Comparison of the clinical and radiographic appearance of the cervical vertebrae with histological and anatomical findings in an eight-month old warmblood stallion suffering from cervical vertebral stenotic myelopathy (CVSM).

BACKGROUND: Cervical vertebral stenotic myelopathy (CVSM) remains one of the most important abnormalities of the cervical spine resulting in neurological deficits in horses. The aim of the following study was to compare the results of the clinical and neurological examination, the results of myelography and the post mortem anatomical and histological appearance of the spinal cord and cervical vertebrae in a horse with CVSM. CASE PRESENTATION: The following study describes a clinical case of an eight-month-old stallion with ataxia. Plain cervical radiographs indicated narrowing of the spinal canal. Conservative therapy using NSAIDs did not result in any improvement in the gait of the horse. Due to economic constraints, surgical intervention was excluded. The owner chose to humanely euthanise the horse. Immediately after euthanasia, post mortem myelography was performed, and measurements of the myelographic dye column were taken. They revealed a 67% DMC reduction and a 64% DD reduction at the C3/C4 level. Afterwards, an anatomical dissection was performed. The cervical vertebrae and vertebral canal were macroscopically inspected and measured and indicated a 44% narrowing of the canal at the C3/C4 level. The spinal cord was removed and underwent histological evaluation after staining. Microscopic lesions were visible at the level of the compression and included axonal degeneration with partial or complete loss of myelin in the white matter of the lateral and dorsal funiculi as well as the formation of dysfunctional so-called "spongy structures". An increase in the number of microglial cells and collagen was also observed. The formation of glial scars was excluded. Immunohistochemical studies revealed a negative transmembrane glycoprotein CD68(-) - monocyte response and a negative tumor necrosis alpha TNFα (-) reaction. CONCLUSIONS: CVSM may be difficult to diagnose, even for experienced veterinary surgeons. Currently, an ex vivo histopathologic examination of the spinal cord is thought to be the gold standard in the diagnosis of CVSM. Our histological examination revealed no CVSM-specific glial scar formation and a CD68(-) negative and TNF-α negative reaction, which have not been previously reported. Histological lesions in CVSM may vary depending show inter-individual variability and on the treatment, which further hinders ex-vivo diagnostics.

Management of high risk pulmonary embolism - a single center experience.

BACKGROUND AND AIM: Patients with acute pulmonary embolism (APE) associated with hemodynamic instability, i.e. high-risk APE (HR-APE), are at risk for early mortality and require urgent reperfusion therapy with thrombolysis or embolectomy. However, a considerable proportion of HR-APE subjects is not reperfused but only anticoagulated due to high bleeding risk. The aim of the present study was to assess the management of HR-APE in a single large-volume referral center. METHODS: A single-center retrospective study of 32 HR-APE subjects identified among 823 consecutive patients hospitalized for symptomatic APE. RESULTS: Out of 32 subjects with HR-APE (19 women, age 69 ± 19 years), 20 patients were unstable at admission and 12 subsequently deteriorated despite on-going anticoagulation. Thrombolysis was applied in 20 (62.5%) of HR-APE subjects, limited mainly by classical contraindications in the remainder. Percutaneous pulmonary embolectomy was performed in 4 patients. In-hospital PE-related mortality tended to be higher, albeit insignificantly, in the patients who developed hemodynamic collapse during the hospital course compared to those unstable at admission (67% vs. 40%, p = 0.14). Also, survival was slightly better in 22 patients treated with thrombolysis or percutaneous embolectomy in comparison to 10 subjects who received only anticoagulation (54% vs. 40%, p = 0.2). Major non-fatal bleedings occurred in 7 of 20 patients receiving thrombolysis (35%) and in 2 (17%) of the remaining non-thrombolysed 12 HR-APE subjects. CONCLUSIONS: Hemodynamically instability, corresponding to the definition of HR-APE, affects about 4% of patients with APE, developing during the hospital course in approximately one-third of HR-APE subjects. As almost 40% of patients with HR-APE do not receive thrombolytic therapy for fear of bleeding, urgent percutaneous catheter-assisted embolectomy may increase the percentage of patients with HR-APE undergoing reperfusion therapy. Further studies are warranted for a proper identification of initially stable intermediate-risk APE subjects at risk of hemodynamic collapse despite appropriate anticoagulation.

[Physicians' contribution in the creation and development of bioethics in Poland]. / Wklad lekarzy w powstanie i rozwój bioetyki w Polsce.

Bioethics was created in the 70s of the last century. From this period come works written by Van Resselaer Potter and André Hellegers, in which the authors raised the issue of the need to discuss the progress and to define a new reflection, they used for the first time in the scientific literature the term bioethics. In the United States of America, as well as in European countries, institutions began to emerge in which scientific bioethical reflection was realized. In Poland in the 1970s, the socio-political situation was not conducive to the integration of scientific circles or the freedom of exchange of views. Significant progress in the field of medicine, emerging new treatment options, posed questions that had to be answered. The questions were formulated in different environments, including by doctors who themselves participated in the progress of medical sciences, introducing, for example, new increasingly aggressive treatments, or saw new challenges emerging in the medical profession, which were not regulated by the hitherto applied principles of classical medical ethics. The article describes the achievements of prof. Jan Nielubowicz, prof. Józef Bogusz and prof. Tadeusz Kielanowski in the creation of Polish bioethical regulations in the field of experimental research in humans, the activity of prof. Kornel Gibinki in the formation of Bioethics Commisions, contribution of prof. Stefan Raszeja and prof. Marek Sych in bioethical education of the society and the role of prof. Krzysztof Szczygiel in the creation of the first Polish scientific bioethical unit.

Expression of hypoxia inducible factor 1α and 2α and its association with vitamin C level in thyroid lesions.

BACKGROUND: Cells adapt to hypoxia by transcriptional induction of genes that participate in regulation of angiogenesis, glucose metabolism and cell proliferation. The primary factors mediating cell response to low oxygen tension are hypoxia inducible factors (HIFs), oxygen-dependent transcription activators. The stability and activity of the α subunits of HIFs are controlled by hydroxylation reactions that require ascorbate as a cofactor. Therefore, deficiency of intracellular vitamin C could contribute to HIFs overactivation. In this study, we investigated whether vitamin C content of human thyroid lesions is associated with HIF-1α and HIF-2α protein levels. METHODS: Expression of HIF-1α and HIF-2α as well as vitamin C content was analyzed in thyroid lesions and cultured thyroid carcinoma cell lines (FTC-133 and 8305c) treated with hypoxia-mimetic agent (cobalt chloride) and ascorbic acid. The expression of HIFs and hypoxia-induced glucose transporters were determined by Western blots while quantitative real-time PCR (qRT-PCR) was performed to detect HIFs mRNA levels. Ascorbate and dehydroascorbate levels were measured by HPLC method. RESULTS: We found an inverse correlation between vitamin C level and HIF-1α but not HIF-2α expression in thyroid lesions. These results agree with our in vitro study showing that vitamin C induced a dose - dependent decrease of HIF-1α but not HIF-2α protein level in thyroid cancer cells FTC-133 and 8305C. The decreased HIF-1α expression was correlated with reduced expression of hypoxia-related glucose transporter 1 (GLUT1) in thyroid cancer cells. CONCLUSION: The results demonstrate that HIF-1α activation is associated with vitamin C content in thyroid lesions. Our study suggests that high tumor tissue ascorbate level could limit the expression of HIF-1α and its targets in thyroid lesions.

[About the introduction the criteria of brain death in Poland in 1984]. / Wokól wprowadzenia kryteriów smierci mózgowej w Polsce w 1984 roku.

Significant recovery of discussion about the need for the formulation of criteria of death was provoked due to the progress of medicine. Development of anaesthesiology and intensive therapy, introduction of new resuscitation techniques and devices, which are increasingly better able to maintain and sometimes even replace functions of the respiratory or cardiovascular system, which are essential for the survival meant that existing for centuries, socially accepted classical criteria of death, based on cessation of breath and circulation, are no longer sufficient. The criteria of brain death developed by the Ad Hoc Committee - 12-experts commission at Harvard University, published in Journal of American Medical Association in August 1968 and were the breaking point. The concept has been adopted and accepted by society in many countries soon, while in Poland the process of formulating and introduction of criteria for brain death took much more time. Based on the available literature and preserved archival materials, this article describing the process of development of new death criteria, acceptance by the National Team of Specialists and publication in 1984 the announcement by the Minister of Health and Social Welfare about the criteria of brain death.

Effect of Glucose on GLUT1-Dependent Intracellular Ascorbate Accumulation and Viability of Thyroid Cancer Cells.

Enhanced glucose requirement of cancer cells is associated with an increased glucose transport across plasma membrane that is mediated by a family of facilitated glucose transporter proteins, named GLUTs. GLUT1 is the main transporter in thyroid cancer cells. Glucose is the principal physiological substrate of GLUT1; however, it is also capable of transporting of oxidized form of vitamin C [i.e., dehydroascorbic acid (DHAA) which inside the cells is reduced to ascorbic acid (AA)]. The objective of this study was to determine the effect of normo-, hypo-, and hyperglycemia conditions on GLUT1-dependent intracellular ascorbate accumulation and viability of thyroid cancer cells. GLUT1 seems to be the main DHAA transporter in thyroid cancer cells because its knockdown by RNAi reduced DHAA accumulation by more than 80%. The results showed that in thyroid cancer cells high glucose inhibits both transport of AA and DHAA. Inhibition of vitamin C transport by glucose had a cytotoxic effect on the cells. However, stabilization of vitamin C in one of 2 forms (i.e., AA or DHAA) abolished this effect. These results suggest that cytotoxic effect is rather associated with extracellular accumulation of vitamin C and changes of its oxidation state than with intracellular level of ascorbate.

[About the genesis of the first Polish local research ethics committee]. / Wokól utworzenia pierwszej polskiej komisji bioetycznej.

From the moment in which the development of medicine became necessary experimental research involving human subjects, the question arose about the ethical limits and limitations of the experiment. The turning point was the year 1947. The Nuremberg Code was formulated after the disclosure of pseudo-medical experiments involving human subjects during the Second World War. In 1964, the medical world accepted the Declaration of Helsinki, which, however, did not prevent abuses and it became necessary to appoint independent ethics committees supervising and enforcing the application of ethics in biomedical experiments. In Poland in the 60's and 70's started a discussion on the ethical rules related to conduct of research involving humans. The initiators of the appointment of bioethics committees were professors of medicine, inspiring experiences of their Western colleagues. It was difficult for reasons of political ideologies to convince the authorities to use the best of western solutions. This paper attempts to describe the circumstances connected with the appointment in 1979 at the Medical University of Gdansk, the first Polish bioethics committee.

[How do first codes of medical ethics inspire contemporary physicians?]. / Czego mozemy sie dowiedziec z lektury pierwszych kodeksów etyki lekarskiej?

First codes of medical ethics appeared between 18th and 19th century. Their formation was inspired by changes that happened in medicine, positive in general but with some negative setbacks. Those negative consequences revealed the need to codify all those ethical duties, which were formerly passed from generation to generation by the word of mouth and individual example by master physicians. 210 years has passed since the publication of "Medical Ethics" by Thomas Percival, yet essential ethical guidelines remain the same. Similarly, ethical codes published in Poland in 19 century can still be an inspiration to modem physicians.

Effects of fermented rapeseed meal on performance, intestinal morphology, the viscosity of intestinal content, phosphorus availability, and egg quality of laying hens.

Fermented rapeseed meal has the potential to partial replace soybean meal in feed mixtures for poultry without a negative impact on the health condition and performance of birds. This is due to the fact that the fermentation process can reduce the amount of antinutritional factors, improve the use of nutrients and impart probiotic properties to rapeseed meal. Therefore, this study was undertaken to investigate the effect of fermented rapeseed meal on the performance, egg quality, intestinal morphometry, the viscosity of intestinal content and total phosphorus availability. A total of 108 Lohmann Brown laying hens at 26 wk of age were used in the 90-day study. All hens were randomly divided into 3 treatment groups, with 12 replicates (cages) each, as follows: control group received no rapeseed meal, the URSM group received 3% unfermented rapeseed meal and the FRSM group received 3% fermented rapeseed meal. In the case of performance, egg traits, sensory evaluation of eggs, the viscosity of intestinal content and the availability of total phosphorus, if the distribution was normal, a 1-way analysis of variance was performed. If the distribution was not normal, the Kruskal-Wallis test was performed. In the case of histomorphometric evaluation of the intestine, if the distribution was normal, the Student t test for independent samples was performed. If not, a Mann-Whitney U test was performed. The performed analyses showed that the supplementation of fermented rapeseed meal had no negative effect on the performance of birds and the quality of eggs. Fermented rapeseed meal was also associated with improved histomorphometric parameters of the small intestine compared to the group receiving unfermented rapeseed meal in the feed. Laying hens from FRSM group were characterized by significantly lower viscosity of intestinal content (P < 0.05) compared to URSM group. Phosphorus in FRSM group was significantly more available to the birds (P < 0.05) compared to URSM group. These results suggest that supplementation with fermented rapeseed meal may be beneficial, especially in times of unstable prices of soybean meal and problems with its availability.

[The role of glucose transporter 1 (GLUT1) in the diagnosis and therapy of tumors]. / Rola transportera glukozy 1 (GLUT1) w diagnostyce i terapii nowotworów.

Malignant cells are known to enhance glucose metabolism, to increase glucose uptake and to inhibit the process of oxidative phosphorylation. Accelerated glycolysis is one of the biochemical characteristics of cancer cells that allow them to compensate the inefficient extraction of energy from glucose in order to continue their uncontrolled growth and proliferation. Upregulation of glucose transport across the plasma membrane is mediated by a family of facilitated glucose transporter proteins named GLUT. Overexpression of GLUTs, especially the hypoxia-responsive GLUT1, has been frequently observed in various human carcinomas. Many studies have reported a correlation between GLUT1 expression level and the grade of tumor aggressiveness, which suggests that GLUT1 expression may be of prognostic significance. Therefore, GLUT1 is a key rate-limiting factor in the transport and glucose metabolism in cancer cells. This paper presents the current state of knowledge on GLUT1 regulation as well as its utility in the diagnosis and therapy of cancers.

Differences in the level of physical fitness and mobility among older women with osteoporosis and healthy women-cross-sectional study.

The main aim of the study was to assess the risk of falls, and physical fitness in the group of women aged 60 to 65 years of age suffering from an identified osteoporosis in comparison to a similar group of healthy women. The main question was: What is the level of physical fitness and risk of fall among women with osteoporosis compared to healthy women? The research included 262 women aged 60 to 65 of age: 135 with osteoporosis and 127 healthy ones, living in the Malopolskie and the Swietokrzyskie Provinces of Poland. To assess the level of physical fitness, the Senior Fitness Test (SFT) was used, while the Tinetti POMA (Performance Oriented Mobility Assessment) and Timed Up&Go test (TUG) were used to asses the risk of fall. Significant statistical differences in average results of physical fitness assessment were noticed as regards the following aspects: flexibility of the lower body part p < 0.001; flexibility of the upper body part p < 0.001. Essential differences were demonstrated in assessing the risk of falling with p < 0.01. Women with osteoporosis are marked by a lower physical fitness than healthy women. A higher percentage of great and serious risk of fall was demonstrated among women with osteoporosis.

The size and shape of parasitic larvae of naiads (Unionidae) are not dependent on female size.

The naiads, large freshwater mussels (Unionida), have very long life spans, are large-bodied, and produce thousands to millions of larvae (glochidia) which typically must attach to host fish tissues to metamorphose into a juvenile mussel. Glochidia develop within a female's marsupial gill demibranch, thus their number is restricted by female size. However, larger mussels acquire more energy, which could be invested in either larger-sized glochidia, in a more glochidia, or a combination of both. The high level of host specialization seen in many naiads may constrain glochidial size and shape around a narrow optimum, while naiads that use a wide range of host fishes may be predicted to possess greater plasticity in glochidial morphology. In this paper, we investigated the relationship between maternal body size and progeny body size and shape, aided by modern digital microscopy. We analyzed the between- and within- species variation of glochidia size and shape relative to female size in four widespread species of European naiads: Anodonta anatina, Anodonta cygnea, Unio crassus and Unio tumidus. Whereas the total reproductive output is collinear with female body size, substantial differences between species in glochidia size were found within genus Anodonta, but not genus Unio where glochidial size is remarkably consistent. The glochidial shape, however, differed within both Unio and Anodonta. We interpret this constant within-species glochidial size in Unio as reflecting a constraint imposed by the likelihood of successful transmission onto and off from a narrow range of hosts, whereas their shape seems to be less constrained. The Anodonta species, inhabiting a wide spectrum of habitats and using more than twice the number of fish hosts than Unio spp., have larger glochidia with greater variation in size and shape. Our results suggest that measures of glochidial variability may also serve as an indicator of host specificity in other naiads.

Local Effects of a 1940 nm Thulium-Doped Fiber Laser and a 1470 nm Diode Laser on the Pulmonary Parenchyma: An Experimental Study in a Pig Model.

The lungs are a common site of metastases from malignant tumors. Their removal with a minimal but safe tissue margin is essential for the long-term survival of patients. The aim of this study was to evaluate the usefulness of a 1940 nm thulium-doped fiber laser (TDFL) and a 1470 nm diode laser (DL) in a pig model of lung surgery that involved the incision and excision of lung tissue. Histopathological analysis was performed on days 0 and 7 after surgery. Neither TDFL nor DL caused significant perioperative or postoperative bleeding. Histological analysis revealed the presence of carbonized necrotic tissue, mixed fibrin-cellular exudate in the superficial zone of thermal damage and bands of deeper thermal changes. The mean total width of thermal damage on day 0 was 499.46 ± 61.44 and 937.39 ± 109.65 µm for TDFL and DL, respectively. On day 7, cell activation and repair processes were visible. The total width of thermal damage was 2615.74 ± 487.17 µm for TDFL vs. 6500.34 ±1118.02 µm for DL. The superficial zone of thermal damage was narrower for TDFL on both days 0 and 7. The results confirm the effectiveness of both types of laser in cutting and providing hemostasis in the lungs. TDFL caused less thermal damage to the lung parenchyma than DL.

Novel Nanohydroxyapatite (nHAp)-Based Scaffold Doped with Iron Oxide Nanoparticles (IO), Functionalized with Small Non-Coding RNA (miR-21/124) Modulates Expression of Runt-Related Transcriptional Factor 2 and Osteopontin, Promoting Regeneration of Osteoporotic Bone in Bilateral Cranial Defects in a Senescence-Accelerated Mouse Model (SAM/P6). PART 2.

PURPOSE: Healing of osteoporotic defects is challenging and requires innovative approaches to elicit molecular mechanisms promoting osteoblasts-osteoclasts coupling and bone homeostasis. METHODS: Cytocompatibility and biocompatibility of previously characterised nanocomposites, i.e Ca5(PO4)3OH/Fe3O4 (later called nHAp/IO) functionalised with microRNAs (nHAp/IO@miR-21/124) was tested. In vitro studies were performed using a direct co-culture system of MC3T3-E1 pre-osteoblast and 4B12 pre-osteoclasts. The analysis included determination of nanocomposite influence on cultures morphology (confocal imaging), viability and metabolic activity (Alamar Blue assay). Pro-osteogenic signals were identified at mRNA, miRNA and protein level with RT-qPCR, Western blotting and immunocytochemistry. Biocompatibility of biomaterials was tested using bilateral cranial defect performed on a senescence-accelerated mouse model, ie SAM/P6 and Balb/c. The effect of biomaterial on the process of bone healing was monitored using microcomputed tomography. RESULTS: The nanocomposites promoted survival and metabolism of bone cells, as well as enhanced functional differentiation of pre-osteoblasts MC3T3-E1 in co-cultures with pre-osteoclasts. Differentiation of MC3T3-E1 driven by nHAp/IO@miR-21/124 nanocomposite was manifested by improved extracellular matrix differentiation and up-regulation of pro-osteogenic transcripts, ie late osteogenesis markers. The nanocomposite triggered bone healing in a cranial defect model in SAM/P6 mice and was replaced by functional bone in Balb/c mice. CONCLUSION: This study demonstrates that the novel nanocomposite nHAp/IO can serve as a platform for therapeutic miRNA delivery. Obtained nanocomposite elicit pro-osteogenic signals, decreasing osteoclasts differentiation, simultaneously improving osteoblasts metabolism and their transition toward pre-osteocytes and bone mineralisation. The proposed scaffold can be an effective interface for in situ regeneration of osteoporotic bone, especially in elderly patients.

[RNA interference as a potential tool for diagnosis and therapy of some human diseases]. / Zastosowanie interferencji RNA w diagnostyce i terapii niektórych chorób czlowieka.

RNA interference is one of the most important discoveries in the field of molecular biology. To date, many studies have been reported which suggest that RNA interference takes part in various biological processes essential for the development of many diseases. On the other hand, owing to its high specificity and efficiency, RNA interference has become a powerful tool in gene therapy. However, introduction of small interfering RNAs (siRNAs) to a target tissue requires overcoming some critical biological barriers. This, in turn, contributes to searching for new effective vectors, which should selectively penetrate the tissue and be able to exclude the innate immune response. This study provides a brief overview of the application of RNA interference in the diagnosis and therapy of some human diseases.

Response of freshwater mussel recruitment to hydrological changes in a eutrophic floodplain lake.

Although eutrophication of freshwaters is a natural process, the human impact often leads to inland waters becoming overloaded with nutrients, impoverishing many valuable and vanishing habitats, such as floodplain lakes. These changes need to be reversed if the occurrence of endangered aquatic species is to be restored. In this paper we analyse the impact of a change in the water regime of a naturally eutrophic floodplain lake, which harbours a large diversity of Unionidae (large freshwater mussels), a globally threatened taxonomic group that provides important ecosystem functions and services. We found that a slight increase in the discharge from this waterbody, following the construction of an additional outflow pipe, positively influenced recruitment in three of the five mussel species inhabiting the lake. We also found that, after the construction of this additional outflow, the niches of juveniles of Anodonta cygnea and Unio spp. changed, revealing differences in their hydrological requirements. Our results suggest that, as in lotic habitats, complex hydraulic parameters are highly significant to unionid mussels in lentic conditions.

Development and Evaluation of a Polyvinylalcohol -Cellulose Derivative-Based Film with Povidone-Iodine Predicted for Wound Treatment.

The aim of this study was to develop and assess a polyvinyl alcohol-cellulose derivatives-based film with incorporated povidone-iodine (PVP-I) predicted for applications in the treatment of periodontitis. Films were fabricated by solvent-casting, and their physical characteristics, such as their surface and structure morphology, mechanical properties, and disintegrating time, were evaluated. For in vitro iodine release studies and evaluation, the antimicrobial activity was tested using a modified disc diffusion method against five microbial strains. For further use, we selected the film with polyvinyl alcohol-hydroxypropyl methylcellulose (PVA/HPMC_B) based on acceptable physicochemical properties. To assess the subacute toxicity of the film composition, the tissue regeneration process was tested in rats and compared to a conventional dressing commonly used in wound healing (Spongostan). Seven days after implantation, dorsal skin sections and blood samples (n = 10, in total n = 30) were examined. The wound area, epithelium, and dermis were evaluated microscopically, while the blood collected from the rats underwent biochemical analysis. The blood biochemistry results were comparable in all three groups. No significant histological differences between the Spongostan and the placebo film developed after subcutaneous implantation were observed. In contrast, the inflammation stage was reduced and the "scar" in the dermis was smaller when PVP-I and PVA/HPMC_B films were used. A smaller local inflammatory response inflicted less tissue damage, leading to the activation of subsequent regeneration phases and restoration of the area to its original state. The results obtained confirmed that PVP-I incorporated into PVA-hydroxypropyl methylcellulose film is a promising drug carrier, working faster and more effectively than the other two dressing materials evaluated. These developments provide a promising alternative in tissue regeneration and the wound healing process.

Intracellular glycoproteins binding galectin-1 in thyroid lesions.

AIMS AND BACKGROUND: The increased expression of galectin-1 on the mRNA and protein level observed in malignant thyroid tumors in comparison with benign lesions suggests that this protein may be associated with malignant transformation of thyroid epithelium. Extracellular and membrane glycoproteins are the main known ligands for this galactose-binding lectin. However, immunofluorescence studies have shown that galectin-1 is found predominantly in the intracellular compartment. The aim of this study was to examine intracellular carbohydrate ligands of galectin-1 in the thyroid, with particular attention to potential differences in their expression levels between benign and malignant lesions. METHODS: Identification of cytosolic and nuclear glycoproteins binding galectin-1 was performed by affinoblotting after separation of proteins in 8% polyacrylamide slab gels and electrotransfer onto Immobilon-P membranes. For semiquantitative analysis of glycoproteins binding galectin-1, an enzyme-linked lectino-solid-phase assay (ELLSA) was used. RESULTS: The predominant cytosolic glycoproteins binding galectin-1 had molecular masses of 50, 55, 59, 64, 85-87, 100, and 133 kDa and nuclear glycoprotein had a molecular mass of 75 kDa. There were no evident differences in glycoprotein patterns between benign and malignant thyroid lesions. The results obtained by ELLSA did not show any significant differences in lectin binding by cytosolic and nuclear proteins of thyroid lesions either. CONCLUSIONS: On the basis of these results it is tempting to suggest that interactions between galectin-1 and intracellular glycoconjugates are not critical for malignant transformation in the thyroid gland.

[RHD variant in RhD/-/ mother with anti-D makes noninvasive fetal RHD genotyping impossible]. / Obecnosc "niemego" genu RHD u RhD ujemnej kobiety ciezarnej z przeciwcialami anty-RhD uniemozliwia okreslenie genotypu RHD plodu metoda nieinwazyjna.

OBJECTIVES: Noninvasive fetal RHD genotyping from maternal plasma of RhD(/-) pregnant women of Caucasian race may be used for predicting the risk of hemolytic disease because the RHD gene is usually absent in such populations. If detected in plasma of such women, the RHD gene originates from the RhD(+) fetus. The number of fetal copies of the gene in maternal plasma is extremely small. In the presented case of the RhD(/-) pregnant woman with anti-D it was impossible to give a fetal RHD result due to mother's RHD(+) genotype. The fetal RHD was determined from amniocytes. AIM: to present the difficulties related to the interpretation of results of invasive and noninvasive procedures. MATERIAL AND METHODS: whole blood, plasma and amniotic fluid of the RhD(-) woman with anti-D (14 week of pregnancy) as well as whole blood of the newborn. RHD and RHCE*c were genotyped by real-time PCR in DNA isolated from maternal plasma and amniocytes and the RHD and d-genotypes were tested by SSP methods in DNA isolated from whole blood and amniocytes. RESULTS: RHD and RHCE*c were detected in DNA isolated from plasma. The high level of RHD suggested its origin from the mother's DNA therefore it was impossible to determine the fetal RHD. The d-little test identified a RHD(IVS3+ 1G>A) variant in the mother's genome. A weak signal of real-time PCR for the RHD was obtained in amniocytes but the RHD was not detected by SSP. The RHCE*c was detected by both methods. Results were inconclusive; the fetal RHD status remained unknown. The child was RhD(-) with RHD in its DNA undetected by either method. CONCLUSIONS: 1/The RHD(IVS3+ 1G>A) variant in the RhD(-) mother precluded formal noninvasive fetal RHD genotyping. 2/Real-time PCR is too sensitive for amniocyte testing and may lead to false results as it detects trace maternal DNA in amniotic fluid. 3/The frequency of RHD(IVS3+1G>A) occurrence in Poland requires further studies.

Intraseasonal asynchrony as a factor boosting isolation within a metapopulation: The case of the clouded apollo.

This article discusses the influence of phenology-related intraseasonal asynchrony on metapopulation dynamics and stability. As the part played by intraseasonal asynchrony is as yet unclear and poorly described, greater account of it should be taken in both metapopulation research and conservation practice. The subpopulations of the Parnassius mnemosyne metapopulation studied here are strongly isolated because of the phenological shift between them, despite the relatively small physical distances between them. This isolation is the result of a significant temporal shift in the species' flight periods in the main metapopulation centers: in some seasons its flight times in the different subpopulations did not overlap at all. The predicted results of such strong intraseasonal asynchrony are not altogether clear. On the one hand, they reduce the vulnerability of the entire metapopulation to the effects of short-term random disasters. On the other, the ever-greater isolation of subpopulations may cause the metapopulation to become a nonequilibrium one, which will have a serious impact on its long-term survival.