Vitamin D and interferon-γ co-operate to increase the ACE-2 receptor expression in primary cultures of human thyroid cells.

BACKGROUND: A more severe course of COVID-19 was associated with low levels of Vitamin D (VitD). Moreover in vitro data showed that VitD up-regulates the mRNA of the Angiotensin Converting Enzyme 2 (ACE-2), the SARS-COV-2 receptor in different type of cells. ACE-2 is expressed in several type of tissues including thyroid cells, on which its mRNA was shown to be up-regulated by interferon-gamma (IFN-γ). The aim of the present study was to investigate if treatment with VitD alone or in combination with IFN-γ would increase ACE-2 both at mRNA and protein levels in primary cultures of human thyrocytes. MATERIALS AND METHODS: Primary thyroid cell cultures were treated with VitD and IFN-γ alone or in combination for 24 h. ACE-2 mRNA levels were measured by Real-time Polymerase Chain Reaction (RT-PCR). The presence of ACE-2 on thyroid cell membrane was assessed by immunocytochemistry basally and after the previous mentioned treatments. RESULTS: ACE-2 mRNA levels increased after treatment with VitD and IFN-γ alone. The combination treatment (VitD + IFN-γ) showed an additive increase of ACE-2-mRNA. Immunocytochemistry experiments showed ACE-2 protein on thyroid cells membrane. ACE-2 expression increased after treatment with VitD and IFN-γ alone and further increased by the combination treatment with VitD + IFN-γ. CONCLUSIONS: VitD would defend the body by SARS-COV2 both by regulating the host immune defense and by up-regulating of the expression of the ACE-2 receptor. The existence of a co-operation between VitD and IFN-γ demonstrated in other systems is supported also for ACE-2 up-regulation. These observations lead to an increased interest for the potential therapeutic benefits of VitD supplementation in COVID-19.

On-line prediction of the feeding phase in high-cell density cultivation of rE. coli using constructive neural networks.

Streptococcus pneumoniae (pneumococcus) is a bacterium responsible for a wide spectrum of illnesses. The surface of the bacterium consists of three distinctive membranes: plasmatic, cellular and the polysaccharide (PS) capsule. PS capsules may mediate several biological processes, particularly invasive infections of human beings. Prevention against pneumococcal related illnesses can be provided by vaccines. There is a sound investment worldwide in the investigation of a proteic antigen as a possible alternative to pneumococcal vaccines based exclusively on PS. A few proteins which are part of the membrane of the pneumococcus seem to have antigen potential to be part of a vaccine, particularly the PspA. A vital aspect in the production of the intended conjugate pneumococcal vaccine is the efficient production (in industrial scale) of both, the chosen PS serotypes as well as the PspA protein. Growing recombinant Escherichia coli (rE. coli) in high-cell density cultures (HCDC) under a fed-batch regime requires a refined continuous control over various process variables where the on-line prediction of the feeding phase is of particular relevance and one of the focuses of this paper. The viability of an on-line monitoring software system, based on constructive neural networks (CoNN), for automatically detecting the time to start the fed-phase of a HCDC of rE. coli that contains a plasmid used for PspA expression is investigated. The paper describes the data and methodology used for training five different types of CoNNs, four of them suitable for classification tasks and one suitable for regression tasks, aiming at comparatively investigate both approaches. Results of software simulations implementing five CoNN algorithms as well as conventional neural networks (FFNN), decision trees (DT) and support vector machines (SVM) are also presented and discussed. A modified CasCor algorithm, implementing a data softening process, has shown to be an efficient candidate to be part of an on-line HCDC monitoring system for detecting the feeding phase of the HCDC process.

An unusual cause of lumbar discitis.

STUDY DESIGN: Case report. OBJECTIVE: To identify an unusual cause of lumbar discitis. SUMMARY OF BACKGROUND DATA: Discitis resulting from a foreign body has not been reported in the English-language literature. METHOD: A case report is reviewed identifying the presence of this foreign object (titanium), erroneously implanted into the disc space at L5-S1. Imaging studies identify the location and surrounding reaction. RESULTS: The surgical exploration of this disc revealed an inflammatory response surrounding the disc with degeneration within the disc space. CONCLUSION: If this gynecologic procedure is performed using bone anchors, care must be take to ensure that the sacrum is identified before insertion.

High-performance liquid chromatographic method for assay of otilonium bromide, diazepam, and related compounds in finished pharmaceutical forms.

A rapid, simple, stability-indicating assay procedure for otilonium bromide, a smooth muscle relaxant agent, and diazepam in composite tablet analysis was developed with high-performance liquid chromatography. The tablet matrix was dissolved with water, and drugs were extracted with acetonitrile containing an internal standard. An aliquot was centrifuged and chromatographed on a 5-microns, reversed-phase column with 0.5 M sodium acetate trihydrate buffer containing 5 mM 1-heptanesulfonic acid monohydrate sodium salt:methanol (30:70; v/v; adjusted to pH 6.0 with glacial acetic acid) as the mobile phase. The selectivity of the chromatographic system was demonstrated by resolving both compounds from various potential degradation products of each compound. The method is linear, quantitative, and reproducible.

Rapid simultaneous determination of tryptophan and tyrosine in synthetic peptides by derivative spectroscopy.

A method for the simultaneous quantitative determination of tryptophan and tyrosine in synthetic peptides by second-order derivative diode-array spectroscopy is reported. The method does not require hydrolysis of the peptides or a derivatization reaction; the sample is dissolved in 0.1 N NaOH and directly scanned between 262 and 264 nm to detect tyrosine and between 304 and 306 nm to detect tryptophan. From these results the peptide content of the synthetic sample can be easily calculated in a very simple and fast way. The results obtained by the described method with several peptides are compared with those obtained by classical high-performance liquid chromatographic analysis of amino acids after peptide hydrolysis. A very good correlation was found both for the tryptophan/tyrosine molar ratio and the peptide content.

Simultaneous determination of otilonium bromide and diazepam by first-derivative spectroscopy.

A rapid, simple assay procedure was developed for simultaneous analysis of otilonium bromide, a smooth-muscle relaxant, and diazepam in tablets containing 20 mg of otilonium bromide and 2 mg of diazepam (20:2 tablets) or 40 mg of otilonium bromide and 2 mg of diazepam (40:2 tablets) by "zero-crossing" first-derivative spectroscopy. The tablets were dissolved in 0.01 N HCl, mixtures were centrifuged at 3500 rpm (2472 x g) for 5 min, and first-derivative spectra were recorded. The absolute values of the derivative were measured at 264 nm for determination of otilonium bromide and between 406 and 408 nm (380 nm for analysis of 40:2 tablets) for determination of diazepam. The method is linear, quantitative, and reproducible and can also be used for the tablet dissolution test. Ten tablets of the same batch were analyzed by the described method and by a high-performance liquid chromatographic method, and the results were in good agreement.

Simultaneous quantitative determination of calcium leucovorin and parabens in lyophilized ampoules by derivative spectroscopy.

A simple and rapid procedure for quantitation of Calcium Leucovorin and parabens simultaneously in lyophilized ampoules formulation by "zero crossing" first-order derivative spectroscopy was developed. The ampoules content was dissolved in a mixture water/ethanol (50/50) and first-derivative spectra were recorded. The absolute values of the derivative at 312 nm for the determination of Calcium Leucovorin and between 244 and 246 nm for the determination of parabens were measured. The method is linear, quantitative and reproducible.

Analysis of platelet-activating factor by gas chromatography-mass spectrometry: low-energy electron impact of the corresponding 3-acetyl-2-tert.-butyldimethylsilyl derivative.

A method for the analysis of platelet-activating factor in platelets employing gas chromatography and selected-ion monitoring mass spectrometry with low-energy electron impact and stable isotope dilution was developed. The procedure involved Bligh and Dyer extraction of the sample followed by thin-layer chromatographic purification. Platelet-activating factor is successively hydrolysed to the corresponding 2-acetyl-1-O-alkylglycerol by digestion with phospholipase C, and the product is allowed to isomerize to the more thermodynamically stable 3-acetyl-1-O-alkylglycerol before column purification and derivatization of the free OH with tert.-butyldimethylchlorosaline-imidazole. This reagent is of common use in platelet-activating factor derivatization, but is made to react with 2-acetyl instead of 3-acetyl isomer. The advantages of using the latter for the final derivatization are discussed and this method is compared with others currently available for gas chromatographic-mass spectrometric analysis of platelet-activating factor.

The etiology and management of genital injuries.

In a 10-year period, we have seen and treated 70 patients with trauma to the genitalia. There were 42 penile, 38 scrotal, and 16 testicular injuries. Patients with severe multisystem or extensive genital wounds were successfully managed by early conservative debridement with delayed definitive repair.

Renal cell carcinoma in a patient with situs inversus totalis.

Situs inversus occurs with a frequency of about 1 in 8,000. We present a case of renal cell carcinoma in a patient with situs inversus totalis treated with nephrectomy. Uneventful operative management was attained by adequate preoperative radiologic evaluation.

Evaluation of nocturnal penile tumescence with PotenTest.

PotenTest is a standardized, hygienically packaged, and reliable stamp test, useful for differentiating organic from psychogenic sexual dysfunction. Fifteen potent control patients wore a PotenTest band on three separate nights, and during a selfinduced erection, while PotenTest reliably induced nocturnal erections and penile rigidity. PotenTest and Snap-Gauge were used to evaluate pre- and postoperative potency in twelve cystectomy patients. The result with each test was essentially the same.

The natural history of traumatic branch renal artery injury.

Between 1979 and 1984, 24 patients with traumatic branch renal artery injuries documented by selective angiography were managed at our center. A total of 7 patients (29 per cent) died of nonurological organ system injury during the initial hospitalization. Of the remaining patients 10 were followed for 1 to 5 years after injury. None of these patients required immediate or delayed surgical intervention for complications, while 2 had hypertension after injury that resolved spontaneously by 1 year. The eventual renal function attained by these patients remained normal, the mean creatinine was 0.9 plus or minus 0.1 mg. per dl. and there were no other urological complications. Patients who sustain branch renal artery injuries should be managed nonoperatively unless they exhibit cardiovascular instability. Excision of ischemic parenchyma is indicated only when intractable hypertension associated with increased renin secretion can be identified.

Buckshot colic.

A patient with a shotgun injury to the abdomen and kidney with delayed spontaneous ureteral passage of a shotgun pellet is reported. This rare complication "buckshot colic" usually can be treated nonoperatively. The term "buckshot colic" is clarified.