Pharmacologic profiling reveals lapatinib as a novel antiviral against SARS-CoV-2 in vitro.

The emergence of SARS-CoV-2 virus has resulted in a worldwide pandemic, but effective antiviral therapies are not widely available. To improve treatment options, we conducted a high-throughput screen to uncover compounds that block SARS-CoV-2 infection. A minimally pathogenic human betacoronavirus (OC43) was used to infect physiologically-relevant human pulmonary fibroblasts (MRC5) to facilitate rapid antiviral discovery in a preclinical model. Comprehensive profiling was conducted on more than 600 compounds, with each compound arrayed across 10 dose points. Our screening revealed several FDA-approved agents that can attenuate both OC43 and SARS-CoV-2 viral replication, including lapatinib, doramapimod, and 17-AAG. Importantly, lapatinib inhibited SARS-CoV-2 RNA replication by over 50,000-fold. Further, both lapatinib and doramapimod could be combined with remdesivir to improve antiviral activity in cells. These findings reveal novel therapeutic avenues that could limit SARS-CoV-2 infection.

Comparison of two bioassay techniques for assessing the acute toxicity of pesticides to chironomid larvae (Diptera: Chironomidae).

Two container and substrate combinations were compared to determine which provided optimal survival of larvae of Chironomus tepperi and Glyptotendipes paripes in the absence of toxicants. Unfed final-stage larvae of G. paripes survived significantly (P < 0.05) better in waxed paper cups with sand substrate (92.8% after 3 days) than in glass tubes with a shredded paper substrate (85.3%). Survival of larvae of C. tepperi over the same period did not differ significantly in the 2 systems. Larvae of C. tepperi were bioassayed against 3 insecticides (technical and formulated imidacloprid, chlorpyrifos, and betacypermethrin) with both container and substrate combinations. Median lethal concentration values (24 h) obtained with waxed cups with sand were 1.8 times higher on average than those obtained with glass tubes with shredded paper (range 1.13-2.65 times). To determine the cause of this variability, solid-phase microextraction was used to measure changes in chlorpyrifos availability over time in the 2 bioassay systems. Chlorpyrifos concentrations in the waxed cups and sand system fell from 7.50 to 3.36 microg/liter over 24 h, probably as a consequence of chemical adsorption to the waxed surfaces. Chlorpyrifos concentrations in the glass tubes and paper system remained unchanged over this period. Excluding substrates from the containers had only a minor effect on chlorpyrifos availability. These results demonstrate that the behaviors of both test organisms and toxicants within bioassay systems need to be understood if the data generated with different systems are to be compared. Understanding how toxicant availability is affected by different container and substrate types is particularly important where bioassays are conducted with nominal concentration values rather than analytically determined exposure concentrations.

Effectiveness of spinal manipulative therapy in the treatment of mechanical thoracic spine pain: a pilot randomized clinical trial.

BACKGROUND: To date, no substantiated studies have been performed to investigate the efficacy of spinal manipulative therapy on thoracic spinal syndromes. OBJECTIVE: To investigate the effectiveness of spinal manipulative therapy in the treatment of mechanical thoracic spine pain. STUDY DESIGN: A single-blind, randomized, comparative, controlled pilot study. SETTING: Technikon Natal Chiropractic Clinic in Durban, South Africa. PARTICIPANTS: Thirty subjects selected from the general population (from 16 to 60 years old) were randomly divided into two different treatment groups of 15 each. METHODS: The objective measurements collected were the thoracic spine ranges of motion with the BROM II goniometer and pain threshold with an algometer. The subjective information required completion of the Oswestry Back Pain Disability Index, short-form McGill Pain Questionnaire, and Numerical Pain Rating Scale-101 Questionnaire by the patient. These 3 forms and objective measurements were collected before the first and final treatment and again at the 1-month follow-up consultation. The data gathered were then statistically analyzed with use of a 95% confidence level. The nonparametric Mann-Whitney U test and the Wilcoxon signed-rank test were used to compare intergroup and intragroup data, respectively. This was conducted at the alpha =.05 level of confidence. Further assessment of the data was conducted by use of power analysis. INTERVENTIONS: The treatment group received thoracic spinal manipulation. The placebo group received nonfunctional ultrasound application only. The research project was carried out so that both groups received 6 treatments over a period of 2 to 3 weeks. A 1-month follow-up appointment was scheduled after the final treatment to assess the relative long-term benefits of the two different treatments. RESULTS: Statistically significant results (P < or = .025) were noted for the percentage of pain experienced (Numerical Pain Rating Scale) and for right and left lateral flexion during intergroup comparison after the final treatment. The final treatment results were maintained at the 1-month follow-up consultation; however, there were no further statistically significant results. It was noted that the power was weak, so the probability of committing type II error (falsely accepting the null hypothesis) for the other measurements was high. The intragroup analysis showed statistically significant improvements in the group that received spinal manipulative therapy in both subjective and objective measurements between the first to final treatment and the first treatment to the 1-month follow-up consultation. The placebo group analysis showed a statistically significant improvement in sensory pain only (subjective measurement) between the first treatment and the final treatment. CONCLUSIONS: This pilot study suggests that spinal manipulative therapy has greater benefits than placebo treatment. The sample size was small, therefore the findings of this trial study should not be considered conclusive but rather should be used as a foundation for planning future studies. In further studies a larger sample size will be necessary to identify subtle changes in measurement parameters and to add to the validity of the results.

Constipation and fecal incontinence in the elderly.

Although constipation and fecal incontinence are common symptoms in the elderly, relatively little research has been done to differentiate physiologic changes in rectoanal function resulting from aging and pathologic changes resulting from diseases occurring as patients age. Certain physiologic changes occur in many older patients and may predispose them to the development of constipation or fecal incontinence. These symptoms need the same thoughtful evaluation and management in the elderly as in younger patients. Results of therapy often can be good, leading to alleviation of suffering and the ability to lead a fuller life.

Review article: the therapy of constipation.

Constipation is a common symptom that may be idiopathic or due to various identifiable disease processes. Laxatives are agents that add bulk to intestinal contents, that retain water within the bowel lumen by virtue of osmotic effects, or that stimulate intestinal secretion or motility, thereby increasing the frequency and ease of defecation. Drugs which improve constipation by stimulating gastrointestinal motility by direct actions on the enteric nervous system are under development. Other modalities used to treat constipation include biofeedback and surgery. Laxatives and lavage solutions are also used for colon preparation and evacuation of the bowels after toxic ingestions.

Diarrhea.

Diarrhea is a common symptom of many conditions, and making an accurate diagnosis can be daunting. Research efforts have expanded the differential diagnosis of chronic diarrhea and have provided improved methods for the evaluation and management of patients with diarrheal diseases. This article discusses some of these trends and presents the emerging consensus regarding appropriate pathways of caring for patients with diarrheal diseases.

Microscopic colitis syndrome: lymphocytic colitis and collagenous colitis.

Microscopic colitis is a syndrome consisting of chronic watery diarrhea, a normal or near-normal gross appearance of the colonic lining, and a specific histological picture described as either lymphocytic colitis or collagenous colitis. Since its initial descriptions a quarter of a century ago, microscopic colitis has become a frequent diagnosis in patients with chronic diarrhea. Understanding of the cause and pathogenesis of microscopic colitis remain incomplete, but potentially important clues have been discovered that shed light on predisposing factors. In particular, specific HLA-DQ genotypes may be permissive for the development of microscopic colitis, and suggest a linkage to the pathogenesis of celiac sprue. Although the differential diagnosis of chronic watery diarrhea is broad, the diagnosis of microscopic colitis is straightforward, involving endoscopic inspection of the colonic mucosa and proper pathologic interpretation of biopsy specimens. As the limitations of drugs ordinarily used for other forms of inflammatory bowel disease are being recognized, new approaches, such as the use of bismuth subsalicylate, are being evaluated. The prognosis of patients with microscopic colitis syndrome remains good, and symptomatic improvement can be expected in most patients.

AGA technical review on the evaluation and management of chronic diarrhea.

This literature review and the recommendations therein were prepared for the American Gastroenterological Association Clinical Practice and Practice Economics Committee. The paper was approved by the committee on September 27, 1998.

Double external direct-current shocks for refractory atrial fibrillation.

Encouraged by preliminary data using double external direct-current (DC) shocks in patients with atrial fibrillation refractory to single external DC shocks, we undertook a prospective study of all patients with atrial fibrillation of > 1-month duration using a shock sequence with (1) 1 shock of 200 J anterior-posterior, (2) 1 shock of 360 J anterior-posterior, (3) 1 shock of 360 J apex-anterior, and (4) double shocks with configurations 2 and 3 delivered almost simultaneously by 2 defibrillators. The double shocks appeared to be safe and restored sinus rhythm in approximately 2 of 3 of patients in whom DC cardioversion failed with single shocks.

Clinical pharmacology and use of laxatives and lavage solutions.

Laxatives and lavage solutions are used in the treatment of constipation and toxic ingestion and also for preparation of the colon before endoscopic or surgical procedures. Several different categories of agents are available for use. These include bulking agents, osmotic agents, secretagogues and agents with direct effects on epithelial nerve or smooth muscle cells, and lubricating agents. Each category has different pharmacologic effects, side effects, and clinical indications. This review summarizes current information about these agents.

Secretory diarrhea.

Diarrhea, defined as loose stools, occurs when the intestine does not complete absorption of electrolytes and water from luminal contents. This can happen when a nonabsorbable, osmotically active substance is ingested ("osmotic diarrhea") or when electrolyte absorption is impaired ("secretory diarrhea"). Most cases of acute and chronic diarrhea are due to the latter mechanism. Secretory diarrhea can result from bacterial toxins, reduced absorptive surface area caused by disease or resection, luminal secretagogues (such as bile acids or laxatives), circulating secretagogues (such as various hormones, drugs, and poisons), and medical problems that compromise regulation of intestinal function. Evaluation of patients with secretory diarrhea must be tailored to find the likely causes of this problem. Specific and nonspecific treatment can be valuable.

A comparison of stool characteristics from normal and constipated people.

In people with constipation, it is not known if decreased frequency of defecation is associated with abnormalities in the weight or in the consistency of stools or if the weight or the consistency of stools correlates with the severity of various discomforts associated with bowel movements. In neither normal nor constipated subjects has the consistency of stools been carefully correlated with their relative contents of water and solids. Our aim was to gain insight into these questions. Twenty subjects with idiopathic chronic constipation and 20 age- and sex-matched control subjects were recruited by advertisement. Stools were collected for one week. After each bowel movement, the subject's perception of various discomforts associated with the bowel movement were recorded. The stools were then analyzed. The results and conclusions were as follows: (1) Stool weight per bowel movement was similar in the two groups but stool weight per week was markedly reduced in constipated subjects. (2) Reduced stool weight per week in constipated subjects was due to a nearly proportional reduction in stool water and stool solids output. (3) Using data from both groups, there was a curvilinear correlation between percent insoluble stool solids and stool hardness, as measured by a texture analyzer; hardness increased only slightly as percent insoluble solids increased between 7 and 20%, but hardness increased dramatically when percent insoluble solids exceeded 25%. (4) Only 6% of stools from constipated subjects (2 of 34) had abnormally high values for percent stool solids and physical hardness. (5) In subjects with constipation, the severity of various discomforts associated with bowel movements (such as straining) correlated poorly with the weight or the hardness of stool that was produced by the bowel movement.

Efficacy and toxicity of 2-Chlorodeoxyadenosine (Cladribine)--2 h infusion for 5 days--as first-line treatment for advanced low grade non-Hodgkin's lymphoma.

2-Chlorodeoxyadenosine (Cladribine) is a new purine analogue with high activity in pretreated low grade non-Hodgkin's lymphoma (NHL). To evaluate the efficacy of this drug in untreated patients with advanced NHL, we performed a prospective multicentre trial. Cladribine (0.12 mg/kg) was administered intravenously daily for 5 consecutive days in an out-patient setting. The treatment was repeated every 28 days for four cycles. Included were patients with a histological diagnosis of low grade NHL according to the Kiel classification and stage III or IV disease. Stage II patients were included when radiotherapy had failed. 55 patients were entered into the study. 50 patients were evaluable. The remission rate was 44/50 (88%; 95% confidence interval 82-100%), including complete remissions (CR) in 14 (28%) patients. Only 2 patients showed progression while on Cladribine treatment. The estimated overall survival, and time to treatment failure (TTF) were 85% and 51%, respectively, after a median observation time of 92 weeks. 11 (22%) patients showed grade 3 or 4 toxicity according to the WHO grading. Haematological toxicity was responsible for 86% of the overall toxicity and 100% of grade 3 and 4 toxicity. 7 patients (14%) had an infection, two of which were opportunistic. 12 (24%) patients did not experience any toxicity during the treatment. The results of this study clearly demonstrate the safety and considerable activity of this regimen. Cladribine is very effective even at lower doses than have been used so far.

Glucose-stimulated sodium transport by the human intestine during experimental cholera.

BACKGROUND & AIMS: Net sodium absorption from oral rehydration solution is increased by both glucose-sodium cotransport and solvent drag. The aim of this study was to measure the relative importance of glucose-sodium cotransport and solvent drag in the stimulation of net sodium absorption by oral rehydration solution. METHODS: Total intestinal perfusion was used in normal subjects with and without intrajejunal cholera toxin using three test solutions containing 100 mmol/L sodium and either 100 mmol/L mannitol (control), 100 mmol/L glucose, or no additional solute (hypotonic solution). The increase in sodium absorption greater than control with hypotonic solution represented sodium absorption stimulated by solvent drag; the further increase in sodium absorption induced by glucose, greater than that noted with the hypotonic solution, represented sodium absorption stimulated by cotransport. RESULTS: Without cholera toxin, solvent drag and cotransport promoted sodium absorption at rates of 62 and 33 mmol/h, respectively. With cholera toxin, solvent drag and cotransport promoted sodium absorption at rates of 44 and 71 mmol/h, respectively. CONCLUSIONS: Net sodium absorption caused by cotransport increased more than twofold after exposure of the intestine to cholera toxin (P < 0.003). This could be mediated by increased cotransport, a change in the stoichiometry of cotransport, or an increase in chloride permeability.

Validation of polyethylene glycol 3350 as a poorly absorbable marker for intestinal perfusion studies.

Polyethylene glycol (PEG) has been used as a poorly absorbable marker in intestinal perfusion studies, but there is controversy about the absorbability of PEG, particularly when glucose-sodium cotransport is occurring. Total intestinal perfusion studies were done in five normal humans using three solutions containing 1 g/liter PEG 3350 and designed to produce low rates of water absorption, high rates of water absorption, or high rates of glucose-sodium cotransport. Water absorption rates were calculated by traditional nonabsorbable marker equations and by a novel balance technique in which absorption was taken as the difference between the volumes of solution infused and recovered during steady-state conditions. Effluent PEG recovery was 99 +/- 4%, 109 +/- 2%, and 104 +/- 6% of the amount infused with each solution. Water absorption rates measured by use of PEG concentrations were similar to those calculated by the balance technique (r = 0.99). The complete recovery of PEG confirms the poor absorbability of PEG 3350, and the excellent agreement between techniques validates PEG as a poorly absorbed marker, even when glucose-sodium cotransport is occurring.

Upper gastrointestinal motility disorders and respiratory symptoms.

The association between upper gastrointestinal (GI) motility disorders and respiratory problems is reviewed. Upper GI motility disorders, such as gastroesophageal reflux disease, gastroparesis, and achalasia, have been associated with respiratory problems, including aspiration, airway obstruction, asthma, bronchospasm, chronic cough, and laryngitis. These associations, which had been based solely on clinical observation, have recently been supported by physiologic studies and treatment trials. The association of reflux disease with asthma has the most support. Up to 80% of persons with asthma have evidence of pathologic gastroesophageal reflux, and in several studies antireflux therapy with prokinetic agents, antisecretory drugs, or fundoplication surgery has been found to reduce asthma symptoms and the need for medication in some patients. Reflux has also been associated with chronic cough and laryngitis, and antireflux therapy can reduce respiratory symptoms. Gastroesophageal reflux, gastroparesis, and achalasia are all associated with aspiration. In addition, in rare instances, the megaesophagus associated with achalasia can produce mechanical airway obstruction. Effective therapy for these GI motility disorders can eliminate complicating respiratory problems.

Evidence for a graft-versus-tumor effect in a patient treated with marrow ablative chemotherapy and allogeneic bone marrow transplantation for breast cancer.

Graft-versus-leukemia (GvL) has been shown to be an important immune-mediated antitumor effect in hematologic malignancies. It is still unknown whether such an immunemediated antitumor effect has clinical implications in patients with solid tumors. A 32-year-old woman with inflammatory breast cancer received a bone marrow transplant (BMT) from her HLA-identical sibling. During graft-versus-host disease (GvHD) cytotoxic T lymphocytes were grown and tested in a chromium-release assay against B and T lymphocytes of the patient and donor and against a panel of breast cancer cell lines. Resolution of liver metastases was observed simultaneously with clinical GvHD in the first weeks after transplant. In addition, minor histocompatibility antigen (MiHA)-specific and major histocompatibility complex (MHC) class I antigen-restricted cytotoxic T lymphocytes recognizing breast carcinoma target cells were isolated from the blood of the patient. Pretreatment of such target cells with tumor necrosis factor (TNF)-alpha but not with interferon (IFN)-alpha or IFN-gamma increased susceptibility of these cells to lysis by cytotoxic T lymphocytes. Clinical course and in vitro results suggest that a graft-versus-tumor (GvT) effect might exist after allogeneic BMT for breast cancer. However, clinical experience on a larger scale would be required to determine the clinical efficacy of GvT effects in patients with solid tumors.