Autonomic balance determines the severity of COVID-19 courses.

COVID-19 has left mankind desperately seeking how to manage dramatically rising infection rates associated with severe disease progressions. COVID-19 courses range from mild symptoms up to multiple organ failure and death, triggered by excessively high serum cytokine levels (IL 1ß, IL 6, TNF α, IL 8). The vagally driven cholinergic anti-inflammatory pathway (CAP) stops the action of nuclear factor κB (NF-κB), the transcriptional factor of pro-inflammatory cytokines. Thus, well-balanced cytokine release depends on adequate vagal signaling. Coronaviruses replicate using NF-κB transcriptional factor as well. By degrading the cytoplasmatic inhibitor of NF-κB subunits (IκB), coronaviruses induce unrestricted NF-κB expression accelerating both, virus replication and cytokine transcription.We hypothesize that CAP detriment due to depressed vagal tone critically determines the severity of COVID-19.

Buffered Romanowsky-Giemsa method for formalin fixed, paraffin embedded sections: taming a traditional stain.

Romanowsky-Giemsa (RG) stains were devised during the 19th century for identifying plasmodia parasites in blood smears. Later, RG stains became standard procedures for hematology and cytology. Numerous attempts have been made to apply RG staining to formalin-fixed paraffin-embedded tissue sections, with varied success. Most published work on this topic described RG staining methods in which sections were overstained, then subjected to acid differentiation; unfortunately, the differentiation step often caused inconsistent staining outcomes. If staining is performed under optimal conditions with control of dye concentration, pH, solution temperature and staining time, no differentiation is required. We used RG and 0.002 M buffer, pH 42, for staining and washing sections. All steps were performed at room temperature. After staining and air drying, sections were washed in 96-100% ethanol to remove extraneous stain. Finally, sections were washed in xylene and mounted using DPX. Staining results were similar to routine hemalum and eosin (H & E) staining. Nuclei were blue; intensity depended largely on chromatin density. RNA-rich sites were purple. Collagen fibers, keratin, muscle cells, erythrocytes and white matter of the central nervous system were stained pinkish and reddish hues. Cartilage matrix, mast cell granules and areas of myxomatous degeneration were purple. Sulfate-rich mucins were stained pale blue, while those lacking sulfate groups were unstained. Deposits of hemosiderin, lipofuscin and melanin were greenish, and calcium deposits were blue. Helicobacter pylori bacteria were violet to purple. The advantages of the method are its close similarity to H & E staining and technical simplicity. Hemosiderin, H. pylori, mast cell granules, melanin and specific granules of different hematopoietic cells, which are invisible or barely distinguishable by H & E staining, are visualized. Other advantages over previous RG stains include shorter staining time and avoidance of acetone.

Use of eriochrome cyanine R for routine histology and histopathology: an improved dichromatic staining procedure.

A modified dichromatic iron-eriocyanine R (Fe-ECR) staining method is described. Staining obtained with this new technique generally was similar to that of hematoxylin and eosin (H & E). Cell nuclei were stained blue. Cardiac, smooth and skeletal muscle, and red blood cells, were stained different shades of red. Collagen fibers were stained different shades of orange, usually faintly. Decalcified bony tissue was stained pinkish violet. Epithelial cells were strongly stained deep shades of red, magenta and violet. Cartilage matrix, and goblet and mast cells were unstained. Although Fe-ECR staining differed too much from standard H & E staining to be a substitute for diagnostic purposes, the dichromatic method described might usefully replace van Gieson or trichrome stains, especially if muscle is of interest. A pH 0.95 staining solution was used to differentiate initially over-stained sections followed by washing in distilled water. This dichromatic technique is easier to perform and more precisely controllable than other ECR dichromatic methods. The entire procedure can be completed in less than 5 min. The technique has the advantages of greater technical simplicity and speed, a larger range of polychromasia, and a longer shelf-life than H & E. ECR also is more reliably available than hematoxylin and usually is less expensive.

Use of eriochrome cyanine R in routine histology and histopathology: is it time to say goodbye to hematoxylin?

Eriochrome cyanine R (ECR) is a synthetic anionic dye that forms complexes with cations such as iron. We found that an iron-ECR (Fe-ECR) mixture provided either nuclear or myelin staining depending on the differentiator used. Selective nuclear staining was obtained by differentiation in an aqueous HCl solution, pH 0.95, followed by a wash in slightly alkaline tap water; the pH difference facilitated control of differentiation. When used with an eosin B counterstain, results were nearly indistinguishable from standard hematoxylin and eosin (H & E) staining. Nuclear staining with Fe-ECR provides tinctorial features similar to regressive aluminum-hemateins as well as resistance to acidic solutions such as those of iron hemateins. Fe-ECR also stained selectively intestinal cells of the diffuse neuroendocrine system (DNES). In addition to its use as an H & E substitute, acid differentiated Fe-ECR produced acid-resistant and selective nuclear counterstaining in combination with Alcian blue, and in the Papanicolaou and van Gieson techniques. With alkali differentiation, Fe-ECR produced selective myelin staining, which was compatible with neutral red counterstaining. Myelin sheaths were stained aqua blue. Fe-ECR could be used for both cytological and histological samples, and was suitable for use in automated tissue stainers. ECR also is less expensive than hematoxylin. Hematoxylin still may be preferred as a nuclear counterstain for some immunostaining methods for which Fe-ECR mixtures probably are too acidic.

Acute toxic effects of single dose dacarbazine: hematological and histological changes in an animal model.

Treatment of advanced soft tissue sarcoma usually includes dacarbazine (DTIC), an alkylating agent that methylates DNA and is active during all phases of the cell cycle. Common side effects of DTIC include nausea, vomiting, impaired liver and kidney function, myelosuppression, and pneumonia. There are no accounts, however, of histological and hematological changes caused by DTIC. We investigated acute hematological and morphological changes in different organs and in tumors that were caused by a single dose of DTIC. Adult Syrian golden hamsters were inoculated with a suspension of tumorigenic baby hamster kidney (BHK) cells by subcutaneous injection. On day 14 after inoculation, doses of 1.4, 1.6, 1.8 or 2.0 g/m(2) DTIC were injected intraperitoneally into the hamsters. Hamsters in the control group were injected with physiological saline in the same way. Seven days after drug or saline injection the animals were sacrificed and samples of blood, heart, kidney, liver, lungs, spleen, small intestine and tumor were excised, processed and analyzed. Mitoses were counted using an ocular extension with engraved frame. Anemia, thrombocytopenia and leukocytosis were found in the control group of hamsters with fibrosarcoma, whereas animals with fibrosarcoma treated with DTIC developed anemia, thrombocytopenia and leukopenia. Severe pneumonia and moderate hepatitis were detected in all DTIC treated groups. Effects of DTIC on tumor cells included rounding and enlargement of nuclei and rarefaction of chromatin. The number of mitoses was reduced with increasing doses of DTIC. Hepatitis, myelosuppression, pneumonia, and dose-related inhibition of tumor cell proliferation were observed after a single dose of DTIC.

Romanowsky-Giemsa as a counterstain for immunohistochemistry: optimizing a traditional reagent.

We describe a detailed protocol for using Romanowsky-Giemsa (RG) counterstaining on formalin fixed, paraffin embedded tissue sections that are stained immunohistochemically (IHC) after antigen retrieval using hot acidic citrate buffer. RG staining is easy to perform and provides consistent results that are similar to hematoxylin and eosin (HE) staining. The counterstaining was applied after a variety of antibodies that used the DAB chromogen and the intensity of IHC stained structures was preserved. Moreover, RG counterstaining provided finer cell detail than HE, methyl green or nuclear fast red. A detailed troubleshooting guide is provided for the RG staining protocol.

Survivin expression in odontogenic keratocysts and correlation with cytomegalovirus infection.

OBJECTIVES: The aim of this study was to investigate the expression of survivin, an inhibitor of apoptosis, in odontogenic keratocysts and to compare it to the findings in non-neoplastic jaw cysts - periapical cysts, as well as to establish a possible relationship between survivin expression and human cytomegalovirus presence within these cysts. MATERIALS AND METHODS: Samples of 10 odontogenic keratocysts (five positive and five negative for the presence of cytomegalovirus, as determined by polymerase chain reaction) and 10 periapical cysts (five positive and five negative for the cytomegalovirus presence) were analysed. The expression of survivin was assessed by immunohistochemical methods, using monoclonal antibody that selectively recognizes the cytoplasmic form of survivin. RESULTS: All 10 odontogenic keratocysts showed immunostaining for survivin, while all 10 periapical cysts were negative for its presence. There was no correlation between cytomegalovirus presence and expression of survivin within odontogenic keratocysts. CONCLUSION: Survivin may contribute to the aggressive behavior of odontogenic keratocysts, and thus support the emerging opinion of their neoplastic nature.

Long-term outcome in Serbian patients with Wilson disease.

BACKGROUND AND PURPOSE: To investigate survival rates, prognostic factors, and causes of death in Wilson disease (WD). METHODS: In the years 1980-2007, a cohort of 142 patients with WD was prospectively registered (54 presented with neurologic symptoms, 49 with hepatic symptoms, 33 had mixed form, and data were missing for six patients). The duration of follow-up for patients alive was 11.1 +/- 8.8 years. RESULTS: After initiation of treatment (d-penicillamine and zinc salts), 79% of patients had a stable or improved course of disease. Despite early diagnosis and appropriate therapy, 15 patients still had a relentlessly progressive course. Thirty patients died. The cumulative probability of survival in a 15-year period for the whole group was 76.7 +/- 4.9%. Better prognosis of WD was associated with male sex, younger age at onset, neurologic form of the disease, and treatment continuity. Causes of death were predominantly related to hepatic failure (16 patients), but also suicide (four patients) and cancer (three patients). CONCLUSION: Despite the relatively early diagnosis and treatment of our patients with WD, mortality was still considerably high.

Microanatomical structure of the human sciatic nerve.

BACKGROUND: Sciatic nerve is the largest peripheral nerve of the human body. It gives motor and sensitive innervation for the most of lower limb. The aim of the present investigation was revealing his fascicular pattern in relation to microanatomic morphometric characteristics of its connective tissue sheaths. METHODS: The material consisted of sciatic nerve slices, excised from 17 cadavers of humans aging 8-93 years. After routine histologic processing and light microscopic examination of the preparations, morphometric analysis was performed at magnifications of 40 and 630x. RESULTS: Sciatic nerve showed to be polyfascicular nerve type, with the group pattern of nerve fascicless distribution. The number of fascicless ranged from 27 to 70, whereas the number of fascicless per square millimeter was 1-4. Morphometric and correlation analysis confirmed the significant increase of whole sciatic nerve cross section area, which was associated with the significant increase of its epi- and perineural connective tissue sheaths. Interfascicular sciatic nerve domains of elderly persons contained more adipose tissue. Moreover, already detected loss and degeneration of the large myelinated nerve fibers within fascicles was accompanied by the significant increase of endoneural connective tissue. CONCLUSIONS: In conclusion, our study revealed comparative connective tissue enlargement of human sciatic nerve in the course of aging. These phenomena might influence on result of injured nerve's surgical reparations. We interpret this finding as non-specific compensatory phenomenon elicited by loss of thickest myelinated nerve fibers, higher vulnerability of remaining ones, and age-dependent decrease of connective tissue elasticity.

[Traumatic rupture of the right hemidiaphragm].

The right hemidiaphragm rupture, caused by blunt force, occurs rarely and often is overlooked because of immediate management of life-threatening injuries. Our paper presents 31-year-old female patient, admitted as emergency case due to traffic accident injuries. Physical examination revealed diminished respiratory mobility of the right hemothorax, along with the impaired respiratory sound to the right infrascapular aspect, diffuse painful sensitivity of the abdomen and limited mobility of the lower extremities. X-ray examination of the chest verified the tight hemidiaphragm elevation without any signs of bone structure fractures and hydro-pneumothorax. Computerized tomography of the abdomen and small pelvis showed the rupture of the right diaphragmatic cupola with the prolaps of the right hepatic lobe, sacral fracture and fracture of both ischial rami. The surgery verified the rupture of the right hemidiaphragm in transversal direction of about 10 cm long, with partial prolaps of liver to the right hemithorax. Surgical intervention included the insertion of thoracic drain and liver mobilization by severing the falciform ligament. The operation was completed by interrupted sutures of the right hemidiaphragm and hematomas in the retroperitoneum were incised. Postoperative course was uneventful, and oral nutrition was initiated on day 4. The patient was discharged 18 days after the surgery. Preoperative diagnostics, attentive intraoperative examination and early surgical treatment significantly reduce morbidity and mortality of patients with traumatic diaphragmatic rupture.

[Endometriosis in cesarean section surgical scar].

INTRODUCTION: Endometriosis is the presence of endometrial glands and stroma outside of uterine cavity. It may occur in the abdominal wall scar after the operation in which uterus was opened. In cesarean section scar it occurs in 0.4%. It is in 2/3 patients characterised with triad of: tumor, periodic pain associated with menses and history of cesarean section. The mechanism of endometriosis occuring in the cesarean scar is felt to be secondary to iatrogenic transplantation of endometrium or extrauterine decidual tissue into the incision during the cesarean section. CASE OUTLINE: Forty years old patient with tumor 4,5x4 cm that appeared in abdominal wall scar one year after second cesarean section, followed by periodic pain and macroscopic changes associated with menses. First diagnosis was granuloma in the surgical scar, but as she had periodic simptoms, diferential diagnosis was endometriosis. Hormonal therapy with contraceptiv drugs was ordered. As it was no improvement she was operated. The surgical excision of the tumor including fascia and muscle tissue was done. Sample revealed endometrium after histopathologic examination. Patient was complitely recoverd and without relepse of simptoms during followup to date. CONCLUSION: When there is a tumor in the cesarean section scar or scar after the operation in which uterus or ovarial tube was opened, followed with periodical pain and macroscopic changes associated with menses, endometriosis should be considered. Surgical excision of the tumor is sufficient and patohistological examination confirms diagnosis.

["I am sorry, I have bad news... "- principles in informing patients on the gravity of a disease and the treatment].

The discovery of the appearance of a serious disease, and the necessity for diagnostics, treatment and rehabilitation, particularly when malignity is involved, represents exceptionally stressful news for the patient and his family. Most often this is a task for the physician. In this paper the authors consider the meaning of bad news in unexpected life events which significantly disturb the individual psycho-social ballance of the patient, as well as the familly dynamics and structure. It presents a review of available literature on the subject of the definition of so-called "bad news", the development of approaches in announcement, and current practical approaches and models which assist physicians in helping suffering patients in professional and humane ways. It points out inadequacies in the education of phisicians for this kind of task, and the variety of approaches. With the aim of improving the education of phisicians, and especially clinicians in surgical fields, the authors recommend a number of educational programs: education in the area od psychological and psychiatric aspects of learning of a serious disease, including acute reactions such as depressive states, reactive psychotic states, consumption of alcohol, etc.; education in the area of the family life cycle, and the effect of the disease as an unexpected life crisis on the family dinamic; education in the area of fundamental principles of psycho-and pharmaco-therapy, as well as the provision of support to the patient and his familly; training in the skills of announcing bad news to the patient and his familly through the explanatory presentation of the case and supervised simulation of sessions.

Lupus band test and disease activity in systemic lupus erythematosus: does it still matter?

BACKGROUND: Lupus band test still has no clearly defined position within either diagnostic or disease activity measuring tools for systemic lupus erythematosus (SLE). OBJECTIVES: We tested the hypothesis that positive LBT correlates with global activity of SLE measured by the SLE Disease Activity Index (SLEDAI) score. METHODS: In total, 90 SLE patients who underwent biopsy of sunprotected non-lesional (SPNL) skin were studied prospectively. The skin specimen was processed for standard direct immunofluorescence. The patients were classified into groups as negative and positive LBT, and the latter were further subdivided on the basis of the type and morphology of the deposits. Every patient was thoroughly examined and assigned a SLEDAI score. The relationship between LBT findings and SLEDAI score was analysed. RESULTS: The disease was significantly more active in patients with positive LBT and in those with a higher number of deposited immunoreactants. Almost all patients with renal involvement had a positive LBT. CONCLUSIONS: LBT on SPNL skin may be a good marker of severe disease at presentation, particularly when three immunoglobulins are found at the dermoepidermal junction.

[Assessment of the degree of maturity in the wall of pancreatic pseudocysts in relation to choice of surgical procedure].

The aim of this study is to present our experience in the diagnosis and treatment of pancreatic pseudocysts. A pancreatic pseudocyst is an incapsulated collection of pancreatic juice, enclosed by nonepithelial elements, containing a high concentration of pancreatic enzymes, bicarbonates and necrotic detritus. It is a common complication of acute pancreatitis and trauma of the pancreas. In the period between 1996 and 2001, 53 surgical procedures were performed for pancreatic pseudocyst at the Institute for Digestive Diseases (First Surgical University Hospital), 35 male patients (67%) and 17 female patients (33%) underwent surgery. In 39 (75%) patients the method of choice was cystojejunostomy by Roux. In 4 cases distal pancreatectomy for pseudocysts localized within the pancreatic tail was performed, complete pseudocyst excision only was performed in one case and complete pseudocyst excision combined with cystojejunostomy was also performed in one case. Cystogastrostomy and drainage in one case and partial cystectomy and drainage also in one case. Surgical internal drainage is the method of choice for the treatment of pancreatic pseudocysts, involving low morbidity and mortality rates.

Adamantiades-Behçet's disease, deep venous thrombosis and anticardiolipin antibodies: report of two cases.

Vascular thrombosis was found in different proportions of patients with Adamantiades-Behçet's disease (ABD), depending on the ethnicity of the population under study. Various thrombophilic factors, including the levels of anticardiolipin antibodies (ACA), were investigated for their role in the thrombotic process with conflicting results. The prevalence of ACA varies considerably in different studies, but their presence has not been associated with increased risk for vascular thrombosis. We present two cases with ABD, deep venous thrombosis (DVT) and elevated levels of ACA that fulfil the criteria for both ABD and antiphospholipid syndrome (APS).

The pharmacologic stability of 35-year old theophylline.

The propensity to preserve and to hoard drugs over the years at home is a well-known phenomenon and offers the possibility for intentional and accidental drug poisoning in man. We report a case of acute theophylline poisoning in an 80-year old women after ingestion of 'Asthmo-Kranit', a 35-year old combined preparation containing theophylline and aminopyrine as the main ingredients. The patient developed the typical clinical picture of a symptomatic theophylline poisoning with flush, tremor, tachycardia, hyperventilation, hypotonia, and hyperglycaemia. The clinical course after treatment with beta-blockers was without complications. The determination of theophylline in tablets showed stability of 90% of the labelled amount of the drug 30 years beyond the expiration date. The case illustrates the prolonged shelf stability and pharmacological potency of some pharmaceuticals and points to the risk of long-outdated prescriptions. Physicians should primarily not underestimate drug toxicity in consequence of old-age pharmaceuticals.

[Autotransplantation of the spleen]. / Autotransplantacija slezine.

Auto transplantation of the spleen can be performed in the patients with traumatic rupture of the spleen, in whom spleen could not be conserved in the other way. The right indication for this method is isolated rupture of the spleen (concvasation or complete devascularisation). This method is not recommended in the endangered patients, patients with previous disease of the spleen as well as in the patients with the perforation of the other abdominal organs at the same time. Auto transplantation was performed in 12 patients with isolated splenic rupture and hematoperitoneum, 11 men and one woman. The majority of patients are younger. In 8 patients, autotransplant was placed into big omentum, in three into lipomatous tissue surrounding left kidney, and in one into anterior abdominal wall. In all the patients from this group, following analysis were taken: MCV (middle volume of erythrocytes), HTC, Hb, Le, Glucose, urea, creatinin, sodium, potassium, alkali phosphatasis, target cells, Howell Jolly's bodies, Heinz's bodies, IgG, IgA, IgM, C3, C4, T3, T4, T8, B, segmentated, eosinophiles, lymphocytes, reticulocytes, thrombocytes, fibrinogen, PT, APTT, aggregation of thrombocytes and aggregation of thrombocytes on collagen. The same parameters were taken in 12 patients with surgery similar to splenectomy and in 12 after splenectomy. After splenectomy, there was decrease of the immunologic defending abilities of the organism because of the loss of the childrens function of the spleen, decreased level of the opsonines and tutsin, which leads to the impaired phagocytosis, decreased concentration of IgM and T and B lymphocytes, while in patients after auto transplantation the results were physiological. The most important thing in the assessment of the function of the autotransplanted spleen is scintigraphic investigation using 99mTc-denaturated red blood cells. In our study, auto transplant function was assessed in 10/12 patients by scintigraphy. Five years after surgery, no one patient was proved to have postsplenectomic sepsis.

Skin lesions--an indicator of disease activity in systemic lupus erythematosus?

Cutaneous manifestations have great diagnostic value for systemic lupus erythematosus (SLE). In this study we tried to establish a correlation between lupus erythematosus LE-specific and LE-nonspecific cutaneous lesions and disease activity measured by the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI). Sixty-six patients with SLE were evaluated. They were divided into three groups having: (1) only LE-specific lesions (38 or 58.46%); (2) only LE-nonspecific lesions (4 or 6.15%); and (3) both types of lesions (23 or 35.38%). Results were analyzed using the Student t-test. Patients with LE-nonspecific skin manifestations had significantly increased disease activity compared to those with only LE-specific lesions. The number of different skin lesion types also correlated with disease activity. It was significantly increased in a group with three different types of lesion, either specific or nonspecific. Patients with only one type of lesion had mild disease. An intermediate disease activity was found in the group with two different lesion types. Lupus-specific skin manifestations serve primarily as an important diagnostic clue. In conclusion, patients with LE-nonspecific lesions have significantly more active SLE than those with LE-specific lesions and may therefore require more intensive therapy and disease monitoring.