysis.(Arch Intern Med 1986;146:1956-1960)

We analyzed the sugar components of the glycoprotein fractions in human middle ear fluid samples by using gas chromatography. All ear effusions included serous, seromucinous and mucoid samples, and contained appreciable amounts of fucose, mannose, galactose, glucose, N-acetyl glucosamine and sialic acid. Total sugar content to protein was significantly higher in mucoid effusions than in serous effusions. Among the sugar components, the contents of sialic acid, N-acetyl glucosamine, fucose and galactose were much higher in the mucoid effusions. The data show new evidence that some glycoproteins having high proportions of sialic acid, N-acetyl glucosamine, fucose and galactose appear in the middle ear fluid of the glue ear.

Amino acid transport in human tonsillar lymphocytes was investigated in 32 patients of various ages and with different tonsillar diseases. Tonsillar lymphocytes appeared to possess at least four different transport systems for neutral amino acids including the activated Na+-dependent A system transport. The transport activity of neutral amino acid was significantly higher in child cases than in adult cases. In adult cases with focal tonsillitis, tonsillar lymphocytes were taken from patients with rheumatoid arthritis (RA) and skin diseases as secondary lesions who showed good improvement of the skin lesions after tonsillectomy. These lymphocytes were revealed to have more activated Na+-dependent transport than those from patients with skin diseases who showed poor improvement of the skin lesions after the tonsillectomy, and those from patients with recurrent tonsillitis. The characteristics of amino acid transport in human tonsillar lymphocytes and changes influenced by age and tonsillar diseases are discussed.

ABSTRACTAlthough previous studies suggest that cancer cachexia is a poor prognostic factor for immune checkpoint inhibitor monotherapy, the impact of cancer cachexia on chemoimmunotherapy is unclear. We investigated the impact of cancer cachexia on the therapeutic outcomes of chemoimmunotherapy for non-small cell lung cancer (NSCLC). We retrospectively analyzed patients’ medical records with NSCLC who received chemoimmunotherapy in 12 institutions in Japan between January and November 2019. We defined cancer cachexia as weight loss exceeding 5% of the total body weight or a body mass index of < 20 kg/m2and weight loss of more than 2% of the total body weight within 6 months before chemoimmunotherapy initiation, with laboratory results exceeding reference values. This study enrolled 235 patients with NSCLC, among whom 196 were eligible for analysis, and 50 (25.5%) met the criteria for cachexia diagnosis. Patients with cancer cachexia had a significantly higher frequency of a programmed death-ligand 1 (PD-L1) expression of ≥ 50% (48%, p= .01) and shorter progression-free survival (PFS; log-rank test: p= .04) than patients without cachexia. There was no significant difference in overall survival (OS) between the cachexia and no-cachexia groups (log-rank test: p= .14). In the PD-L1 ≥ 50% population, there was no significant difference in PFS and OS (log-rank test: p= .19 and p= .79, respectively) between patients with NSCLC in the cachexia or no-cachexia groups. Cancer cachexia might be a poor prognostic factor in patients with NSCLC receiving chemoimmunotherapy.

TO THE EDITOR. —Although D-penicillamine is of demonstrable value in the treatment of Wilson's disease and rheumatoid arthritis, its use has been limited by side effects. Recently, it has been debated whether a cytotoxic effect or an immunologic mechanism is responsible for the neutrophilic agranulocytosis appearing in patients treated with D-penicillamine. Corcos et al1 reported that the presence of antibodies to D-penicillamine or penicillin could not be demonstrated by the Coombs' test, and that no leukocyte agglutinins were found in serum at the time agranulocytosis developed or during the recovery phase. This suggests a cytotoxic effect of D-penicillamine. These findings are supported by evidence of D-penicillamine—induced interference with the synthesis of DNA, RNA, protein, collagen, and mucopolysaccharides.2 On the other hand, a positive result from a direct Coombs' test in a rheumatoid patient receiving D-penicillamine3 indicates that some immunologic mechanism may be responsible for D-penicillamine—induced neutrophilic agranulocytosis.

Diamond Blackfan Anemia (DBA) is a congenital syndrome characterized by decreased production of red blood cells and developmental abnormalities. This syndrome is caused by mutations in ribosomal genes (RPs), most frequently in RPS19, RPL11, and RPL5. Previously, we created RPS19 deficient zebrafish using morpholinos and demonstrated that p53 family members mediate the effects of RPS19 deficiency on development and hematopoiesis through upregulation of p53-related proteins. In this study, we found that RPL11 mutations, similar to RPS19 deficiency, leads to dysregulation of p53, its related family members, and downstream signaling pathways in zebrafish. Expression of both total p53 and Δ113Np53 is strongly upregulated in mutants with more modest upregulation of ΔNp63. The p53 protein also induced p21 and G1 cyclin expression, which resulted in cell cycle arrest. The pro-apoptotic genes, puma and bax, were also upregulated. We observed that the expression of several genes involved in hematopoiesis was altered, for example, globin gene expression was decreased. The ΔNp63 protein, which increases expression of adenosine deaminase (ADA) and is upregulated in >70% of DBA patients, was also upregulated in RPL11 zebrafish mutants. Interestingly, we also observed increased expression of the proto-oncogenes c-fos, c-jun, and c-myc in RPL11 mutants. Increased expression of proto-oncogenes may contribute to the mechanisms of transcriptional activation by p53 family members in response to ribosomal protein deficiency. Furthermore, aberrant expression of oncogenes could provide a possible mechanism for the increased risk of malignancies in DBA patients. We propose that the zebrafish is a model system to study developmental and hematologic defects in addition to signaling pathways in the pathogenesis of DBA.

Diamond Blackfan Anemia (DBA) is a congenital syndrome characterized by decreased production of red blood cells and developmental abnormalities. This syndrome is caused by mutations in ribosomal genes (RPs), most frequently in RPS19, RPL11, and RPL5. Previously, we created RPS19 deficient zebrafish using morpholinos and demonstrated that p53 family members mediate the effects of RPS19 deficiency on development and hematopoiesis through upregulation of p53-related proteins. In this study, we found that RPL11 mutations, similar to RPS19 deficiency, leads to dysregulation of p53, its related family members, and downstream signaling pathways in zebrafish. Expression of both total p53 and Δ113Np53 is strongly upregulated in mutants with more modest upregulation of ΔNp63. The p53 protein also induced p21 and G1 cyclin expression, which resulted in cell cycle arrest. The pro-apoptotic genes, puma and bax, were also upregulated. We observed that the expression of several genes involved in hematopoiesis was altered, for example, globin gene expression was decreased. The ΔNp63 protein, which increases expression of adenosine deaminase (ADA) and is upregulated in >70% of DBA patients, was also upregulated in RPL11 zebrafish mutants. Interestingly, we also observed increased expression of the proto-oncogenes c-fos, c-jun, and c-myc in RPL11 mutants. Increased expression of proto-oncogenes may contribute to the mechanisms of transcriptional activation by p53 family members in response to ribosomal protein deficiency. Furthermore, aberrant expression of oncogenes could provide a possible mechanism for the increased risk of malignancies in DBA patients. We propose that the zebrafish is a model system to study developmental and hematologic defects in addition to signaling pathways in the pathogenesis of DBA.

TO THE EDITOR. —Ankylosing spondylitis (AS) is a chronic inflammatory arthritis with a predilection for involvement of cartilagenous joints of the axial skeleton. Pain may be localized to the lumbar region or sacroiliac joints, with stiffness pronounced after prolonged inactivity in the morning, and relieved by movement or heat. In addition, this disease is characterized by the absence of both rheumatoid factor and subcutaneous nodules, and the presence of extraskeletal manifestations such as acute anterior iridocyclitis, atrioventricular block, pulmonary fibrocystic disease in the upper lobes, spinal articular involvement, and nerve root involvement. Thomas et al1 have demonstrated that neurologic profiles in AS can fall into five major categories as follows: multiple sclerosis, the cauda equina syndrome, focal epilepsy, vertebrobasilar insufficiency, and peripheral nerve lesions. With regard to clinical management, relief of articular pain and discomfort and long-range planning for correction or prevention of deformity are of first importance. Actually,