New approach for reproductive toxicity assessment: chromatoid bodies as a target for methylmercury and polychlorinated biphenyls in prepubertal male rats.

This study investigated the reproductive toxicity of methylmercury (MeHg) and Aroclor (Sigma-Aldrich), alone or in combination, following exposure of prepubertal male rats considering the chromatoid body (CB) as a potential target. The CB is an important molecular regulator of mammalian spermatogenesis, primarily during spermatid cytodifferentiation. Male Wistar rats were exposed to MeHg and/or Aroclor , according the following experimental design: control group, which was administered in corn oil (vehicle) only; MeHg-treated group, which was administered 0.5mg kg-1 day-1 MeHg; Aroclor-treated group, which was administered 1mg kg-1 day-1 Aroclor; Mix-LD, group which was administered a low-dose mixture of MeHg (0.05mg kg-1 day-1) and Aroclor (0.1mg kg-1 day-1); and Mix-HD group, which was administered a high-dose mixture of MeHg (0.5mg kg-1 day-1) and Aroclor (1.0mg kg-1 day-1). MeHg was diluted in distilled water and Aroclor was made up in corn oil (volume 1mL kg-1). Rats were administered the different treatments from PND23 to PND53 by gavage, . The morphophysiology of CBs was analysed, together with aspects of steroid hormones status and regulation, just after the last treatment on PND53. In addition, the long-term effects on sperm parameters were assessed in adult animals. MeHg exposure increased mouse VASA homologue (MVH) protein levels in seminiferous tubules, possibly affecting the epigenetic status of germ cells. Aroclor produced morphological changes to CB assembly, which may explain the observed morphological defects to the sperm flagellum and the consequent decrease in sperm motility. There were no clear additive or synergistic effects between MeHg and Aroclor when administered in combination. In conclusion, this study demonstrates that MeHg and Aroclor have independent deleterious effects on the developing testis, causing molecular and morphological changes in CBs. To the best of our knowledge, this is the first study to show that CBs are targets for toxic agents.

High-Resolution Melting Assay for Genotyping Variants of the CYP2C19 Enzyme and Predicting Voriconazole Effectiveness.

Voriconazole is a triazole antifungal agent recommended as primary treatment for invasive aspergillosis, as well as some other mold infections. However, it presents some pharmacokinetic singularities that lead to a great variability intra- and interindividually, nonlinear pharmacokinetics, and a narrow therapeutic range. Most experts have recommended tracing the levels of voriconazole in patients when receiving treatment. This azole is metabolized through the hepatic enzyme complex cytochrome P450 (CYPP450), with the isoenzyme CYP2C19 being principally involved. Allelic variations (polymorphisms) of the gene that encodes this enzyme are known to contribute to variability in voriconazole exposure. Three different allelic variants, CYP2C19*17, CYP2C19*2, and CYP2C19*3, could explain most of the phenotypes related to the voriconazole metabolism and some of its pharmacokinetic singularities. We designed a rapid molecular method based on high-resolution melting to characterize these polymorphisms in a total of 142 samples, avoiding sequencing. Three PCRs were designed with similar cycling conditions to run simultaneously. The results showed that our method represents a fast, accurate, and inexpensive means to study these variants related to voriconazole metabolism. In clinical practice, this could offer a useful tool to individually optimize therapy and reduce expenses in patients with fungal infections.

Development of Anastrepha fraterculus (Diptera: Tephritidae) Related to the Phenology of Blueberry, Blackberry, Strawberry Guava, and Surinam Cherry Fruits.

Anastrepha fraterculus (Wiedemann, 1830) is the main pest of temperate climate orcharding. The study investigated the development of A. fraterculus related to phenological stage of blueberry, blackberry, strawberry guava, and Surinam cherry trees. The phenological stages I (green fruits), II (intermediate ripening stage of fruits), and III (fruits close to harvesting) were determined, and they are from 8th, 10th, and 11th week; 6th, 8th, and 9th week; 8th, 13th, and 16th week; and 5th, 6th, and 7th week after the first flowering of blueberry, blackberry, strawberry guava, and Surinam cherry trees, respectively. We collected fruits from orchards to determine the infestation index using the formula: number of pupa/fruit weight. To investigate the development of A. fraterculus, we determined the following biological parameters: egg-to-adult period, weight of pupae, oviposition period, fecundity, number of pupae, and number of infested fruits. The infestation index for the fruits collected in the field was greater in strawberry guava and Surinam cherry fruits. In the laboratory, the development of A. fraterculus occurred in stage III of blueberry. In blackberry, besides stage III, we also observed the development in stage II, however, at lower infestation. In strawberry guava, the development of A. fraterulus occurred in stages II and III, and the development in both stages was similar. For Surinam cherry, the development occurred in the three phenological stages with similar values for biological parameters. Overall, of the four hosts studied, the strawberry guava and Surinam cherry fruits allowed a better biological development of A. fraterculus, corroborating its preference for fruits native to Brazil.

Implementing a coronary CT angiography protocol based on the body mass index: Radiation dose reduction, image quality, and diagnostic performance.

OBJECTIVES: To evaluate the relation between the coronary calcium score and the posterior choice of kilovoltage according to radiologists' criteria in a standard coronary CT angiography protocol to rule out coronary disease. To quantify the reduction in ionizing radiation after linking kilovoltage to patients' body mass index in a low-dose protocol with iterative model reconstruction. To evaluate the image quality and diagnostic performance of the low-dose protocol. MATERIAL AND METHODS: We compared anthropometric characteristics, calcium score, kilovoltage levels, size-specific dose estimates (SSDE), and the dose-length product (DLP) between a group of 50 patients who were prospectively recruited to undergo coronary CT angiography with a low-dose protocol and a historical group of 50 patients who underwent coronary CT angiography with the standard protocol. We correlated these parameters, the number of coronary segments that could not be evaluated with and without temporal padding, the attenuation, and the signal-to-noise ratio in the ascending aorta in the low-dose protocol with excellent imaging quality according to a semiquantitative scale. To calculate the diagnostic performance per patient, we used 24-month clinical follow-up including all tests as the gold standard. RESULTS: In the standard protocol, the presence of coronary calcium correlated with the selection of high kilovoltage (p = 0.02); this correlation was not found in the low-dose protocol (p = 0.47). Median values of SSDE and DLP were significantly (p < 0.001) lower and less dispersed in the low-dose protocol [9.22 mGy (IQR 7.84-12.1 mGy) vs. 26.5 mGy (IQR 21.3-36.3 mGy) in the standard protocol] and [97 mGy cm (IQR 78-134 mGy cm) vs. 253 mGy cm (IQR 216-404 mGy cm) in the standard protocol], respectively. The overall quality of the images obtained with the low-dose protocol was considered good or excellent in 96% of the studies. The parameters associated with image quality in a multivariable model (C statistic = 0.792) were heart rate (estimated coefficient, -0,12 [95% confidence interval: -0.2, -0.04]; p < 0.01) and the SSDE (estimated coefficient, -0,26 [95% confidence interval: -0.51, -0.01]; p < 0.05). The CAD-RADS modifier for a not fully evaluable or diagnostic study was used on two occasions (4%); the final measures for the diagnosis of coronary disease were sensitivity 100%, specificity 94%, and efficacy 94%. CONCLUSIONS: In the standard protocol, the radiologist selects higher kilovoltage for CT angiography studies for patients whose previous calcium score indicates the presence of coronary calcium. In the low-dose protocol, linking kilovoltage with body mass index enables the dose of radiation to be reduced by 65% while obtaining excellent or good image quality in 96% of studies and excellent diagnostic performance.

Assay of acetylcholinesterase activity by potentiometric monitoring of acetylcholine.

An acetylcholine-selective electrode based on a plasticized polymeric membrane has been developed. The electrode exhibited good selectivity for acetylcholine (ACh) over choline and some common ions, low drift, and a fast response to ACh. The response was linear over an ACh concentration range of 1×10(-6) to 1×10(-3) M with a slope of 59.1±0.1 and a detection limit of 1.5×10(-7)±1.2×10(-8) M. The electrode was used to monitor enzymatic ACh hydrolysis catalyzed by acetylcholinesterase (AChE) at different substrate and enzyme concentrations. A kinetic data analysis permitted the determination of the Michaelis-Menten constant of the enzymatic hydrolysis and AChE activity in the range of 2×10(-5) to 3.8×10(-1)U ml(-1).

Voltammetric behaviour and square-wave voltammetric determination of the potent antioxidant and anticarcinogenic agent ellagic acid in foodstuffs.

The voltammetric behaviour of ellagic acid (EA) is investigated by cyclic, differential pulse and square-wave voltammetry (CV, DPV and SWV, respectively). Based on the anodic oxidation peak at approximately 0.42V in acetic/acetate buffer (pH 5.5) a robust and a highly reliable square-wave voltammetric method is presented for the determination of EA. The oxidation peak current was linearly dependent on the concentration of EA in the range of 1.0×10(-7)-1.5×10(-6)mol/L (r=0.9997), with a detection limit of 1.0×10(-8)mol/L (S/N=3) and a quantification limit of 3.4×10(-8)mol/L (S/N=10), good reproducibility and a satisfactory level of selectivity towards others polyphenols. The proposed method was applied to the determination of free and total EA in fruits, nuts and juices with good analytical results being obtained.

Influence of the host diet on the performance of Doryctobracon areolatus (Hymenoptera: Braconidae).

Fitness in parasitoids is generally influenced by host quality. We evaluated the development parameters of Doryctobracon areolatus (Hymenoptera: Braconidae) parasitoid on Anastrepha fraterculus (Diptera: Tephritidae) larvae, reared in artificial diets with different protein sources. Larvae of A. fraterculus were reared in the diets based on 1) raw wheat germ (control); 2) whole rice flour; 3) corn flour; and, 4) whole wheat flour + soybean meal. The larvae were used for the development of D. areolatus, to evaluate the number of offspring/female, emergence rate, sex ratio, egg-adult period, adult weight posterior tibia length and longevity. Larvae of A. fraterculus reared in the diets based on wheat germ and corn flour obtained a greater number of offspring, with shorter duration of egg-adult period for males and females. In diets with rice flour and corn flour, A. fraterculus larvae generated adults of D. areolatus with greater weight, longer tibia length and higher sex ratio, whereas larvae reared in wheat flour + soybean meal diet had a lower sex ratio. The diets based on rice flour and corn are the most appropriate for multiplication of the parasitoid.

Development and reprotuction of Spodoptera eridania on natural hosts and artificial diet.

The development and reproductive potential of Spodoptera eridania (Stoll, 1782) (Lepidoptera, Noctuidae) was compared on natural and artificial diets. Its biology was determined under laboratory conditions, providing an artificial diet, leaves of white clover and the peach cultivar 'Chimarrita' and fruits and leaves of the apple cultivar 'Gala'. Larvae of S. eridania could not complete their biological cycle when fed on apple fruits or peach leaves. The artificial diet provided the shortest development time, with a cycle of 28.5±0.14 days from egg to adult and 37.3% total viability. Apple leaves led to a longer development time from egg to adult (62.8±1.22 days) and lower total viability (23%) and white clover provided the highest total viability (54.3%). The results showed that S. eridania can use white clover and apple leaves to develop normally in apple orchards. The reduction of food sources such as white clover used by the larvae results in infestation and damage to fruits from "test bites".

Rearing two fruit flies pests on artificial diet with variable pH.

Fruit flies (Diptera: Tephritidae) are considered the main fruit pests worldwide. In Brazil, two species are predominant: the South American fruit fly, Anastrepha fraterculus and the Mediterranean fruit fly, Ceratitis capitata. In this study, we evaluated the effect of artificial diets with variable pH in their larval development and adult performance. The experiments were carried out in the laboratory at 25 ± 2 °C, 70 ± 10% RH and 12:12h (L:D) photoperiod. Semisolid diets with pH values of 6.0, 5.0, 4.0, 3.0, 2.0, 1.5, and 1.0, adjusted by adding hydrochloric acid were tested. Results indicated that the diet with pH 6.0 did not support larval development of both species of fruit fly. Diets with greater acidic pH values did not allow egg, larvae or pupae development and adult reproduction of A. fraterculus. For C. capitata , the pH of artificial diet exerts greater influence compared to A. fraterculus on the duration and viability of the larval stage, number of pupae, sex ratio and longevity of males.

An Accurate Data Preparation Approach for the Prediction of Mortality in ACLF Patients using the CANONIC Dataset.

The incidence of chronic liver disease has increased in Europe and can lead to Acute on Chronic Liver Failure (ACLF) which is associated with high levels of mortality due to multisystem organ failure. The characteristics of the ACLF patients can change very rapidly within a short period of time. Continuous assessment of their recovery status is critical for clinicians to adjust and deliver effective treatment. The aim of this paper is to validate the usefulness of a data preparation approach by combining different criteria to replace missing values, balance target-class variables, select useful patient characteristics and optimise hyperparameters of machine learning models for the prediction of ACLF associated mortality rates. A key step in the data preparation is a feature selection Mutual Information (MI) based multivariate approach to build smaller, and yet equally and in some cases more informative, subsets of patient characteristics than those frequently proposed for the prediction of mortality, from patients with ACLF in the CANONIC dataset. The usefulness of the data preparation approach proposed to predict mortality was evaluated by training the XGBoost and Logistic Regression models with the prepared data. Evaluations of the models trained using a test set provided evidence of an overall high accuracy in the prediction of the mortality rates of patients for days after their diagnosis, and in some cases even higher when reduced and more informative subsets of patient characteristics were found.

Population Dynamics of Drosophila suzukii (Diptera: Drosophilidae) in Berry Crops in Southern Brazil.

Drosophila suzukii (Matsumura) is an invasive species originating in Southeast Asia and considered a severe pest in berry crops in several countries of the Northern Hemisphere and Europe. In South America, the species was first detected in 2013. The objective of the study was to monitor the seasonal activity of D. suzukii in commercial crops of blackberry, strawberry guava, surinam cherry, blueberry, and strawberry during two consecutive harvests and in three properties in the Southern region of Brazil during the 2015/2016 and 2016/2017 harvests, with the aid of traps baited with apple cider vinegar. The highest population peaks were observed during late spring to mid-fall in all areas and plant species studied. It was verified that temperature is the factor that most influenced the seasonal activity of D. suzukii in the field, promoting low catches of the species during winter. However, even during periods of low temperatures (winter period), the presence of D. suzukii in the crops was verified, demonstrating the species' ability to stay in place from year to year, surviving in alternative hosts such as Eriobotrya japonica, a common species in the region. The information on the time of the highest occurrence of the pest in different hosts presented in this study provides the basis for decision-making in relation to the management of D. suzukii, to avoid further economic damage.

Oviposition of fruit flies (Diptera: Tephritidae) and its relation with the pericarp of citrus fruits.

Fruit flies (Diptera: Tephritidae) represent a threat to fruit growing worldwide, mainly the citrus culture, however, biological studies show that fruit flies are not perfectly adapted to this host. This study investigated oviposition of Anastrepha fraterculus (Wiedemann, 1830) and Ceratitis capitata (Wiedemann, 1824) and its relation with the pericarp of citrus fruits. We evaluated the relationship between depth of oviposition of A. fraterculus and C. capitata and epicarp thickness of orange [Citrus sinensis (L.) Osbeck)] 'Navelina' and tangerine [C. reticulata (L.)] 'Clemenules' and the influence of fruit mesocarp of tangerine 'Clemenules' on oviposition of these species. The study was conducted under controlled conditions of temperature (25 ± 2 °C), relative humidity (70 ± 10% RH) and photophase (12 h). A. fraterculus and C. capitata laid their eggs in the flavedo region of orange 'Navelina' and between the albedo and flavedo of tangerine 'Clemenules'. When fruits with mesocarp exposed were offered, there was no oviposition by both fruit fly species. The results show that epicarp thickness of citrus fruits did not influence oviposition of A. fraterculus and C. capitata as oviposition did not occur only in the presence of the mesocarp, suggesting that other factors are involved in oviposition of these species.

Strategies for Establishing a Rearing Technique for the Fruit Fly Parasitoid: Doryctobracon brasiliensis (Hymenoptera: Braconidae).

Anastrepha fraterculus (Wiedemann) (Diptera: Tephritidae) is the most important pest in South American orchards. When control measures are not adopted, this pest can cause losses of up to 100%. Doryctobracon brasiliensis (Szépligeti) (Hymenoptera: Braconidae) is a larval-pupal endoparasitoid that can be used as a native biological control agent against A. fraterculus. This study aimed to develop a rearing technique for D. brasiliensis in larvae of A. fraterculus. Trials were carried out to: 1) determine the optimal instar for parasitism, 2) define the exposure time of larvae to parasitoids, 3) determine the density requirements of A. fraterculus larvae offered to each parasitoid, and 4) evaluate the effect of diet on adults of D. brasiliensis. In all trials, we evaluated the number of offspring, parasitism rate, and sex ratio. Moreover, in the experiment to investigate the effects of diet, we determined the longevity of males and females. In both choice and nonchoice parasitism tests, the parasitoids preferred third-instar larvae of A. fraterculus over second- and first-instar larvae. An exposure time of 12 h of A. fraterculus larvae produced larger numbers of parasitoids and higher parasitism rates. The density of 15 larvae of A. fraterculus to each female of D. brasiliensis produced a larger number of offspring. A supply of honey solution (20 and 50%) to the parasitoids yielded the highest number offspring and resulted in greater longevity. Our findings can be used to support the development of a mass rearing protocol for D. brasiliensis.

Antioxidant content of and migration from commercial polyethylene, polypropylene, and polyvinyl chloride packages.

Antioxidants commonly used in polyolefins were studied in commercial food packages made of low- and high-density polyethylene (LDPE and HDPE), polypropylene (PP), polyvinylchloride (PVC), and polyethylene terephtalate (PET) and in a LDPE film extruded at the laboratory. The phenolic antioxidants BHA, BHT, AO 2246, AO 425, Ethanox 330, Irganox 1010, and Irganox 1076 were studied together with the phosphite Irgafos 168 and their two degradation products, phosphate and DBP. Antioxidants were extracted from polyolefins using microwave energy and analyzed using high-performance liquid chromatography (HPLC) to determine the antioxidant content in the diverse commercial films. Irganox 1010 and Irganox 1076 were found in the majority of the samples generally together with the phosphite Irgafos 168 and its oxidized product (phosphate). Specific migration levels of each antioxidant were determined by HPLC after pretreatment with solid-phase extraction (SPE) in aqueous food simulants and after their dilution with tetrahydrofuran (THF) in fatty food simulant. These levels were much lower than limits allowed by legislation.

Different Clinical Features of Acral Abortive Hemangiomas.

Some infantile hemangiomas called in literature "minimal or arrested growth hemangiomas" or "abortive hemangiomas" are present at birth and have a proliferative component equaling less than 25% of its total surface area. Often, they are mistaken for vascular malformation. We present five patients (three girls and two boys) with abortive hemangiomas diagnosed between January 2010 and December 2015 localized in acral part of the extremities. They were congenital lesions resembling precursor of hemangiomas but did not show proliferation phase. Immunohistochemical Glut-1 was performed in all of them as a way to confirm the abortive hemangioma diagnosis. The most common appearance was a reticulated erythematous patch with multiple fine telangiectasias on the surface. We remark that one of them presented a segmental patch with two different morphologies and evolutions. The proximal part showed pebbled patches of bright-red hemangioma and presented proliferation and the distal part with a reticulated network-like telangiectasia morphology remained unchanged. We detected lower half of the body preference and dorsal region involvement preference without ventral involvement. The ulceration occurred in three patients with two different degrees of severity.

Track Analysis of the North, Central, and South American Species of the Epicauta maculata Group (Coleoptera: Meloidae).

We undertook a panbiogeographic analysis of 23 species of the Epicauta maculata group of America-Epicauta abeona Pinto, Epicauta adspersa (Klug), Epicauta andersoni Werner, Epicauta atomaria (Germar), Epicauta apache Pinto, Epicauta cavernosa (Courbon), Epicauta dilatipennis Pic, Epicauta fulvicornis (Burmeister), Epicauta horni Champion, Epicauta jeffersi Pinto, Epicauta koheleri Denier, Epicauta lizeri Denier, E. maculata (Say), Epicauta magnomaculata Martin, Epicauta minutepunctata Borchmann, Epicauta nigropunctata (Blanchard), Epicauta normalis Werner, Epicauta ocellata (Dugès), Epicauta pardalis LeConte, picauta phoenix Werner, Epicauta pluvialis Borchmann, Epicauta proscripta Werner, Epicauta rubella Denier, and Epicauta ventralis Werner-with the purpose of analyzing the distributional data for taxa, to establish patterns of distribution of an ancestral biota and areas where these groups have interacted. Based on the overlap of 20 individual tracks, four generalized tracks constituted by different numbers of species were identified; two of them are located in the Nearctic region and the Mexican transition zone (tracks "A" and "B"), and the other two are distributed in the Neotropical region and the South America transition zone ("C", "D"). Six nodes were recognized: Two of them are included in the Nearctic Region, node 'I' located in northern USA and node 'II' located in southwestern USA, both at the intersection of the tracks "A" and "B". The other four are included in the Neotropical Region at the intersection of the tracks "C" and "D": Node 'III' is located in Chaco province; node 'IV' is located in Parana Forest province; node 'V' is located in the northwest of Argentina in Puna province, and node 'VI' is located in Monte province.

Biology and fertility life table of Eriopis connexa, Harmonia axyridis and Olla v-nigrum (Coleoptera: Coccinellidae).

The coccinellids Eriopis connexa (Germar), Harmonia axyridis (Pallas) and Olla v-nigrum (Mulsant) are important natural biological control agents. The purpose of this paper was to study the biology and create a fertility life table of these three coccinellid species. For the biology study, 50 insects/species were used and kept in groups of 10 in glass vials (2300 cm3). For the three species studied, the viability of the total cycle varied from 45 to 50%. O. v-nigrum was the species which presented the longest oviposition period. However, H. axiridis demonstrated the best reproductive performance and ability of population growth in each generation. In conclusion, the use of commercially obtained pollen and A. kuenhiella eggs enables the development of coccinellids E. connexa, H. axyridis and O. v-nigrum under laboratory conditions, since the insects completed their biological cycle and originated adults with good reproductive performance.

Influence of the host diet on the performance of Doryctobracon areolatus (Hymenoptera: Braconidae) / Influência da dieta do hospedeiro na performance de Doryctobracon areolatus (Hymenoptera: Braconidae)

Abstract Fitness in parasitoids is generally influenced by host quality. We evaluated the development parameters of Doryctobracon areolatus (Hymenoptera: Braconidae) parasitoid on Anastrepha fraterculus (Diptera: Tephritidae) larvae, reared in artificial diets with different protein sources. Larvae of A. fraterculus were reared in the diets based on 1) raw wheat germ (control); 2) whole rice flour; 3) corn flour; and, 4) whole wheat flour + soybean meal. The larvae were used for the development of D. areolatus, to evaluate the number of offspring/female, emergence rate, sex ratio, egg-adult period, adult weight posterior tibia length and longevity. Larvae of A. fraterculus reared in the diets based on wheat germ and corn flour obtained a greater number of offspring, with shorter duration of egg-adult period for males and females. In diets with rice flour and corn flour, A. fraterculus larvae generated adults of D. areolatus with greater weight, longer tibia length and higher sex ratio, whereas larvae reared in wheat flour + soybean meal diet had a lower sex ratio. The diets based on rice flour and corn are the most appropriate for multiplication of the parasitoid.

Resumo O custo adaptativo de parasitoides é geralmente influenciado pela qualidade do hospedeiro. Nós avaliamos parâmetros de desenvolvimento do parasitoide Doryctobracon areolatus (Hymenoptera: Braconidae) em larvas de Anastrepha fraterculus (Diptera: Tephritidae) criadas em dieta artificial com diferentes fontes proteicas. Larvas de A. fraterculus foram criadas em dietas a base de: 1) germe de trigo cru (testemunha); 2) farinha de arroz integral; 3) farinha de milho; e, 4) farinha de trigo integral + farelo de soja. As larvas foram usadas para o desenvolvimento de D. areolatus para avaliar o número de descendentes/fêmea, taxa de emergência, razão sexual, duração do período ovo-adulto, peso de adultos, comprimento da tíbia posterior e longevidade. Larvas de A. fraterculus criadas nas dietas a base de germe de trigo e farinha de milho, permitiram obter um maior número de descendentes do parasitoide, com menor duração do período de desenvolvimento ovo-adulto, para ambos os sexos. Nas dietas com farinha de arroz e farinha de milho, larvas de A. fraterculus geraram adultos de D. areolatus com maior peso e tamanho de tíbia de machos e fêmeas e uma maior quantidade de fêmeas. A dieta contendo farinha de trigo + farelo de soja afetou a geração de descendentes, causando uma menor razão sexual. As dietas a base de farinha de arroz e milho são as mais adequadas para a multiplicação do parasitoide.

Distribution of luteinizing hormone-releasing hormone in the nervus terminalis and brain of the mouse detected by immunocytochemistry.

Immunoreactive luteinizing hormone-releasing hormone (LHRH) was localized in a relatively large number of ganglion cells and fibers of the nervus terminalis of neonatal and adult mice, indicating that this nerve is a substantial source of LHRH in the mouse brain. Whole-head specimens of neonatal mice, prior to calcification of the cranium, revealed an extensive distribution of LHRH neurons and fine fibers throughout the peripheral, intracranial, and central parts of the nervus terminalis. The most striking difference between the neonatal and adult animals, in the nervus terminalis, was the increase in immunoreactive axons that made up the fiber bundles of this nerve. In the adult mouse, the intracranial and central projections were composed of thick fascicles of immunoreactive axons, ensheathed by glial cells and accompanied by ganglia that contained both LHRH-reactive and nonimmunoreactive neurons. LHRH-immunoreactive cells and axons were seen in a branch of the nervus terminalis that coursed along the medial, posterodorsal aspect of the olfactory bulb and in branches of this nerve that accompany the vomeronasal nerves to the accessory olfactory bulb. A few LHRH neurons and many immunoreactive processes were seen in the accessory and main olfactory bulbs. LHRH-reactive neurons were seen in the hypothalamus and extrahypothalamic structures. Examination of adult mouse brains revealed a pattern of distribution and number of immunoreactive neurons similar to that seen in the neonate. However, many more LHRH-reactive axons were seen in all areas of the brain of the mature animal.