Search

ABSTRACTIntroductionSunitinib still represents a milestone in the treatment for progressive, well-differentiated, advanced panNETs.Areas coveredWe performed an evidence reappraisal to critically discuss its safety profile. We included nine studies, five clinical trials and four real-world (RW) studies. Within non-real-world (NRW) studies, diarrhea was the most frequent clinical AE. With regard to G3-4 AEs, fatigue and hypertension were the two most frequent, while neutropenia was the most recurrent hematological one. Considering four real-world trials, hand-foot-syndrome (HFS) was the most frequent clinical any-grade AE of any grade and neutropenia was the most common G3-4. Alongside to the AEs rate, the discontinuation rate of sunitinib due to TRAEs was variable among all the nine selected studies, ranging from 10% to 35% in the NRW setting and from 7% to 31% in the RW setting. Conversely, temporary interruption is an accepted strategy to reduce toxicity, even though not specifically tested in pan-NET.Expert opinionTill now, sunitinib continues to be one of the main therapeutic options for patients with well differentiated advanced panNETs, potentially covering any line of treatment. Therefore, tolerability plays a crucial role to increase adherence to therapy and maximize QoL.

Purpose: Grade 3 neuroendocrine tumor (NET G3) is a novel pathologic category within gastro-entero-pancreatic (GEP) neuroendocrine neoplasms (NENs) but its clinical behavior and therapeutic management still remain challenging. Prognostic and predictive factors aiding NET G3 management are needed. Patients and methods: We performed a retrospective analysis from 2015 to 2020 of all patients with > 20% Ki-67, well-differentiated NETs evaluated within our NEN-dedicated multidisciplinary team. We divided the sample according the timing of NET G3 diagnosis, the radiotracers distribution and Ki-67. We analyzed the correlation between these NET G3 features and clinical outcomes. Results: Among 3238 multidisciplinary discussion reports, we selected 55 patients, 48 from GEP and 7 from an occult GEP origin. In 45 patients, NET G3 diagnosis occurred at the beginning of clinical history (upfront-NET G3), whereas in 10, during the NET G1-G2 clinical history (late-NET G3). Conclusion: Our findings suggested that Ki-67 cutoff, functional imaging and the timing to NET G3 diagnosis may help clinicians in more accurate selection of NET G3 management. Prospective studies are needed.

Primary neuroendocrine tumors of the thymus are extremely rare. In patients with advanced disease, tumor growth control, and sometimes also syndrome control are the main goals of systemic therapy. Unfortunately, no standard therapies are available in clinical practice; therefore, clinical studies are strongly recommended. Axitinib (AXI) is a tyrosine kinase inhibitor, currently under investigation in an international phase II/III trial including thymic neuroendocrine tumors. Over the past 5 months, the entire world has been facing a devastating medical emergency brought about by a pandemic due to a novel coronavirus known as severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), which emerged in Wuhan, China, in late 2019. Since then, health professionals have been expending all their efforts on trying to provide the best available treatments for patients involved. Patients with cancer, especially those with thoracic involvement, are at higher risk of coronavirus disease 19 (COVID-19) and its complications because of their immunosuppressive status caused by the cancer and the anticancer therapies. As it remains unclear how to optimally manage such patients, we wished to report our experience with a patient with a metastatic neuroendocrine tumor of the thymus infected with SARS-CoV-2 in the hope that it may provide some insights and reflections on the management of cancer patients during this challenging time in our history.

The Southern Ocean occupies 14% of the Earth’s surface and plays a fundamental role in the global carbon cycle and climate. It provides a direct connection to the deep ocean carbon reservoir through biogeochemical processes that include surface primary productivity, remineralization at depth and the upwelling of carbon-rich water masses. However, the role of these different processes in modulating past and future air–sea carbon flux remains poorly understood. A key period in this regard is the Antarctic Cold Reversal (ACR, 14.6–12.7 kyr bp), when mid- to high-latitude Southern Hemisphere cooling coincided with a sustained plateau in the global deglacial increase in atmospheric CO2. Here we reconstruct high-latitude Southern Ocean surface productivity from marine-derived aerosols captured in a highly resolved horizontal ice core. Our multiproxy reconstruction reveals a sustained signal of enhanced marine productivity across the ACR. Transient climate modelling indicates this period coincided with maximum seasonal variability in sea-ice extent, implying that sea-ice biological feedbacks enhanced CO2sequestration and created a substantial regional marine carbon sink, which contributed to the plateau in CO2during the ACR. Our results highlight the role Antarctic sea ice plays in controlling global CO2, and demonstrate the need to incorporate such feedbacks into climate–carbon models.

Low atmospheric carbon dioxide (CO2) concentration during the Little Ice Age has been used to derive the global carbon cycle sensitivity to temperature. Recent evidence confirms earlier indications that the low CO2was caused by increased terrestrial carbon storage. It remains unknown whether the terrestrial biosphere responded to temperature variations, or there was vegetation re-growth on abandoned farmland. Here we present a global numerical simulation of atmospheric carbonyl sulfide concentrations in the pre-industrial period. Carbonyl sulfide concentration is linked to changes in gross primary production and shows a positive anomaly during the Little Ice Age. We show that a decrease in gross primary production and a larger decrease in ecosystem respiration is the most likely explanation for the decrease in atmospheric CO2and increase in atmospheric carbonyl sulfide concentrations. Therefore, temperature change, not vegetation re-growth, was the main cause of the increased terrestrial carbon storage. We address the inconsistency between ice-core CO2records from different sites measuring CO2and δ13CO2in ice from Dronning Maud Land (Antarctica). Our interpretation allows us to derive the temperature sensitivity of pre-industrial CO2fluxes for the terrestrial biosphere (γL= −10 to −90 Pg C K−1), implying a positive climate feedback and providing a benchmark to reduce model uncertainties.

Imidazoles and benzimidazoles are privileged heterocyclic bioactive compounds used with success in the clinical practice of innumerous diseases. Although there are many advancements in cancer therapy, microtubules remain as one of the few macromolecular targets validated for planning active anti-cancer compounds, and the design of drugs that modulate microtubule dynamics in unknown sites of tubulin is one of the goals of the medicinal chemistry. The discussion of the role of new and commercially available imidazole and benzimidazole derivatives as tubulin modulators is scattered throughout scientific literature, and indicates that these compounds have a tubulin modulation mechanism different from that of tubulin modulators clinically available, such as paclitaxel, docetaxel, vincristine and vinblastine. In fact, recent literature indicates that these derivatives inhibit microtubule formation binding to the colchicine site, present good pharmacokinetic properties and are capable of overcoming multidrug resistance in many cell lines. The understanding of the mechanisms involved in the imidazoles/benzimidazoles modulation of microtubule dynamics is very important to develop new strategies to overcome the resistance to anti-cancer drugs and to discover new biomarkers and targets for cancer chemotherapy.

The objective of this study is to investigate bone metabolism, density, and quality in patients with diabetes type 2 using DEXA and spinal deformity index (SDI), a surrogate index of bone quality. Fifty-six patients with type 2 diabetes were studied; exclusion criteria were diseases and medications that affect bone and mineral metabolism. Mean age was 65 ± 7 years. Mean diabetes duration was 10 ± 7 years and mean HbA1C was 6.6 ± 0.5 %. BMI was 30 ± 4. Fifty-six sex, age, and BMI matched served as controls. All subjects underwent a clinical and biochemical examination. Spinal and femoral neck BMD were measured by DEXA, and a spine radiography was performed to assess vertebral fractures and to calculate SDI. Mean serum 25-OH vitamin D levels were 19.6 ± 3.7 ng/ml in patients and 30 ± 14 ng/ml in controls (p< 0.01). PTH serum levels were 47.9 ± 40 pg/ml in patients versus 37 ± 5.3 pg/ml in controls (p< 0.01). At lumbar spine there was a significant difference between patients and controls only for T-score (p= <0.01), while at femoral neck there was a difference in BMD (p< 0.01) and in T-score (p< 0.01). Radiological vertebral fractures were found in 46 % of patients and 17 % of controls (p< 0.05). SDI was higher in patients than in controls (p< 0.05). The percentage of fractures with T-score BMD greater than −2.5 was 69 % in patients and 10 % in controls (p< 0.05). As a conclusion, BMD was similar in patients and in controls, while SDI value was higher in patients; therefore, SDI was more specific than BMD for the diagnosis of osteoporosis due to metabolic diseases.

The type I receptor tyrosine kinases (RTKs) are involved in various aspects of cell growth, survival, and differentiation. Among the known RTKs, the epidermal growth factor receptor (EGFR) and ErbB-2 (HER-2) are two widely studied proteins that are prototypic members of the ErbB family which also includes ErbB-3 (Her-3) and ErbB-4 (Her-4). Overexpression of ErbB-2 and EGFR has been associated with aggressive disease and poor patient prognosis in a range of human tumour types (e.g. breast, lung, ovarian, prostate, and squamous carcinoma of head and neck). Disruption of signal transduction of these kinases has been shown to have an antiproliferative effect. Various approaches have been developed to target the ErbB signalling pathways including monoclonal antibodies (trastuzumab/ Herceptin™ and cetuximab/Erbitux™ ) directed against the receptor, and synthetic tyrosine kinase inhibitors (gefitinib/Iressa™ and erlotinib/Tarceva™). Since many tumours overexpress ErbB receptors, simultaneous targeting of multiple ErbB receptors therefore becomes a promising approach to cancer treatment. Lapatinib (Tykerb™), a potent dual EGFR/ErbB-2 inhibitor, was approved for the treatment of ErbB-2-positive breast cancer. Despite years of intensive research on EGFR inhibitors, there is a surprising dearth of chemically distinct small inhibitors with a high degree of selectivity. There is also a need for new scaffolds due to the recent finding of EGFR mutations which render the kinase resistant to gefinitib and erlotinib. The structures under study will be quinazolines with different substituents. The structure-activity relationships and biological evaluation of compounds published during the last four years will be reviewed herein.

The purine ring is undoubtedly among the most ubiquitous of all the heterocyclic compounds. This arises not only from the universal occurrence of adenine and guanine in DNA and RNA and of additional modified derivatives in the various tRNAs but also from the subsidiary uses of the ring system in a great number of biochemical systems. The development of new methods for metal-mediated coupling with aryl or heteroaryl halide substrates has expanded the range of synthetically accessible arylpurine derivatives. Aryl boronic acids have proved to be extremely available reagents for metal-mediated C-C and C-N coupling reactions. Coupling reactions resulting in C-C bond formation are catalyzed by palladium and nickel catalysts at positions C2, C6 and C8. The reactivity at C6 position has been demonstrated using fluoro-, chloro-, bromo-, iodo-, sulfanyl, sulfonyl, sulfonyloxy- and azole-substrates. The ease of obtaining the activated purine substrates is an important factor when selecting appropriate coupling conditions. Copper-mediated N-arylations occur at positions N7 and N9. These methods are also applicable using solid-supported purine substrates and provide convenient access to structurally unique derivatives with applications in drug discovery. Continuing advances in this field can be expected to result through improved mechanistic understanding and the development of new catalysts and ligands.

The application of the neighboring-group participation involving the oxygen atom of O,O- or O,N-acetals can be very fruitful. For instance, the naphthoate ester derivatives of (Z)-hex-3-ene-1,5-diyne were used to generate biradicals via -oxo ketene acetal intermediates, and a synthesis of tricyclic 9-crown-3 ethers bearing a chiral oxathiane ring was achieved by utilizing nucleophilic displacement of a triflic ester leaving-group assisted by neighbouring-group participation of a 1,3-dioxolane function. In a different field, the reaction between o-(hydroxymethyl)phenoxyacetaldehyde dimethyl acetals, or (/-)-3-methoxy-2,3-dihydro-5H-1,4-benzodioxepins with 5-fluorouracil was studied. The intramolecular cyclization may be explained through a neighbouring-group attack to give a 2-(5- fluorouracil-1-yl)oxyranium ion that can be attacked by the silylated benzylic hydroxyl group to yield the benzannelated sevenmembered O,N-acetals. Before carrying out the synthesis of 7- or 9-substituted (/-)-2-methoxy-2,3-dihydro-5H-1,4-benzodioxepins it is necessary to protect the phenolic hydroxy group of the 2-hydroxybenzyl alcohol. Among other functionalities, the 2- methoxyethoxylmethyl (MEM) group was developed as a protective group of alcohols and phenols. Accordingly, it was decided to use the MEM group for the preparation of 7- or 9-substituted (/-)-2-methoxy-2,3-dihydro-5H-1,4-benzodioxepins and their ten-membered benzo-fused analogs. The importance of the ten-membered O,O-acetals sheds light on the mechanism of reaction in which the neighbouring- group participation plays a pivotal role. Transannular versions of the reaction result in the facile ring contraction of 12-membered intermediates to the 10- and to 7-membered benzene-fused O,O-acetals. Acetals with several oxygen atoms in their side-chains have been used in the preparation of highly oxygenated cyclic ether compounds.

The issue of drug chirality is now a major theme in the design and development of new drugs, underpinned by a new understanding of the role of molecular recognition in many pharmacologically relevant events. In general, three methods are utilized for the production of a chiral drug: the chiral pool, separation of racemates, and asymmetric synthesis. Although the use of chiral drugs predates modern medicine, only since the 1980’s has there been a significant increase in the development of chiral pharmaceutical drugs. An important commercial reason is that as patents on racemic drugs expire, pharmaceutical companies have the opportunity to extend patent coverage through development of the chiral switch enantiomers with desired bioactivity. Stimulated by the new policy statements issued by the regulatory agencies, the pharmaceutical industry has systematically begun to develop chiral drugs in enantiometrically enriched pure forms. This new trend has caused a tremendous change in the industrial small- and large-scale production to enantiomerically pure drugs, leading to the revisiting and updating of old technologies, and to the development of new methodologies of their large-scale preparation (as the use of stereoselective syntheses and biocatalyzed reactions). The final decision whether a given chiral drug will be marketed in an enantiomerically pure form, or as a racemic mixture of both enantiomers, will be made weighing all the medical, financial and social proficiencies of one or other form. The kinetic, pharmacological and toxicological properties of individual enantiomers need to be characterized, independently of a final decision.

Photooxygenation of non-aromatic heterocycles and cyclic compounds containing non-usual heteroatoms, namely silicon, germanium and tellurium has been reviewed. All three types of photooxygenation (Types I-III) can take place. Moreover the heteroatom can be frequently involved endorsing electron-transfer reactions which turn out to be the main pathways, even in singlet oxygen oxygenation. A vast collection of novel and unexpected products are often formed, sometimes in a stereocontrolled manner.

A systematic analysis of efficiency, reproducibility and accuracy of cryogenic purification of CO2 from air samples for isotopic analyses is presented. The technical characteristics of the cryogenic line are given in detail. To study the cryogenic process, three different operating parameters are considered: flow rate of the gas entering the line, pressure of the gas in the line, and CO2‐trap shape. Experimental results demonstrate that efficiency, reproducibility and accuracy strongly depend on the CO2trap shape. Moreover, a dependence of reproducibility and accuracy on the flow rate of the gas is found, but not on its pressure. High precision (≤0.02‰ for δ13C and ≤0.05‰ for δ18O) and good accuracy (<0.09‰ for δ13C and <0.14‰ for δ18O) is achieved after applying the N2O correction. Copyright © 2005 John Wiley & Sons, Ltd.

Surgical management of congenital malformation of the mitral valve (MV) in the pediatric age group remains a therapeutic challenge for the wide spectrum of the morphological abnormalities and the high incidence of associated cardiac anomalies. We reviewed our experience so as to assess whether MV conservative surgery is always advisable and its results are superior to MV replacement.

The molecular mechanisms ensuring directionality of endocytic membrane trafficking between transport vesicles and target organelles still remain poorly characterized. We have been investigating the function of the small GTPase Rab5 in early endocytic transport. In vitro studies have demonstrated a role of Rab5 in two membrane fusion events: the heterotypic fusion between plasma membrane-derived clathrin-coated vesicles (CCVs) and early endosomes and in the homotypic fusion between early endosomes. Several Rab5 effectors are required in homotypic endosome fusion, including EEA1, which mediates endosome membrane docking, as well as Rabaptin-5 x Rabex-5 complex and phosphatidylinositol 3-kinase hVPS34. In this study we have examined the localization and function of Rab5 and its effectors in heterotypic fusion in vitro. We report that the presence of active Rab5 is necessary on both CCVs and early endosomes for a heterotypic fusion event to occur. This process requires EEA1 in addition to the Rabaptin-5 complex. However, whereas Rab5 and Rabaptin-5 are symmetrically distributed between CCVs and early endosomes, EEA1 is recruited selectively onto the membrane of early endosomes. Our results suggest that EEA1 is a tethering molecule that provides directionality to vesicular transport from the plasma membrane to the early endosomes.

Background Straddling tricuspid valve, despite extensive investigation, remains an incompletely understood form of complex congenital heart disease. Methods A morphometric study of 19 postmortem cases of straddling tricuspid valve was performed, and the results were compared with 32 normal control heart specimens. Results In straddling tricuspid valve, marked malalignment of the ventricles was always found relative to the atria. The angle between the ventricular septum and the atrial septum in the short-axis projection averaged 61^o +/- 24^o, the normal ventriculoatrial septal angle averaging 5^o +/- 2^o (P < .001). The right ventricular sinus (inflow tract) was significantly smaller than the left (P < .01). A ventricular septal defect was present in 79%: atrioventricular canal type in 42%, atrioventricular canal type confluent with a conoventricular defect in 26%, and a conoventricular defect in 11%. When the straddling tricuspid valve adhered to the crest of the muscular ventricular septum (n = 4 cases, 21%), the 2 salient findings were (1) an intact ventricular septum and (2) double-outlet right atrium. The nonstraddling part of the tricuspid valve opened into the small right ventricle. The straddling part of the tricuspid valve opened into the larger left ventricle. The mitral valve also opened into the left ventricle. Hence hearts with double-outlet right atrium had 3 atrioventricular valves. Congenital mitral stenosis was present in 26% of this series. Conclusion Straddling tricuspid valve was always characterized by marked ventriculoatrial malalignment, indicated by an abnormally large ventriculoatrial septal angle, best seen in the short-axis projection. (Am Heart J 1999;138:1184-95.)

Abstract:  To define factors predictive of failure to respond to nucleoside reverse-transcriptase inhibitors in human immunodeficiency virus type-1 (HIV-1)-infected subjects pretreated with zidovudine (ZDV), three groups of subjects shifted to double therapy with ZDV plus didanosine (ddI, n=13), zalcitabine (ddC, n=14), or lamivudine (3TC, n=12) were retrospectively evaluated, with respect to addition of the second NRTI, at week 0 and week 24. Factors considered included duration of ZDV pretreatment, CD4+ cell counts, plasma HIV-1 RNA load, peripheral blood mononuclear cell HIV-1 DNA load, and HIV-1 DNA genotypic resistance to nucleoside reverse-transcriptase inhibitors. The three groups were well matched for baseline characteristics and did not differ significantly in virological and immunological response to the different combination treatments. Drug-specific resistance mutations were selected in more than half the cases by 3TC, but not by ddI and ddC. Low-level and substantial genotypic resistance to ZDV was detected 13 (33.3%) and in 19 (48.7%) patients at baseline, respectively, and evolved through week 24 in several patients. When subjects were divided into responders and nonresponders to the second nucleoside reverse-transcriptase inhibitor on the basis of a decrease of more than 0.5 log10 (n=15) or less than 0.5 log10 (n=21) in HIV-1 RNA, respectively, baseline genotypic ZDV resistance was the only independent predictor of failure in a logistic regression model (P=0.003 or P=0.024, depending on whether low-level resistance was considered or not, respectively). Thus, selection of ZDV resistance mutations may impair subsequent use of different nucleoside reverse-transcriptase inhibitor compounds.

Abstract:: Although case series reports are the most frequent type of article published in surgical periodicals, their scientific value has recently been judged questionable for assessing the effectiveness of a treatment. It has also been suggested that strict rules, such as those published for randomised controlled trials (the CONSORT statement) could improve their quality. Forty papers on surgery for colorectal tumours were reviewed and a check list of items considered useful for reporting a case series was drawn up. Some items were borrowed from the CONSORT statement. The final list was checked against the 40 articles reviewed. Virtually all the published papers were defective in one or more aspects (selection of patients, details of clinical, pathological and technical data, methods of follow-up and statistical analysis). The quality of observational studies can be improved. Data collection is easier in prospective studies, and strict follow-up and appropriate statistical analysis may render a case series more reliable.

Anti-Jkawas detected by solid-phase red cell adherence(SPRCA) anti-body detection and identification tests in the plasma of a 9-month-old female intant during a roatine presurgical evaluation. the pation and her nonidentical twin sister who also had anti-Jkain her plasma, were products at an uncomplicated in vitro fertilization, full -term preganancy, and vaginal delivery Neither twin had been tranfered recently infected. or treated with medication. their mother had no prior pregnancies or transfusions. Red blood cells (RBCs) from the patient and her sister typed as Jk(a-b+) by direct hemagghutination and this phenotype was confirmed by negative adsorption and elution studies. Both infants plasma samples were strongly reactive with 20 examples of Jk(a+) RBCs and nonreactive with 20 examples of Jk(a-) RBCs by SPRCA assays. Anti-Jkawas not detected in either twins plasma by Indirect antiglobulin tests by tube method in lowionie-strength saline solution or polyethylene glycol, or with ficin- or papain-treated RBCs. Monocyte monolayer assays using Jk(a+) RBCs sensitized by either twins’ serum were nonreactive (0%). RBCs from both parents typed as Jk(a+b+). Both parents’ antibody detection test results by SPRCA assay were negative. The absence of a history of exposure to allogeneic RBCs or possible passive transfer of maternal or other alloantibody classifies these antibodies as natually-occurring anti-Jka. Immunohemalology1999;15: 159-162.

The ability of surgeons to make a reproducible and reliable evaluation of the nutritional status of patients was assessed. Three independent observers assigned 64 patients to one of four nutritional states — normal nourishment, and mild, moderate and severe malnutrition — using a questionnaire and clinical examination. Interobserver variability was analysed using the K statistic and reliability by assessing the correlation between the assigned class and total serum protein, haemoglobin, albumin, transferrin and cholesterol levels, weight loss and the results of immunological skin tests. Concordance was observed in 49 of the 64 judgements (77 per cent) and partial agreement in the remainder (K = 0·74, P < 0·001). Clinical judgement correlated significantly with albumin, transferrin and cholesterol levels and weight loss (R2= 0·73, P < 0·001). Surgeons are able to assess nutritional status using clinical judgement.

The contrasting results of treatment of patients with postoperative enterocutaneous fistulae reflect the heterogeneity of the disease and depend on the patient's condition and the characteristics of the fistulae. For this reason, the use of a prognostic index, which enables such patients to be classified according to their risk of death, could be useful. In this study we propose a prognostic index based on a logistic regression analysis, obtained by using two (APACHE II score and serum albumin concentration) of the eight risk factors that have been retrospectively analysed in a series of 70 patients with postoperative enterocutaneous fistulae treated in our surgical department since 1981. The logistic regression equation indicates that patients with a probability of dying of less than 0·35 have a good prognosis, with a sensitivity of 90 per cent, a specificity of 90 per cent, a negative predictive value of 79 per cent, a positive predictive value of 96 per cent and an accuracy of 90 per cent. The predictive performance of the index has also been evaluated in a group of 17 patients studied prospectively, and this confirms the sensitivity and specificity of the model. This postoperative enterocutaneous fistulae index could be a helpful tool in clinical trials and surgical audit.

Intralipid, a fat emulsion widely used in parenteral nutrition, can produce marked functional changes of the mononuclear phagocyte system. We investigated the effect of Intralipid administration on the generation of procoagulant activity by rabbit mononuclear phagocytes. Two groups of ten rabbits given either a single infusion of Intralipid 10% or a similar volume of sterile saline were studied before and after infusion. Procoagulant activity was measured on isolated blood mononuclear cells after incubation with and without endotoxin, using a one-stage clotting assay. Cells from animals infused with Intralipid produced significantly more procoagulant activity than controls (P less than .01). Results were similar when freshly collected whole blood was incubated with and without endotoxin, and procoagulant activity was measured on subsequently isolated mononuclear cells (P less than .01). In addition, when rabbits were given a single injection of endotoxin, blood and spleen mononuclear cells harvested 50 to 60 minutes after the injection from animals pretreated with Intralipid expressed five to seven times more procoagulant activity than did cells from animals pretreated with saline. In all instances, procoagulant activity was identified as tissue factor. These findings suggest that Intralipid may cause functional changes in mononuclear phagocytes, resulting in increased production of tissue factor on incubation in short-term culture in vitro and in response to endotoxin in vivo.

Microscopic features of lepromatoid lesions in a chimpanzee inoculated with bovine leukemia virus consisted of diffuse dermal infiltrations of foamy histiocytes, except for a subepidermal “clear zone” devoid of infiltrating cells. Acid-fast organisms were in histiocytes, dermal nerves and a branch of the left radial nerve. The organisms were more intensely acid-fast with the Fite-Faraco than with the Ziehl-Neelsen stain. The organisms did not oxidize 3,4-dihydroxyphenylalanine (dopa) to indole but their acid-fast staining quality was removed by pretreatment with pyridine. Human leprosy patients, inoculated simultaneously with chimpanzee and human lepromin, reacted similarly to both lepromins. Organisms, when inoculated into mouse footpads, multiplied in 6 months to a quantity that was compatible with their identification as Mycobacterium leprae.

In a preliminary cross‐sectional analysis of 109 human immunodeficiency virus type 1 (HIV‐1)‐infected subjects the presence of 2‐long terminal repeat (LTR) unintegrated circular HIV‐1 DNA in peripheral blood mononuclear cells (PBMC) was found to be associated with both symptomatic infection (P= 0.0037) and low CD4 counts (P= 0.0004). To investigate the prognostic significance of the presence of 2‐LTR HIV‐1 DNA, a subset of 23 2‐LTR‐negative and 25 2‐LTR‐positive asymptomatic individuals were followed up for 12–24 months. The two groups did not differ in terms of baseline CD4 counts, zidovudine (ZDV) therapy, and duration of HIV‐1 infection. Longitudinal analysis of CD4 values did not indicate a significantly different CD4 outcome between the two groups. However, when only ZDV‐treated subjects were considered, a significant (P= 0.042) decrease in CD4 counts was found at month 24 with respect to baseline in 2‐LTR‐positive (n = 12) but not in 2‐LTR‐negative (n = 11) patients. Moreover, when >40% CD4 loss from baseline and/or development of CDC stage B or C symptoms were considered as indicators of disease progression, there was a significantly higher number of events in the whole 2‐LTR‐positive group than in the whole 2‐LTR‐negative group (P= 0.0197 at month 12, P= 0.0299 at month 18, P= 0.0373 at month 24). Thus, the presence of 2‐LTR‐HIV‐1 DNA in PBMC merits further investigation as a simple, qualitative, molecular predictor of disease progression and decreased response to antiretroviral therapy. J. Med. Virol. 52:20–25, 1997. © 1997 Wiley‐Liss, Inc.

Abstract: Changes in sinapic acid when exposed to aqueous alkaline conditions were elucidated. Sinapic acid was exposed to a volatile buffer (pH 8.5) for 24 h, lyophilized, acidified, extracted, and characterized using nuclear magnetic resonance and mass spectroscopy. The product obtained was identified as the lignan thomasidioic acid. This identification was confirmed by comparison with a synthesized authentic sample of thomasidioic acid. Conversion of sinapic acid to thomasidioic acid under alkaline conditions previously has not been reported. Thomasidioic acid was present after exposure of sinapic acid to pH 8.5 for as few as 6 h. Thomasidioic acid also was formed at pH 7.

Previous studies from other laboratories, using rabbit reticulocyte lysate filtered through Sephadex G-25 or G-50, have demonstrated that glucose 6-phosphate is required to maintain active rates of translation, but its mechanism of action is currently unsettled. We have tested whether glucose 6-phosphate is required to prevent activation of the hemin-controlled translational repressor and the phosphorylation of the smallest or alpha subunit of eukaryotic initiation factor 2 (eIF-2). We have found that antibody to the hemin-controlled translational repressor can completely restore protein synthesis in reticulocyte lysate, filtered through Sephadex G-25, that is incubated in the absence of hemin and presence of glucose 6-phosphate, but cannot restore protein synthesis in such lysate incubated in the presence of hemin and absence of glucose 6-phosphate. We have also found, using a modification of the method of Matts and London [1984) J. Biol. Chem. 259, 6708-6711) to measure the ability of gel-filtered lysate to dissociate and exchange GDP from eIF-2.GDP, that this endogenous eIF-2B activity is reduced to the same low level in the presence of hemin and absence of glucose 6-phosphate as it is in the absence of hemin and presence of glucose 6-phosphate. Although there is a low level of phosphorylation of eIF-2 alpha in gel-filtered lysate given hemin but no glucose 6-phosphate, it cannot account for the loss of eIF-2B activity, since this phosphorylation is removed by antibody to the hemin-controlled translational repressor or isocitrate, which do not restore protein synthesis or eIF-2B activity, and not by fructose 1,6-diphosphate, which does partially restore protein synthesis and eIF-2B activity. These findings suggest that sugar phosphates may exert a direct effect on eIF-2B and may be required for its proper function. Additional support for this conclusion is our finding that protein synthesis and eIF-2B activity in partially hemin-deficient lysate can be restored by high levels of glucose 6-phosphate or fructose 1,6-diphosphate without a reduction in the level of phosphorylated eIF-2 alpha, suggesting that such levels of sugar phosphate may permit restoration of normal function with a limiting amount of eIF-2B.

Changes in sinapic acid when exposed to aqueous alkaline conditions were elucidated. Sinapic acid was exposed to a volatile buffer (pH 8.5) for 24 h, lyophilized, acidified, extracted, and characterized using nuclear magnetic resonance and mass spectroscopy. The product obtained was identified as the lignan thomasidioic acid. This identification was confirmed by comparison with a synthesized authentic sample of thomasidioic acid. Conversion of sinapic acid to thomasidioic acid under alkaline conditions previously has not been reported. Thomasidioic acid was present after exposure of sinapic acid to pH 8.5 for as few as 6 h. Thomasidioic acid also was formed at pH 7.

Recent observations have indicated that eukaryotic initiation factor (eIF)-2 and GTP or GDP normally bind to 60 S ribosomal subunits in rabbit reticulocyte lysate and that when eIF-2 alpha is phosphorylated and polypeptide chain initiation is inhibited, eIF-2 × GDP accumulates on 60 S subunits due to impaired dissociation that is normally mediated by the reversing factor (eIF-2B). Current findings now indicate that inhibition due to phosphorylation of eIF-2 alpha is mediated, at least in part, by the inability to dissociate eIF-2 × GDP from the 60 S subunit of complete initiation complexes. At the onset of inhibition, there is an accumulation of Met-tRNA(f) and eIF-2 on the polysomes, despite a marked reduction in Met-tRNA(f) bound to 40 S subunits and Met-peptidyl-tRNA bound to the polysomes. This initial effect is not associated with the formation of “half-mers” (polysomes containing an extra unpaired 40 S subunit), and the 40 S × Met-tRNA(f) complexes, though reduced, still sediment at 43 S. When inhibition is maximal and the polysomes are largely disaggregated, there is an accumulation of 48 S complexes consisting of a 40 S subunit and Met-tRNA(f) bound to globin mRNA as well as small polysomal half-mers, such that residual protein synthesis occurs to about the same degree on “1 1/2”s and “2 1/2”s as on mono-, di-, and triribosomes. Exogenous eIF-2B increases protein synthesis on mono-, di-, and triribosomes and decreases that on half-mers. This is associated with reduced binding of Met-tRNA(f) and eIF-2 to ribosomal particles sedimenting at 80 S and greater and a shift from 48 S to 43 S complexes. These results suggest that eIF-2B must normally promote dissociation of eIF-2 × GDP from the 60 S subunit of complete initiation complexes before they can elongate but cannot when eIF-2 alpha is phosphorylated, resulting in the accumulation of these complexes, some of which dissociate into Met-tRNA(f) × 40 S × mRNA and 60 S × eIF-2 × GDP.

The measurement of estrogen as well as progesterone receptors has been applied clinically to predict the effectiveness of endocrine therapy in patients with breast or endometrial carcinoma. The presence of cytosolic estrogen receptors in human colorectal carcinomas has been reported by several different groups during the past 10 yr. The aim of our current study was to evaluate the estrogen binding activity in the nuclear and cytosolic fractions of these carcinomas, as well as in surrounding noncancerous colonic tissue. Twenty-six patients, operated on for colorectal carcinoma, were studied: 16 were men and 10 were postmenopausal women, mean age 61 yr (range 43–78 yr). In neoplastic tissue, cytosolic estradiol receptors were detected in 42.3% of the patients (women 40%, men 43.7%). The values for cytosolic estrogen receptor ranged from 118 to 1214 fmol/g wet colon. Nuclear estrogen receptors were detectable in 46.1% of the patients (women 40%, men 50%) and their values displayed a range from 3.4 to 2554 fmol/g wet colon. In 30.7% of the patients, both nuclear and cytosolic receptors were demonstrated. In 38.4% of the patients, receptors were found in neither cytosol nor nuclei. Receptor positivity in men was most frequently associated with tumors removed from the rectum, and those with Dukes' classification of C1. In the surrounding noncancerous tissue cytosolic estrogen receptors were also detected (range 133–1105 fmol/g wet colon) and were present in 34.6% of the patients (women 30%, men 37.5%). Nuclear estrogen receptors (range 225–1105 fmol/g wet colon) were detected in 57.6% of the patients (women 40%, men 68.7%). In 23% of the patients, both nuclear and cytosolic receptors were demonstrated. In 30.7% of the patients, no receptors were found in either cytosol or nucleus. Therefore, the presence of cytosolic or nuclear estrogen receptors, or both, in 61.6% of human colorectal cancer specimens emphasizes the need to evaluate both forms of these receptors for studies of potential hormone dependence in these tumors.

To facilitate the preoperative diagnosis and surgical management of visceral heterotaxy and asplenia, 72 postmortem cases were reviewed with particular attention focused on the systemic and pulmonary venous connections. The superior vena cava was bilateral in 51 cases (71%), but in 9 cases one of the superior venae cavae was partly or totally atretic. Patent bilateral superior venae cavae were found in 42 cases (58%) and the superior vena cava was unilateral in 21 (29%). Although the inferior vena cava was never interrupted, a prominent azygos vein was found in 6 cases (8%). Some hepatic veins drained separately from the inferior vena cava in 20 cases (28%). An intact coronary sinus was rare (2 cases, 3%). Anomalous pulmonary venous connection to a systemic vein was total in 42 (58%) of 72 and partial in 2 (3%) of 72, with obstruction in 24 (55%) of 44. Abnormal pulmonary artery branches (severe hypoplasia, localized stenosis, or discontinuity) were present in 21 (29%), and these obstructive arterial anomalies were associated with a significantly higher prevalence of anomalous pulmonary venous connection (p < 0.01) and of pulmonary venous obstruction (p < 0.01). Cardiac pulmonary venous connections were found in 28 (39%), with the pulmonary veins and the inferior vena cava entering the same atrium in 10 (36%) of 28. (J T HORAC CARDIOVASC SURG 1995; 110: 641-50)

A retrospective study of 87 patients, subjected to colostomy closure between 1976 and 1987, was conducted in order to evaluate the role of 8 potential risk factors on morbidity and mortality. Possible risk factors were age >65 years, presence of hypoalbuminaemia (<3.0 gr%), anaemia (Hb<10 gr%), operative technique, duration of colostomy, site of colostomy, underlying disease and presence of subcutaneous drainage. Apart from hypoalbuminaemia, no clear risk factor was identified, although an interval of more than 90 days between construction and closure of colostomy appears to be safer than shorter intervals. A comparison was also made between two different periods from 1976 to 1982 and from 1983 to 1987 which resulted in important changes in patient management in the second period including: type of antibiotic prophylaxis, type of anastomosis and suture material, site of colostomy and mean duration of colostomy. Four post-operative deaths (4.6%) (two for myocardial insufficiency and two for sepsis), 11 major (13%) and 25 (29%) minor complications were recorded. The analysis of the two different periods showed a strong reduction in both mortality and morbidity in the second period, which could be related to a better management of this type of patient. In conclusion, the incidence of mortality and morbidity in colostomy closure cannot be underestimated and therefore the same skill and meticulous approach are required for this operation as for any major surgical procedure on the colon.

To determine the sensitivity of a nested PCR procedure for detecting human immunodeficiency virus type 1 DNA in clinical specimens, 553 peripheral blood mononuclear cell samples obtained from 268 human immunodeficiency virus type 1-seropositive subjects were assayed by use of two independent primer sets for each sample. Overall, 1,088 of 1,106 (98.37%) reactions were positive. Investigation of the negative reactions showed that a low viral burden in some infected subjects, rather than primer-template mismatches, was the primary cause for the false-negative PCR results.

A highly sensitive nested polymerase chain reaction (PCR) protocol was used to detect human immuno-deficiency virus type 1 (HIV-1) DNA in peripheral blood mononuclear cells from 271 HIV-1-seropositive patients, 240 HIV-1-seronegative subjects at increased risk for HIV-1 infection, 51 serologically indeterminate individuals, and 120 healthy blood donors. PCR was carried out in a multiplex nested configuration with pol and env region primer sets. HIV-1 DNA was detected in all of the HIV-1 seropositive patients. In contrast, HIV-1 DNA was not detected in any of the either seronegative or serologically indeterminate subjects. Only one of 37 seronegative regular sexual partners of HIV-1-infected patients who were followed longitudinally was found to seroconvert to HIV-1. However, HIV-1 DNA and antibody results were concordant in the four samples obtained from this subject prior to and after seroconversion. These results show an excellent concordance between HIV-1 DNA and antibody detection for diagnosis of HIV-1 infection and suggest that long-term HIV-1 infection in the absence of detectable antibody is likely to occur at a very low frequency.

We recently reported that the translational control of protein synthesis by glucose 6-phosphate in gel-filtered, rabbit recitulocyte lysate is exerted on the activity of eukaryotic initiation factor (eIF)-2B, the factor that catalyzes the exchange of GTP for GDP bound to eIF-2, by a mechanism that is independent of the phosphorylation of eIF-2 (α subunit). We now demonstrate that two other conditions regulate the activity of eIF-2B in rabbit reticulocyte lysate: polyamines (spermidine and spermine) and amino acid deficiency. In the absence of added polyamines, protein synthesis in gel-filtered lysate is reduced to about 70% and eIF-2B activity to about 35% of optimal. The former is likely a result of the latter, since we find that reticulocyte lysate has about twice the eIF-2B necessary to recycle the eIF-2.GDP generated under conditions of optimal protein synthesis. In contrast, the reduction in eIF-2B activity (to about 50% of optimal) occurring in the absence of added amino acids in unfractionated or gel-filtered lysate is insufficient, by itself, to slow the rate of protein synthesis, and the inhibition of protein synthesis that does occur with amino acid deficiency is exerted on polypeptide chain elongation, not initiation. The reduction in eIF-2B activity occurring with amino acid deficiency cannot be reversed by adding more glucose 6-phosphate or polyamines nor can the reduced eIF-2B activity seen with polyamine deficiency be overcome by increasing the glucose 6-phosphate, suggesting that these three components regulate eIF-2B activity by different mechanisms.