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The objectives of this study are to identify the factors that influence the decision of rubber farmers to convert rubber plants to oil palm which consists of internal factors including plant age (years), land area (ha), number of dependents (org), and external factors include the distance from home to land (km), marketing channels, selling price (Rp/kg); The method used in this study was the survey method. The sampling method used in this randomized uneven study with a sample of 21 farmers who did not convert plants (rubber farmers) and 21 farmers who converted (oil palm farmers). The results of this research used the Binary logistic data processing method and showed that the factors that significantly affected farmers' decisions in converting rubber crops to oil palm were plant age, land area, number of dependents, and gap distance, while those that did not significantly affect the marketing channel. These results indicate that internal factors are more significantly influential than external factors in the decision-making of farmers to convert rubber plants to oil palms. The results of the t-test analysis indicate that tcount 27,581> ttable 2,021, which means rejecting H0 or there is a significant difference in income, between the income of farmers who convert crops (oil palm farmers) is greater than farmers who do not convert crops (rubber farmer) with the difference in an average income of Rp 8,925,519/ha/year.

The increasing of cropping index (CI) has contributed to an increase in rice production and regional food availibiality. The increasing of cropping index, especially in rainfed rice fields, has opened up opportunities for rice farmers to increase their income. However, there are still obstacles, so that CI 200 has not applied by all of rice farmers yet. The purpose of this research is to analyze rice farmers’ perceptions fields in implement of IC 200. The research method used was a survey method while the sampling method used was Disproportionated Stratified Random Sampling where each level of stratification was represented by 50 farmers who applied CI 200 and 50 farmers who did not apply. The results of the study found that three indicators were in the agreed criteria, consisting of indicators of conformity with technical aspects, social factors, and benefits from economic aspects. On the other hand, the implementation risk indicator based on the results of a survey conducted was in the disagree criteria.

The volume of beef imports from Australia increased enormously, while domestic beef production in few slow growth. This study aimed to analyze the demand for beef imports from Australia as well as domestic beef production Indonesia. The type of data that will be used in this research is data sequential time (time series) in the period 1990-2014. The price data obtained from the Ministry of Trade and the FAO and the Ministry of Agriculture. Imports of beef from the authority of Australia (Australian Bureau of Statistics, Meat Livestock Association, and the Australian Bureau of Agricultural and Resource Economics and Sciences). This research was conducted by econometric analysis. Imports of beef from Australia, influenced by the price of Australian beef, cattle imports from Australia, imports of beef origin other than Australia, and the price of beef imports from Australia. While domestic beef production or in the country affected by the imports of Australian beef and domestic beef consumption.

The construction of the Kayu Agung-Palembang-Betung (abbreviated as Kapal Betung) toll road has an impact on changes in the income of rice farmers. The aim of this research was to analyze the changes of farmers’ income after converting their non-tidal swamp land to toll road. The research method used in this study is the survey method. In this study the random sampling method used by incidentally. The samples were 60 persons. There was a change in farmers' income before and after converting their rice fields. The livelihoods of post-conversion farmers also experience an increase in the variety of occupations. Even though the value of farmers' income after conversion of paddy fields has increased; but this condition needs special attention for policy makers, because of the decreasing number of farmers who will have an impact on national food sovereignty.

The increased role of the sub-optimal land to support food security continue to be encouraged in Indonesia, given the more limited expansion for potential land. But until recently, development of sub-optimal land becomes not an easy thing. Ecological and technical barriers became the main issue. A series of these issues resulted in a high number of underemproleymeny and poverty in agriculture region. Technological inovation of agriculture and the business diversification can be seen be the solution to those issues. This research aims to analyze the impact of the technological innovation and business diversification on underemployment, working time, household income and also sustainable livelihoods of farmers on the sub-optimal land. The research was carried out in Pemulutan District, Ogan Ilir Regency, South Sumatra Province, Indonesia. The objects of research are farmers which adopter and non adopter technological innovation, and also work outside of paddy farming (business diversification). The research method is the survey. Method of sampling is stratified random sampling. Data obtained in the field analyses using descriptive statistics and inferesia. The results showed there are positive impact of technological innovation on the allocation of working time farmer households, the numbers underemployment, household income and livelihood sustainability. Determinant factors for farmers in applying technology and business diversification are paddy farming income, off-farm income, and age. The use of technology and business diversification proves to be one of the positive scenarios for sustainable livelihood of farmers in sub-optimal land.

The research aim is to use sea water as nutrient source of plant especially soybean, so it reduces amount of anorganic fertilizer usage. The research held in green house of Ecology and Plant Production Laboratory of Animal Husbandry and Agriculture Faculty, Diponegoro University Semarang. Sea water as sample was taken from Semarang Marina Beach. Soybean which choosen is local bean of Grobogan. The design arranged was completely randomized design with factorial design 4 x 2 in 4 replications. First factor was sea water salinity level, L0 = without sea water and without mulch, L1 = sea water EC 1 mmhos/cm and water hyacinth mulch of 8 tons/ha, L2 = sea water EC 2 mmhos/cm and water hyacinth mulch of 8 tons/ha, L3 = sea water EC 3 mmhos/cm and water hyacinth mulch of 8 tons/ha. The second factor was Rhizobium inoculation, R1 = without rhizobium inoculation, and R2 = with Rhizobium inoculation. The parameters were length plant, number of leaves, fresh weight production of shoot and dry matter production. Based on variant analysis, showed no influenced between sea water salinity level and rhizobium inoculation againt to length plant, number of leaves, fresh weight production of shoot and dry matter production. Sea water and rhizobium inoculation tended to increase growth and production of soybean. Key words : soybean, nutrient of sea water, rhizobium, growth, production.

ABSTRAK. Air laut mengandung nutrisi yang dibutuhkan oleh tanaman. Tujuan penelitian adalah untuk mendapatkan informasi manfaat air laut untuk pertanaman kedelai. Penelitian dilaksanakan di lahan percobaan Fakultas Peternakan dan Pertanian Universitas Diponegoro Semarang. Materi adalah air laut dari Pantai Marina Semarang, dan kedelai lokal Grobogan. Penelitian menggunakan rancangan acak kelompok monofaktor delapan perlakuan empat ulangan. Perlakuan tingkat salinitas/EC (Electrical Conductivity) air penyiraman yaitu J0 = penyiraman air tawar; J1, J2, J3, J4, J5, J6, dan J7 berturut-turut adalah penyiraman air laut 1, 2, 3, 4, 5, 6, dan 7 mmhos/cm. Semua petak perlakuan menggunakan 8 ton/ha mulsa eceng gondok dan diinokulasi bakteri rhizobium. Parameter yang diamati panjang tanaman, jumlah daun, produksi berat segar dan bahan kering jerami, produksi biji, kadar protein kasar dan serat kasar jerami kedelai. Analisis ragam, menunjukkan penyiraman air laut hingga EC 7 mmhos/cm belum menunjukkan pengaruh yang signifikan pada pertumbuhan, produksi, maupun kualitas jerami kedelai dibandingkan penyiraman dengan air tawar. Penyiraman air laut EC 7 mmhos/cm menghasilkan rerata panjang tanaman 26,53 cm, jumlah daun 245,25 helai petiole, produksi berat segar jerami 2029,5 g/petak, produksi bahan kering jerami 235,05 g/petak, produksi biji kedelai 515,22 g/petak, kadar protein kasar jerami 12,63 %, dan kadar serat kasar jerami 48,78 %. Growth, production, and quality of soybean straw at various levels of sea water to support ruminant feed ABSTRACT. Seawater contains nutrients needed by plant. The research aim was to get information the benefit of seawater for soybean. The research held in research field of Animal Husbandry and Agriculture Faculty, Diponegoro University Semarang. Seawater was taken from Semarang Marina Beach. Soybean was local bean of Grobogan. The experimental design used was a randomized block design eight treatment with four block.The factor was EC(electrical conductivity) level of watering diluted seawater namely J0 = Without sea water (fresh water); J1, J2, J3, J4, J5, J6, and J7 were EC level of watering diluted seawater of 1, 2, 3, 4, 5, 6, and 7 mmhos/cm respectively. All treatments using 8 ton/ha of water hyacinth mulch and inoculated of rhizobium bacteria. The parameters were plant length, number of leaves, seed production, fresh weight and dry matter production of shoot, crude protein and crude fibre content of soybean straw. Variant analysis, showed that watering sea water up to EC level 7 mmhos/cm has not shown significant influence on the growth, seed and straw production and quality of soybean straw compared to watering with fresh water. Level of watering diluted seawater EC 7 mmhos/cm yields plant length of 26,53 cm, number of leaves 245,25 pieces of petiole, weight production of fresh straw 2029,5 g/plot, weight production of dry straw 235,05 g/plot, weight production of seed 515.22 g/plot, 12.63% crude protein content, and a crude fiber content of 48.78%.

(The quality and absorption efficiency of N at Centrosema pubescens (centro) and Pueraria phaseoloides (puero) cause add of iodine fertilizer) ABSTRACT. The aimed of the experiment was to know the effect of iodine fertilizer when applied to the soil on the quality and the efficiency of Nitrogen absorption of legume. Centrosema pubescens (Centro) and Pueraria phaseoloides (Puero), soil from Tenbalang, manure, an organic fertilizer ( N,P,K and KI ). H2SO4 solution, aquades, HCL, NaOH, indikator MR+MB, and blanko solution. The experiment also used 24 pot for plantation with 10 kg capacity , analytic balance etc. The experiment used 4 x 2 factorial completely randomized design. The first factor were Centro (Centrosema pubescens)- (L1) and Puero (Pueraria phaseoloides)- (L2) and the level of iodine fertilizer (I0) without iodine, I1 (iodine 5 kg/ha), I2 (iodine 10 kg/ha) and I3 (iodine 15 kg/ha) were the second factor. Application of iodine fertilizer has no significant result, to nitrogen absorption, nitrat reductase activity, crude protein contain, and fibre. In the other hand, the interaction between legume and iodine has significantly result on the efficient and absorption of nitrogen, crude protein. The conclusion showed that puero was more responsive than centro, however centro was more potential in efficiency and absorption of N and crude protein.

Researchs related to purun (Heleocharisfistula) economic development should be encouraged for peatland sustainability. For long time, purun crafting was marketed traditionally only to traders who already have closebonding to the crafters. So, the research objective were (1) to evaluate the marketing performance, and (2) to design the marketing model’s plan of purun. This research was conducted in Pedamaran Area, Ogan Komering Ilir on September 2017. The result showed the performance of marketing of purun agroindustry in Pedamaran area has not yet all have a level of performance that corresponds to the importance of all marketing actors. There are gaps between performances and importances of marketing performance for atbributes, i.e. commitment in cooperation, profit, sales, prices, information, and skills improvement. The marketing model’s plan of purun are (1) building cooperative institution formally, (2) improving inputs for increasing profit, and (3) introducing e-commerce.

We analyzed the transcriptional signatures of mouse bone marrow-derived macrophages at different times after infection with promastigotes of the protozoan parasite Leishmania major. Ingenuity Pathway Analysis revealed that the macrophage metabolic pathways including carbohydrate and lipid metabolisms were among the most altered pathways at later time points of infection. Indeed, L. major promastiogtes induced increased mRNA levels of the glucose transporter and almost all of the genes associated with glycolysis and lactate dehydrogenase, suggesting a shift to anaerobic glycolysis. On the other hand, L. major promastigotes enhanced the expression of scavenger receptors involved in the uptake of Low-Density Lipoprotein (LDL), inhibited the expression of genes coding for proteins regulating cholesterol efflux, and induced the synthesis of triacylglycerides. These data suggested that Leishmania infection disturbs cholesterol and triglycerides homeostasis and may lead to cholesterol accumulation and foam cell formation. Using Filipin and Bodipy staining, we showed cholesterol and triglycerides accumulation in infected macrophages. Moreover, Bodipy-positive lipid droplets accumulated in close proximity to parasitophorous vacuoles, suggesting that intracellular L. major may take advantage of these organelles as high-energy substrate sources. While the effect of infection on cholesterol accumulation and lipid droplet formation was independent on parasite development, our data indicate that anaerobic glycolysis is actively induced by L. major during the establishment of infection.

The highly anticipated hearing in the Celmer case took place on 1 June 2018 before the Grand Chamber of the ECJ. The stakes are undoubtedly high. On the one hand, the efficiency of the European Arrest Warrant mechanism is clearly at risk — a risk which could lead to broader consequences for the whole architecture of mutual trust and recognition. On the other hand, the Celmer dispute goes to the heart of the problems surrounding the current Polish judicial reforms, and to the ensuing concerns about judicial independence. Taking into account the present negotiations between the Polish government and the Commission, Celmer is unquestionably both political and delicate in the extreme.

Headache is a significant and debilitating health problem, affecting more than half of the population worldwide. Migraine is a type of headache that is strongly associated with women and accounts for the high number of years lived with disability among women. The pathophysiology of migraine attacks may begin with a premonitory phase, followed by an aura phase and migraine headache. In women, many factors influence the prevalence of migraine, and sex hormone fluctuations around the menstruation cycle were believed to impact the pathogenesis of migraine. The International Classification of Headache Disorders, 3rd edition identifies menstrual migraine as pure menstrual migraine without aura and menstrually related migraine without aura. While migraine without aura (MwoA) was clearly associated with menstruation, migraine with aura (MwA) was generally unrelated to menstruation. Studies suggested that estrogen withdrawal is a trigger for MwoA, but high estrogen states are a trigger for MwA. During pregnancy, the increase in estrogen hypothetically prevents migraine attacks. There are several strategies for managing menstrual migraine, from acute/abortive, mini-preventive, and continuous preventive treatment. Managing migraine during pregnancy follows a similar strategy, but the drugs' safety profile should be considered., Competing Interests: The authors have declared that no competing interests exist., (Copyright © 2023, Barus et al.)

BackgroundIn high-income countries, hepatitis E virus (HEV) infection is mainly a zoonosis. However, it is also transfusion-transmissible and some countries, but not Italy, have introduced HEV screening for blood donations.AimWe assessed HEV infection prevalence and risk factors in a nationwide sample of Italian blood donors.MethodsWe selected 107 blood establishments (BE) distributed in the 20 Italian regions by a stratified two-stage design and invited them to participate in the study. Donors were tested for anti-HEV IgG and IgM and HEV RNA. Sociodemographic data and risk factors were collected through a questionnaire.ResultsOverall, 60 BE from 60 provinces in 19 Italian regions joined the study. We assessed HEV markers in 7,172 blood donors, of whom 6,235 completed the questionnaire. Overall crude and adjusted anti-HEV IgG prevalences were 8.3% and 5.5%, respectively. Overall anti-HEV IgM prevalence was 0.5%, while no blood donor was HEV RNA-positive. Anti-HEV IgG prevalence varied widely among regions (range: 1.3%-27.20%) and hyperendemic prevalences (> 40%) were detected in some provinces in two regions. Older age (AOR = 1.81; 95% CI: 1.36-2.41), foreign nationality (AOR = 2.77; 95% CI: 1.06-7.24), eating raw pork liver sausages (AOR = 2.23; 95% CI: 1.55-3.20) and raw homemade sausages (AOR = 3.63; 95% CI: 2.50-5.24) were independent infection predictors.ConclusionItalian blood donors showed a low to moderate HEV seroprevalence. High levels in some regions and/or provinces were mainly attributable to eating habits. Prevention should include avoiding consumption of raw or undercooked meat and safe production of commercial pork products.

Background: Melasma is present in 40% of cases in Southeast Asia. The condition is often unresponsive to therapy; treatment has variable success rates, and melasma has high recurrence rates. Lycopene-rich tomato extract is needed to avoid oxidative stress due to ultraviolet rays that cause melasma through the melanogenesis pathway. The aim of this study was to evaluate the effectiveness of an oral tomato extract supplement as an adjuvant for melasma therapy., Methods: The study recruited 62 subjects with melasma to a true-experimental clinic with a double-blind, randomized, pre and post-test control design over 12 weeks at the Diponegoro National Hospital, Indonesia. The subjects received an oral tomato extract supplement contains lycopene 30 mg (placebo). All subjects applied topical sunscreen and hydroquinone-4%-cream. Subjects were assessed by superoxide dismutase (SOD) and melasma area and severity index (MASI)., Results: Fifty-nine patients completed the research. The serum SOD levels in the treatment group (tomato extract supplementation) were higher than in the control group given the placebo, with delta SOD (P<0.05). The difference in MASI Scores after therapy in the treatment group had a significant decrease compared to the control group, with statistical review results suggesting that the difference was significant (P<0.05)., Conclusions: Supplementation of tomato extract as an adjuvant therapy can increase serum SOD levels and improve melasma severity.

Background: We investigated the presence of anti-SARS-CoV-2 antibodies in Italian plasma pools and intravenous immunoglobulins sent to our Institute (Italian National Institute of Health - Istituto Superiore di Sanità) in the context of the Official Control Authority Batch Release. The plasma pools were made up from donations collected in several different Italian regions from May 2017 to October 2020, i.e. in the pre-pandemic and pandemic periods., Materials and Methods: All plasma pools were initially tested for the qualitative detection of anti-SARS-CoV-2 antibodies against the nucleocapsid protein using the Roche Elecsys ® Anti-SARS-CoV-2 test kit. Plasma pools positive for these antibodies were further tested using the Roche Elecsys ® Anti-SARS-CoV-2 S test kit for the quantitative detection of antibodies against SARS-CoV-2 spike receptor binding domain. All plasma pools showing reactivity to these antibodies were tested undiluted for the presence of SARS-CoV-2 RNA using the Grifols Procleix SARS-CoV-2 transcription-mediated amplification assay. Intravenous immunoglobulins were tested using both test kits to determine the presence of anti-SARS-CoV-2 antibodies., Results: All plasma pools made up from donations collected in the pre-pandemic period were negative for anti-SARS-CoV-2 antibodies against the nucleocapsid protein. Of the plasma pools made up from donations collected from December 2018 to March 2020, only 1 pool out of 68 (1.4%), that was made up from donations from the Lombardy region, was reactive for these antibodies. Interestingly, 105 out of 174 (60.3%) of the plasma pools made up from donations collected from November 2018 to October 2020 showed the presence of these antibodies. All plasma pools positive for these antibodies were tested for antibodies against SARS-CoV-2 spike receptor binding domain and were confirmed positive., Discussion: None of these plasma pools tested were reactive for SARS-CoV-2 RNA. In the case of intravenous immunoglobulins, 20 out of 25 (80%) batches showed the presence of both anti-SARS-CoV-2 antibodies, reflecting the concentration in the plasma pools used for their production.

Objective: We report our 10-year experience in the endovascular treatment of acute traumatic thoracic aorta rupture at the Hospital Clinic., Methods: We reviewed 20 patients with an acute traumatic thoracic aorta lesion treated with a thoracic endograft between August 1997 and July 2007. All patients had multi-trauma resulting from high-velocity accidents or accidents with great impact. The diagnosis of aortic injury was made on a clinical basis and conventional imaging, confirmed by computed tomographic angiography. The following parameters were studied: age, sex, type and site of the lesion, type of endovascular graft, endovascular operation time, length of stay in the intensive care unit, length of stay in the hospital, immediate and perioperative complications, and mortality. Follow-up data were recorded, consisting of clinical visits, computed tomographic angiography, and plain chest radiographs at regular intervals (3, 6, and 12 months and every subsequent year). The mean follow-up was 58 months., Results: All endovascular procedures were technically successful, and the mean operating time for the endovascular procedure was 74 minutes (range, 55-130 minutes). We recorded an external iliac lesion during the procedure as an unique immediate complication, and it was corrected by an iliofemoral bypass. The only perioperative death (perioperative mortality rate of 5%) was unrelated to the aortic rupture or stent placement. There was no intervention-related mortality during the follow-up. Postoperative data showed no severe endovascular graft- or procedure-related morbidity. We recorded 2 cases of stent fracture, diagnosed by chest radiograph and computed tomographic angiography, without clinical impact or signs of endoleak., Conclusion: The short- and mid-term results of immediate endovascular repair of traumatic aortic injuries are promising, especially when compared with open surgical treatment, indicating that endovascular therapy is preferable in patients with multi-trauma and traumatic ruptures of the thoracic aorta. Nevertheless, long-term follow-up data are necessary to assess the overall durability of this procedure, considering the young age of these patients. The long-term follow-up results will determine whether endovascular treatment should replace open surgery as first-line therapy in thoracic aortic injuries.

Assessment of the viral load in hepatitis C virus (HCV) genotype 1-infected patients is critical before, during, and after antiviral therapy. In patients achieving a rapid virological response at week 4 of treatment, the viral load at the baseline is considered a predictive criterion of a sustained virological response 24 weeks after the discontinuation of treatment. A >or=2-log(10) drop in the viral load at week 12 of treatment (early virological response) triggers the continuation of therapy. We organized a multicenter study (MS) for diagnostic laboratories involved in the quantification of HCV RNA. Commercial assays, including two based on real-time reverse transcription-PCR (TaqMan system), and in-house methods, were used by the 61 participants. The overall reproducibility of the commercial quantitative nucleic acid amplification techniques (qNAT) was acceptable. As the intermethod variability among commercial qNAT for HCV RNA was still present, the manufacturers of these test kits should join efforts to harmonize the means of quantification of HCV RNA. This study also shows that caution should be exercised when the baseline viral load is evaluated and when the 2-log(10) reduction after 12 weeks of therapy is interpreted. Finally, this MS confirms the higher sensitivity of the commercial qNAT based on the TaqMan system, making them the elective assays for the monitoring of therapy.

Introduction and Objectives: To review experience at our center with the use of hybrid techniques for treating complex aneurysms of the thoracic aorta., Methods: The medical records of 41 patients (40 male) with complex aortic aneurysms affecting supra-aortic or visceral vessels who underwent hybrid procedures between 1998 and 2007 were reviewed retrospectively. All patients were in American Society of Anesthesiologists category IV. They were divided in two groups: group A comprised 32 patients with aneurysms involving the aortic arch and its branches (2 ascending aorta replacements, 1 arch repair, 13 carotid-carotid bypasses, 12 carotid-subclavian bypasses, and 4 with total arch debranching); and group B comprised 9 patients with thoracoabdominal aneurysms and visceral vessel transpositions (4 partial and 5 total debranching). The mean age in group A was 69.3+/-1.3 (range, 62-73) years and in group B, 71.5+/-5.0 (range, 68-74) years., Results: The mean hospital stay was 18+/-7.1 days (range, 5-35) and 12+/-8.2 days (range, 2-15) in groups A and B, respectively. Overall mortality was 12.2% (3.4% and 44.4% in groups A and B, respectively) and neurological morbidity was 3.4% and 11.1% in the two groups, respectively., Conclusions: The results obtained with hybrid treatment of the aortic arch area were comparable with those of conventional surgery. However, thoracoabdominal repair with complete revascularization of the visceral branches was associated with high mortality. Consequently, hybrid treatment is a valid option for high-risk patients who do not require complete revascularization in the visceral area.

Peptides derived from the sequence of a single-chain, recombinant, antiidiotypic antibody (IdAb; KT-scFv) acting as a functional internal image of a microbicidal, wide-spectrum yeast killer toxin (KT) were synthesized and studied for their antimicrobial activity by using the KT-susceptible Candida albicans as model organism. A decapeptide containing the first three amino acids (SAS) of the light chain CDR1 was selected and optimized by alanine replacement of a single residue. This peptide exerted a strong candidacidal activity in vitro, with a 50% inhibitory concentration of 0.056 microM, and was therefore designated killer peptide (KP). Its activity was neutralized by laminarin, a beta1-3 glucan molecule, but not by pustulan, a beta1-6 glucan molecule. KP also competed with the binding of a KT-like monoclonal IdAb to germinating cells of the fungus. In a rat model of vaginal candidiasis, local, postchallenge administration of KP was efficacious in rapidly abating infections caused by fluconazole-susceptible or -resistant C. albicans strains. In systemic infection of BALB/c or SCID mice preinfected intravenously with a lethal fungal load, KP caused a highly significant prolongation of the median survival time, with >80% of the animals still surviving after >60 days, whereas >90% of control mice died within 3 to 5 days. KP is therefore the first engineered peptide derived from a recombinant IdAb retaining KT microbicidal activity, probably through the interaction with the beta-glucan KT receptor on target microbial cells.

The protective roles of different lymphocyte subsets were investigated in a rat vaginal candidiasis model by adoptive transfer of vaginal lymphocytes (VL) or sorted, purified CD3(+) T cells, CD4(+) or CD8(+) T cells, or CD3(-) CD5(+) B cells from the vaginas of naïve or immune rats following three rounds of Candida albicans infection. The adoptive transfer of total VL from nonimmune animals did not alter the course of vaginal candidiasis of the recipient rats. In contrast, the animals receiving total VL or CD3(+) T cells from immune rats showed a highly significant acceleration of fungus clearance compared with animals which received nonimmune VL. The animals with vaginal CD3(-) CD5(+) B cells transferred from immune rats also had fewer Candida CFU than the controls, but fungal clearance was significantly retarded with respect to the animals administered immune T cells. Sorted, purified CD4(+) and CD8(+) vaginal T cells from immune rats were also adoptively transferred to naïve animals. Although both populations were seen to accelerate the clearance of the fungus from the vagina, CD4(+) T cells were much more effective than CD8(+) T cells. Overall, there was no difference between the antifungal effects of immune vaginal CD4(+) T cells and those achievable with the transfer of whole, immune VL. Histological observations of the vaginal tissues of rats with adoptively transferred immune T cells demonstrated a remarkable accumulation of lymphocytes in the subepithelial lamina propria and also infiltrating the mucosal epithelium. These results strongly suggest that distinct vaginal lymphocyte subsets participate in the adaptive anti-Candida immunity at the vaginal level, with the vaginal CD4(+) T cells probably playing a major role.

Oophorectomized, estrogen-treated rats were immunized by the intravaginal or intranasal route with a mannoprotein extract (MP) or secreted aspartyl proteinases (Sap) of Candida albicans, with or without cholera toxin as a mucosal adjuvant. Both routes of immunization were equally effective in (i) inducing anti-MP and anti-Sap vaginal antibodies and (ii) conferring a high degree of protection against the vaginal infection by the fungus. These data suggest that appropriate fungal antigens and adjuvant can be used to protect against candidal vaginitis, by either route.

Humoral (antibody [Ab]) and cellular Candida-specific immune responses in the vaginas of pseudoestrus rats were investigated during three successive infections by Candida albicans. After the first, protective infection, Abs against mannan and aspartyl proteinase antigens were present in the vaginal fluid, and their titers clearly increased during the two subsequent, rapidly healing infections. In all animals, about 65 and 10% of vaginal lymphocytes (VL) were CD3(+) (T cells) and CD3(-) CD5(+) (B cells), respectively. Two-thirds of the CD3(+) T cells expressed the alpha/beta and one-third expressed the gamma/delta T-cell receptor (TCR). This proportion slightly fluctuated during the three rounds of C. albicans infection, but no significant differences between infected and noninfected rats were found. More relevant were the changes in the CD4(+)/CD8(+) T-cell ratio, particularly for cells bearing the CD25 (interleukin-2 receptor alpha) marker. In fact, a progressively increased number of both CD4(+) alpha/beta TCR and CD4(+) CD25(+) VL was observed after the second and third Candida challenges, reversing the high initial CD8(+) cell number of controls (estrogenized but uninfected rats). The CD3(-) CD5(+) cells also almost doubled from the first to the third infection. Analysis of the cytokines secreted in the vaginal fluid of Candida-infected rats showed high levels of interleukin 12 (IL-12) during the first infection, followed by progressively increasing amounts of IL-2 and gamma interferon during the subsequent infections. No IL-4 or IL-5 was ever detected. During the third infection, VL with in vitro proliferative activity in response to an immunodominant mannoprotein antigen of C. albicans were present in the vaginal tissue. No response to this antigen by mitogen-responsive blood, lymph node, and spleen cells was found. In summary, the presence of protective Ab and T helper type 1 cytokines in the vaginal fluids, the in vitro proliferation of vaginal lymphocytes in response to Candida antigenic stimulation, and the increased number of activated CD4(+) cells and some special B lymphocytes after C. albicans challenge constitute good evidence for induction of locally expressed Candida-specific Ab and cellular responses which are potentially involved in anticandidal protection at the vaginal level.

A clear understanding of the pathogenesis of fungal disease remains elusive. While technological advances in molecular biology and microbial genetics have provided scientists with major new insights into both microbial virulence factors as well as host susceptibility to infection, there is currently no substitute for animal models in elucidating microbe-host interactions. Animal models are also essential for the evaluation of new antimicrobial agents, including studies of efficacy, adverse reactions and pharmacokinetics. The single most important advance in animal models in the last decade, has been the availability of genetically unique strains of animals as alternative to animals treated with immunosuppressive drugs for use in studies on microbial virulence and host defence mechanisms. These unique strains of test animals also enhance our understanding of the modes of action of antifungal drugs and their metabolism. Some of these advances will be discussed in this symposium.