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Aim: Intraoperative radiotherapy with electrons (IOERT) may represent a viable choice for partial breast re-irradiation (rePBI) after repeat quadrantectomy for local recurrence (LR) for primary breast cancer (BC) in lieu of mastectomy., Materials and Methods: A database collecting data on rePBI with IOERT from 8 Italian centres was set up in 2016- 2018, providing data on cumulative incidence (CumI) of 2 nd LR nd survival with a long follow-up (FU) RESULTS: From 2002 to 2015, 109 patients underwent the conservative retreatment. The median primary BC -1stLR interval was 11.1 years (range: 2.4-27.7). The median 1stLR size was 0.9 cm (range: 0.3-3.0) and 43.6% were Luminal A. Median IOERT dose was 18 Gy (range: 12-21) and median collimator was 4 cm (range: 3-6). Median FU was 11.7 years (interquartile range: 7.7-14.6). The 2ndLR CumI was 12.2% (95% CI: 6.8-19.2) at 5 years and 32.3% at 10 years (95% CI: 22.8-42.2), occurring in the same site as the 1stLR in about half of the cases. HER2 status and collimator size were independent LR predictors. The 5- and 10-year overall survival were 95.2% and 88.3%, respectively, while 5- and 10-year BC specific survival were 98% and 94.5%. The development of a 2ndLR significantly reduced BCSS (HR=9.40, P<0.001). Grade ≥3 fibrosis was 18.9%. Patient-reported cosmesis was good/excellent in 59.7% of the cases., Conclusion: 2ndLR CumI was within the range of the literature, but higher than expected, opening questions on radiation field extension and fractionation schedule. Since a 2ndLR worsened the outcome, salvage modality must be carefully planned., (Copyright © 2024. Published by Elsevier Inc.)

Channel networks are key to coastal wetland functioning and resilience under climate change. Vegetation affects sediment and hydrodynamics in many different ways, which calls for a coherent framework to explain how vegetation shapes channel network geometry and functioning. Here, we introduce an idealized model that shows how coastal wetland vegetation creates more complexly branching networks by increasing the ratio of channel incision versus topographic diffusion rates, thereby amplifying the channelization feedback that recursively incises finer-scale side-channels. This complexification trend qualitatively agrees with and provides an explanation for field data presented here as well as in earlier studies. Moreover, our model demonstrates that a stronger biogeomorphic feedback leads to higher and more densely vegetated marsh platforms and more extensive drainage networks. These findings may inspire future field research by raising the hypothesis that vegetation-induced self-organization enhances the storm surge buffering capacity of coastal wetlands and their resilience under sea-level rise., (© 2023. The Author(s).)

Introduction: A peripheral rim of globules represents a marker of the horizontal growth phase in nevi and is a common feature in children and adolescents. The observation of melanocytic lesions with peripheral globules (MLPGs) in adulthood deserves more attention, since melanoma may exhibit this feature, albeit rarely. Risk-stratified management recommendations considering a global clinical approach are still missing., Objectives: To analyze current knowledge on MLPGs and propose an integrated management algorithm stratified for age groups., Methods: We conducted a narrative review of current published data on MLPGs, analyzing clinical dermoscopic and confocal distinguishing features of melanoma from benign nevi., Results: The risk of finding a melanoma when removing an MLPG increases with age, especially in people >55 years old, and is significantly higher in the extremities, head/neck and in case of a single asymmetrical lesion, ≥6 mm in diameter. Dermoscopic features associated with melanoma diagnosis include atypical peripheral globules, asymmetrical distribution, multiple rims as well as the reappearance of globules after prior loss. In addition, wide blue-grey regression areas, atypical networks, eccentric blotches, tan structureless peripheral areas and vascularization are atypical dermoscopic features. Confocal worrisome findings are represented by pagetoid cells within the epidermis, architectural disarrangement and atypical cells of the dermo-epidermal junction with irregular peripheral nests., Conclusion: We proposed a multi-step age-stratified management algorithm integrating clinical, dermoscopic and confocal findings that may increase the early recognition of melanoma and avoid surgical excision of benign nevi.

Polysaccharides strongly affect the sensorial properties of foods, which is partly related to the lubrication they provide. We investigated the lubrication performance of polysaccharides with different characteristics (i.e. molecular weight, conformation, stiffness, and charge density) to identify the most relevant structural features related to lubrication ability. The effect of viscosity was excluded using isoviscous systems. Conformation resulted to be a key factor in determining lubrication. Rigid rod-like polysaccharides (xanthan) showed higher lubricating capacity and better resistance to changes in pH and ionic strength compared to semi-flexible (pectin and carboxymethyl cellulose) and flexible polysaccharide (guar gum). The friction behavior of flexible polysaccharides was highly dependent on concentration and sliding speed, for which a model including parameters of friction, concentration, and speed was developed. The concentration-dependency of the lubrication was related to the shear-thinning behavior, and possible mechanisms to explain these differences in terms of molecular properties of the polysaccharides are proposed., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

This research aimed to investigate the effect of starch gelatinization and swelling degree on the lubrication properties of starch aqueous suspensions. Three types of maize starch with different amylose content of 70% (HAS), 25% (NS), and <1% (WS) were used to vary the swelling capacities. The granule suspension of NS showed the highest swelling factor (SF) of 26.5, and gave the best lubrication capacity by decreasing friction by 78%. WS was only able to decrease friction by 50% due to a lower swelling capacity. The leached-out amylose increased friction of highly swollen granules (SF = 26.5) but decreased friction of stiff granules (SF = 2.5). Adding unstimulated human saliva to starch suspensions with native and limited swollen granules reduced friction and masked differences in friction coefficients among starch types. Both the salivary layer on the contact surfaces and the salivary proteins in the bulk phase played a role in determining lubrication properties., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

In this study we investigated the individual contribution of different ingredients to the lubrication properties of dairy-based model beverages containing whey protein (native or aggregated), micellar casein and emulsified oil in different combinations. In single-component systems, whey protein isolate (WPI) solutions showed the lowest friction coefficients of all protein samples. Whey protein aggregates (WPA, ∼247 nm, irregular morphology) led to higher friction coefficients than micellar casein isolate (MCI) with comparable size (∼207 nm, near spherical shape). When protein particles were combined, i.e. WPA and MCI, lubrication was not much affected. However, when WPI was added to either WPA or MCI, higher friction coefficients were observed compared to single-component systems. Emulsions (droplet size ∼ 440 nm) provided better lubrication than the protein samples. Oil droplets stabilized by soy lecithin (SL) were more efficient at reducing friction than those stabilized by WPI. The friction coefficient of SE strongly increased in the presence of WPI. In comparison, lubrication of WE was less affected by the addition of WPI. These results show that different proteins affect the lubrication properties of emulsions stabilized with different emulsifiers in a different way. Our research also indicates that multicomponent systems are complex, and that control over the lubrication properties requires a better understanding of the contribution of individual components., (Copyright © 2022 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

The continuous improvement of life expectancy of patients with chronic lymphocytic leukemia (CLL) has resulted in increased risk of second primary malignancy that potentially may affect survival and quality of life of CLL patients. We performed a systematic review to assess the risk and the clinical-pathological features and prognosis of cutaneous squamous cell carcinoma (cSCC) in patients with CLL. We searched PubMed, Embase, and Cochrane Central Register of Control Trials databases for articles published from database inception to December 31, 2019. English-language studies reporting original data on patients with a specific diagnosis of CLL and cSCC were included. Data were extracted using a standardized extraction form, and any discordance was resolved by consensus. Descriptive data were generated by pooling patients from eligible studies. Of the 4588 non-duplicate records identified, 55 articles met our inclusion criteria. These studies reported that CLL patients have a 3.2% prevalence of cSCC, with an 11.5% cSCC-related lethality and an overall risk of metastasis of 5.7% (7.3% for regional lymph node involvement and 3.8% for distant metastasis). The quality of evidence was limited by the high heterogeneity in the design, populations, and objectives of the included studies. This systematic review suggests that cSCC in CLL patients tends to behave less aggressively compared with the solid organ transplant recipients but has a higher morbidity and mortality than in the general population. Future prospective studies are needed to increase the quality of evidence and to determine the best treatment modalities and screening intervals for these patients., (© 2021 The Authors. International Journal of Dermatology published by Wiley Periodicals LLC on behalf of the International Society of Dermatology.)

Besides affecting 8% of the general population, nightmares are one of the most frequent symptoms of traumatized individuals. This can be a significant factor in the treatment of post-traumatic disorders; indeed, several studies demonstrated its strong predictive and prognostic value. Sleep disorders, nightmares in particular, could be very distressing for individuals and need targeted interventions, especially if they are associated with a PTSD diagnosis. To date, the best technique for the treatment of traumatic sleep disturbances seems to be Imagery Rehearsal Therapy (IRT), an empirically supported method. Through a review of the literature on this matter, this article aims to outline the incidence and consequences of nightmares in PTSD, illustrate how IRT could prove useful in their treatment, and investigate its clinical applications., 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. The handling editor declared a shared affiliation with one of the authors ML at time of review., (Copyright © 2022 Albanese, Liotti, Cornacchia and Mancini.)

Undernutrition is prevalent in the older adult population. Oral nutritional supplements (ONS) are a clinically effective nutritional intervention, however, patient acceptance of ONS can be limited by their palatability. While sensory attributes such as sweetness and mouthfeel have been investigated, the contribution made by aroma to the perceived flavour of ONS has not been studied. Firstly, this research aimed to identify the aroma active compounds within a commonly prescribed ONS using estimated odour activity values (OAV) and gas chromatography olfactometry mass spectrometry (GC-O-MS). Secondly, age related differences in olfactory detection were explored. Eight aroma active compounds were identified within the ONS, including diacetyl (sweet), isoamyl acetate (banana), dimethyl trisulfide (sulfur) and methanethiol (sulfur). When compared with younger adults (n = 24, 18-44 years), older adults (n = 24, 62-80 years) had higher detection thresholds for all aroma compounds and this was significant for isoamyl acetate (sweet, fruity) and methanethiol (sulfur) (p = 0.01 and p = 0.03, respectively). Thus, a decline in olfactory sensitivity was present in the older subjects included in the study, and this reduced detection sensitivity was aroma specific. Thus, older adults' flavour perception of ONS likely depends on the combined effect of product factors (the aroma profile) along with age related consumer factors (the degree of impairment in perception). This is a fundamental study which will aid future research into how the aroma profile, and associated age related impairments in perception, shape the global perception of ONS for nutritionally at risk older individuals., (© 2021. The Author(s).)

Many older adults fail to meet their daily protein requirements, potentially due to social, physical and medical factors, including sensory and appetite changes. Additionally, our previous research has identified potential sulfurous off-flavours, originating from heat-treatment of protein ingredients, which could play a role in consumer acceptance. This study aims to determine the hedonic impact of these potential off-flavours when added to a dairy beverage, identify the specific off-flavour concentrations which cause rejection by consumers, and lastly investigate difference in acceptance between older and younger consumers. A rejection threshold (RjT) protocol was used, in combination with Best Estimate Thresholds (BET), whereby sulfurous flavours (dimethyl sulfide, dimethyl disulfide and dimethyl trisulfide), and diacetyl were added to create a range of concentrations. 95 participants (younger n = 49, 18-38 years; older n = 46, 60-79 years) tasted 7 pairs of samples (one blank and one with ascending off-flavour concentration) and selected their preferred samples. Sulfurous flavours negatively impacted consumer acceptance, however, the extent to which they impart a negative effect differs between age groups. Younger adults rejected samples containing low concentrations of sulfurous off-flavours (1.55 ppb), however, older adults rejected samples with concentrations over 3 times higher (5.08 ppb). When combined with sulfurous flavours, diacetyl increased the rejection threshold for both groups. In conclusion, these observations imply that a greater quantity of off-flavour may be present before acceptance is reduced in the older consumer group. Moreover, diacetyl demonstrates partial masking abilities of sulfurous off-flavours, and BET gave a more conservative estimate of acceptability. This knowledge will help guide sensory innovation of high-protein beverages for older consumers to support product acceptance and optimal intake., Competing Interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2021 The Authors.)

Background: There is lack of studies on the diagnostic accuracy of dermoscopy and reflectance confocal microscopy (RCM) for dark pigmented lesions., Objective: To assess the diagnostic accuracy of dermoscopy plus confocal microscopy for melanoma diagnosis of dark pigmented lesions in real life., Methods: Prospective analysis of difficult dark lesions with clinical/dermoscopic suspicion of melanoma referred for RCM for further analysis. The outcome could be excision or dermoscopic digital follow-up., Results: We included 370 clinically dark lesions from 350 patients (median age, 45 y). Because of the clinical/dermoscopic/RCM approach, we saved 129 of 213 unnecessary biopsies (specificity of 60.6%), with a sensitivity of 98.1% (154/157). The number needed to excise with the addition of RCM was 1.5 for melanoma diagnosis., Limitations: Single institution based; Italian population only., Conclusions: This study showed that RCM coupled with dermoscopy increases the specificity for diagnosing melanoma, and it helps correctly identify benign lesions. Our findings provide the basis for subsequent prospective studies on melanocytic neoplasms belonging to patients in different countries., (Copyright © 2020 American Academy of Dermatology, Inc. Published by Elsevier Inc. All rights reserved.)

Background: No large studies have defined the best treatment of actinic cheilitis., Methods: We conducted a systematic review to define the best therapies for actinic cheilitis in clinical response and recurrences., Results: We first identified 444 papers, and 49 were finally considered, including 789 patients and 843 treated areas. The following therapies were recorded in order of frequency: laser therapy, photodynamic therapy (PDT), 3% diclofenac in 2.5% hyaluronic acid, PDT + 5% imiquimod, aminolevulinic acid-laser or methyl-aminolevulinic acid-laser, 5% imiquimod, fluorouracil, partial surgery, 0.015% ingenol mebutate, 50% trichloroacetic acid, and laser + PDT. Concerning the primary outcome, complete clinical response was achieved in 76.5% of patients, and 10.2% had clinical recurrences. Partial surgery and laser therapy showed the highest complete response rates (14 of 14 [100%] and 244 of 260 [93.8%], respectively) with low recurrences. Only a limited number of patients were treated with other therapies, with the exception of PDT, with 68.9% complete responses and 12.6% of recurrences. Interestingly, when combined with 5% imiquimod, the efficacy of PDT was significantly enhanced., Limitations: Heterogeneity across studies., Conclusion: Laser therapy appears the best option among nonsurgical approaches for actinic cheilitis, and PDT showed higher efficacy when sequentially combined with 5% imiquimod. Larger studies are needed to confirm these data., (Copyright © 2019 American Academy of Dermatology, Inc. Published by Elsevier Inc. All rights reserved.)

Global climate change is expected to impact hydrodynamic conditions in stream ecosystems. There is limited understanding of how stream ecosystems interact and possibly adapt to novel hydrodynamic conditions. Combining mathematical modelling with field data, we demonstrate that bio-physical feedback between plant growth and flow redistribution triggers spatial self-organization of in-channel vegetation that buffers for changed hydrological conditions. The interplay of vegetation growth and hydrodynamics results in a spatial separation of the stream into densely vegetated, low-flow zones divided by unvegetated channels of higher flow velocities. This self-organization process decouples both local flow velocities and water levels from the forcing effect of changing stream discharge. Field data from two lowland, baseflow-dominated streams support model predictions and highlight two important stream-level emergent properties: vegetation controls flow conveyance in fast-flowing channels throughout the annual growth cycle, and this buffering of discharge variations maintains water depths and wetted habitat for the stream community. Our results provide important evidence of how plant-driven self-organization allows stream ecosystems to adapt to changing hydrological conditions, maintaining suitable hydrodynamic conditions to support high biodiversity.

Recently, 5-fluorouracil 0.5%/salicylic acid 10% (5-FU/SA) topical solution has been included in the National Italian portfolio for lesion-directed treatment of grade I/II actinic keratosis (AKs) located on the face or scalp. To describe the utility of dermoscopy and RCM in treatment response monitoring of a series of AKs treated with 5-FU/SA as lesion-directed therapy. Consecutive patients were prospectively treated for a maximum of 12 weeks with 5-FU/SA for AKs located on the face or scalp. Clinical, dermoscopic, and confocal images of one index AK were acquired at each visit and pre-specified criteria were evaluated. Clinical, dermoscopic, and confocal responses were evaluated at last follow-up visit. Fourteen patients were enrolled, of which five were treated for 12 weeks, seven for 8, and two for 4 weeks. At a median follow up of 30 weeks, 64.3% (9/14) index AKs achieved complete clinical, 50% (7/14) complete dermoscopic and 42.9% (6/14) complete confocal clearance. Local skin reaction was mild and significantly decreased during therapy administration. Although the small number of cases, our study underlines the utility of both dermoscopy and in-vivo RCM in 5-FU/SA treatment response monitoring for AKs located on the face or scalp., (© 2020 Wiley Periodicals LLC.)

Spatial heterogeneity plays a crucial role in the coexistence of species. Despite recognition of the importance of self-organization in creating environmental heterogeneity in otherwise uniform landscapes, the effects of such self-organized pattern formation in promoting coexistence through facilitation are still unknown. In this study, we investigated the effects of pattern formation on species interactions and community spatial structure in ecosystems with limited underlying environmental heterogeneity, using self-organized patchiness of the aquatic macrophyte Callitriche platycarpa in streams as a model system. Our theoretical model predicted that pattern formation in aquatic vegetation - due to feedback interactions between plant growth, water flow and sedimentation processes - could promote species coexistence, by creating heterogeneous flow conditions inside and around the plant patches. The spatial plant patterns predicted by our model agreed with field observations at the reach scale in naturally vegetated rivers, where we found a significant spatial aggregation of two macrophyte species around C. platycarpa. Field transplantation experiments showed that C. platycarpa had a positive effect on the growth of both beneficiary species, and the intensity of this facilitative effect was correlated with the heterogeneous hydrodynamic conditions created within and around C. platycarpa patches. Our results emphasize the importance of self-organized patchiness in promoting species coexistence by creating a landscape of facilitation, where new niches and facilitative effects arise in different locations. Understanding the interplay between competition and facilitation is therefore essential for successful management of biodiversity in many ecosystems., (© 2018 The Authors Ecology published by Wiley Periodicals, Inc. on behalf of Ecological Society of America.)

The amount of heat-denatured whey protein is typically determined by pH 4.6 precipitation. Using this method, a significant amount of nondenatured protein was reported even after long heating times. Apparently, a fraction of the unfolded protein refolds into the "native" state rather than form aggregates. This fact is known and has been explained using kinetic models. How the conditions affect the refolding and aggregation is, however, not fully understood. Therefore, this study investigates the unfolding, refolding, and aggregation process of β-lactoglobulin using circular dichroism and size-exclusion chromatography to characterize different folding variants and to quantify their content. The proteins remaining in solution at pH 4.6 were confirmed to be native-like. The nonaggregated fraction contains proteins with a native-like and two types of non-native-like conformations. The nonaggregated fraction increased with decreasing temperature (60-90 °C) and concentration (1-50 g/L) and increasing electrostatic repulsion (pH 7-8; 0-50 mM). The native-like fraction in the nonaggregated fraction was independent of pH, ionic strength, and concentration but increased with decreasing temperature.

Processing beverages containing high concentrations of globular proteins represents a technological challenge due to their instability during heating caused by protein aggregation and gelation. Aggregation of whey protein mixtures was investigated in aqueous model systems at pH 3.5, 4.0, and 4.5 at heating conditions resembling conventional industrial treatment (90 °C for 30 s). The extent of aggregation progressively decreased moving away from the pI. Protein aggregates became smaller and had a more open structure compared to higher pH values. Significant loss of protein dispersibility occurred at pH 4.0 and 4.5 above the denaturation T of whey protein (∼70 °C), at which aggregation was caused by intermolecular hydrophobic interactions. Accessible thiol groups were detected in the heat-treated systems, with a higher intensity at higher pH and increasing extent of aggregation. Intermolecular -S-S- bonding played only a minor role in the aggregation at all conditions studied.

Liposoluble bioactive compounds are often included in foods in emulsified lipid carriers. In the present study, the impact of the physical state of the lipid carrier and the interfacial composition of oil-in-water emulsions on the stability of β-carotene was studied. Emulsions with hydrogenated palm kernel oil (HPKO) concentration of 10% (w/w) dispersing 0.05% (w/w) β-carotene, and a water phase at pH 7 containing 30% (w/w) sucrose, were stabilized by 1%, 1.5%, 2%, and 3% (w/w) whey protein isolate (WPI). Crystallization and melting behavior of emulsified and bulk HPKO were studied by differential scanning calorimetry. The hysteresis of emulsified HPKO crystallization (onset approximately 10 °C; endset approximately 6 °C) and melting (onset approximately 17 °C; endset approximately 45 °C) allowed us to operate at 15 °C on systems with identical compositions but different physical states of the same lipid phase. Surface protein coverage of emulsions was calculated and size of the dispersed particles was characterized by dynamic light scattering. β-Carotene contents of the emulsions during storage at 15 °C was analyzed spectrophotometerically. Results highlighted an impact of the phase of the lipid carrier and of the concentration of WPI on β-carotene degradation. β-Carotene loss showed zero-order kinetics. A liquid dispersed phase resulted in a low degradation rate but a high concentration of protein on a solid lipid carrier was likewise effective for β-carotene protection. Practical Application:  The inclusion of lipophilic bioactive compounds, such as carotenoids, is a current trend in the production of functional foods aiming to enhance health and well-being. However, the use of functional ingredients in food products is complicated because of the sensitivity of the active molecules to physical and chemical factors to which they are exposed during processing, storage, and consumption. The present work gives indications of the influence of the lipid carrier physical state and surface structure on ß-carotene stability in formulated oil-in-water liquid food models, suggesting possible strategies for an enhanced stabilization of lipophilic labile compounds., (© 2011 Institute of Food Technologists®)

Inclusion of liposoluble bioactive compounds in fortified foods represents a complex challenge due to the labile nature of such compounds and the instability of oil-in-water emulsion-based delivery systems. In the present study, dispersions prepared with 10% (w/w) sunflower oil (SO) or hydrogenated palm kernel oil (HPKO) containing 0.05% (w/w) β-carotene were stabilized by various concentrations of whey protein isolate (WPI) or sodium caseinate (NaCas) (0.1 to 2.0% w/w) in 30% (w/w) sucrose aqueous solutions. Physicochemical characterization of emulsions was done considering the particle size, the particle surface protein coverage, and the physical state of continuous and dispersed phases. Physical stability of the systems and their protection properties on β-carotene were compared. The lipid carrier type and interfacial structure were investigated as the two key factors which regulate the stability of labile lipophilic bioactive molecules in food model systems. Our results showed high β-carotene stability when O/W systems were stable (protein concentration ≥0.8% w/w.) A (partially) solid lipid carrier (HPKO) enhanced protection compared to the liquid carrier (SO) as the bioactive molecules were entrapped in isolated domains within the solid lattice and kept apart from reactive species in the surroundings. NaCas provided a better barrier than WPI due to the different amino acid composition and interface structure which significantly reduced β-carotene degradation rate.

Thirty-four 'normal' preterm newborns were tested at 40 weeks postconceptional age. To separate electromagnetic artifacts from cochlear potentials and subsequent auditory brainstem responses, a test was given using an insert earphone at 90, 70, 50, 30 dB nHL. The detectability of receptor potentials, waves I, III, V, as a function of stimulus intensity is described: at 90 dB nHL exclusively, we could always identify these peaks because of the better morphological distinctiveness of each potential. When trying to evaluate the acoustic pathway in premature newborns, we suggest that brainstem response audiometry should be performed at 90 dB nHL with an insert earphone.

Phosphatidylcholines (PC) are secreted into the bile via a membrane transport protein(s). Recently, evidence for ATP-dependent mdr2-encoded PC transport as well as for carrier-mediated PC transport had been reported. Therefore, we investigated whether mdr2 P-glycoprotein is involved in the transport of a water-soluble short chain phosphatidylcholine analogue L-alpha-dibutyroyl-PC (diC4PC) induced by expression of liver mRNA in Xenopus laevis oocytes. Expression of mouse and rat mdr2 cRNA did not result in diC4PC net uptake in Xenopus laevis oocytes. By contrast oocytes showed a similar carrier-mediated uptake activity for diC4PC after injection of mouse, rat and human liver total mRNA (Km 7.7, 9.6, and 11.6 mM). Antisense inhibition of mdr2 mRNA expression increased diC4PC uptake induced by total liver mRNA from mouse and rat. The present data prove the existence of a specific mRNA for a non-mdr2-coded cell membrane PC carrier in mouse, rat, and human liver which exhibits similar transport affinity for diC4PC as the PC carrier in rat liver canalicular membranes.

The gene coding for the prepolypeptides of alpha and beta, obtained as a 429 bp fragment from chromosomal DNA of Rhodospirillum rubrum S1 by polymerase chain reaction amplification, were cloned in tandem into the high-level expression vector pOTSNco 12 for expression in Escherichia coli. The vector pOTSNco12 is a derivative of the pAS vector system, which contains the strong lambda PL promotor and is under tight control by the cI857 repressor encoded by the expression strain AR58. Induction of transcription from the lambda PL promotor is achieved by shifting the growth temperature from 32 to 42 degrees C. Expression of the gene products was monitored by sodium dodecylsulfate polyacrylamide gel electrophoresis and western blotting. The expressed B875 light-harvesting prepolypeptides were located in the E. coli inner membrane and could not be removed by washing with high salt. The amount of expressed B875 light-harvesting prepolypeptides was estimated to be about 0.1% of the total soluble protein.

Adult swine (n = 18) were studied to compare the effects on neuronal morphology of hypothermic circulatory arrest with hypothermic very-low-flow cardiopulmonary bypass. Animals were anesthetized with halothane and prepared in a standard manner for nonpulsatile cardiopulmonary bypass. Monitored variables included mean arterial pressure, arterial blood gases, the processed electroencephalogram, and subdural brain temperature. Bypass was initiated with pump flows of 100 ml.kg-1.min-1, and mean arterial pressure was kept above 50 mm Hg at all times. Animals were cooled to 18 degrees C, using a heat exchanger, and were randomly assigned to one of three groups. Group 1 animals were control animals who underwent 1 hour of hypothermic cardiopulmonary bypass. Group 2 animals underwent 1 hour of circulatory arrest. Group 3 animals underwent 1 hour of very-low-flow cardiopulmonary bypass (10% of normal). At the end of the 1 hour of hypothermic bypass, very-low-flow bypass, or arrest period, animals were rewarmed to 37 degrees C with normal bypass flows, and normothermic perfusion continued for 1 additional hour. Animals were then perfusion fixed with formalin and the brains were removed for electron microscopic analysis. Electron microscopic analysis was used to determine the effects of treatment and was limited to 20 neurons of the CA1 sector of the hippocampus in each animal. Golgi bodies were identified and classified as normal, mildly affected, or severely affected. Animals subjected to either very-low-flow bypass or circulatory arrest had significantly more severely affected and significantly fewer normal Golgi bodies than control animals (p < 0.001). Animals maintained with very-low-flow bypass, however, had significantly more severely affected and fewer normal Golgi bodies than animals subjected to circulatory arrest (p < 0.001). We conclude that under the conditions of this experiment very-low-flow hypothermic cardiopulmonary bypass is associated with significantly greater neuronal Golgi abnormalities than total circulatory arrest.

This anomaly is protean in its presentations and often hard to diagnose. A case report focuses on embryologic as well as diagnostic and therapeutic aspects.

Peroxisomes, participants in lipid metabolism, have been shown to be altered in liver in two metabolic diseases in which long-chain fatty acids accumulate in tissues: Zellweger's syndrome and neonatal adrenoleukodystrophy (ALD). The intestine also plays a role in lipid metabolism, and we have had the opportunity to compare peroxisomes in normal intestinal epithelium with those from patients with Zellweger's syndrome and neonatal ALD at the electron microscopic level by using the combined techniques of cytochemistry and stereological analysis. Peroxisomes were numerous in intestinal epithelium of the normal individuals. They were ellipsoidal in shape with average diameters of 0.37 by 0.56 micron and filled with coarsely granular, DAB+ content. Peroxisomes in the intestinal epithelium of the ALD patient were similar in appearance and number but smaller in size (0.28 by 0.44 micron). Peroxisomes of normal appearance were absent from the intestinal epithelium of patients with Zellweger's syndrome; DAB+ content, however, was observed in rare, membrane-bound structures of much smaller size (0.12 by 0.19 micron). In liver of patients with Zellweger's syndrome, peroxisomes are lacking; in neonatal ALD they are abnormal in appearance and greatly reduced in number. The presence of rare minute peroxisomes in the intestinal epithelium in Zellweger's syndrome and of small peroxisomes in this epithelium in neonatal ALD indicate that peroxisomes in the intestinal epithelium are affected in these diseases, but to a lesser extent than in the liver. In the ALD intestinal epithelium, DAB+ material was also seen in long, sinuous, tubular or cisternal elements intermingled and occasionally in continuity with peroxisomes. It is suggested that these represent the early stages of peroxisome formation, the peroxisomal reticulum as originally envisioned by Lazarow, while the rare structures seen in Zellweger's represent rudiments of such a reticulum. Lamellar inclusions and clear spaces occurred in the cytoplasm adjacent to these structures indicating either that material accumulated there had been extracted during fixation or that these regions are more susceptible to autolysis. Mitochondria are also involved in lipid metabolism and have been reported to be abnormal in Zellweger's tissue. No qualitative differences were observed in the mitochondria of the intestinal epithelia examined in this study. Although quantitation revealed a greater mean volume, number, and surface density of mitochondria in the intestinal epithelia of neonatal ALD, it was not a statistically significant difference in all cases.

35 neonates (mean gestational age: 34.9 +/- (SD) 3.5 weeks; mean birth weight: 2,180 +/- 890 g) treated with amikacin were examined for possible ototoxicity and nephrotoxicity. Audiometric tests were performed at 14.2-30.0 months postconceptional age by cross-checking behavioural with brain stem-evoked response audiometry. Only 1 infant was found with a mild hearing loss with brain stem audiometry, which, however, could not be attributed to amikacin with certainty. The effect of the aminoglycoside on the kidney was studied both by monitoring serum creatinine and the urinary elimination of the lysosomal enzyme N-acetylglucosaminidase (NAG). No significant difference was found in serum creatinine between the group under investigation and a control group of untreated premature neonates. A transient elevation was found in NAG urinary excretion in the neonates treated with amikacin. These results lead to the conclusion that amikacin at the recommended dosages (7.5 mg/kg every 12 h) causes a subclinical and reversible tubular damage in the neonate despite the high serum concentrations of the drug. The clinical relevance of this finding is discussed.

COSAS, an acronym for COmputerized Stereological Analysis System, is software designed to facilitate the process of quantitative morphological analysis. It is a flexible, reliable, and easy to use software system which provides the biologist with a simple means for performing stereological analysis. It provides for estimation of volume and surface density and allows for direct calculation of diameter information from planimetric data. The latter capability is unique to COSAS and provides the ability to calculate longest and shortest diameters for convex profiles, enabling calculation of number density. Access to all intermediate calculations is possible, so the process of transformation of raw data into the stereological descriptors can be completely exposed. This serves two purposes: (1) it provides a means for tracing results; (2) the intermediate values can be utilized independently. This system was used to analyze peroxisomes in tissues from patients with Zellweger's disease and neonatal-onset adrenoleukodystrophy. The system was also used to facilitate an immunocytochemical analysis of cytochrome P450 topology in the endoplasmic reticulum membrane.