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Abstract We study the solvability of a system of second-order differential equations with Dirichlet boundary conditions and non-local terms depending upon a parameter. The main tools used are a dual variational method and the topological degree.

Ever since agriculture started, plants have been bred to obtain better yields, better fruits, or sustainable products under uncertain biotic and abiotic conditions. However, a new way to obtain products from plant cells emerged with the development of recombinant DNA technologies. This led to the possibility of producing exogenous molecules in plants. Furthermore, plant chemodiversity has been the main source of pharmacological molecules, opening a field of plant biotechnology directed to produce high quality plant metabolites. The need for different products by the pharma, cosmetics agriculture and food industry has pushed again to develop new procedures. These include cell production in bioreactors. While plant tissue and cell culture are an established technology, beginning over a hundred years ago, plant cell cultures have shown little impact in biotechnology projects, compared to bacterial, yeasts or animal cells. In this review we address the different types of bioreactors that are currently used for plant cell production and their usage for quality biomolecule production. We make an overview of Nicotiana tabacum , Nicotiana benthamiana , Oryza sativa , Daucus carota , Vitis vinifera and Physcomitrium patens as well-established models for plant cell culture, and some species used to obtain important metabolites, with an insight into the type of bioreactor and production protocols., Competing Interests: FVN and JMC both work for BIONET. The company had no influence on the design or ideas described in the review. The remaining 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 author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2023 Verdú-Navarro, Moreno-Cid, Weiss and Egea-Cortines.)

Ticks are vectors of diseases that affect humans and animals worldwide. Tick vaccines have been proposed as a cost-effective and environmentally sound alternative for tick control. Recently, the Rhipicephalus microplus Subolesin (SUB)-Anaplasma marginale MSP1a chimeric antigen was produced in Escherichia coli as membrane-bound and exposed protein and used to protect vaccinated cattle against tick infestations. In this research, lipidomics and proteomics characterization of the E. coli membrane-bound SUB-MSP1a antigen showed the presence of components with potential adjuvant effect. Furthermore, vaccination with membrane-free SUB-MSP1a and bacterial membranes containing SUB-MSP1a showed that bacterial membranes enhance the immunogenicity of the SUB-MSP1a antigen in animal models. R. microplus female ticks were capillary-fed with sera from pigs orally immunized with membrane-free SUB, membrane bound SUB-MSP1a and saline control. Ticks ingested antibodies added to the blood meal and the effect of these antibodies on reduction of tick weight was shown for membrane bound SUB-MSP1a but not SUB when compared to control. Using the simple and cost-effective process developed for the purification of membrane-bound SUB-MSP1a, endotoxin levels were within limits accepted for recombinant vaccines. These results provide further support for the development of tick vaccines using E. coli membranes exposing chimeric antigens such as SUB-MSP1a., (Copyright © 2015 Elsevier GmbH. All rights reserved.)

Test equating is a method of making the test scores from different test forms of the same assessment comparable. In the equating process, an important step involves continuizing the discrete score distributions. In traditional observed-score equating, this step is achieved using linear interpolation (or an unscaled uniform kernel). In the kernel equating (KE) process, this continuization process involves Gaussian kernel smoothing. It has been suggested that the choice of bandwidth in kernel smoothing controls the trade-off between variance and bias. In the literature on estimating density functions using kernels, it has also been suggested that the weight of the kernel depends on the sample size, and therefore, the resulting continuous distribution exhibits bias at the endpoints, where the samples are usually smaller. The purpose of this article is (a) to explore the potential effects of atypical scores (spikes) at the extreme ends (high and low) on the KE method in distributions with different degrees of asymmetry using the randomly equivalent groups equating design (Study I), and (b) to introduce the Epanechnikov and adaptive kernels as potential alternative approaches to reducing boundary bias in smoothing (Study II). The beta-binomial model is used to simulate observed scores reflecting a range of different skewed shapes., Competing Interests: Declaration of Conflicting Interests: The author(s) declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article.

Diseases transmitted by arthropod vectors such as ticks greatly impact human and animal health. In particular, many diseases of dogs and cats are potentially transmissible to people by arthropod vectors and therefore their control is important for the eradication of vector-borne diseases (VBD). Vaccination is an environmentally friendly alternative for vector control that allows control of several VBD by targeting their common vector. Recent results have shown that it is possible to use vector protective antigens for the control of arthropod vector infestations and pathogen infection. However, as reviewed in this paper, very little progress has been made for the control of ectoparasite infestations and VBD in pets using vaccination with vector protective antigens. The growing interaction between pets and people underlines the importance of developing new interventions for the monitoring and control of VBD., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Background: The control of vector-borne diseases is important to improve human and animal health worldwide. Malaria is one of the world's deadliest diseases and is caused by protozoan parasites of the genus Plasmodium, which are transmitted by Anopheles spp. mosquitoes. Recent evidences using Subolesin (SUB) and Akirin (AKR) vaccines showed a reduction in the survival and/or fertility of blood-sucking ectoparasite vectors and the infection with vector-borne pathogens. These experiments suggested the possibility of using AKR for malaria control., Methods: The role of AKR on Plasmodium berghei infection and on the fitness and reproduction of the main malaria vector, Anopheles gambiae was characterized by evaluating the effect of akr gene knockdown or vaccination with recombinant mosquito AKR on parasite infection levels, fertility and mortality of female mosquitoes., Results: Gene knockdown by RNA interference in mosquitoes suggested a role for akr in mosquito survival and fertility. Vaccination with recombinant Aedes albopictus AKR reduced parasite infection in mosquitoes fed on immunized mice when compared to controls., Conclusions: These results showed that recombinant AKR could be used to develop vaccines for malaria control. If effective, AKR-based vaccines could be used to immunize wildlife reservoir hosts and/or humans to reduce the risk of pathogen transmission. However, these vaccines need to be evaluated under field conditions to characterize their effect on vector populations and pathogen infection and transmission.

Tuberculosis (TB) remains a pandemic affecting billions of people worldwide, thus stressing the need for new vaccines. Defining the correlates of vaccine protection is essential to achieve this goal. In this study, we used the wild boar model for mycobacterial infection and TB to characterize the protective mechanisms elicited by a new heat inactivated Mycobacterium bovis vaccine (IV). Oral vaccination with the IV resulted in significantly lower culture and lesion scores, particularly in the thorax, suggesting that the IV might provide a novel vaccine for TB control with special impact on the prevention of pulmonary disease, which is one of the limitations of current vaccines. Oral vaccination with the IV induced an adaptive antibody response and activation of the innate immune response including the complement component C3 and inflammasome. Mycobacterial DNA/RNA was not involved in inflammasome activation but increased C3 production by a still unknown mechanism. The results also suggested a protective mechanism mediated by the activation of IFN-γ producing CD8+ T cells by MHC I antigen presenting dendritic cells (DCs) in response to vaccination with the IV, without a clear role for Th1 CD4+ T cells. These results support a role for DCs in triggering the immune response to the IV through a mechanism similar to the phagocyte response to PAMPs with a central role for C3 in protection against mycobacterial infection. Higher C3 levels may allow increased opsonophagocytosis and effective bacterial clearance, while interfering with CR3-mediated opsonic and nonopsonic phagocytosis of mycobacteria, a process that could be enhanced by specific antibodies against mycobacterial proteins induced by vaccination with the IV. These results suggest that the IV acts through novel mechanisms to protect against TB in wild boar.

Background: Ticks represent a significant health risk to animals and humans due to the variety of pathogens they can transmit during feeding. The traditional use of chemicals to control ticks has serious drawbacks, including the selection of acaricide-resistant ticks and environmental contamination with chemical residues. Vaccination with the tick midgut antigen BM86 was shown to be a good alternative for cattle tick control. However, results vary considerably between tick species and geographic location. Therefore, new antigens are required for the development of vaccines controlling both tick infestations and pathogen infection/transmission. Tick proteins involved in tick-pathogen interactions may provide good candidate protective antigens for these vaccines, but appropriate screening procedures are needed to select the best candidates., Methods: In this study, we selected proteins involved in tick-Anaplasma (Subolesin and SILK) and tick-Babesia (TROSPA) interactions and used in vitro capillary feeding to characterize their potential as antigens for the control of cattle tick infestations and infection with Anaplasma marginale and Babesia bigemina. Purified rabbit polyclonal antibodies were generated against recombinant SUB, SILK and TROSPA and added to uninfected or infected bovine blood to capillary-feed female Rhipicephalus (Boophilus) microplus ticks. Tick weight, oviposition and pathogen DNA levels were determined in treated and control ticks., Results: The specificity of purified rabbit polyclonal antibodies against tick recombinant proteins was confirmed by Western blot and against native proteins in tick cell lines and tick tissues using immunofluorescence. Capillary-fed ticks ingested antibodies added to the blood meal and the effect of these antibodies on tick weight and oviposition was shown. However, no effect was observed on pathogen DNA levels., Conclusions: These results highlighted the advantages and some of the disadvantages of in vitro tick capillary feeding for the characterization of candidate tick protective antigens. While an effect on tick weight and oviposition was observed, the effect on pathogen levels was not evident probably due to high tick-to-tick variations among other factors. Nevertheless, these results together with previous results of RNA interference functional studies suggest that these proteins are good candidate vaccine antigens for the control of R. microplus infestations and infection with A. marginale and B. bigemina.

Background: Despite the use of chemical acaricides, tick infestations continue to affect animal health and production worldwide. Tick vaccines have been proposed as a cost-effective and environmentally friendly alternative for tick control. Vaccination with the candidate tick protective antigen, Subolesin (SUB), has been shown experimentally to be effective in controlling vector infestations and pathogen infection. Furthermore, Escherichia coli membranes containing the chimeric antigen composed of SUB fused to Anaplasma marginale Major Surface Protein 1a (MSP1a) (SUB-MSP1a) were produced using a simple low-cost process and proved to be effective for the control of cattle tick, Rhipicephalus (Boophilus) microplus and R. annulatus infestations in pen trials. In this research, field trials were conducted to characterize the effect of vaccination with SUB-MSP1a on tick infestations and the prevalence of tick-borne pathogens in a randomized controlled prospective study., Methods: Two cattle and two sheep farms with similar geographical locations and production characteristics were randomly assigned to control and vaccinated groups. Ticks were collected, counted, weighed and classified and the prevalence of tick-borne pathogens at the DNA and serological levels were followed for one year prior to and 9 months after vaccination., Results: Both cattle and sheep developed antibodies against SUB in response to vaccination. The main effect of the vaccine in cattle was the 8-fold reduction in the percent of infested animals while vaccination in sheep reduced tick infestations by 63%. Female tick weight was 32-55% lower in ticks collected from both vaccinated cattle and sheep when compared to controls. The seroprevalence of Babesia bigemina was lower by 30% in vaccinated cattle, suggesting a possible role for the vaccine in decreasing the prevalence of this tick-borne pathogen. The effect of the vaccine in reducing the frequency of one A. marginale msp4 genotype probably reflected the reduction in the prevalence of a tick-transmitted strain as a result of the reduction in the percent of tick-infested cattle., Conclusions: These data provide evidence of the dual effect of a SUB-based vaccine for controlling tick infestations and pathogen infection/transmission and provide additional support for the use of the SUB-MSP1a vaccine for tick control in cattle and sheep.

Tick-borne pathogens cause diseases that greatly impact animal health and production worldwide. The ultimate goal of tick vaccines is to protect against tick-borne diseases through the control of vector infestations and reducing pathogen infection and transmission. Tick genetic traits are involved in vector-pathogen interactions and some of these molecules such as Subolesin (SUB) have been shown to protect against vector infestations and pathogen infection. Based on these premises, herein we characterized the efficacy of cattle vaccination with tick proteins involved in vector-pathogen interactions, TROSPA, SILK, and Q38 for the control of cattle tick, Rhipicephalus (Boophilus) microplus infestations and infection with Anaplasma marginale and Babesia bigemina. SUB and adjuvant/saline placebo were used as positive and negative controls, respectively. The results showed that vaccination with Q38, SILK and SUB reduced tick infestations and oviposition with vaccine efficacies of 75% (Q38), 62% (SILK) and 60% (SUB) with respect to ticks fed on placebo control cattle. Vaccination with TROSPA did not have a significant effect on any of the tick parameters analyzed. The results also showed that vaccination with Q38, TROSPA and SUB reduced B. bigemina DNA levels in ticks while vaccination with SILK and SUB resulted in lower A. marginale DNA levels when compared to ticks fed on placebo control cattle. The positive correlation between antigen-specific antibody titers and reduction of tick infestations and pathogen infection strongly suggested that the effect of the vaccine was the result of the antibody response in vaccinated cattle. Vaccination and co-infection with A. marginale and B. bigemina also affected the expression of genes encoding for vaccine antigens in ticks fed on cattle. These results showed that vaccines using tick proteins involved in vector-pathogen interactions could be used for the dual control of tick infestations and pathogen infection., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Diseases transmitted by arthropod vectors such as mosquitoes, ticks and sand flies greatly impact human and animal health, and therefore, their control is important for the eradication of vectorborne diseases (VBD). Vaccination is an environmentally friendly alternative for vector control that allows control of several VBD by targeting their common vector. Recent results have suggested that subolesin (SUB) and its orthologue in insects, akirin (AKR) are good candidate antigens for the control of arthropod vector infestations and pathogen infection. SUB was discovered as a tick-protective antigen in Ixodes scapularis. Vaccination trials with recombinant SUB/AKR demonstrated effective control of arthropod vector infestations in various hard and soft tick species, mosquitoes, sand flies, poultry red mites and sea lice by reducing their numbers, weight, oviposition, fertility and/or moulting. SUB/AKR vaccination also reduced tick infection with tickborne pathogens, Anaplasma phagocytophilum, A. marginale, Babesia bigemina and Borrelia burgdorferi. The effect of vaccination on different hosts, vector species, developmental stages and vectorborne pathogen infections demonstrated the feasibility of SUB/AKR universal vaccines for the control of multiple vector infestations and for reduction in VBD., (© 2013 Blackwell Verlag GmbH.)

Tick-borne encephalitis (TBE) is a growing zoonotic disease caused by tick-borne encephalitis virus (TBEV) infection. Although effective vaccines for TBEV are available, on-going vaccination efforts are insufficient to prevent increase in TBE cases annually. Vaccination with arthropod vector antigens to reduce vector infestations and vector capacity allows control of several vector-borne diseases by targeting their common vector. Subolesin (SUB) is a tick protective antigen that has a role in tick innate immunity and other molecular pathways and has been shown to protect against tick infestations and infection by vector-borne pathogens. However, SUB expression and the effect of SUB immunization have not been evaluated for tick-borne viruses. Herein, we showed that SUB expression is downregulated during Ixodes ricinus tick feeding but induced in ticks infected with TBEV, thus supporting a role for this molecule in tick innate immune response to virus infection. Immunization with recombinant SUB reduced SUB mRNA levels in nymphs co-feeding with infected females and suggested and effect on tick infestations in mice. However, SUB immunization did not reduce tick infection with TBEV nor protect mice against TBE. These results suggested that SUB is not a good candidate antigen for vaccination against TBEV and support the characterization of tick-pathogen interactions to identify mechanisms that could be targeted to reduce TBEV infection and transmission by ticks., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Diseases transmitted by arthropod vectors such as mosquitoes, ticks and sand flies greatly impact human and animal health and thus their control is important for the eradication of vector-borne diseases (VBD). Vaccination is an environmentally friendly alternative for vector control that allows control of several VBD by targeting their common vector. Recent results have suggested that subolesin/akirin (SUB/AKR) is good candidate antigens for the control of arthropod vector infestations. Here we describe the comparative effect of vaccination with SUB, AKR and Q38 and Q41 chimeras containing SUB/AKR conserved protective epitopes on tick, mosquitoes and sand flies vector mortality, molting, oviposition and/or fertility. We demonstrated that SUB vaccination had the highest efficacy (E) across all vector species (54-92%), Q41 vaccination had the highest vaccine E in mosquitoes (99%) by reducing female survival and fertility, and Q38 vaccination had the highest effect on reducing mosquito (28%) and sand fly (26%) oviposition. The effect of vaccination on different developmental processes in several important arthropod vectors encourages the development of SUB/AKR universal vaccines for the control of multiple vector infestations and reduction of VBD., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Red deer (Cervus elaphus) and white-tailed deer (Odocoileus virginianus) are hosts for different tick species and tick-borne pathogens and play a role in tick dispersal and maintenance in some regions. These factors stress the importance of controlling tick infestations in deer and several methods such as culling and acaricide treatment have been used. Tick vaccines are a cost-effective alternative for tick control that reduced cattle tick infestations and tick-borne pathogens prevalence while reducing the use of acaricides. Our hypothesis is that vaccination with vector protective antigens can be used for the control of tick infestations in deer. Herein, three experiments were conducted to characterize (1) the antibody response in red deer immunized with recombinant BM86, the antigen included in commercial tick vaccines, (2) the antibody response and control of cattle tick infestations in white-tailed deer immunized with recombinant BM86 or tick subolesin (SUB) and experimentally infested with Rhipicephalus (Boophilus) microplus, and (3) the antibody response and control of Hyalomma spp. and Rhipicephalus spp. field tick infestations in red deer immunized with mosquito akirin (AKR), the SUB ortholog and candidate protective antigen against different tick species and other ectoparasites. The results showed that deer produced an antibody response that correlated with the reduction in tick infestations and was similar to other hosts vaccinated previously with these antigens. The overall vaccine efficacy was similar between BM86 (E=76%) and SUB (E=83%) for the control of R. microplus infestations in white-tailed deer. The field trial in red deer showed a 25-33% (18-40% when only infested deer were considered) reduction in tick infestations, 14-20 weeks after the first immunization. These results demonstrated that vaccination with vector protective antigens could be used as an alternative method for the control of tick infestations in deer to reduce tick populations and dispersal in regions where deer are relevant hosts for these ectoparasites., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Vaccines containing the Rhipicephalus (Boophilus) microplus BM86 and BM95 antigens protect cattle against tick infestations. Tick subolesin (SUB), elongation factor 1a (EF1a) and ubiquitin (UBQ) are new candidate protective antigens for the control of cattle tick infestations. Previous studies showed that R. microplus BM95 immunogenic peptides fused to the Anaplasma marginale major surface protein (MSP) 1a N-terminal region (BM95-MSP1a) for presentation on the Escherichia coli membrane were protective against R. microplus infestations in rabbits. In this study, we extended these results by expressing SUB-MSP1a, EF1a-MSP1a and UBQ-MSP1a fusion proteins on the E. coli membrane using this system and demonstrating that bacterial membranes containing the chimeric proteins BM95-MSP1a and SUB-MSP1a were protective (>60% vaccine efficacy) against experimental R. microplus and Rhipicephalus annulatus infestations in cattle. This system provides a novel, simple and cost-effective approach for the production of tick protective antigens by surface display of antigenic protein chimera on the E. coli membrane and demonstrates the possibility of using recombinant bacterial membrane fractions in vaccine preparations to protect cattle against tick infestations., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The ultimate goal of vector vaccines is the control of vector infestations while reducing pathogen infection and transmission to protect against the many diseases caused by vector-borne pathogens. Previously (Vaccine 2011;29:2248-2254), we demonstrated that subolesin vaccination and release of tick larvae after subolesin knockdown by RNA interference (RNAi) were effective for the control of cattle tick, Rhipicephalus (Boophilus) microplus infestations in cattle. In this study, we used the fact that these animals were naturally infected with Anaplasma marginale and Babesia bigemina to evaluate the effect of subolesin vaccination and gene knockdown on tick infection by these cattle tick-transmitted pathogens. Ticks fed on vaccinated cattle had lower subolesin mRNA levels when compared to controls, resembling RNAi results. A. marginale and B. bigemina infection was determined by PCR and decreased by 98% and 99%, respectively in ticks fed on vaccinated cattle and by 97% and 99%, respectively after subolesin knockdown. These results demonstrated that targeting subolesin expression by vaccination or RNAi results in lower subolesin mRNA and pathogen infection levels, probably due to the effect of subolesin downregulation on tick feeding, gene expression and gut and salivary glands tissue development and function. These results suggested that subolesin vaccines could be used for the dual control of tick infestations and pathogen infection, a result that could be relevant for other vectors and vector-borne pathogens., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Diseases caused by arthropod-borne pathogens greatly impact on human and animal health. Recent research has provided evidence that tick protective antigens can be used for development of vaccines with the dual target of controlling arthropod infestations and reducing their vector capacity for pathogens. As reviewed herein, protective antigens such as subolesin/akirin, which are highly conserved across vector species, show promise for use in development of a universal vaccine for the control of arthropod infestations and the reduction of pathogen transmission. However, further research is needed in critical areas towards achieving this goal., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Tick subolesin was shown in immunization trials using the recombinant protein to protect hosts against tick infestations. In this study, we demonstrated that subolesin vaccination and release of ticks after subolesin knockdown by RNA interference (RNAi) could be used for the control of Rhipicephalus (Boophilus) microplus tick infestations in cattle and suggested that the combination of these methods could increase the efficacy of cattle tick control under some circumstances. The greatest tick control was obtained when both release of ticks after subolesin knockdown and vaccination were used concurrently. However, modeling results suggested that vaccine efficacy could be increased if at least 80% of the ticks infesting cattle correspond to subolesin-knockdown ticks. The results of this proof-of-concept trial demonstrated the efficacy of the sterile acarine technique (SAT) through production of subolesin-knockdown larvae by dsRNA injection into replete females for the control of R. microplus tick infestations, alone or in combination with subolesin vaccination., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The control of arthropod vectors of pathogens that affect human and animal health is important for the eradication of vector-borne diseases. Recent evidences showed a reduction in the survival and/or fertility of mosquitoes, sand flies and poultry red mites fed in vitro with antibodies against the recombinant Aedes albopictus akirin. These experiments were the first step toward the development of a multi-target arthropod vaccine. In this study, we showed that the oviposition of A. albopictus and Phlebotomus perniciosus fed on mice vaccinated with recombinant A. albopictus akirin was reduced by 17% and 31%, respectively when compared to controls. However, Aedes aegypti mosquitoes were not affected after feeding on vaccinated mice. These results showed that recombinant A. albopictus akirin could be used to vaccinate hosts for the control of mosquito and sand fly infestations and suggested new experiments to develop improved vaccine formulations., (Copyright © 2010 Elsevier Ltd. All rights reserved.)

Objective: To conduct an opinion survey on osteoporosis in Spanish internists., Method: Survey sent by mail and by personal visit to members of the Spanish Internists Society. Collection of data on opinion on the disease, diagnostic and therapeutic attitude and means available (general laboratory analyses, conventional radiology, biochemical markers of bone remodeling, densitometry and ultrasounds) and preference when choosing a certain treatment., Results: A total of 538 internists answered. More than 90% of those surveyed consider that osteoporosis is a disease that should be treated by internists. A total of 93% consider that osteoporosis is a prevalent disease. More than 80% have access to densitometry., Conclusions: The majority of Spanish internists consider that osteoporosis is a disease that should be treated by internists and that it is a disease that enters into their action scope. In general, they have the means necessary for its study and treatment. Bisphosphonates constitute the drug of choice and calcium and vitamin D supplements are indicated in almost all the cases.

The role of ultrasound scanning (USS) in patients complaining of a breast lump where the clinical examination (CE) is normal is not clearly defined. To determine this in greater detail, all patients complaining of a breast lump underwent CE. Where no lump could be found, but was still reported by the patient, an USS was performed. All lesions underwent biopsy and/or aspiration, as well as mammography in suspicious cases or those over 40 years of age. This cohort represented 5% of all referrals in the study period. Four hundred and twenty women were prospectively studied in this way. Median follow up is 3.4 years (range 2.5-4.2 years). Twenty two had solid lumps (of which 3 were cancers) and 48 had cysts. Nineteen patients re-presented with symptoms in the same breast (median time = 12 months (range 4.5-20 months), all of which were imaged on USS: 15 cysts and 4 further cancers (3 in the same quadrant as the original lump, one contralateral) were identified. Women with symptomatic breast lumps and a normal CE can be considered a reliable indication that cancer is very unlikely to be present (negative predictive value = 0.98). Ultrasound may be a suitable complimentary investigation, which will relieve symptoms in those with cysts and can detect small clinically--and sometimes mammographically--occult breast cancers.

Background: Elderly patients represent a unique surgical challenge because of the associated complex comorbidity and diminished cardiopulmonary reserve. Therefore, minimally invasive surgery in the elderly may have a larger impact compared to the younger population. The aim of this study was to prospectively evaluate the experience of laparoscopic surgery in patients >or=70 years of age in our unit., Methods: Two hundred and thirty-two patients (34 females and 98 males) older than 70 years who underwent various elective and emergency laparoscopic procedures between 1992 and 1997 were assessed prospectively. Preoperative comorbidity, operative results, and postoperative outcomes were analyzed., Results: The median age of the patients was 76 years. The majority of patients were ASA class II. The mean hospital stay was 3.4 days. The overall morbidity and mortality rates were 10.8% and 3.4% respectively, and the conversion rate was 4.3%., Conclusions: Our experience suggests that laparoscopic surgery in the elderly is safe, is associated with short hospital stay, and produces less morbidity and mortality. Therefore, it should be adopted widely if the expertise in the area of laparoscopic surgery is available for this group of patients.

Objective: To ascertain the incidence of osteoporosis and epidemiological factors in patients admitted to an Internal Medicine Service of a University General Hospital in Madrid, Spain. We also took into account the reasons for admission, associated diseases, and the preventive and therapeutic diagnostic implications., Patients and Methods: The analytical, descriptive, retrospective and transversal study of one hundred patients over fifty years of age admitted to an internal medicine service during 1997, obtaining in addition to the clinical discharge report, epidemiological data with regard to the radiological diagnosis of osteoporosis, the reason for admission, past history, the prevention and treatment of the initial complaint as well as the osteoporosis previous diagnosis., Results: 26% of those patients included in the survey had radiologic criteria of osteoporosis although only in 1 per cent of them had osteoporosis been diagnosed previously. The main risk factors, apart from the age and sex, are the associated illnesses which cause malnutrition and immobilization, and the use of drugs, especially heparin., Conclusions: Osteoporosis is a condition common in patients above 50 especially women, who are admitted to an Internal Medicine Service. A quarter of those patients admitted have osteoporosis. Very few are diagnosed and treated. In this group of patients apart from the habitual risk factors, are added those for the associated disease generally chronic, and the treatment they receive. During their stay in the hospital osteoporotic patients are three times more likely to receive heparin and other drugs which aggravate osteoporotic, than non osteoporotic patients. For that reason the risk of fractures is greater and the need for prospective diagnostic studies is even more necessary in this group of patients.

The chemical-nutritional composition and some indexes of biological quality of sorghum (Sorghum saccharum, var. sugar drip) unwashed whole flour were studied. The culture and selection of sorghum were assayed in the "San Roque" Experimental Station of the Agriculture Department, Undersecretary of Agricultural Affairs, San Luis, Argentina. The obtained protein content was 7.5% (Nx6.25). The values for net protein utilization (NPU); true digestibility (tD); net protein ratio (NPR) and and relative net protein ratio (RNPR) were 12.4 +/- 0.6, 22.0 +/- 0.2, 0.8 +/- 0.1 and 24%, respectively. Deficient nitrogen utilization constitutes a limitation for the use of sorghum. However, considering its abundance and low cost, sorghum could become a profitable feeding resource if conveniently supplemented without increasing costs. Sorghum flour was supplemented with protein recovered from ricotta serum (50:50), discarded as industrial waste, in order to compensate for lysine and threonine deficiency in this cereal. Studies performed on this mixture gave RNPR values of 85.7%. This might permit us to infer that despite its low biological value, sorghum can be used as food resource, potentially for poultry, with adequate supplementation.

The study of Atriplex suberecta leaf protein concentrate has been included within our work line entitled "Biological value of the leaf protein of Atriplex" amino acid profile and chemical score. In order to evaluate nitrogen retention, net protein utilization (NPU), true digestibility (TD) and biological value (BV) data were obtained. Weight gain was studied after determining net protein ratio (NPR) and relative net protein ratio (RNPR) with the following results: CS = 96, calculated for sulphur amino acids; NPU = 60 +/- 4.00; TD = 76 +/- 5.20; BV = 87; NPR = 3.34 +/- 2.50 and RNPR = 88%, which enable us to infer that its protein is of good quality. These data, therefore, should indicate that the product studied could be a good complement for cereals, considering its high lysine content (CS superior to 100).

An account is given of some considerations concerning the chemical recognition and evaluation by biological methods of the quality of the flour obtained from Amaranthus quitensis seeds, submitted to grinding and sifting. The protein content of the flour (21.70g/100g) can be considered as very important, as well as that of the available lysine (5.20g/16gN). The calcium content was also remarkable (500mg/100g), and the starch ratio reached 45.00g/100g. The amount of nitrates found was 20.00mg/100g, which is an acceptable limit. The research for organic nitro compounds was negative. As for the biologic quality of the protein, the experiments revealed that it has quite an acceptable availability, as demonstrated by the following values: NPU = 42.50 +/- 6.10, D = 68.50 +/- 5.30, VB = 62, NPR = 2.10 +/- 1.80 and RNPR = 42.

The chemical and biological value of the flour from Cassia aphylla was determined. The chemical study showed that this flour contains a good amount of protein, calcium, phosphorus, iron, ascorbic acid, niacin and thiamine. It is also a good source of lysine and sulfur amino acids. Tests concerning nutritive value carried out were: net protein utilization (NPU), 54.7 +/- 2.45; digestibility, 71.00 +/- 0.30; biological value, 77.00, and protein efficiency ratio (PER) 1.74, corrected with respect to casein. Toxicological tests were performed with rats during a period of 30 days. The criteria used in these tests were: hematological data, organ weights, liver composition and histopathology of the liver and kidney. These tests revealed no signs of pathological damage under the experimental conditions used by us. Based on these results, it can be concluded that this product has a potential value as a feedstuff.

The present research was carried out to determine the nutritive quality of the flour and protein concentrate from Melilotus albus (white clover) seeds. The flour was studied first. The protein analysis showed methionine to be the first limiting amino acid with a chemical score of 25, with threonine as the second. The biological value obtained was 27. Supplementation studies were performed with different levels of methionine and it was found that the 0.3% level resulted in the best net performance. The biological value obtained under these conditions was 60. The protein efficiency ratio (PER) was also determined, with a value of 1.40 after being corrected with respect to casein. The protein was isolated after studying the pH optimum solubility and precipitation conditions until the flour coumarin compounds were eliminated. Further biological experiments were carried out with the supplemented isolated protein. Under these conditions, a PER value of 2.4 and a biological value of 69 were obtained. No toxicity was observed in rats of both sexes by administration of the protein concentrate during 60 days, at least in the parameters studied during this period.

The Atriplex lampa, chenopodiáceae, is a very abundant bush in the arid and semiarid regions of our Province. This work was undertaken to characterize from the chemical point of view, both the fresh leaves and the protein concentrate (PC) obtained from them. A second purpose was to determine the biological value of the protein, and to evaluate its capacity as a complement to wheat flour (WF). Through this study, the leaf was shown to have a high content of ashes, sodium, potassium and silica, what accounts for its low palatability. The PC obtained has a protein concentration of 59.37 g/100 g and its biological value is limited by the sulfur amino acids, with a chemical score of 85.70. Since the protein is poorly utilized, this results in a relatively low value of its net protein utilization (NPU), a fact which may be attributed to its low true digestibility. The studies on the complementary effects of PC on WF were made at three levels, i.e., 30, 50 and 60%, respectively. The results revealed that the best complementation, measured by the nitrogen utilization, happened to be when 50% of the protein contribution was provided by the A. lampa protein concentrate.

Plant leaf proteins have acquired great relevance during the last two decades because of their well-balanced amino acid composition. A study was therefore undertaken to evaluate the biological quality of the leaf protein of Atriplex numularia. The protein content of the fresh leaves from this plant was found to be 4.70 g/100, with a dry matter content of 18.70 g/100 g. A protein concentrate (PC) from the same material was then obtained by macerating the leaf in a 2% sodium sulfite solution at a pH of 10 and subjecting it to filtration and pressing. The product thus obtained had a dark greenish color and contained 55.42 g/100 g of protein. The amino acid analysis revealed that its protein has a balance similar to that of animal origin proteins, with a lysine and methionine content of 8.5 g/16 g N and 3.0 g/16 g N, respectively. Biological assays were then carried out to evaluate nitrogen utilization, with the following resulting values: net protein utilization (NPU) = 48.3 +/- 2.7; digestibility (D) = 58.0 +/- 1.4, and biological value (BV) = 83. As inferred from the NPU value, nitrogen utilization was low. Therefore, to improve digestibility values, the action of papain on the PC was assayed. The material thus treated was again submitted to biological trials, obtaining, this time a D of 75.4 +/- 1.05, and thus, an improved new NPU value of 54.8 +/- 1.1 (p less than 0.01).