Immunology and Biosynthesis of the Mammalian Pyruvate Dehydrogenase Complex

The mammalian pyruvate dehydrogenase complex, located in the inner membrane-matrix compartment of mitochondria, is a large multi-molecular aggregate, Mr 8.5 x 10e6, containing multiple copies of its three constituent enzymes, pyruvate dehydrogenase (E1), dihydrolipoyl acetyltransferase (E2) and lipoamide dehydrogenase (E3). High-titre, monospecific, polyclonal antibodies against the native complex and its individual components are characterised and employed to study the events involved in the biosynthesis and import of the components of this complex into mitochondria. Antisera are examined with respect to their ability to inhibit the overall activity of the complex and of the intrinsic protein kinase. Monospecificity of all antisera is demonstrated when challenged with crude cell extracts and subsequent immunoblotting analysis. The antiserum to native PDC exhibits high reactivity against all components of the complex, except E3, which cannot be explained by a low content of E3 in the purified antigen or by its inaccessibility to the immune system. The significance of the low immunogenicity of E3 is probably related to the conservation of its primary sequence and tertiary structure during evolution. A major observation from this study is that ox heart PDC contains an additional polypeptide, M 51,000+/-1,000 of unknown function. This protein is called component X and constitutes approx. 6% of the total complex. Detailed immunological studies suggest that component X is a normal cellular component, located in the mitochondrial compartment and does not represent a fragment of subunit E2 or the intrinsic protein kinase of the complex. Additional studies on the subcellular localisation of PDC suggest that this enzyme is associated with the inner membrane of rat liver and ox heart mitochondria. These and other studies also indicate that component X is an integral component of the complex and not a membrane protein, which becomes associated with the PDC during its isolation. The individuality of protein X is also demonstrated by comparison of the peptide maps of the 125 I and 14C-labelled subunits E2 and X obtained with several proteases. Protein X seems to be tightly-associated with the E2 core of the complex and is at least partially exposed on the surface of the native assembly, as it is accessible to proteases and to antibodies directed against it. Further studies reveal that, after incubation of the complex in the presence of [2-14C] pyruvate, 14C-label, probably in the form of acetyl groups is incorporated rapidly into both E2 and component X. Phosphorylation of the complex causes a parallel decrease in the acetylation of both proteins, indicating the involvement of the E1 component in the acetylation of these groups. Similarly, the transfer of acetyl groups from E2 and X onto CoA is observed to occur in a parallel fashion from both proteins. Studies on the effects of NEM on the incorporation of 14C-labelled acetyl groups also reveal interesting features of the acetylation reaction, which suggest the existence of secondary NEM-sensitive acetylation sites on the complex. Additional studies on the nature of the 'acetylatable' group in protein X and the physiological acceptor of acetyl groups are required to clarify the function of this component in the acetylation reactions of the complex. Studies on cultured bovine kidney, rat liver and pig kidney cells, 35 incubated with [35S] methionine in the presence of uncouplers of oxidative phosphorylation, demonstrate the accumulation of larger M precursor polypeptides to the subunits E2, E3, E1 alpha and E1 beta of the complex. Precursor forms for the individual subunits of the PDC are identified by immunoprecipitation techniques using antisera against the native complex and its SDS-denatured subunits followed by fluorographic analysis. These precursors, possessing M values 2,000-8,000 larger than their mature counterparts in the mitochondria, are relatively stable in the cytoplasm of the cells when monolayers are incubated in the presence of uncouplers for several hours. Removal of the uncoupler and subsequent chase shows that these precursors are processed into their mature forms with a similar lag time. Complete processing is observed within 30 min

Perencanaan Sistem Drainase Di Kawasan Pusat Kota Amurang

Kawasan pusat kota Amurang yang terletak di ibukota kabupaten Minahasa Selatan merupakan kawasan yang dipadati oleh fasilitas-fasilitas seperti pertokoan, rumah makan, pasar tradisional, hotel, bank termasuk pemukiman warga. Genangan air yang terjadi setiap kali turun hujan memberikan dampak yang negatif antara lain kerusakan jalan, serta terganggunya aktivitas warga di kawasan tersebut. Untuk menanggulangi masalah genangan yang sering terjadi perlu perencanaan sistem drainase yang baik di kawasan pusat kota. Untuk mengidentifikasi masalah genangan air, dilakukan observasi langsung di daerah penelitian, kemudian dilanjutkan dengan desain rencana tata letak sistem drainase. Dilakukan analisis hidrologi untuk mendapatkan debit rencana berdasarkan data curah hujan yang telah diperoleh,dilanjutkan dengan analisis hidrolika untuk mencari kapasitas eksisting saluran yang relevan dengan rencana tata letak sistem drainase. Tata letak rencana sistem drainase direncanakan dengan menentukan saluran interceptor di sisi selatan jalan trans Sulawesi terlebih dahulu sehingga pembebanan aliran dari luar lokasi penelitian tidak masuk ke lokasi tinjauan. Dari hasil analisis, perlu dilakukan Perubahan tata letak sistem drainase. Terdapat 48 ruas saluran eksisting yang masih relevan dengan rencana tata letak sistem drainase dan 10 gorong- gorong yang masih sesuai. Untuk saluran eksisting, 9 ruas saluran tidak memenuhi sehingga dilakukan Perubahan dimensi saluran. Sedangkan untuk gorong- gorong terdapat 4 ruas yang tidak memenuhi. Rekomendasi untuk saluran yang baru berjumlah 17 ruas dan 5 gorong- gorong

Comparison of the diagnostic yield of routine versus indicated flowmetry, ultrasound and cystoscopy in women with recurrent urinary tract infections

INTRODUCTION AND HYPOTHESIS: To quantify and compare the outcomes of routine vs. urologist-requested diagnostic testing for recurrent urinary tract infections (rUTI). METHODS: A retrospective cohort study of patients with rUTI referred to a large non-academic teaching hospital between 2016 and 2018 (Hospital A) and a university hospital between 2014 and 2016 (Hospital B). Electronic medical records were reviewed for baseline and diagnostic data. Women underwent the following assessments routinely: urinalysis, voiding diary, flowmetry in Hospital A and urinalysis, voiding diary, flowmetry, ultrasound, abdominal x-ray and cystoscopy in Hospital B. All other diagnostics were performed by indication in each hospital. RESULTS: We included 295 women from Hospital A and 298 from Hospital B, among whom the mean age (57.6 years) and mean UTI frequency (5.6/year) were comparable, though more were postmenopausal in Hospital A. We identified abnormalities by flowmetry or post-void residual volumes in 134 patients (Hospital A: 79; Hospital B: 55), cystoscopy in 14 patients (Hospital A: 6; Hospital B: 8) and ultrasound in 42 patients (Hospital A: 16; Hospital B: 26), but these differences were not significant. Diagnostics altered treatment in 117 patients (e.g., pelvic floor muscle training, referral to another specialist, surgical intervention), mostly due to flowmetry and post-void residual volume measurement. The retrospective design and absence of follow-up data limit these results. CONCLUSIONS: The routine use of cystoscopy and ultrasound in female patients with rUTIs should not be recommended as they yield few abnormalities and lead to additional costs

Performances of Solid Oxide Cells with La0.97_{0.97}Ni0.5_{0.5}Co0.5_{0.5}O3−δ_{3-\delta} as Air-Electrodes

Based on previous studies of perovskites in the quasi-ternary system LaFeO$_{3}$–LaCoO$_{3}$–LaNiO$_{3}$, La$_{0.97}$Ni$_{0.5}$Co$_{0.5}$O$_{3}$ (LNC) is chosen as the most promising air-electrode material in the series for solid oxide cells (SOCs). The properties of the material itself have been investigated in detail. However, the evaluation of LNC97 air electrodes in practical SOCs is still at a very early stage. In the present study, SOCs were prepared based on LNC97 air electrodes. The I-U performance of the SOCs in both solid oxide fuel cell (SOFC) and solid oxide electrolysis cell (SOEC) modes, i.e. reversible SOCs (r-SOCs), was investigated systematically for different air-electrode designs, temperatures and fuel gases. In general, the performance of the r-SOCs tested in the present study is higher than the published results of other LaFeO$_{3}$–LaCoO$_{3}$–LaNiO$_{3}$-based SOCs and is comparable to or even better than state-of-the-art La$_{1-x}$Sr$_{x}$Fe$_{1-y}$Co$_{y}$O$_{3}$ (LSCF)-based SOCs. Mid-term operation of about 1000 h for SOCs in both SOFC and SOEC modes primarily proved the stability of LNC97-based air electrodes. Impedance spectra were systematically applied to understand the polarization processes of the SOCs

Quality appraisal of clinical guidelines for recurrent urinary tract infections using AGREE II:a systematic review

INTRODUCTION AND HYPOTHESIS: Recommendations for preventing and diagnosing recurrent urinary tract infection (UTI) tend to vary between clinical practice guidelines (CPGs) because of low-quality scientific evidence, potentially leading to practice variation and suboptimal care. We assessed the quality of existing CPGs for recurrent UTI. METHODS: A systematic search was performed from January 2000 to June 2021 in PubMed and EMBASE for CPGs on recurrent UTI prevention or hospital diagnostics in Dutch, English, and Spanish. Each CPG was assessed by four appraisers in a multidisciplinary review team, using the Appraisal of Guidelines, Research, and Evaluation II (AGREE II) instrument. RESULTS: We identified and assessed eight CPGs published between 2013 and 2021. The scope and purpose (mean and standard deviation: 67.3 ± 21.8) and clarity of presentation (74.8 ± 17.6) domains scored highly. However, issues with methods, patient participation, conflict of interests, and facilitators and barriers were common and resulted in lower scores for the rigour of development (56.9 ± 25.9), applicability (19.6 ± 23.4), stakeholder involvement (50.4 ± 24.6), and editorial independence (62.1 ± 23.1) domains. Overall, two CPGs were recommended, three were recommended with modifications, and three were not recommended. CONCLUSIONS: Significant room for improvement exists in the quality of CPGs for recurrent UTI, with most displaying serious limitations in the stakeholder involvement, rigour of development, and applicability domains. These aspects must be improved to decrease diagnostic and therapeutic uncertainty. Developers could benefit from using checklists and following guidelines when developing de novo CPGs

Treatment Patterns and Use of Immune Checkpoint Inhibitors Among Patients with Metastatic Bladder Cancer in a Dutch Nationwide Cohort

Since 2017, two immune checkpoint inhibitors (ICIs) have become the standard of care for the treatment of metastatic urothelial carcinoma in Europe: pembrolizumab as second-line therapy and avelumab as maintenance therapy. Our aim was to describe the use of ICIs as first and later lines of treatment in patients with metastatic bladder cancer (mBC) in the Netherlands. We identified all patients diagnosed with primary mBC between 2018 and 2021 in the Netherlands from the Netherlands Cancer Registry (NCR). NCR data were supplemented with data from the Dutch nationwide Prospective Bladder Cancer Infrastructure (ProBCI) collected from medical files, with follow-up until death or end of data collection on January 1, 2023. A total of 1525 patients were diagnosed with primary mBC between 2018 and 2021 in the Netherlands. Of these, 34.7% received at least one line of systemic treatment with chemotherapy or ICI. After first-line platinum-based chemotherapy, 34.1% received second-line ICI and 3.9% received maintenance ICI. Among patients who completed or discontinued first-line cisplatin- or carboplatin-based chemotherapy after approval of maintenance ICI in the Netherlands, 40.7% and 19.7% received second-line ICI, and 9.3% and 14.1% received maintenance ICI, respectively. ICI use for mBC treatment has not increased considerably since their introduction in 2017. Future research should assess whether the introduction of maintenance avelumab (available since April 2021 in the Netherlands) has led to increases in the proportion of patients with mBC patients receiving systemic treatment and the proportion receiving ICI. Patient summary: We assessed the rate of immunotherapy use for patients with metastatic bladder cancer in the Netherlands. Since its introduction, immunotherapy has been used in a minority of patients, mostly as second-line treatment after platinum-based chemotherapy.</p

Magnetic Resonance Imaging-targeted Prostate Biopsy Compared with Systematic Prostate Biopsy in Biopsy-naïve Patients with Suspected Prostate Cancer

BACKGROUND: It remains uncertain whether transrectal ultrasound (TRUS)-guided systematic biopsies can be omitted and rely solely on multiparametric magnetic resonance imaging–targeted biopsies (MRI-TBx) in biopsy-naïve men suspected of prostate cancer (PCa). OBJECTIVE: To compare PCa detection in biopsy-naïve men between systematic biopsy and MRI-TBx. DESIGN, SETTING, AND PARTICIPANTS: A prospective cohort study was conducted in a Dutch teaching hospital. Consecutive patients with suspected PCa, no history of biopsy, and no clinical suspicion of metastasis underwent both TRUS-guided systematic biopsies and MRI-TBx by multiparametric magnetic resonance imaging (mpMRI)-ultrasound fusion, including sham biopsies in case of negative mpMRI. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: Clinically significant PCa (csPCa), defined as group ≥2 on the International Society of Urological Pathology grading, was detected. RESULTS AND LIMITATIONS: The overall prevalence of csPCa, irrespective of biopsy technique, was 37.4% (132/353) in our population. MRI-TBx were performed in 263/353 (74.5%) patients with suspicious mpMRI (Prostate Imaging Reporting and Data System [PI-RADS] ≥3). The detection rates for csPCa were 39.5% for MRI-TBx and 42.9% for systematic biopsies. The added values, defined as the additional percentages of patients with csPCa detected by adding one biopsy technique, were 8.7% for the systematic biopsies and 5.3% for MRI-TBx. In patients with nonsuspicious mpMRI, five cases (6%) of csPCa were found by systematic biopsies. CONCLUSIONS: This study in biopsy-naïve patients suspected for PCa showed that systematic biopsies have added value to MRI-TBx alone in patients with mpMRI PI-RADS >2. PATIENT SUMMARY: We studied magnetic resonance imaging (MRI)-guided prostate biopsy for diagnosing prostate cancer and compared it with the standard method of prostate biopsy. Standard systematic biopsies cannot be omitted in patients with suspicious MRI, as they add to the detection of significant prostate cancer