BRAIN AND EYE AS POTENTIAL TARGETS FOR IONIZING RADIATION IMPACT. PART V - ORGANIC AND FUNCTIONAL CHANGES CORRELATION ON THE EXAMPLE OF CERTAIN DISEASES. / ГОЛОВНИЙ МОЗОК ТА ОРГАН ЗОРУ ЯК ПОТЕНЦІЙНІ МІШЕНІ ДЛЯ ВПЛИВУ ІОНІЗУЮЧОГО ВИПРОМІНЮВАННЯ: ЧАСТИНА V ­ СПІВВІДНОШЕННЯ ОРГАНІЧНИХ І ФУНКЦІОНАЛЬНИХ ЗМІН НА ПРИКЛАДІ ОКРЕМИХ ЗАХВОРЮВАНЬ.

The question about correlation between organic and functional changes in persons, exposed to radiation is still insufficiently studied. Dynamics of morbidity for different forms and classes of non-tumour diseases periodisation, proposed by epidemiologists, suggests the identification of three main periods: «early¼ (the first 6 post-Chornobyl accident years); «distant¼ (12-21 years) and «late¼ (22-30 years). However, the correspondence this periodisation to the results of epidemiological data, without taking into account the clinical features of the diseases, may contribute to the impression, that in the first period after a radiation disaster functional disorders (or autonomic regulation disorders, etc.) prevail in all cases. Meanwhile, the data from ophthalmological and neurological studies, which we aim to demonstrate in this paper, rather indicate the presence of a significant morphological basis for the development of functional disorders in early period after a radiation disaster. The objective of this work is analyse modern experimental, epidemiological and clinical data on the correlation between organic and functional changes, characteristic of radiation cerebro-ophthalmological effects - radiation cataracts, age-related macular degeneration, cerebral small vessel disease, and neurocognitive deficits. Materials and methods. The criteria for inclusion in the analytical review were peer-reviewed publications in PubMed/MEDLINE, Scopus, Web of Science, and manually selected papers; the results of our own research were also used. An additional analysis of the results of examinations conducted in 1991-2004 was performed, which included a total of 11 123 persons irradiated as the result of catastrophy at the Chornobyl NPP. Results. In the first period of radiation cataract development, which is a specific consequence of radiation exposure, morphological changes (lens opacities) are observed, which lead to a decrease in visual function only later. Analysing the correlation between organic and functional changes in the development of diseases, for which ionising radiation exposure is a significant risk factor, we observe a similar picture. For example, CSVD associated with arterial hypertension may be a manifestation of accelerated aging associated with exposure to ionising radiation.Similarly, the initial signs of age-related macular degeneration in radiation-exposed individuals are usually manifested by changes in the morphology of the retina, choroid, and pigment epithelium in the macular area, while functional disorders in the form of decreased central vision and metamorphopsia, etc., occur later.

BRAIN AND EYE AS POTENTIAL TARGETS FOR IONIZING RADIATION IMPACT. PART IV - FETAL RADIATION-INDUCED BINOCULAR VISION DISORDERS: A PILOT STUDY. IMMUNOHISTOCHEMICAL CHANGES OF NON/IRRADIATED EYEBALL MUSCLES AFFECTED BY STRABISMUS. EVIDENCE OF A POSSIBLE MECHANISM OF THE STRABISMUS DEVELOPMENT IN PERSONS EXPOSED TO FETAL RADIATION. / GOLOVNYI MOZOK TA ORGAN ZORU IaK POTENTsIINI MIShENI DLIa VPLYVU IONIZUIuChOGO VYPROMINIuVANNIa. ChASTYNA IV - ROZLADY BINOKULIaRNOGO ZORU PLODA, SPRYChYNENI RADIATsIINYM VPLYVOM IN UTERO: PILOTNE DOSLIDZhENNIa. IMUNOGISTOKhIMIChNI ZMINY NEOPROMINENYKh M'IaZIV OChNOGO IaBLUKA PRY KOSOOKOSTI. MOZhLYVYI MEKhANIZM ROZVYTKU KOSOKOSTI V OSIB, IaKI ZAZNALY PRENATAL'NOGO OPROMINENNIa.

Strabismus is a significant cause of a decrease in the functional capabilities of the organ of vision, additionally, it leads to the deteriorating quality of life. Orbital factors and nervous system changes may play an important role in strabismus pathogenesis. There are few reports on binocular vision disorders in radiation-exposed persons.Functions of the external eyeball muscles might be disturbed based on the changed coordination process of subcortical nerve structures and due to altered metabolism. A carefully conducted research is necessary to clarify the possible pathogenesis of binocular vision disorders in radiation-exposed persons. OBJECTIVE: to assess the peculiarities of the development of strabismus and binocular vision disorders in people who were exposed to ionizing radiation in utero; to investigate the changes in distribution and appearance of myosin, dystrophin, and collagen IV between non-irradiated persons with normal binocular vision and patients with strabismus. MATERIALS AND METHODS: 583 persons, irradiated in utero because of the Chornobyl disaster were examined (at the time of examination average age was 11.3 ± 0.1 years). The control group - 808 people - Kyiv residents. Overall, 15 non-irradiated eyeball muscle samples were examined. 10 were from strabismus patients and 5 were controls. To evaluate morphological structure haematoxylin and eosin staining were used. For the detection of myosin, dystrophin, and collagen IV biotin-avidin (IMH) immunohistochemistry method was performed. Semi-quantitative grading method was used for the evaluation of immunoreactive structure appearance and local distribution. RESULTS: An increased frequency of divergent strabismus (p = 0.04190) and heterophoria (p = 0.002603) was found in the group exposed to prenatal (fetal) radiation because of the Chornobyl disaster compared to the control group. The relative risk of heterophoria was 5.08 (1.42 - 18.13). A decrease in dystrophin, myosin, and collagen IV positive structures was observed in non-irradiated strabismus-affected eyeball muscles compared to the control group. CONCLUSIONS: Detected changes indicate an increased probability in the development of strabismus in persons who had been exposed to fetal radiation. Non-irradiated strabismus-affected eyeball muscles are characterized by diminished myosin, dystrophin, and collagen IV immunohistochemical structures. Additionally, determined qualitative morphological changes in skeletal striated muscle fibers lead to the changed structural organization, indicating possible muscular dystrophy. Thereby, the presence of dystrophic processes in the eyeball muscles may play a significant role in the morphopatogenesis of strabismus. Further morphological studies are necessary to clarify the development of binocular vision disorder and the methods of their correction. These studies would be especially important to populations that were exposed to radiation.

Expression of anti-inflammatory markers IL-2, IL-10, TGF-ß1, ßDEF-2, ßDEF-3 and Cathelicidin LL37 in dairy cattle milk with different health status of the udder.

Great economic losses to the dairy industry are associated with bovine mastitis, which results in poor milk quality and high treatment costs. Anti-inflammatory proteins play an important role in the suppression of the immune response against invading pathogenic microorganisms and are therefore being studied for possible use in the early diagnosis of mastitis. In our study, we used milk samples from 15 cows of Holstein Friesian breed with different health status (5 healthy, 5 subclinical, and 5 clinical animals), and tested them using immunohistochemical (IHC) analysis to evaluate the presence of IL-2, IL-10, TGF-ß1, ßDEF-2, DEF-3, and Cathelicidin LL37 proteins. The calculation of positively and negatively stained cells for each biomarker was performed using the semiquantitative counting method. We found the presence of all factors with the exception of Cathelicidin LL37, which was almost absent in milk samples of all animal groups. The significant decrease of IL-10, ß-def2, and ß-def3 expression levels within the 3 days of sampling, found in the milk of animals with sub- and clinical mastitis, indicates the loss of antiinflammatory protection of the affected cow's udder. In contrast, the stable increase of IL-2 and TGF-ß1 positive cells observed in the milk of mastitis-affected cows, and the similar expression of these factors in the milk of healthy animals, indicate the possible lack of involvement of these cytokines at an early stage of udder inflammation.

Morphologic comparison of blood vessels used for coronary artery bypass graft surgery.

BACKGROUND: The aim of this study was to evaluate morphologic features of healthy saphenous vein and internal thoracic artery, blood vessels used in coronary artery bypass graft (CABG) surgery, and compare results. MATERIALS AND METHODS: Ten specimens of saphenous veins and ten of internal thoracic arteries used for CABG were obtained from 20 patients. Histological routine and immunohistochemical staining was performed with: endothelin (ET), tissue inhibitor of metalloproteinase 2 (TIMP2), metallomembranoproteinase 2 (MMP2), transforming growth factor beta (TGFß), hepatocyte growth factor (HGF), vascular endothelial growth factor (VEGF), protein gene product 9.5 (PGP9.5), vascular cell adhesion molecule (VCAM), intercellular adhesion molecule (ICAM). A semiquantitative evaluation method was used. RESULTS: There was found: a moderate number of endothelin-positive cells in both blood vessel types; a moderate number of MMP2-positive cells and moderate in number to numerous TIMP2-positive cells in veins. In arteries - occasionally marked positive MMP2 cells and negative TIMP2; moderate in number to numerous VEGF-positive endothelial cells on small blood vessels in vein wall and occasionally in artery wall; numerous TGFß-positive structures in veins and abundance of VCAM- and ICAM-positive cells, few in arteries; few HGF-positive structures in veins, negative in arteries; In veins, few PGP9.5-positive nerve fibres, in arteries - moderate. Moderate TUNEL reaction-positive apoptotic cells in veins and few to moderate in arteries. CONCLUSIONS: Vena saphena magna grafts are characterised by increased plasticity when it comes to modelling. Number of VEGF, VCAM and ICAM found in vena saphena magna proves the possible tendency of graft failure on basis of local blood supply intensification. Appearance of endothelin positive cells indicate the similar homeostasis condition in endotheliocytes in both - vein and artery grafts.

Age-related lung tissue remodeling due to the local distribution of MMP-2, TIMP-2, TGF-ß and Hsp70.

Lung tissue remodeling requires complex interactions of matrix metalloproteinases (MMPs), tissue inhibitors of metalloproteinases (TIMPs), transforming growth factor (TGF) family and heat shock protein 70 (Hsp70). We evaluated the appearance and distribution of MMP-2, TIMP-2, TGF-ß1 and Hsp70 in lung tissue using immunohistochemistry. Stained structures were graded semiquantitatively. Overall, more MMP-2, TIMP-2, TGF-ß1 and Hsp70 were observed in bronchial cartilage, bronchial and alveola repithelium, and among alveolar macrophages. We evaluated mostly alveolar macrophages, bronchial epithelial cells and mucosal fibroblasts stained for TGF-ß1, MMP-2 and TIMP-2. We also assessed strong or moderate correlations between numbers of cells containing TGF-ß1, MMP-2, TIMP-2 in patients ≥ 60 years old. The presence of less TGF-ß1 and more MMP-2, TIMP-2 and Hsp70 containing cells in all tissue groups indicated that local regulation was more dependent on MMP-2, TIMP-2 and Hsp70 distribution. Fewer TIMP-2, Hsp70 and TGF-ß1 immunoreactive cells in younger individuals and increased expression of Hsp70 in elderly individuals demonstrated the influence of aging in lung remodeling. Findings of MMP-2, TIMP-2 and TGF-ß1 immunoreactive cells in elderly individuals indicate lung remodeling due to aging.

Strontium and strontium ranelate: Historical review of some of their functions.

The review covers historical and last decade's scientific literature on the biological and clinical role of strontium (Sr) and strontium ranelate (Sr RAN). It enrols the description of the main effects of Sr on supportive tissue, its proven and possible morphopathogenetical mechanisms and the interaction with the bone, and especially focuses on the Sr ability to inhibit osteoclasts and affect the programmed cell death. The main experimental and clinical experience regarding the Sr RAN influence in the treatment of osteoporosis and the search for correct doses is also highlighted. The review gives insight into the role of Sr/Sr RAN on stem cells, apoptosis, animal and clinical research.

Alternative for improving gut microbiota: use of Jerusalem artichoke and probiotics in diet of weaned piglets.

The aim of the study was to determine the effect of Jerusalem artichoke and probiotics on defence activity of intestinal cells of weaning pigs. One hundred eighty piglets (7 weeks old) were fed with basal feed supplemented with Jerusalem artichoke, Lactobacillus reuteri and Pediococcus pentosaceus. After 5 weeks, the piglets were slaughtered and the gastrointestinal contents and intestine samples were taken for analysis. Results demonstrated that in pigs fed basal diet with both probiotics and Jerusalem artichoke (5% of basal diet) (T3 group) had less (P<0.05) faecal Enterobacteriaceae microorganisms and coliforms and had more (P<0,05) faecal Lactobacillus than in pigs from other groups. Increase by 2% of Enterobacteriaceae and E. coli levels were seen only in control piglets (T1 group). E. coli O157 was found at the closing stage in the piglets fed basal diet with only Jerusalem artichoke powder (T2 group), but Salmonella enteritidis - only in T1 group. In jejunum of T2 group piglets, large deterioration of crypts, a moderate inflammation process and plasmocytes were seen, but in jejunum of T3 group piglets - branching of apical surface of villi, moderate degeneration and mitosis of enterocytes were observed. A moderate number of apoptotic cells in T2 group was found mainly in colon inflammation cells and plasmocytes, but for T3 group piglets--both in jejunum enterocytes and migrating cells. Our study indicated that beta-defensin 2 and 3 expression in jejunum and colon segments were incresed in T1 and T2 groups. Findings suggest that feeding with probiotics and Jerusalem artichoke significantly improves the microbial contents, defence and regeneration processes in the intestine of pigs.

[Morphological changes in the lumbar dorsal root ganglion of the domestic porcine after pulsed radiofrequency stimulation].

Pulsed radiofrequency (PRF) is a percutaneous minimal invasive procedure that can be used when conservative pain therapy methods have been ineffective. The effectiveness of PRF was demonstrated in various good quality randomized control studies, but mechanisms of action are still unclear. The aim of our study is to analyse the histological effects of PRF on the domestic porcine dorsal root ganglion (DRG), and evaluate the expression of biomarkers in gangliocytes. 3 domestic porcines were investigated. Under general anaesthesia and X-ray control, DRG PRF was performed. Four lumbar DRGs (L1, L2, L3, L4) were randomly treated. The opposite side DRGs was used as control. One month after the procedure the animal was euthanized. The lumbar region of the spine was placed in 10% formaldehyde for a month. After this fixation DRG samples were prepared for slide analysis. They were embedded in paraffin in order to obtain 3 microm thick sections, which were then cut by microtome and collected on slide glasses. Using standard immunohistochemical reactions, the materials were tinted to define biomarkers NF, GFAP, Hsp-70 expression and apoptosis by TUNEL kit. The number of cells with NF (26.0 +/- 3.0 vs 16.1 +/- 3.3; p < 0.05), GFAP (12.0 +/- 1.3 vs 3.2 +/- 0.9; p < 0.05) and Hsp-70 (10.0 +/- 1.6 vs 4.2 +/- 1.0; p < 0.05) expression, were larger in the PRF side comparing with the control side. Additionally, glial cells in spinal ganglia of both sides demonstrated immunoreactivity. The instances of apoptosis were not significantly different, in statistical terms, between the control and experimental sides (18.0 +/- 4.0 vs 20.0 +/- 4.0; p = 0.35). PRF in spinal gangliocytes of lumbar region increases neural tissue cytoskeleton factors like NF and GFAP suggesting about active regeneration processes into the cells 1 month after the procedure. Spinal gangliocytes one month after PRF treatment notably increases Hsp-70 expression suggesting about activation of cellular activity and inhibitory role reducing of oxidative stress. Similar number of apoptotic cells in spinal ganglia of lumbar region after PRF and control side suggests about inhibitory role of PRF on programmed cell death and stimulation of cell survival.

Histopathologic and immunohistochemical lesions in liver of mink infected with Aleutian disease virus.

Parvovirus of Aleutian disease causes mainly damage to kidneys, but immune complexes deposition and damage may occur also in other organs. In mink farms of Latvia the liver dystrophy or hepatic lipidosis of mink is widely distributed. The goal of this study was to examine probability of liver damage and regeneration of mink infected with Aleutian disease virus. Liver injury was assessed histologically. The mink liver demonstrated inflammation of liver parenchyma and foci of fatty liver. In immunohistochemistry, during liver regeneration the matrix metalloproteinases MMP-9, vascular endothelial growth factor and beta-defensin 2 expressions were lower, but MMP-2 and nerve growth factor receptor p75 expression was increased.

Postsplenectomy sepsis in splenectomized, partially splenectomized and non-splenectomized rats after Streptococcus pneumoniae challenge.

INTRODUCTION: Purpose of the study was the evaluation of the role of 1/3 of the spleen in host defense after a challenge with Streptococcus pneumoniae. MATERIALS AND METHODS: Forty Wistar rats divided into four groups underwent splenectomy (SPR), partial splenectomy (PSR) or sham operation (SOR). Healthy rats were used as controls (CGR). Operations were performed under general anesthesia. Ten weeks after operation the rats were challenged with 6 × 107 cfu/ml Streptococcus pneumoniae administered intravenously. All surviving animals were sacrificed 12 days after intravenous injection Interleukin-10, tumor necrosis factor-α and human ß-defensin-2 containing cells were detected in the parenchymatous organs (spleen, lungs, liver and kidneys) of all groups. Kaplan-Meier and Mann-Whitney tests were used for statistical analysis. RESULTS: Survival after Streptococcus pneumoniae challenge was longer in animals with a greater amount of splenic tissue, with mortality increased proportionately to the reduction in splenic tissue. In the SOR group survival was 11.6 ± 1.3 days (10% mortality). In the PSR group survival was 6.0 ± 2.5 days (90% mortality). In the SPR group survival was 1.6 ± 0.8 days (100% mortality). In splenic tissue the levels of HßD-2, IL-10 and TNF-α-containing cells did not differ statistically (z=5.021; p<0.01) and were higher than in other parenchymatous organs (PSR, SOR, CGR). Levels of IL-10-containing cells were higher in parenchymatous organs of the SPR group (z=7.919; p<0.001), similar in the PSR and SOR groups (z=1.020; p=0.308) and lower in the CGR group (z=4.366; p<0.01). There were no statistically significant differences in the levels of IL-10 containing cells in the lungs of all group rats with spleen (z=4.266; p<0.01). Levels of TNF-α-containing cells were similar in PSR and SOR groups (z=1.004; p=0.315). Relative levels of HßD-2 in kidney differed between all groups (z=2.916; p=0.004). CONCLUSIONS: All of the splenectomized animals (100%) and 90% of the partially splenectomized animals died. Partial splenectomy (with 1/3 of splenic tissues remaining) does not offer full protection against Streptococcus pneumoniae sepsis. In all groups, the amounts of HßD-2, IL-10 and TNF-α-containing cells in the spleen were higher than in other parenchymatous organs (lungs, liver and kidneys).

Comparative studies on bone structure in dairy cows with different feeding conditions.

The bone belongs to the dynamic tissues and its structure in domestic cows is still not completely understood in correlation to the impact of different food components. The aim of our work was a histomorphometrical and immunohistochemical research on bone morphology and factors influencing it in healthy dairy cows fed with self-produced grain and with rapeseed cakes. The bone of self-produced grain-fed cows demonstrated statistically significant difference in the number of osteocytes from the bone of rapeseed cakes-fed cows. The rapeseed cakes-fed cows didn't show any bone cell positive for BMP2/4, while FGFR1 increased significantly in their supportive tissues. The number of bFGF- and apoptosis-containing structures varied in cows of both groups. MMP2 expression showed statistically significant difference between both animals' groups with domination in bone of cows fed with self-produced grain. Defensin-, osteopontin- and osteocalcin-containing cells showed tendency to increase in bone of cows on rapeseed cakes diet. Conclusions. The rapeseed-fed cow's long bones demonstrate significant decrease of osteocytes per mm2 and selective increase of FGFR1, suggesting the (compensatory) growth stimulation in supportive tissue. The statistically significant selective absence of MMP2 with a slight tendency of increase in osteopontin and osteocalcin in rapeseed-fed cow's long bones indicates the persistence of seemingly still compensated qualitative changes in bone (beginning of disturbances in mineralization, metabolism etc.) proved also by a slight increase of the bone antimicrobial peptide.

Investigation of K+ channel expression in human peripheral lymphocytes of healthy donors by means of flow cytometry.

Evaluation of different types of K+ channel expression was performed in resting and PHA (phytohemagglutinine)-activated human peripheral lymphocytes (HPL) of healthy donors by means of flow cytometry. In resting peripheral lymphocytes, the application of kaliotoxin (a selective blocker for voltage-dependent K+ (K(V)) channels), K(V) resulted in pronounced depolarization of lymphocyte membrane potential, with further promotion in the presence of thapsigargin (compound discharging Ca(i) from endoplasmic reticulum). In activated HPL, the expression of various types of K+ channels was estimated utilizing cell-cycle analysis data. In contrast to the resting cells, kaliotoxin-induced depolarization of membrane potential in PHA-activated lymphocytes of the G0/G1 phase was not enhanced by thapsigargin and in PHA-activated lymphocytes of the S and G2/M phases we were able to observe repolarization of membrane potential after kaliotoxin-induced depolarization of membrane potential. Substitution of kaliotoxin for charybdotoxin (a non-selective drug blocking both K(V) and K(Ca) channels) abrogated the above effects in PHA-activated lymphocytes. Thus, K(V) channels are active in both resting and activated HPLs and K(Ca) channel expression occurs with cell-cycle progress on PHA-induced activation of peripheral lymphocytes.

Changes in neuroendocrine elements in bronchial mucosa in chronic lung disease in adults.

BACKGROUND: It is not clear whether there is any association between metaplasia of the bronchial epithelium and changes in the distribution of neuroendocrine cells. This study examined, by immunohistological techniques, the distribution of neuroendocrine cells and juxtamucoscal nerve fibres in bronchial biopsies showing metaplastic changes. METHODS: Bronchial biopsies from 12 subjects with epithelial metaplasia associated with bronchiectasis and diffuse pulmonary fibrosis were examined by conventional light microscopy and immunohistological techniques for protein gene product 9.5 (PGP), chromogranin A and B (CAB), serotonin, vasoactive intestinal peptide (VIP), substance P (SP), calcitonin gene-related peptide (CGRP), calcitonin (CT), and gastrin releasing peptide (GRP). RESULTS: Regions of non-metaplastic epithelium contained numerous PGP and serotonin immunoreactive cells. Sub-populations of these cells displayed CAB, CGRP, CT, and GRP immunoreactivity. Metaplastic epithelium contained only a few weakly stained PGP, serotonin, CAB, GRP, CT and CGRP immunoreactive cells in six cases. Metaplastic epithelium was characterised by a high number of CAB-containing cells in six cases and in these biopsies prominent PGP-containing nerve bundles were seen in the subepithelial layer beneath the metaplastic epithelium. CONCLUSIONS: The distribution patterns of neuroendocrine cells and neuronal elements vary between areas of normal and metaplastic epithelium and within areas of metaplastic epithelium. Neuronal hyperplasia was associated with an increase in the number of CAB-containing cells within the metaplastic epithelium.

[The localization of protein gene product 9.5 in the lungs of children with bronchitis and bronchiectasis]. / Lokalizatsiia proteingenprodukta 9,5 v legkikh u detei s bronkhitom i bronkhoéktazami.

Human respiratory tract has a rich innervation with nerve fibers. Presence and distribution of nerve elements labeled with protein gene product 9.5 (PGP) were studied in the lungs of children with bronchitis and bronchiectasis. Smooth myocytes, blood vessels and mixed glands of the walls of bronchi principales were surrounded with single PGP-positive nerve fibers in perichondrium grows smaller with age. Nerve fibers also surround lymph nodules of the bronchi principales mucosa. Small or medium number of them in nerve plexuses and surround basically blood vessels. As neuropeptide function in respiratory tract vary, they may be of certain importance in pathogenesis of chronic lung diseases. Morphological base of this suggestion was given.

Basement membrane components of bronchial epithelium in humans suffering from chronic nonspecific lung diseases.

Collagen IV and laminin are important constituents of the basement membrane (BM). By use of immunocytochemistry we examined the occurrence and distribution of these two components in the BM beneath normal, mucoid and metaplastic epithelium of large bronchi in 22 adults suffering from chronic nonspecific lung diseases. Both collagen IV and laminin were expressed as a thin and continuous layer beneath the epithelium in most tissue specimens with normal epithelium. In a few specimens the layer showed interruptions with a patchy distribution of the immunoreactivity. Three patterns of distribution of BM components were found under the metaplastic epithelium. Total absence of immunoreactive collagen IV and laminin was the most common variant. Weak and scarce staining for both proteins in the BM characterized the second pattern. The third variant showed strong collagen IV immunoreactivity but lack of laminin. The BM beneath the mucoid epithelium was characterized by irregular distribution of collagen IV and laminin. We suggest that the occurrence and distributional pattern of the BM components are related to the type of overlying epithelium and connected with an altered synthesis of these components.