Bone mass in women with celiac disease: role of exercise and gluten-free diet

Background and aimCeliac patients report fatigue and reduced social activities, which may decrease physical activity. This study investigated the physical activity, fatigue and bone mineral density in celiac women at diagnosis and during diet.Materials and methodsThe first group (n = 48) had the bone mineral density measured at diagnosis and after 2 years of a gluten-free diet; in the second group (n = 47) bone mineral density was measured at diagnosis and after 5 years of a gluten-free diet. Both groups completed a physical activity questionnaire and visual analogue scale for the perception of fatigue at diagnosis and follow-up. Data about smoking habits, alcohol use, presence of gastrointestinal symptoms, drug therapy and body mass index were also collected.ResultsAt diagnosis, the two groups were similar for all considered variables. At follow-up, the mean body mass index and physical activity questionnaire's score were similar to baseline. The bone density increased in both groups, whilst the physical activity questionnaire and visual analogue scale remained unchanged.ConclusionThe improvement in bone density following a gluten-free diet was significant after 2 years; physical activity is frequently low and plays a minor role in determining the changes in bone mineral density.     

Synthetic progress toward the marine natural product zamamiphidin A

An asymmetric synthetic approach to the octahydrofuro[3,4-b]pyridine framework of marine natural product zamamiphidin A has been described. The key steps include an asymmetric Michael addition of (R)-N-tert-butanesulfinyl imidate with enamidomalonate to install the C10 stereocenter, an intramolecular alkoxide exchange/Michael addition/hydrogenation sequence to construct the bicyclic ring system.

An asymmetric synthetic approach to the octahydrofuro[3,4-b]pyridine framework of marine natural product zamamiphidin A has been described.

Marine natural products have been recognized as an important source of potential lead compounds.¹ Recently, the growing attention on innovative drug discovery has propelled the identification of numerous marine-derived bioactive compounds with superior chemical novelty.² In 2013, Kobayashi and co-workers isolated a new macrocyclic diamine alkaloid zamamiphidin A (1, Fig. 1A) from an Okinawan marine sponge Amphimedon sp. (SS-1231),³ along with its biogenetically related manzamine alkaloids such as ircinic acid (2) and manzamine A (3). In the isolation work, the authors also demonstrated that compound 1 exhibited moderate antibacterial activity against Staphylococcus aureus (MIC, 32 μg mL⁻¹).³ Architecturally, zamamiphidin A was characterized by an unprecedented heptacyclic framework, comprising a highly fused pentacyclic caged core, two 11-membered rings and seven contiguous stereocenters including one all-carbon quaternary center. More specifically, the fusion of 7-oxa-2-azabicyclo[2.2.1]heptane (4, Fig. 1B), azabicyclo[3.3.1]nonane (5) and octahydrofuro[3,4-b]pyridine (6) fragments as well as the presence of quaternary ammonium salt render zamamiphidin A unique among other macrocyclic diamine alkaloids.⁴ To the best of our knowledge, the chemical synthesis of 1 has yet to be disclosed. Herein, we report our recent synthetic efforts toward zamamiphidin A (1).Open in a separate windowFig. 1Structures of (A) zamamiphidin A (1), its related manzamine alkaloids (2 and 3) and (B) fragments 4–6 of 1.From a retrosynthetic perspective (Scheme 1), we envisioned that zamamiphidin A would arise from the advanced intermediate 7 through late-stage formation of two 11-membered rings.^4,5 Central to the synthesis of the target molecule (1) would be the construction of the pentacyclic and cage-like backbone 7. Structural analysis suggested the diazapentacyclic core of 7 could be simplified as a fully substituted tetrahydrofuran moiety containing two amine functionalities at C3 and C10 (Scheme 1, in blue), which thus could be traced back to the precursor 10. Specifically, disconnecting the C1–N2 bond in 7 revealed ketone 8; and the azabicyclo[3.3.1]nonane unit in 8 could be formed by a Mannich reaction and an intramolecular Dieckmann condensation from bicycle 9. Disassembly of the piperidine ring in 9 through an intramolecular Michael addition led back to the functionalized tetrahydrofuran 10. Based on our previous studies,⁶10 could be easily traced back to the chiral malonate 12 bearing a pendant N-tert-butanesulfinyl imidate motif, relying on an intramolecular alkoxide exchange to forge the tetra-hydrofuran ring (11 to 10) and a Davis oxidation to install the C9 hydroxyl group (12 to 11). In turn, 12 could be accessed via a diastereoselective Michael addition of N-tert-butanesulfinyl imidate 13 to enamidomalonate 14.Open in a separate windowScheme 1Retrosynthetic analysis of zamamiphidin A.Our synthetic approach started from the preparation of the chiral malonate 12 (Scheme 2). First, condensation of the known primary amine 15⁷ with dimethyl methoxymethylenemalonate (16) in refluxing toluene delivered enamine 17 (92% yield), which was subsequently protected with Boc to give compound 14. The key asymmetric Michael addition of (R)-N-tert-butanesulfinyl imidate 13 to enamidomalonate 14 proceeded smoothly by employing LiHMDS as base,^6,8–10 providing adduct 12 as the major diastereomer in 72% yield (d.r. = 7 : 1 at C10). We supposed that the addition would preferentially take place through Ts-18 to avoid the significant steric repulsion between the tert-butyl group of 13 and the N-Boc side chain of 14, thus favoring the generation of 12. The configuration of the newly generated C10 stereocenter in 12 could be confirmed later by further transformations (vide infra). As far as we knew, this example represents the first asymmetric conjugate addition of N-tert-butanesulfinyl metalloenamines by using enamidomalonate as the Michael acceptor, which would be useful for the preparation of β-amino acid.¹¹Open in a separate windowScheme 2Synthesis of the chiral malonate 12.With malonate 12 available, we next sought to construct the octahydrofuro[3,4-b]pyridine ring system of zamamiphidin A (Scheme 3). To this end, our first task was to introduce a hydroxyl group at C9 in 12. After exploring various oxidative conditions such as Mn(OAc)₃/AcOH,¹² IBX/DMSO/H₂O,¹³ and O₂/I₂/NaOAc,¹⁴ we found that a Davis protocol¹⁵ (Davis'' oxaziridine/NaH/THF, −50 °C) was successful to convert 12 to 11 with 82% yield. Formation of the tetrahydrofuran ring from 11 proceeded in the presence of Et₃N/toluene at reflux temperature via an intramolecular alkoxide exchange, resulting in 19 (71% yield). Desilylation of 19 with HF afforded the primary alcohol 20 (85% yield). The latter then underwent a two-step synthetic sequence including oxidation of the hydroxyl group to aldehyde and Wittig olefination of the resulting aldehyde with methyl(triphenylphosphoranylidene)acetate (21), providing enoate 10 in 64% overall yield. Upon treating 10 with NaH in the presence of HMPA at −20 °C, an intramolecular Michael addition reaction occurred to enable the coupling of C4 and C4a, leading to the formation of the bicyclic intermediate 22 (72% yield). The structure of 22 was determined through extensive interpretation of the NMR data of its deprotected derivative 23. Unfortunately, we found that the configuration of the newly formed C4a stereocenter was opposite to that in the natural product zamamiphidin A. Inversion of the C4a stereochemistry through generation of the C4–C4a double bond (22 to 24) followed by hydrogenation (24 to 25) was investigated. When compound 22 was treated with LDA and PhSeCl followed by oxidation and spontaneous elimination under the conditions of H₂O₂/pyridine/CHCl₃, the above reaction gave a complex mixture, with only 5–10% yield of the desired product 24 isolated. We attempted to improve this transformation by employing various bases (e.g., LiHMDS, NaHMDS, KHMDS, LiTMP) and different oxidants (e.g., m-CPBA, NaIO₄, AcOOH), which all proved to be unsuccessful. These failures impeded further transformations.Open in a separate windowScheme 3Synthesis of the tricyclic compound 28.On the other hand, we investigated the construction of the oxazolidine ring starting from the bicyclic intermediate 22 (Scheme 3). Thus, reduction of the imine double bond in 22 with borane dimethyl sulfide afforded sulfinamide 26 in 55% yield. Subjecting 26 to the conditions of HCl/MeOH¹⁶ followed by treatment of the resulting primary amine with 2-hydroxypyridine in refluxing toluene¹⁷ furnished an unexpected tricyclic compound 28. The structure of 28 was determined unambiguously through X-ray crystallographic analysis, which again confirmed the C4a stereochemistry generated in the aforementioned intramolecular Michael addition reaction (10 to 22). Formation of the undesired 28 was due to the lactamization occurred at C18 rather than at C1 that was initially envisaged to establish the oxazolidine ring. These results indicated that the correct configuration of C4a might be crucial for the subsequent synthesis.In our revised synthetic approach, a propiolate group was employed to replace the originally used enoate for the Michael addition. We envisioned that hopefully reduction of the double bond generated in the Michael addition of the corresponding propiolate would secure the right configuration at the C4a position. As shown in Scheme 4, the known propargyl alcohol 29 was prepared on a decagram scale from 3-butyl-1-ol over two steps.¹⁸ After acylation of 29 and subsequent deprotection of the TBS group with TBAF, the resulting primary alcohol 30 was converted into phthalimide 31via a Mitsunobu reaction in the presence of phthalimide/PPh₃/DIAD (64% overall yield for three steps). Deacylation of 31 followed by silylation of the resulting primary alcohol with TBSCl/Et₃N/DMAP provided 32 in 36% yield over two steps. Exposure of 32 to hydrazine hydrate in refluxing methanol resulted in removal of the phthalimide group and delivered a free amine, which was directly condensed with 16 to afford the secondary enamine 33 in 80% overall yield. Further protection of the amino group in 33 with Boc produced intermediate 34. Next, compound 34 underwent a similar three-step transformation to that performed on enamidomalonate 14, involving asymmetric 1,4-conjugate addition with imidate 13, Davis oxidation to install the C9 hydroxyl group (35 to 36), and cyclization with Et₃N to assemble the tetrahydrofuran ring, yielding the intermediate 37 smoothly. To reach propiolate 39, the precursor of the key Michael addition for forming C4–C4a bond, further functional group manipulations of 37 were conducted. These included desilyation with HF (37 to 38), Dess–Martin oxidation, Pinnick oxidation, and esterification (38 to 39). Gratifyingly, upon treating 39 with LDA and HPMA in THF at −20 °C, compound 24 was formed with 42% yield through an intramolecular Michael addition and spontaneous migration of the double bond from C4a–C8a to C4–C4a.Open in a separate windowScheme 4Synthesis of compound 24.With compound 24 in hand, selective reduction of the C4–C4a double bond was carried out under catalytic hydrogenation conditions (RANEY® Ni/H₂) and furnished 25 (70% yield) as a single diastereomer with correct C4 and C4a configurations (Scheme 5). Ensuing reduction of the imine group in 25 using borane dimethyl sulfide delivered the tert-butyl sulfonamide 40 with excellent diastereoselectivity (single isomer, 72% yield). The structures of both 25 and 40 were extensively interpreted by NMR spectroscopy. As a consequence, the configurations at C3, C4, C4a and C10 were found to be consistent with the ones in zamamiphidin A.¹⁹ Furthermore, transformations of 25 or 40 into the core of the target zamamiphidin A were investigated. Subjecting 25 to various conditions (NaH, NaOMe, t-BuOK, LDA, LiHMDS, AlCl₃, TiCl₄, etc.) for a Dieckmann condensation failed to construct the C1–C8a bond and give the desired product 41. Instead, ring-opening of the tetrahydrofuran unit in 25 was observed under some circumstances, leading to the byproduct 42via a retro-Michael reaction.Open in a separate windowScheme 5Synthesis of 25 and attempted further transformations.Meanwhile, attempts to synthesize the tricyclic β-keto ester 43 or lactam 44 from compound 40 were also unsuccessful. Most of these experiments suffered from either no reaction or decomposition of the starting materials.In conclusion, we have established a synthetic approach to the octahydrofuro[3,4-b]pyridine core of the complex natural product zamamiphidin A. The synthesis features an intermolecular asymmetric Michael addition (compounds 34 to 35) to form the first chiral center at C10 and a Michael addition/hydrogenation sequence (compounds 39 to 25) to secure the correct configurations at C4 and C4a. The highly functionalized bicyclic intermediate (i.e., 40) prepared in the present work contains four contiguous stereocenters and all the requisite heteroatoms at the right positions. While further transformations of related intermediates to zamamiphidin A were unfruitful, efforts to develop a feasible strategy for total synthesis of the target molecule are ongoing in our laboratory.     

Simulation-guided nanofabrication of high-quality practical tungsten probes

Micro/nanoscale tungsten probes are widely utilized in the fields of surface analysis, biological engineering, etc. amongst several others. This work performs comprehensive dynamic simulations on the influences of electric field distribution, surface tension and the bubbling situation on electrochemical etching behaviors, and then the tip dimension. Results show that the etching rate is reliant on the electric field distribution determined by the cathode dimension. The necking position lies in the meniscus rather than at the bottom of the meniscus. A bubble-free condition is mandatory to stabilize the distribution of OH⁻ and WO₄²⁻ ions for a smooth tungsten probe surface. Such simulation-guidance enables the nanofabrication of probes with a high aspect ratio (10 : 1), ultra-sharp tip apex (40 nm) and ultra-smooth surface. These probes have been successfully developed for high-performance application with Scanning Tunneling Microscopy (STM). The acquired decent atomic resolution images of epitaxial bilayer graphene robustly verify the feasibility of the practical level application of these nanoscale probes. Therefore, these nanoscale probes would be of great benefit to the development of advanced analytical science and nano-to-atomic scale experimental science and technology.

Dynamic simulation is employed to reveal the mechanism of electrochemical nanofabrication of nanoscale probes for atomic resolution imaging in STM.     

Positive end-expiratory pressure during laparoscopy: cardiac and respiratory effects

Study ObjectiveTo determine the effect of positive end-expiratory pressure (PEEP) on the respiratory system and on cardiac function.DesignProspective randomized study.SettingOperating room.Patients60 ASA physical status 1 women scheduled for pelvic laparoscopic surgery.InterventionsPatients were ventilated normally during surgery; PEEP was modified depending on patient group allocation. Group A was the control group and did not receive PEEP. Group B received PEEP 5 cmH₂O and Group C received PEEP 10 cmH₂O.MeasurementsRespiratory parameters measured were partial pressure of arterial oxygen (PaO₂), partial pressure of carbon dioxide (PaCO₂), and end-tidal carbon dioxide tension (ETCO₂). Cardiac parameters measured were left ventricular end-diastolic volume index (LVEDVI), ie, ratio of LVEDV/body surface area (BSA; [LVEDVI = end-diastolic volume [EDV]/BSA); left ventricular (LV) systolic function, tricuspid annular plane systolic excursion (TAPSE), right ventricular (RV) fractional area change (FAC), RV dimensions in the apical 4-chamber view, tracing basal and mid-cavity minor dimensions and longitudinal dimension, cardiac index, systolic pulmonary artery pressure (PASP), and systolic RV pressure (RVSP). Respiratory and cardiac measurements were recorded at T0 (baseline); T1 (after anesthesia induction, before pneumoperitoneum induction); at 10 (T2), 20 (T3), and 30 (T4) minutes after CO₂ insufflation; and at the end of surgery (T5).Main ResultsVentilation with PEEP at 10 cm H₂O led to significant improvement in both respiratory and cardiac parameters. A reduction in pulmonary vascular resistance and enhanced washout of expiratory CO₂ occurred. Ten and, to a lesser extent, 5 cm H₂O of PEEP decreased LV stroke work.ConclusionsVentilation with PEEP (up to 10 cm H₂O) recruits the hypoventilated areas of the lungs and reduces cardiac afterload.     

Adipose-derived stem cells and rabbit bone regeneration: histomorphometric, immunohistochemical and mechanical characterization

Background

In the last few years, several attempts have been made to treat large bone loss, including the use of tissue engineering with osteoinductive scaffolds and cells. This study highlights the role of mesenchymal stem cells from adipose tissue (ASCs; adipose-derived stem cells) in a rabbit bone regeneration model.

Methods

We compared the neoformed bone tissues achieved by treating critical tibial defects with either hydroxyapatite alone (HA, group I) or hydroxyapatite–autologous ASC constructs (ASCs-HA, group II), investigating their histomorphometric, immunohistochemical and biomechanical properties.

Results

After eight weeks of follow-up, we observed advanced maturation and a spatial distribution of new bone that was more homogeneous in the inner parts of the pores in group II, not just along the walls (as seen in group I). The new tissue expressed osteogenic markers, and biomechanical tests suggested that the newly formed bone in group II had a higher mineral content than that in group I. Although variability in differentiation was observed among the different cell populations in vitro, no differences in bone healing were observed in vivo; the variability seen in vitro was probably due to local microenvironment effects.

Conclusions

Tibial defects treated with rabbit ASCs-HA showed an improved healing process when compared to the process that occurred when only the scaffold was used. We suggest that implanted ASCs ameliorate the bone reparative process either directly or by recruiting resident progenitor cells.