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We aim to bring awareness of allergies to excipients such as carboxymethylcellulose as “hidden dangers” that can be easily missed in diagnosis, leading to severe effects on patient health, and falsely limit the drug treatments that a patient can receive.

Background. Limited information is available regarding outcomes of islet cell isolation (ICI) and transplantation (ITx) using medical assistance in dying (MAiD) donors. We aimed to assess the feasibility and outcomes of ICI and ITx in MAiD donors. Methods. ICI and ITx from MAiD were compared with donation after circulatory death (DCD) type III between 2016 and 2023. Differences of isolated islet equivalents (IEQs), numeric viability and other quantitative in vitro metabolic measures were assessed. Results. Overall, 81 ICIs were available of whom 34 (42%) and 47 (58%) from MAiD and DCD-III, respectively. There were no differences of pancreas and digested tissue weight and islets viability among the 2 groups; however, cold ischemic time was longer in MAiD (11.5 versus 9.1 h; P = 0.021). The IEQ (P

This article presents a case of aortic root rupture after transfemoral aortic valve replacement, successfully treated by a percutaneous approach, with a good evolution of the patient both during hospitalization and in the long term, being asymptomatic at the cardiovascular level after 18 months of follow-up. (Level of Difficulty: Advanced.)

Renal artery aneurysms involving segmental branches pose a technical challenge to repair. Both endovascular and open repair techniques have been described. This case illustrates the clinical presentation of a patient with a symptomatic renal artery aneurysm with thromboembolic renal infarction managed with laparoscopic nephrectomy, ex vivo aneurysm resection, renal artery reconstruction, and autotransplantation.

Background: Rigorous training may lead to increased rates of knee osteoarthritis and arthroplasties in military service members. Given the large numbers of arthritis and the increasing need for total joint replacements, access to appropriate care can be difficult for this population based on insurance restrictions. The aim of this study was to evaluate access to total knee arthroplasty for TRICARE patients in contracted civilian medical facilities. Methods: Orthopedic surgeons contracted to perform total knee replacements in the state of Florida were identified via TRICARE’s website. Investigators used a secret shopper methodology with a standardized script to request an appointment for their family member for a total knee arthroplasty using either TRICARE Select or BlueCross preferred provider organization. The appointment acceptance rates, wait times, call duration, and accuracy of the physician listing were collected. Results: A total of 228 offices that perform total knee arthroplasties in Florida were successfully contacted. Overall, 43.1% of the clinics had an inaccurate online listing, and 207 (91%) were able to schedule an appointment with TRICARE, compared to 93% for BlueCross Blue Shield (P = .06). The average wait for TRICARE patients was 24 days and 18 days for BlueCross (P < .01). Call times for TRICARE patients averaged 7.2 minutes, compared to 5.2 minutes for BlueCross (P < .01). Conclusions: TRICARE patients encountered longer waiting periods and inaccurate provider listings when accessing orthopedic care. Our results suggest a disparity in healthcare access for patients using TRICARE, which may result in negative health outcomes from receiving delayed care.

Basidiobolomycosis is a rare but emerging fungal infection that causes severe and occasionally fatal illness. We present the case of a child with an abdominal mass and leukocytosis at the expense of eosinophilia, who underwent exploratory laparotomy showing a mass involving the transverse colon and ileum. With histopathological report of hyphae with few septa, highlighted by PAS and Grocott stains and surrounded by eosinophilic and hyaline material (Splendore-Höeppli phenomenon). Treatment with amphotericin B was started, however, he presented jejunal anastomosis dehiscence, with poor evolution, for which he died days later.

Paraneoplastic neurologic syndromes are a rare and heterogeneous group of immune-mediated syndromes caused by underlying solid and nonsolid tumors. We present a case of 8-year-old female with long history of mild headaches and central instability who presented multiple poorly defined signal abnormalities at the subcortical white matter of both cerebral hemispheres and cerebellar atrophy on brain magnetic resonance imaging. Further studies revealed a posterior mediastinum ganglioneuroma derived from a mature ganglioneuroblastoma that was treated with surgery. Two paraneoplastic neurologic syndromes were considered: Anti-N-Methyl-D-Aspartate Receptor (NMDAR) encephalitis due to the resolution of subcortical signal abnormalities after mediastinal mass resection and opsoclonus-myoclonus-ataxia syndrome due to cerebellar atrophy. Intertnational guideline established the criteria for definite diagnosis of paraneoplastic neurologic syndromes and detection of onconeural antibodies is not mandatory for their diagnosis. Paraneoplastic neurologic syndromes may appear several years before the tumor is detected.

Purpose: Lacerations to the ulnar and median nerve in the volar forearm have demonstrated considerable long-term clinical and socioeconomic impacts on patients. The purpose of this study was to evaluate the outcomes of complex volar forearm lacerations involving one or more major peripheral nerves in an economically disadvantaged patient population. Methods: In this study, a retrospective analysis of 61 patients who sustained lacerations to the median nerve, ulnar nerve, or both with volar wrist lacerations was performed. Each patient’s preinjury and postinjury occupation, dominant extremity, and demographic variables were evaluated. Sensation recovery, motor recovery, Disabilities of the Arm, Shoulder, and Hand scores, visual analog scale scores, cold intolerance, and return to work were evaluated at 3, 6, and 12 months after the injury. Results: Patients with isolated median nerve injuries demonstrated improved motor recovery compared with patients with isolated ulnar nerve injuries. Patients with combined nerve injuries had worse sensation recovery and motor recovery, and lower rates of return to work than either group of patients with isolated nerve injuries. Manual laborers had worse motor recovery and lower rates of return to work than did patients who were office workers. Conclusions: Patients with combined median and ulnar nerve injuries have worse functional recovery and lower rates of return to work than do patients with isolated median or isolated ulnar nerve injuries at 1 year. Manual laborers demonstrated worse functional recovery and lower rates of return to work compared with office workers at 1 year. Type of study/level of evidence: Therapeutic III. Key words: Nerve laceration, Nerve repair, Return to work, Spaghetti wrist, Volar laceration

Septic arthritis is an orthopedic emergency requiring surgical intervention for treatment. We describe the first case of native subtalar septic arthritis at our institution which was managed with open irrigation and debridement using the lateral sinus tarsi approach. The early diagnosis with magnetic resonance imaging (MRI) and immediate intervention allowed reduction of morbidity and preservation of the joint. Subtalar septic arthritis is a rare diagnosis and requires a high index of clinical suspicion for diagnosis. Advanced imaging can help localize source of infection and expedite treatment. Early surgical intervention can help decrease morbidity and preserve joint function.

Glomus tumors are rare, mostly benign, and often superficial tumors that commonly occur on the upper and lower extremities. Whereas other locations have been described in the literature, we report the first case of glomus tumor in the abdominal aorta in a 21-year-old patient. Keywords: Glomus tumor, Vessel tumor, Aortic disease

Introduction: Hip fracture is a common and devastating event in older adults causing increased dependence, comorbidity, and mortality. Since new surgical techniques have not significantly improved the mortality rate, a better understanding of patient risk factors could improve the treatment algorithm and outcomes. This prospective study aimed to document the 1-year survival rate of patients with intertrochanteric fracture treated surgically in Latin America and to investigate risk factors associated with 1-year mortality. Patients and Methods: Between January 2013 and March 2015, 199 patients were prospectively enrolled. Inclusion criteria were aged 60 years or older, isolated intertrochanteric fracture (AO/OTA 31-A), and time to surgery within 10 days after injury. The follow-up period was 1 year. The association between mortality and patient demographics, comorbidity, surgical details, and preoperative laboratory parameters was assessed using log-rank tests. Results: Twenty patients died by 365 days after surgery (including 5 that died within 30 days of surgery) resulting in a 1-year survival rate of 89.8% (95% confidence interval = 84.6-93.3). The 1-year mortality was significantly associated with age (≥85 years old, P = .032), existing comorbidity ( P = .002), preinjury mobility level ( P = .026), mental state (Mini-Mental State Examination > 23, P = .040), low preoperative plasma albumin level ( P = .007), and high preoperative blood C-reactive protein level (CRP; P = .012). At the 1-year follow-up, patients on average did not regain their preinjury hip function and mobility, although the self-assessed quality of life was equal or better than before the injury. Discussion: As a prospective study, the current patient population had clear inclusion and exclusion criteria and was relatively homogeneous. The resulting associations between 1-year postoperative mortality and preoperative hypoalbuminemia and preoperative elevated CRP level are therefore especially notable. Previously identified risk factors such as male gender and time to surgery showed no significant association with 1-year mortality—the overall favorable condition of the current population or the lack of statistical power maybe responsible for this observation. Conclusion: The current results showed that under the condition of optimal surgical treatment and low surgery-related complication, preinjury health status as indicated by the blood level of albumin and CRP has a direct and significant impact on 1-year mortality rate.

1 Canrenone, the major active metabolite of spironolactone, decreased [3H]-progesterone binding to isolated uterine cytosolic progesterone receptors. The inhibition was concentration-dependent. 2 Canrenone did not alter [3H]-oestradiol binding to isolated uterine cytosolic oestrogen receptors. 3 Canrenone inhibition of progesterone binding to isolated cytosolic receptors was strictly competitive: Kd (apparent dissociation constant for progesterone binding) was increased in a concentration-dependent manner by canrenone, whereas Bmax (maximal number of progesterone binding sites/mg cytosolic protein) was unaltered. There was marked cooperativity in progesterone binding at high canrenone and low progesterone concentrations. The implication is that canrenone alters the subunit interaction of the receptor protein. 4 Kd for progesterone was 3.2 × 10(-9)M. Ki (the inhibition constant for canrenone with respect to progesterone binding) was 300 × 10(-9)M. Reports in the literature suggest that, following spironolactone administration, canrenone may rise to concentrations sufficiently high to inhibit progesterone binding. This action may contribute to the effect of spironolactone in inducing menstrual disturbances in female patients. [ABSTRACT FROM AUTHOR]

Acute coronary syndrome (ACS) is a common cause of death in individuals older than 55 years. Although younger individuals are less frequently seen with ACS, this clinical event has increasing incidence trends, shows high recurrence rates and triggers considerable economic burden. Young individuals with ACS (yACS) are usually underrepresented and show idiosyncratic epidemiologic features compared to older subjects. These differences may justify why available risk prediction models usually penalize yACS with higher false positive rates compared to older subjects. We hypothesized that exploring temporal framing structures such as prediction time, observation windows and subgroup-specific prediction, could improve time-dependent prediction metrics. Among individuals who have experienced ACS (n global&#95;cohort  = 6341 and n yACS  = 2242), the predictive accuracy for adverse clinical events was optimized by using specific rules for yACS and splitting short-term and long-term prediction windows, leading to the detection of 80% of events, compared to 69% by using a rule designed for the global cohort., (© 2023. The Author(s).)

Cellular and humoral immune responses are essential for COVID-19 recovery and protection against SARS-CoV-2 reinfection. To date, the evaluation of SARS-CoV-2 immune protection has mainly focused on antibody detection, generally disregarding the cellular response, or placing it in a secondary position. This phenomenon may be explained by the complex nature of the assays needed to analyze cellular immunity compared with the technically simple and automated detection of antibodies. Nevertheless, a large body of evidence supports the relevance of the T cell's role in protection against SARS-CoV-2, especially in vulnerable individuals with a weakened immune system (such as the population over 65 and patients with immunodeficiencies). Here we propose to use CoVITEST (Covid19 anti-Viral Immunity based on T cells for Evaluation in a Simple Test), a fast, affordable and accessible in-house assay that, together with a diagnostic matrix, allows us to determine those patients who might be protected with SARS-CoV-2-reactive T cells. The method was established using healthy SARS-CoV-2-naïve donors pre- and post-vaccination ( n= 30), and further validated with convalescent COVID-19 donors ( n= 51) in a side-by-side comparison with the gold standard IFN-γ ELISpot. We demonstrated that our CoVITEST presented reliable and comparable results to those obtained with the ELISpot technique in a considerably shorter time (less than 8 hours). In conclusion, we present a simple but reliable assay to determine cellular immunity against SARS-CoV-2 that can be used routinely during this pandemic to monitor the immune status in vulnerable patients and thereby adjust their therapeutic approaches. This method might indeed help to optimize and improve decision-making protocols for re-vaccination against SARS-CoV-2, at least for some population subsets., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Egri, Olivé, Hernández-Rodríguez, Castro, De Guzman, Heredia, Segura, Fernandez, de Moner, Torradeflot, Ballús, Martinez, Vazquez, Costa, Dobaño, Mazza, Mazzotti, Pascal, Juan, González-Navarro and Calderón.)

This paper aims to study the evolution of CO 2 concentrations and emissions on a conventional farm with weaned piglets between 6.9 and 17.0 kg live weight based on setpoint temperature, outdoor temperature, and ventilation flow. The experimental trial was conducted during one transition cycle. Generally, the ventilation flow increased with the reduction in setpoint temperature throughout the cycle, which caused a reduction in CO 2 concentration and an increase in emissions. The mean CO 2 concentration was 3.12 g m -3 . Emissions of CO 2 had a mean value of 2.21 mg s -1 per animal, which is equivalent to 0.195 mg s -1 kg -1 . A potential function was used to describe the interaction between 10 min values of ventilation flow and CO 2 concentrations, whereas a linear function was used to describe the interaction between 10 min values of ventilation flow and CO 2 emissions, with r values of 0.82 and 0.85, respectively. Using such equations allowed for simple and direct quantification of emissions. Furthermore, two prediction models for CO 2 emissions were developed using two neural networks (for 10 min and 60 min predictions), which reached r values of 0.63 and 0.56. These results are limited mainly by the size of the training period, as well as by the differences between the behavior of the series in the training stage and the testing stage.

Listeria monocytogenes is an important food-borne pathogen and its bacteriophages are promising tools for its control in food and surfaces. Listeria bacteriophages belonging to the genus Pecentumvirus of the family Herelleviridae are strictly lytic, have a contractile tail and a large double stranded DNA genome (mean of 135.4 kb). We report the isolation and genome sequences of two new Pecentumvirus bacteriophages: vB&#95;Lino&#95;VEfB7 and vB&#95;Liva&#95;VAfA18. Twenty-one bacteriophages of this genus have been described and their genomes were used for the study of Pecentumvirus evolution. Analyses showed collinear genomes and gene gain and loss propensity and recombination events were distinctly found in two regions. A large potential recombination event (≈20 kB) was detected in P100 and vB&#95;Liva&#95;VAfA18. Phylogenetic analyses of multi-gene alignments showed that diversification events formed two groups of species distantly related., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Objectives: Traditional risk scores improved the definition of the initial therapeutic strategy in acute coronary syndrome (ACS), but they were not designed for predicting long-term individual risks and costs. In parallel, attempts to directly predict costs from clinical variables in ACS had limited success. Thus, novel approaches to predict cardiovascular risk and health expenditure are urgently needed. Our objectives were to predict the risk of major/minor adverse cardiovascular events (MACE) and estimate assistance-related costs., Methods: We used a 2-step approach that: (1) predicted outcomes with a common pathophysiological substrate (MACE) by using machine learning (ML) or logistic regression (LR) and compared with existing risk scores; (2) derived costs associated with noncardiovascular deaths, dialysis, ambulatory-care-sensitive-hospitalizations (ACSH), strokes, and MACE. With consecutive ACS individuals (n = 1089) from 2 cohorts, we trained in 80% of the population and tested in 20% using a 4-fold cross-validation framework. The 29-variable model included socioeconomic, clinical/lab, and coronarography variables. Individual costs were estimated based on cause-specific hospitalization from the Brazilian Health Ministry perspective., Results: After up to 12 years follow-up (mean = 3.3 ± 3.1; MACE = 169), the gradient-boosting machine model was superior to LR and reached an area under the curve (AUROC) of 0.891 [95% CI 0.846-0.921] (test set), outperforming the Syntax Score II (AUROC = 0.635 [95% CI 0.569-0.699]). Individuals classified as high risk (>90th percentile) presented increased HbA1c and LDL-C both at <24 hours post-ACS and 1-year follow-up. High-risk individuals required 33.5% of total costs and showed 4.96-fold (95% CI 3.71-5.48, P < .00001) greater per capita costs compared with low-risk individuals, mostly owing to avoidable costs (ACSH). This 2-step approach was more successful for finding individuals incurring high costs than predicting costs directly from clinical variables., Conclusion: ML methods predicted long-term risks and avoidable costs after ACS., (Copyright © 2020 ISPOR–The Professional Society for Health Economics and Outcomes Research. Published by Elsevier Inc. All rights reserved.)

Purpose: To determine the frequency of limitations on life support techniques (LLSTs) on admission to intensive care units (ICU), factors associated, and 30-day survival in patients with LLST on ICU admission., Methods: This prospective observational study included all patients admitted to 39 ICUs in a 45-day period in 2011. We recorded hospitals' characteristics (availability of intermediate care units, usual availability of ICU beds, and financial model) and patients' characteristics (demographics, reason for admission, functional status, risk of death, and LLST on ICU admission (withholding/withdrawing; specific techniques affected)). The primary outcome was 30-day survival for patients with LLST on ICU admission. Statistical analysis included multilevel logistic regression models., Results: We recruited 3042 patients (age 62.5 ± 16.1 years). Most ICUs (94.8%) admitted patients with LLST, but only 238 (7.8% [95% CI 7.0-8.8]) patients had LLST on ICU admission; this group had higher ICU mortality (44.5 vs. 9.4% in patients without LLST; p  < 0.001). Multilevel logistic regression showed a contextual effect of the hospital in LLST on ICU admission (median OR = 2.30 [95% CI 1.59-2.96]) and identified the following patient-related variables as independent factors associated with LLST on ICU admission: age, reason for admission, risk of death, and functional status. In patients with LLST on ICU admission, 30-day survival was 38% (95% CI 31.7-44.5). Factors associated with survival were age, reason for admission, risk of death, and number of reasons for LLST on ICU admission., Conclusions: The frequency of ICU admission with LLST is low but probably increasing; nearly one third of these patients survive for ≥ 30 days., Competing Interests: This study was approved by our Institutional Research Ethics Committee de la Fundació UNIO CATALANA HOSPITALS (reference number: CEIC 11/23). It has also approved by the local ethics committees, in accordance with Spanish regulations.All authors read and approved the final manuscript. This study was approved by our Institutional Department of Investigation in accordance with Spanish regulations.The authors declare that they have no competing interests.Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Background: Modern knee designs have popularized its use in younger patients due to its better performance. There remains uncertainty whether higher demands of these patients can affect implant survivorship., Purpose: To assess whether modern knee designs have provided similar results in patients younger than 70 years versus older patients., Methods: We included 203 consecutive patients (236 knees) who underwent knee replacement for osteoarthritis with a mean follow-up of 11.4 years (range: 8.8 to 12). The mean age was 70 years (range: 31 to 85). Knee replacements were stratified into two groups: 109 were younger than 70 years and 127 were older than 70 years (70 years of age is the mandatory retirement age)., Results: There were no significant pre-operative differences between groups with regards to knee alignment, alpha or beta angles, knee score or function score. Fourteen implants were radiographically loose at last follow up visit. Groups were matched in terms of demographic data. We found that patients older than 70 years had significantly better mean survivorship at 12 years. (97% vs. 88%; P=0.010). Patients under 70 years presented with a higher rate of polyethylene wear which was further associated with radiolucent lines in the femur and tibia as well as the presence of osteolysis. There was also an association between migration and presence of osteolysis., Conclusions: Patients over 70 years old undergoing cemented total knee replacement for osteoarthritis showed better implant survivorship versus patients under 70 years old.

The influence of the type of farming on harmful gas exposures to carbon dioxide (CO(2)), ammonia (NH(3)) and hydrogen sulfide (H(2)S) was assessed from the perspective of animal welfare and occupational hygiene. Summer data of H(2)S, NH(3) and CO(2) concentrations and of environmental parameters were collected from 31 farms. The indices of exposure for long-term exposures to NH3 suggest the lowest acceptability of exposure was observed on poultry farms. CO(2) had the highest dependence on production activity. For H(2)S, no differences were found based on farming activity. Both the stocking density and volume of air available affected the daily exposure and the index of exposure to CO(2). Significant differences were observed between hourly CO(2) concentrations, depending on the level of activity inside the building. A positive correlation was found between gas concentrations and temperature increase. All values of daily and short-term exposures were below exposure limit values, which suggests that exposure conditions were appropriate for workers' health during the measurement period. Analysis of the working hours and average hourly concentration of gases during the times of day, with presence of workers inside the farm buildings, revealed significant differences for CO(2).

A previously described Taenia saginata HDP2 DNA sequence, a 4-kb polymorphic fragment, was previously used as the basis for developing PCR diagnostic protocols for the species-specific discrimination of T. saginata from T. solium and for the differentiation of T. saginata from T. asiatica. The latter was shown subsequently to lack the required specificity, so we undertook genetic studies of the HDP2 sequence from T. saginata and T. asiatica to determine why, and to develop a novel HDP2-PCR protocol for the simultaneous unambiguous identification of human taeniids. Sequencing and further analysis of the HDP2 DNA fragments of 19 Asiatic isolates of T. saginata and T. asiatica indicated that the HDP2 sequences of both species exhibited clear genomic variability, due to polymorphic variable fragments, that could correspond to the non-transcribed region of ribosomal DNA. This newly observed polymorphism allowed us to develop a novel, reproducible and reliable HDP2-PCR protocol which permitted the simultaneous discrimination of all T. saginata and T. asiatica isolates examined. This species-specific identification was based on, and facilitated by, the clear size difference in amplicon profiles generated: fragments of 1300 bp, 600 bp and 300 bp were produced for T. asiatica, amplicons of 1300 bp and 300 bp being obtained for T. saginata. Control T. solium samples produced one amplicon of 600 bp with the HDP2-PCR protocol. The assay has the potential to prove useful as a diagnostic tool in areas such as South East Asia where T. saginata, T. asiatica and T. solium coexist.

Background: To estimate the proportion of diabetic patients (DPts) with peripheral vascular disease treated at a primary health care site after an endocrinologist-based intervention, who meet ATP III and Steno targets of metabolic control, as well as to compare the outcome with the results of the patients treated by endocrinologists., Methods: A controlled, prospective over 30-months period study was conducted in area 7 of Madrid. One hundred twenty six eligible diabetic patients diagnosed as having peripheral vascular disease between January 2003 and June 2004 were included in the study. After a treatment period of three months by the Diabetes team at St Carlos Hospital, 63 patients were randomly assigned to continue their follow up by diabetes team (Group A) and other 63 to be treated by the family physicians (FP) at primary care level with continuous diabetes team coordination (Group B). 57 DPts from Group A and 59 from Group B, completed the 30 months follow-up period. At baseline both groups were similar in age, weight, time from diagnosis and metabolic control. The main outcomes of this study were the proportion of patients meeting ATP III and Steno goals for HbA1c (%), Cholesterol, HDL cholesterol, LDL cholesterol, triglycerides, blood pressure, albumine-to-creatinine excretion ratio (ACR), body mass index (BMI), waist circumference (WC), anti-aggregation treatment and smoking status., Results: At the end of the follow up, no differences were found between the groups. More than 37% of diabetic patients assigned to be treated by FP achieved a HbA1c < 6.5%, more than 50% a ACR < 30 mg/g, and more than 80% reached low risk values for cholesterol, LDL cholesterol, triglycerides, diastolic blood pressure and were anti-aggregated, and 12% remained smokers. In contrast, less than 45% achieved a systolic blood pressure < 130 mm Hg, less than 12% had a BMI < 25 Kg.m-2 (versus 23% in group A; p < 0.05) and 49%/30% (men/women) had a waist circumference of low risk., Conclusion: Improvements in metabolic control among diabetic patients with peripheral vascular disease treated at a primary health care setting is possible, reaching similar results to the patients treated at a specialized level. Despite such an improvement, body weight control remains more than poor in both levels, mainly at primary care level. General practitioner and endocrinologist coordination care may be important to enhance diabetes management in primary care settings., Trial Registration: Clinical Trial number ISRCTN75037597.

Background: Antibodies to hepatitis C virus (anti-HCV) have typically been detected by enzyme immunoassay (EIA). A chemiluminescence assay (CA) for anti-HCV is now commercially available., Methods: We compared the positive rate for a CA in a HCV screening program for veterans with historical rates obtained with EIA. We also compared results in 2824 samples tested by both methods and assessed the significance of low signal-to-cutoff (S/C) ratios., Results: The frequency of CA-positive results was significantly lower than with EIA (12.6% vs 16.0%; P <0.0001). The frequency of low S/C ratios was also significantly lower with CA (11.5% vs 20.0%; P <0.0001). Among low-positive values, samples positive by CA were significantly less likely to be recombinant immunoblot assay (RIBA)-negative (64% vs 84%; P <0.0005). In parallel testing, results for 111 samples (3.9%) were discrepant between the two assays; all but 6 had low S/C ratios, and confirmatory testing was performed on all but 8 samples. Of 56 EIA-positive, CA-negative samples tested by RIBA, only 1 was positive. Of 24 CA-positive, EIA-negative samples, 62% were RIBA-negative. Using a negative RIBA result as an indication of false-positive anti-HCV results, the positive predictive value of EIA was 93% compared with 98% with CA. HCV RNA was positive in 90% of samples high-positive by both CA and EIA. Only 2 of 30 (7%) low-positive CA samples were RNA-positive., Conclusions: CA produces fewer false-positive and fewer low-positive results that require confirmatory RIBA testing. The S/C ratio remains useful for characterizing positive results.

Background: Tests for hepatitis C antibodies (anti-HCV enzyme immunoassays) are usually described as positive or negative. Several studies, mainly in blood donors, have found that specimens with low signal/cutoff (S/C) ratios are commonly negative when tested with a recombinant immunoblot assay (RIBA) or for HCV RNA., Methods: We retrospectively reviewed 17 418 consecutive anti-HCV results from a screening program for high-risk veterans; 2986 (17.1%) samples were anti-HCV-positive, and 490 (16.4%) had S/C ratios

We studied the flood tolerance of five tree species growing in the flooded forest adjacent to the Mapire river, in SW Venezuela. Mean photosynthetic rate and leaf conductance were 11 &mgr;mol m(-2) s(-1) and 700 mmol m(-2) s(-1), respectively. Xylem water potential ranged from -0.08 to -1.15 MPa. Based on leaf gas exchange as a criterion of tolerance to flooding, two response patterns were identified: (1) decreasing photosynthetic rate with increasing flooding and leaf conductance (Psidium ovatifolium Berg. ex Desc., Campsiandra laurifolia Benth., Symmeria paniculata Benth. and Acosmium nitens (Vog.) Benth); and (2) independence of photosynthesis and leaf conductance from flooding (Eschweilera tenuifolia (Berg.) Miers.). In the first response pattern, declining photosynthetic rate with flooding may be interpreted as a sign of reduced flood tolerance, whereas the second response pattern may indicate increased flood tolerance. An increase in xylem water potential with depth of water column was found for all species (with the possible exception of P. ovatifolium), indicating that flooding does not cause water stress in these trees. Submerged leaves that had been under water for between four days and four months generally had photosynthetic rates and leaf conductances similar to those of aerial leaves, indicating maintenance of photosynthetic capacity under water. Daily positive oscillations in glucan content in submerged leaves of P. ovatifolium and C. laurifolia suggest that submerged leaves do not represent a sink for photosynthates produced by aerial leaves.

During the import of the precursor for the acyl carrier protein (ACP) into chloroplasts, apoACP is converted to holoACP by the attachment of a phosphopantetheine group transferred from coenzyme A (CoA) by a chloroplast holoACP synthase [Fernandez, M. and Lamppa, G. (1990) Acyl carrier protein import into chloroplasts does not require the phosphopantetheine: evidence for a chloroplast holoACP synthase, Plant Cell 2, 195-206]. Here it is shown that exogenous addition of CoA to intact chloroplasts in the import assay stimulates the conversion of apoACP to holoACP. If adenosine 3',5'-bisphosphate [Ado(3',5')P2], the byproduct of the transfer reaction, was also included the extent of conversion was greatly reduced. CoA has its effect after ACP precursor (pre-ACP) import and proteolytic removal of the transit peptide, thus indicating that the chloroplast holoACP synthase resides in the stroma where fatty acid synthase is found. When Ado(3',5')P2 was added alone to the import assay, it inhibited the synthesis of holoACP. Inhibition of the conversion of apo- to holoACP with Ado(3',5')P2 made it possible to examine whether the holoform of preACP could be imported into chloroplasts. Pre-apoACP was synthesized in Escherichia coli and shown to be competent for import in an ATP- and temperature-dependent manner. A partially purified chloroplast holoACP synthase converted 60-90% of the pre-apoACP to pre-holoACP. Pre-holoACP incubated with chloroplasts in the presence of Ado(3',5')P2 yielded > 60% holoACP, whereas the control reaction with pre-apoACP gave primarily apoACP. Hence the phosphopantetheine prosthetic group of ACP does not block precursor movement through the translocation apparatus of the chloroplast envelope.

Recently a chloroplast holo-acyl carrier protein (holoACP) synthase activity was identified which attached the phosphopantetheine prosthetic group to acyl carrier protein, producing holoACP (Fernandez and Lamppa (1990) Plant Cell 2, 195-206). Here we show that the mature form of ACP (apoACP), after entry into the chloroplast and removal of the transit peptide, is a substrate for modification by the holoACP synthase. Modification occurs optimally at 37 degrees C and is inhibited by 5 mM 3',5'-ADP and 2 mM EDTA. An ACP construct (matACP) lacking the transit peptide was also converted to the holoACP form in an organelle-free assay, independent of precursor cleavage. The matACP construct was used to monitor the chromatographic separation of the holoACP synthase from the transit peptidase. Superose 12 gel filtration analysis indicates that the holoACP synthase has an apparent Mr of approximately 50,000. Using fractions enriched for the holoACP synthase it was demonstrated that the precursor of ACP is also modified in the presence of CoA and subsequently can be proteolytically processed directly to holoACP. Kinetic analysis, however, indicates that removal of the transit peptide is a much faster reaction than phosphopantetheine addition, suggesting that apoACP is the primary substrate for the chloroplast holoACP synthase in vivo.

Import of the acyl carrier protein (ACP) precursor into the chloroplast resulted in two products of about 14 kilodalton (kD) and 18 kD when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Time course experiments indicate that the latter is a modification derivative of the 14-kD peptide after the removal of the transit peptide. Substitution of serine 38 by alanine, eliminating the phosphopantetheine prosthetic group attachment site of ACP, produced a precursor mutant that gave rise to only the 14-kD peptide during import, showing that the modified form depends on the presence of serine 38. Furthermore, these results demonstrate that the prosthetic group is not essential for ACP translocation across the envelope or proteolytic processing. Analysis of the products of import by nondenaturing, conformationally sensitive gels showed reversal of the relative mobility of the 14-kD peptide and the modified form, raising the possibility that the modification is the addition of the phosphopantetheine. Proteolytic processing and the modification reaction were reconstituted in an organelle-free assay. The addition of coenzyme A to the organelle-free assay completely converted the 14-kD peptide to the modified form at 10 micromolar, and this only occurred with the wild-type substrate. Reciprocally, treatment of the products of a modification reaction with Escherichia coli phosphodiesterase converted the modified ACP from back to the 14-kD peptide. These results strongly support the conclusion that there is a holo-ACP synthase in the soluble compartment of the chloroplast capable of transferring the phosphopantetheine of coenzyme A to ACP.

We saw two cases of pure squamous cell carcinoma of the breast, one of which is associated with dermatomyositis. Electron microscopy of appropriately fixed tissue obtained from one tumor confirmed the squamous nature of the polyhedral and spindle-shaped tumor cells. One tumor was assayed for estrogen and progesterone receptor proteins and was found to be lacking both. Test results for mucin and carcinoembryonic antigen (CEA) were negative in both tumors. In contrast, a similarly examined case of invasive ductal carcinoma with areas of squamous metaplasia had slightly elevated concentrations of estrogen and progesterone receptor proteins, and test results for mucin and CEA were positive.