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Male pseudohermaphroditism due to primary 5 alpha-reductase deficiency: variation in gender identity reversal in seven Mexican patients from five different pedigrees.

In the present study, we describe the clinical, endocrinological, psychosexual and biochemical features of 7 Mexican male pseudohermaphrodites with primary 5 alpha-reductase deficiency in whom heterogeneity in the pattern of gender identity change at puberty was observed. The patients belonged to 5 different pedigrees from diverse locations in Mexico. Six of them were admitted to the Hospital during or after puberty. The one prepubertal subject was the sibling of a previously studied patient. Basal serum gonadotropins were determined by double antibody radioimmunoassay. Basal and choriogonadotropin (CG)-stimulated concentrations of androstenedione (A), testosterone (T) and dihydrotestosterone (DHT) were determined by radioimmunoassay after extraction and separation by celite chromatography. Urinary aetiocholanolone, androsterone and C19 and C21 5 beta/5 alpha metabolite ratios were analyzed by capillary gas chromatography. Enzyme activity and androgen receptors were studied in fibroblasts cultured from genital skin. Psychological assessment was performed using the Bender-Gestalt Wechsler Adult Intelligence Scale, the Rorschach Ink Blot and the Thematic Apperception Tests. All 7 patients were unambiguously reared as females; three spontaneously changed their gender identity and role from female to male after puberty, another one changed during psychotherapy at the end of puberty. Two patients (one prepubertal and the other pubertal) have been under therapy during 1.5 years, but due to familial and social factors a female gender has prevailed. The remaining patient consulted at age 15 because of virilization; her female gender identity did not change after more than one year of treatment and due to the fact she was depressed and had suicidal tendencies, the penis and testes were removed.(ABSTRACT TRUNCATED AT 250 WORDS)

3-Oxo-5-alpha-Steroid 4-Dehydrogenase↗

[Diffused traumatic dirt and decorative tattooing. Removal by Q-switched lasers].

BACKGROUND AND OBJECTIVE: Pigment fanning or spread is one complication of decorative tattooing, but is also seen after traumatic tattoos. The reason for this spreading remains unclear. While excision of the diffused pigment was previously considered the treatment of choice, today destruction of the pigment with Q-switched laser systems is the therapy with the highest efficiency and lowest rate of side effects. Therefore areas of pigment spread should be excised only in rare exceptional cases. PATIENTS/METHODS: 4 patients with pigment fanning after permanent make up and traumatic tattooing of the periorbital region were treated with the Q-switched ruby (694 nm) and Q-switched Nd:YAG (1064 nm) lasers. RESULTS: All patients showed a significant (70-80%) clearance of the spread pigment; two had complete clearing. Side effects such as hyper- or hypopigmentation, scarring or ink darkening were not seen. CONCLUSIONS: The Q-switched ruby- and Q-switched Nd:YAG-lasers are a therapeutic modality for pigment fanning with high efficiency and low rate of side effects. Attempts of explanation for pigment spread after tattoos are given, but further histological and electron microscopical investigations are needed to find the pathogenetic mechanism.

Child↗

[Reticular lentigo].

We studied the clinical and histopathologic features in five patients with reticulated lentigo. This peculiar pigmented skin lesions has previously been described under the designations "acquired reticulated lentigo", ink spot lentigo, "reticulated melanotic macule" or "reticulated lentigo". Each of the 4 males and 1 females in our study (age range: 16 to 57 years; mean age: 34 years) had a single, 4-6 mm large, black macule with irregular, fingerlike extensions at the periphery. All lesions were situated on the upper back and surrounded by numerous sun-induced freckles. Dermatoscopic examination revealed irregularly formed "meshes" and confirmed the reticulated pattern seen clinically. Reticulated lentigo presented mostly in individuals with skin type 1, red to blond hair, and blue eyes. The clinical diagnosis in 4 out of 5 cases was melanoma in situ. Histopathologically, reticulated lentigo was mainly characterized by a sharply circumscribed hyperpigmentation of the lower epidermis with accentuation at the tips of elongated and clubbed rete ridges. In addition, a normal or slightly increased melanocyte number in the basal layer and an infiltrate of melanophages in the upper dermis was noted. Reticulated lentigo shows histopathologic features similar to those of melanotic macules on volar skin and mucous membranes. Because of its characteristic clinical, dermatoscopic and histopathologic features reticulated lentigo can be regarded as a distinctive clinicopathologic entity.

Adolescent↗

[Topographic-anatomic study of tissue transplants of the inguinal region].

We performed 25 fresh cadaver dissections to describe the anatomy of the superficial and deep circumflex iliac artery and the superficial inferior epigastric artery to determine how they could least to used as donor vessels for the free transfer of groin flaps and living iliac bone. With injection of ink the capillary region of these vessels was stained in (iliac crest) bone, the internal oblique muscle and skin of the groin and thigh. The superficial and deep circumflex iliac artery were shown to be the main supply vessels of the groin and thigh and could be found in 96%-100% of cases. The venous drainage of this region followed from a superficial system (superficial circumflex iliac veins) and a deep venous system (Vv. comitantes accompanying arterial branches). Both venous systems could always be found. The superficial circumflex iliac artery was shown to only supply the skin and was the main donor vessel to the skin and soft tissue transplants. The deep circumflex iliac artery supplied the pelvic bone, the internal oblique muscle and a small constant area of skin above the iliac crest. Bone, muscle and bone, muscle and skin transplants could be obtained with this donor vessel, with enlargement of the skin area possible by combining two arterial branches combined in one transplant. With an average vessel diameter of 1.5 mm (superficial circumflex iliac artery) and 3 mm (deep circumflex iliac artery) both vessels could be used satisfactorily for microvascular transplantation. Different tissues including muscle, bone and skin could be obtained in adequate amounts to replace composite defects in the upper aerodigestive tract.

Arteries↗

[Cryptococcal meningoencephalitis in HIV infected immigrants. Diagnostic and therapeutic aspects with reference to evidence-based medicine"--an overview].

Cryptococcal meningitis is one of the most frequent forms of meningoencephalitis in AIDS patients. Before the introduction of triazole antibiotics like difluconazole, the frequency of this meningitis among AIDS patients was 5-10%, yet declined during the last years. Clinically, nonspecific signs of a meningeal inflammation predominate while focal neurological signs are rare. Rapid institution of high-dose antimycotic therapy is the keystone to improve prognosis and decrease mortality. Antimycotics of choice are amphotericin B, fluconazole, and flucytosin. Diagnosis is established by detection of cryptococcal antigen in cerebrospinal fluid, microscopic demonstration of cryptococci using India-ink, and cryptococcal cultures. Recent developments aim to improve antimycotic therapies. During the last years, lifelong secondary prevention with difluconazole was established. Cryptococci which are resistant against fluconazole and amphotericin B are a special challenge, yet fortunately are rare. Longterm outcome of patients is determined by the progression of the underlying immunosuppression. Therefore, combination of secondary prophylaxis with modern antiviral substances is important. Clinical decision analysis in patients with suspected cryptococcal meningitis is presented using methods from evidence-based medicine.

AIDS-Related Opportunistic Infections↗

Sulfide pulsing as the controlling factor of spinae production in Chlorobium limicola strain UdG 6038

Chlorobium limicola UdG 6038, a green sulfur bacterium, was isolated from anoxic sediments. Cells were gram-negative, non-motile, ovoid shaped, and contained chlorobactene and bacteriochlorophyll c as the main photosynthetic pigments. The DNA G+C content was 56.4 mol%. Ultrastructural studies revealed the presence of abundant spinae (45-110 spinae per cell) attached to the cell wall. India-ink-stained cells observed under the optical microscope were surrounded by a large capsule (5-11 &mgr;m total diameter). The presence of this capsule was coincident with the presence of a large number of spinae (> 30 spinae per cell). The mucilaginous capsule was attached to the spinae without penetrating it. In batch culture, the synthesis of spinae in strain UdG 6038 was not affected by changes in temperature, pH, salt concentration, or illumination at physiological ranges and hence, the cells remained spined. The control of spinae production was experimentally confirmed using a semicontinuous batch culture refed by sulfide pulsing. The culture remained at a low spination level (> 30 spinae per cell) only when the duration of sulfide starvation between pulses was less than 5 h. After longer sulfide starvation periods, the cells remained spined (more than 38 ± 6.3 spinae per cell). This observation supports the idea that the duration of sulfide limitation in the culture plays a key role in controlling the spination process in strain C. limicola UdG 6038. Chlorobium spinae may play an eco-physiological role in buoyancy capacity and adhesion of sulfur globules to the cells in natural environments where sulfide concentrations are expected to be highly variable.

Journal Article↗

Health hazards due to the inhalation of amorphous silica.

Occupational exposure to crystalline silica dust is associated with an increased risk for pulmonary diseases such as silicosis, tuberculosis, chronic bronchitis, chronic obstructive pulmonary disease (COPD) and lung cancer. This review summarizes the current knowledge about the health effects of amorphous (non-crystalline) forms of silica. The major problem in the assessment of health effects of amorphous silica is its contamination with crystalline silica. This applies particularly to well-documented pneumoconiosis among diatomaceous earth workers. Intentionally manufactured synthetic amorphous silicas are without contamination of crystalline silica. These synthetic forms may be classified as (1) wet process silica, (2) pyrogenic ("thermal" or "fumed") silica, and (3) chemically or physically modified silica. According to the different physicochemical properties, the major classes of synthetic amorphous silica are used in a variety of products, e.g. as fillers in the rubber industry, in tyre compounds, as free-flow and anti-caking agents in powder materials, and as liquid carriers, particularly in the manufacture of animal feed and agrochemicals; other uses are found in toothpaste additives, paints, silicon rubber, insulation material, liquid systems in coatings, adhesives, printing inks, plastisol car undercoats, and cosmetics. Animal inhalation studies with intentionally manufactured synthetic amorphous silica showed at least partially reversible inflammation, granuloma formation and emphysema, but no progressive fibrosis of the lungs. Epidemiological studies do not support the hypothesis that amorphous silicas have any relevant potential to induce fibrosis in workers with high occupational exposure to these substances, although one study disclosed four cases with silicosis among subjects exposed to apparently non-contaminated amorphous silica. Since the data have been limited, a risk of chronic bronchitis, COPD or emphysema cannot be excluded. There is no study that allows the classification of amorphous silica with regard to its carcinogenicity in humans. Further work is necessary in order to define the effects of amorphous silica on morbidity and mortality of workers with exposure to these substances.

Air Pollutants, Occupational↗

Improved incidental memory with nicotine after semantic processing, but not after phonological processing.

RATIONALE: A number of lines of evidence suggest that a nicotinic cholinergic system is mediating attentional processing. However, the evidence is less clear for a nicotinic system being involved in mnemonic processing. OBJECTIVES: The present study investigated the effects of nicotine on memory using a depth of processing paradigm. METHODS: A double-blind design was used with participants (n = 40) smoking either a nicotine containing cigarette (n = 20) and a denicotinized cigarette (n = 20). After smoking, each set of these participants was further subdivided into two groups (n = 10 for each). One group were presented with a series of trials each beginning with the presentation of a "decision word" which they had to say whether it represented something which was living or non-living (semantic-orienting). The second group had to say whether the word had one syllable or two syllables (phonological or non-semantic orienting condition). This decision was followed by a word in coloured ink whose colour participants were required to name as quickly as possible. On completion of the whole task the participants were given an unexpected free recall test. RESULTS: The nicotine-containing cigarette reduced the latencies for decision-making and colour naming in comparison with the denicotinized cigarette. The free recall test showed that nicotine-containing cigarette increased the number of words remembered, but only for the semantic-orienting condition and not the non-semantic condition. CONCLUSIONS: There is a nicotinic cholinergic system that mediates effortful processing. It can be deployed for attentional processing, including the associative processing required for memory encoding.

Adolescent↗

Development and characterisation of disposable gold electrodes, and their use for lead(II) analysis.

There is an increasing need to assess the harmful effects of heavy-metal-ion pollution on the environment. The ability to detect and measure toxic contaminants on site using simple, cost effective, and field-portable sensors is an important aspect of environmental protection and facilitating rapid decision making. A screen-printed gold sensor in a three-electrode configuration has been developed for analysis of lead(II) by square-wave stripping voltammetry (SWSV). The working electrode was fabricated with gold ink deposited by use of thick-film technology. Conditions affecting the lead stripping response were characterised and optimized. Experimental data indicated that chloride ions are important in lead deposition and subsequent analysis with this type of sensor. A linear concentration range of 10-50 microg L(-1) and 25-300 microg L(-1) with detection limits of 2 microg L(-1) and 5.8 microg L(-1) were obtained for lead(II) for measurement times of four and two minutes, respectively. The electrodes can be reused up to 20 times after cleaning with 0.5 mol L(-1) sulfuric acid. Interference of other metals with the response to lead were also examined to optimize the sensor response for analysis of environmental samples. The analytical utility of the sensor was demonstrated by applying the system to a variety of wastewater and soil sample extracts from polluted sites. The results are sufficient evidence of the feasibility of using these screen-printed gold electrodes for the determination of lead(II) in wastewater and soil extracts. For comparison purposes a mercury-film electrode and ICP-MS were used for validation.

Acetic Acid↗

Amperometric sensor for L-ascorbic acid determination based on MnO2 bulk modified screen printed electrode.

A simple biosensor constructed by bulk-modification of carbon ink with manganese dioxide as a mediator was investigated for its ability to serve as amperometric detector for L-ascorbic acid in hydrodynamic mode. The sensor could be operated at pH 5.0 (0.05 M phosphate buffer) and exhibited excellent reproducibility and stability. Optimization of measurement parameters such as applied working potential and pH value were studied in detail. The screen printed electrode exhibited a linear amperometric increase with the concentration of L-ascorbic acid from 50 mg L(-1) to 250 mg L(-1) and gave a (LOD = 3sigma) detection limit of 0.2 mg L(-1) (1.172 micromol L(-1)). The manganese dioxide modified screen printed electrode shows long term stability.

Ascorbic Acid↗

Topochemical investigation of ancient manuscripts.

Various modern instrumental techniques for surface analysis were applied for the non-destructive physicochemical examination of works of art. As samples, pieces of ancient manuscripts endangered by iron-gall ink corrosion were used. Surface characterisation of the morphology of the cellulose fibres within corroded and non-corroded parts of the manuscript performed by scanning electron microscopy (SEM) showed seriously damaged cellulose fibres in the written parts. The elemental composition of selected parts of the manuscript was determined by energy dispersive X-ray fluorescence analysis (EDX). A more detailed study of the paper surface was then performed by electron probe microanalysis (EPMA). This technique yields the morphological characteristics of the surface as well as element distribution maps over the written area of the investigated manuscript.

Journal Article↗

Covalent strategy for immobilization of DNA-microspots suitable for microarrays with label-free and time-resolved optical detection of hybridization.

Sequence-specific detection and quantification of nucleic acids are central steps in many molecular biology procedures which have also been transferred to chip-based procedures. Hybridization-based assays can be used to quantify and discriminate between DNA target sequences down to the level of single base mismatches. Arrays of DNA probes immobilized on a support enable simultaneous testing of multiple sequences of a single sample. DNA arrays can be produced either by in-situ synthesis of oligonucleotides or by immobilization of pre-assembled DNA probes. Covalent and directed immobilization improves the reproducibility and stability of DNA arrays. This is especially interesting with repeated use of transducers or chips. Procedures are described for effective covalent immobilization of pre-assembled amino-linked oligonucleotides, by use of ink-jet techniques, on a modified and heated glass surface, with addressable surface areas ranging from 0.01 mm2 to a few mm2. Almost immediate evaporation of the spotted droplets on the heated surfaces leads to very high surface hybridization capacities. The surfaces are suitable for use with a label-free detection method - reflectometric interference spectroscopy (RIfS). It is shown that hybridization capacity and non-specific interaction at these DNA-surfaces can be characterized by use of RIfS. With a consumption of less than 80 ng mm(-2) oligonucleotide and a specific hybridization capacity of more than 300 fmol mm(-2), the activated aminodextran procedure was usually suitable for setting up a DNA array with label-free detection. Non-specific interactions with random oligomers or protein (ovalbumin) were low. Up to 150 repeated regenerations (stripping) of the surfaces by acid treatment and denaturing agents, and 50 days of storage, have been possible without significant loss of hybridization capacity.

DNA Probes↗

Intracortical connections between motor cortical zones controlling antagonistic muscles in the cat: a combined anatomical and physiological study.

Experiments were done on nine cats anaesthetized with pentobarbitone to determine whether motor cortical zones controlling antagonistic muscles are synaptically interconnected. Motor cortical zones controlling wrist flexors, or extensors, were identified by microstimulation and intramuscular electromyographic recordings (microstimulation: 11 pulses at 333 pulses/s, current 10-40 microA). The position of each zone of interest was marked by a small ink spot on the surface of the cortex and on a scaled drawing of the cortical surface (cruciate region). Following the identification of wrist flexor and extensor zones the anterograde tracer biocytin was injected into one, or two, wrist extensor zones at three depths (400, 800 and 1500 microm) from the cortical surface. A small injection of horseradish peroxidase (HRP)--producing a dark brown spot of approximately 300-500 microm--was made in layer II-III of one or more wrist flexor zones. Similar HRP injections were made in the deep layers of wrist extensor zones that were not labelled by biocytin. The HRP injections served to mark the position of potential targets of biocytin-labelled fibres. In some experiments the biocytin was injected into a wrist flexor zone and HRP was deposited in one or more wrist extensor zones. Biocytin-labelled fibres (blue) were found throughout the expanse of the forelimb representation zone, as has been previously reported. More specifically, in all animals biocytin-labelled fibres were found in identified cortical zones controlling the same muscle(s) as well as in zones controlling an antagonist(s). Club-like swellings, indicative of synaptic boutons, were observed on these fibres. The density of labelled fibres was greater in the upper cortical layers (II-III), but a large number of terminals were also present in the lower cortical layers (V-VI). We conclude that there exist intracortical circuits linking motor cortical zones controlling antagonistic muscles. Elucidating the nature and function of these circuits is likely to be important for understanding the mode of operation of the motor cortex.

Animals↗

A novel phantom design for emission tomography enabling scatter- and attenuation-"free" single-photon emission tomography imaging.

A newly designed technique for experimental single-photon emission tomography (SPET) and positron emission tomography (PET) data acquisition with minor disturbing effects from scatter and attenuation has been developed. In principle, the method is based on discrete sampling of the radioactivity distribution in 3D objects by means of equidistant 2D planes. The starting point is a set of digitised 2D sections representing the radioactivity distribution of the 3D object. Having a radioactivity-related grey scale, the 2D images are printed on paper sheets using radioactive ink. The radioactive sheets can be shaped to the outline of the object and stacked into a 3D structure with air or some arbitrary dense material in between. For this work, equidistantly spaced transverse images of a uniform cylindrical phantom and of the digitised Hoffman rCBF phantom were selected and printed out on paper sheets. The uniform radioactivity sheets were imaged on the surface of a low-energy ultra-high-resolution collimator (4 mm full-width at half-maximum) of a three-headed SPET camera. The reproducibility was 0.7% and the uniformity was 1.2%. Each rCBF sheet, containing between 8.3 and 80 MBq of 99mTcO4- depending on size, was first imaged on the collimator and then stacked into a 3D structure with constant 12 mm air spacing between the slices. SPET was performed with the sheets perpendicular to the central axis of the camera. The total weight of the stacked rCBF phantom in air was 63 g, giving a scatter contribution comparable to that of a point source in air. The overall attenuation losses were <20%. A second SPET study was performed with 12-mm polystyrene plates in between the radioactive sheets. With polystyrene plates, the total phantom weight was 2300 g, giving a scatter and attenuation magnitude similar to that of a patient study. With the proposed technique, it is possible to obtain "ideal" experimental images (essentially built up by primary photons) for comparison with "real" images degraded by photon scattering and attenuation losses. The method can serve as a tool for experimental validation and intercomparison of attenuation and scatter correction methods. Moreover, the large flexibility of this phantom design will allow investigations of arbitrary activity distributions and autoradiography or other imaging techniques such as PET, x-ray computed tomography or magnetic resonance imaging.

Brain↗

Endoscopic management of polypoid early colon cancer.

Endoscopic management of polypoid early colonic cancer (malignant polyps and polypoid carcinomas) is no longer controversial. When the endoscopist is satisfied that excision is complete and histology is "favorable" (a resection margin of 2 mm and well or moderately well differentiated tumor), surgery is unnecessary. When histology show "unfavorable" characteristics (which a few histologists still take to include invasion into lymphatics), surgical or laparoscopic resection may be indicated, providing the patient is considered at suitable risk. Surgery kills some patients without finding residual cancer and cannot save others with metastases, so it should be recommended only with due clinical consideration. Sessile or broad-based polyps, especially those in the rectum, are more likely to be "high risk" and merit specialist management if local removal is to be attempted and to allow proper histologic assessment. Endoscopic approaches such as saline injection polypectomy, india-ink tattooing, and use of the argon beam coagulator are applicable in some cases. New approaches that still require trials include ultrasonographic probes, which occasionally clarify the degree of invasion, and prototype stapling devices to allow full-thickness histologic specimens to be obtained.

Colonic Polyps↗

Transplantation of the proximal fibula based on the anterior tibial artery. Anatomical study and clinical application.

The purpose of this study was to investigate the feasibility of microvascular transplantation, based on the anterior tibial artery, of the epiphysis and the proximal half of the diaphysis of the fibula. India ink injections into the anterior tibial artery were performed in 13 unembalmed adult cadavers, following which transverse frozen sections were taken. Dye was observed in the fibula, both within the intramedullary cavity and at the level of the periosteum, in all specimens. The average distal limit reached by the dye, measured from the tip of the epiphysis, was 10.4 cm in the intramedullary cavity and 11.8 cm in the periosteum. Based on these anatomical findings one case of humeral reconstruction has been performed with a fibular transplant, including both the epiphysis and a segment of diaphysis, vascularized solely by the anterior tibial artery.

Adult↗

Microvascularisation of lips in ageing edentulous subjects.

Macroscopic changes of the lips in the edentulous elderly subject are accompanied by variations in their microvascularisation. This study was undertaken on histological or clarified sections from totally edentulous subjects whose vascular system was injected with Indian ink agar. A reduction in density and size of the superficial (papillary or mucosal) and deep (reticular or submucosal) networks is observed with age. These changes are to be seen in relation to imaging and to the reduction in occlusal potential of the lips.

Aged↗

Arterial supply of the temporo-medial region of the brain: significance for preoperative vascular occlusion testing.

The mesiobasal limbic system is of particular significance in the surgical treatment of temporo-medial tumors and epilepsy. It consists of the uncus, amygdaloid body, hippocampus, dentate gyrus, subiculum, fasciolar gyrus and the parahippocampal gyrus. Knowledge of the vascular microanatomy is a key to the surgical treatment of pathologies in the region. The anterior choroidal artery was selectively injected in fresh brain specimens: 50 specimens with a gelatinous ink mixture to demonstrate vascular territories in stereotactic brain slices, and 50 with a Biodur resin to obtain casts for microanatomical evaluation. The cast technique was also applied to 35 specimens injected into the posterior cerebral artery. The rostral third of the temporomedial region is mainly supplied by branches of the anterior choroidal artery. The occipital two thirds are supplied by hippocampal branches, the posteromedial choroidal artery and the inferior temporal branches of the posterior cerebral artery. Important vessel variations with significant implications for the preoperative Wada-test are presented.

Anterior Cerebral Artery↗