Primary pigmented nodular adrenocortical disease (PPNAD) as an underlying cause of symptoms in a patient presenting with hirsutism and secondary amenorrhea: case report and literature review

Abstract

Hypercortisolemia in females may lead to menstrual cycle disturbances, infertility, hirsutism and acne. Herewith, we present a 18-year-old patient, who was diagnosed due to weight gain, secondary amenorrhea, slowly progressing hirsutism, acne and hot flashes. Thorough diagnostics lead to a conclusion, that the symptoms was the first manifestation of primary pigmented nodular adrenocortical disease (PPNAD). All symptoms of Cushing syndrome including hirsutism and menstrual disturbances resolved after bilateral adrenalectomy. Our report indicates that oligo- or amenorrhea, hirsutism, acne in combination with weight gain, growth failure, hypertension and slightly expressed cushingoid features in a young woman requires diagnostics towards hypercortisolemia. Despite PPNAD is a very rare cause of ACTH-independent Cushing syndrome, it has to be taken into consideration, especially when adrenal glands appear to be normal on imaging and paradoxical rise in cortisol level in high-dose dexamethasone test is observed. Unlike in our patient, in vast majority of patients, PPNAD is associated with Carney complex (CC). Therefore, these patients and their first-degree relatives should be always carefully screened for symptoms of PPNAD, CC and genetic mutations of PRKAR1A, PDE11A, and PDE8B genes.     

Neurogenic factors in the impaired healing of diabetic foot ulcers

BACKGROUND: We hypothesize that the reduced innervation of skin can be observed both in clinically neuropathic and non-neuropathic diabetic foot ulcers and can contribute to low inflammatory cell infiltration. MATERIALS AND METHODS: Twenty patients with type 2 diabetes and active foot ulcers, without clinical evidence of peripheral sensory neuropathy (n = 12) and with sensory neuropathy (n = 8) were involved in this study. Biopsies from ulcer margin were examined immunohistochemically. RESULTS: Studies revealed presence of protein gene product 9.5 (PGP9.5)+ nerve endings only in reticular dermis in 3 of 12 non-neuropathic subjects, however, regenerating GAP-43+ endings were seen in dermis of almost all specimens. Lack of substance P+ nerve endings was characteristic for both groups. The reduced distribution of calcitonin gene-related peptide+ nerves in epidermis and dermis was seen mainly in neuropathic group. In neo-epidermis lack of nerve growth factor expression was observed in both groups, whereas neurotrophin 3 immunostaining was characteristic for neuropathic specimens (P < 0.03). Expression of trkA and trkC receptors did not differ significantly between groups. Low inflammatory cell infiltration and moderate presence of fibroblasts was characteristic for all studied specimens. CONCLUSIONS: The observed reduction of foot skin innervation and neurogenic factors expression can be correlated with low inflammatory cell accumulation and subsequently leads to the observed chronicity of diabetic foot ulcer healing process in both neuropathic and non-neuropathic patients.     

[99mTc]O2-AMD3100 as a SPECT tracer for CXCR4 receptor imaging

PurposeCXCR4 plays an important role in HIV infection, tumor progression, neurogenesis, and inflammation. In-vivo imaging of CXCR4 could provide more insight in the role of this receptor in health and disease. The aim of this study was to investigate [^99mTc]O₂-AMD3100 as a potential SPECT tracer for imaging of CXCR4.MethodAMD3100 was labelled with [^99mTc]pertechnetate. A cysteine challenge assay was performed to test the tracer stability. Heterologous and homologous receptor binding assay and internalization assay were performed in CXCR4 expressing Jurkat-T cells. Ex vivo biodistribution was studied in healthy mice at 30, 60, and 120 min after tracer injection. Tumor uptake of the tracer was determined by microSPECT imaging in nude mice xenografted with human PC-3 prostate tumor. Specificity of tracer uptake was determined by blocking studies using an excess of unlabelled AMD3100.ResultsAMD3100 was labelled with technetium-99 m with a radiochemical yield of > 98%. The tracer was stable in PBS and mouse plasma for at least 6 h at 37 °C. Heterologous and homologous binding assays with AMD3100 showed IC₅₀ values of 240 ± 10 μM, and 92 ± 5 μM for [¹²⁵I]SDF-1α and [^99mTc]O₂-AMD3100 respectively, with negligible receptor internalisation. The tracer showed high uptake in liver, lungs, spleen, thymus, intestine and bone. Blocking dose of AMD3100.8HCl (20 mg/kg) decreased the uptake in these organs (p < 0.05). [^99mTc]O₂-AMD3100 showed specific tumor accumulation in mice bearing PC-3 xenografts model. Time activity curves (TAC) in AMD3100 pre-treated animals tracer showed 1.7 times less tumor uptake as compared to control animals (p < 0.05).Conclusion[^99mTc]O₂-AMD3100 is readily labelled, is stable in plasma and displays a favourable binding affinity for the CXCR4 receptors. [^99mTc O₂-AMD3100 shows specific binding in organs with high CXCR4 expression and in CXCR4 positive tumors. These results justify further evaluation of this radiopharmaceutical as a potential biomarker for the non-invasive imaging of CXCR4 receptors.     

Fully Parametric Sleep Staging Compatible with the Classical Criteria

We present an open system for sleep staging, based explicitly on the criteria used in visual EEG analysis. Slow waves, theta and alpha waves, sleep spindles and K-complexes are parameterized in terms of time duration, amplitude, and frequency of each waveform by means of the matching pursuit algorithm. It allows the detection of these structures using mostly the criteria from visual EEG analysis. For example, within this framework for the first time we compute directly the relative duration of slow waves, which is a basic parameter in recognition of deep sleep stages. Performance of the system is evaluated on 20 polysomnographic recordings, scored by experienced encephalographers. Seven recordings were scored by more than one expert. Proposed system gives concordance with visual staging close to the inter-expert concordance. The algorithm is implemented in a user-friendly software system for display and analysis of polysomnographic recordings, freely available with complete source code from .     

Isobolographic characterization of interactions of levetiracetam with the various antiepileptic drugs in the mouse 6 Hz psychomotor seizure model

The aim of this study was to characterize the anticonvulsant effects of levetiracetam (LEV) in combination with the various antiepileptic drugs (clonazepam [CZP], oxcarbazepine [OXC], phenobarbital [PB], tiagabine [TGB], and valproate [VPA]), in the mouse 6 Hz psychomotor seizure model.Limbic (psychomotor) seizure activity was evoked in albino Swiss mice by a current (32 mA, 6 Hz, 3 s stimulus duration) delivered via ocular electrodes and isobolographic analysis for parallel and non-parallel dose–response effects was used to characterize the consequent anticonvulsant interactions between the various drug combinations. Potential concurrent adverse-effect profiles of interactions between LEV and CZP, OXC, PB, TGB, and VPA at the fixed-ratio of 1:1 were evaluated in the chimney (motor performance), passive avoidance (long-term memory), and grip-strength (muscular strength) tests.LEV administered singly was associated with a dose–response relationship curve (DRRC) that was parallel to that for CZP and non-parallel to that for OXC, PB, TGB and VPA. With isobolography for parallel DRRCs, the combination of LEV with CZP at three fixed-ratios of 1:3, 1:1 and 3:1 was additive in nature. With isobolography for non-parallel DRRCs the combinations of LEV with OXC, TGB and VPA at the fixed-ratio of 1:1 were also additive. In contrast, the isobolography for non-parallel DRRCs revealed that the interaction for the combination of LEV with PB at the fixed-ratio of 1:1 was supra-additive (synergistic). None of the combinations were associated with any concurrent adverse effects with regards to motor coordination, long-term memory or muscular strength.LEV is associated with favorable anticonvulsant synergism with PB and is additive with regards to CZP, OXC, TGB and VPA in the mouse 6 Hz psychomotor seizure model.     

A simple and rapid liquid chromatographic method for the determination of metronidazole and its hydroxymetabolite in plasma and cutaneous microdialysates

A rapid, accurate, simple and low-cost method for quantitative determination of metronidazole and its hydroxymetabolite, in plasma and microdialysate samples, using tinidazole as an internal standard, has been developed. Metronidazole is widely used as a valuable agent for antiprotozoal as well as antibiotic therapy when anaerobic organisms are involved. Separation of the compounds studied was performed on a 120 x 4 mm analytical column, filled with LiChrosorb RP-8, 5 microm, the mobile phase consisted of 0.05 mol/L aqueous solution of potassium dihydrogen phosphate, adjusted to pH = 3.5 with orthophosphoric acid, methanol and acetonitrile (40:2:3. v/v/v). Detection was performed at 320 nm. Intra- and interserial precision was below 4.6% and 7.9%, respectively, for both the compounds studied. The presented method is suitable for pharmacokinetic studies.     

Intrathecal bradykinin administration: opposite effects on nociceptive transmission

Injected intrathecally, bradykinin (BK) produced either hyperalgesia (0.15 microg) or antinociception (6.0 microg) in rats when thermal noxious stimuli were used. Similarly, des-Arg(9)-BK at the lower dose (0.15 microg) decreased, whereas at the higher dose (6.0 microg) it increased the threshold to thermal noxious stimuli; however, these effects were less pronounced than those of BK. The antinociception induced by BK was abolished by HOE 140, a B(2) receptor antagonist, injected intrathecally at a dose of 1.3 ng and was markedly attenuated by des-Arg(10)-HOE 140, a B(1) receptor antagonist (1.15 ng i.t.). The results obtained in this study showed that--depending on the dose used--BK and des-Arg(9)-BK could produce pro- as well as antinociceptive actions. Both B(2) and B(1) receptors are involved in the action of intrathecally applied BK.     

Nutrients intake of the adolescents in Poland--ten years literature review (1990-2000)

The dietary survey among adolescents (11-15 years old) performed during last decade in Poland are not representative for national level. The analysis of data obtained by 24-h recall in different regions showed that contribution of dietary fat to energy intake was too high (37-42%), while contribution of proteins (10-11%) and carbohydrates (47-52%) too low. The intake of proteins, fibre, calcium, magnesium, copper and iron (especially among girls), vitamin C and niacin did not meet the recommended levels. The results suggest that girls are more likely to have risk of nutritional deficiencies than boys as well as adolescents from rural area than from towns. Inadequate intake of nutrients during adolescence can increase risk of dietary related diseases in later life.     

Transplants as a therapy after spinal cord injury

This review describes shortly phenomena that take place in different parts of central nervous system (CNS) after the spinal cord injury: 1/ due to axotomy many of neurones present outside the cavity of lesion (even those of supraspinal origin) can atrophy or die as an effect of necrosis or apoptosis; 2/ at the injury site itself, the primary and secondary effects lead to increased cell loss and the scar or cyst formation that are the mechanical barrier for regenerating axons; 3/ due to abolished conduction across the injury site the spinal cord circuitry below the lesion is deprived of supraspinal inputs. Then this review presents the new therapeutic strategies that were developed recently to obtain at least partial recovery of motor functions after spinal cord injury. The cell body can be rescued by applying various factors that increase intrinsic neural repair (e.g. neurotrophins or anti-apoptotic agents). To enhance the axonal regrowth through the lesion cavity, the scar and cyst formation can be reduced by constructing the bridges using e.g. the Schwann cells, fetal tissue, stem cells, olfactory ensheating glial cells, or by application of macrophages. To induce partial restoration of some functions that are controlled by neural circuitry below the lesion the various methods for enhancing the plasticity in segmental circuitry were developed (e.g. rehabilitation by locomotor training or intraspinal transplantation of monoaminergic cells). As a consequence of the great unpredictability of effects obtained after injury at different parts of the spinal cord the various strategies for repair need to be coordinated for optimal recovery.     

Die Eignung von trypsiniertem Nagelkeratinstaub, Stratumcorneum-Partikeln oder Glutamat als Kohlenstoff-Stickstoff-Quellen für Trichophyton mentagrophytes und Trichophyton rubrum

• 1 Im Rahmen vergleichender Untersuchungen zum Keratinabbau durch Dermatophyten wird über Wachstum, Stoffwechsel, physiologische Prinzipien und Intensität der Keratinolyse, die Ausscheidung von Enzymen und das Verhalten in der autolytischen Phase unter Verwendung von trypsiniertem Nagelkeratinstaub, Stratum-corneum-Partikeln und Glutamat als C-N-Quellen berichtet.
• 2 Schonende Trypsinierung (0,01 % Trypsin; 0,043 M Phosphatpuffer; pH 8,0) führt bei Nagelkeratinstaub, der eine große relative Oberfläche besitzt, innerhalb von 5 Tagen bei 23° C zu Gewichtsverlusten von 23 bis 35 %. Von Stratum-corneum-Partikeln werden unter gleichen Bedingungen bei geringer relativer Oberfläche 84 % zu löslichen Oligo- und Monomeren abgebaut.
• 3 Dermatophyten (Trichophyton mentagrophytes, Tr. rubrum) können trypsinierten Nagelkeratinstaub als vollwertige C-N-Quelle nutzen und erzielen dabel einen Keratinabbau von 80 bis 90 %. Die Geschwindigkeit der Keratinolyse; nicht aber ihr endlich erreichtes Ausmaß, werden durch den Trypsinierungsgrad negativ, durch die relative Oberfläche der Keratinpartikel positiv beeinflußt.
• 4 Ein wichtiges physiologisches Prinzip der mikrobiellen Keratinolyse ist neben dem poteniellen Sulfitmechanismus die aktive Alkalisierung des Substrates (Terrains). Sie erfolgt seitens der Pilze durch Ammoniaksekretion nach intracellulärer oxydativer Desaminierung und durch Ammoniakfreisetzung mittels extracellulärer Säureamidspaltung (Asparaginase).
• 5 Stratum-corneum-Partikel, die nach den Ergebnissen der Trypsinierungsversuche reichlich Nichtkeratinproteine und in geringeren Mengen Präkeratine bzw. trypsinlabile Keratine enthalten, werden als C-N, Quelle von Tr. mentagrophytes in kurzer Zeit praktisch vollständig und zwar ebenso schnell wie, wenn nicht sogar schneller als, Glutamat umgesetzt. Stratum corneum ist demnach als “ideale” Nährstoffquelle für Dermatophyten zu bewerten.
• 6 Das “Keratinase-Problem” wird eingehend diskutiert und dahingehend beantwortet. daß es unseres Erachtens keine streng keratinspezifischen Proteinasen bei Hautpilzen gibt, wohl aber im Komplex der keratinolytischen Prinzipien proteolytische Enzyme, die an die beim Keratinabbau vorliegenden komplizierten Bedingungen angepaßt sind, d. h. relativ hohe Sulfit-, Sulfat-, Ammoniaktiter und entsprechende pH-Werte tolerieren.
• 7 Obwohl in der autolytischen Phase Verluste an Biomasse bis zu 40 % der Myzelhöchstgewichte eintreten, so bleiben doch — jedenfalls in vitro; und warum sollte es in vivo prinzipiell anders sein (?) — über lange Zeit (mit Glutamat mindestens 140 Tage) Thallussegmente vital und ektoenzymatisch aktiv. Diese Befähigung zum langdauernden Überleben wird als Merkmal mit positivem Selektionswert beurteilt.