AU2016101768B4 - Treatment for Irritable Bowel Syndrome - Google Patents

Abstract

Methods of treating irritable bowel syndrome, irritable bowel disease and/or symptoms similar to those observed then using high doses of a molybdenum(II) glycinate chelate supplement, a low sulfite diet and a soy protein isolate supplement.

Description

1 2016101768 06 Oct 2016

TREATMENT FOR IRRITABLE BOWEL SYNDROME TECHNICAL FIELD

The Discipline of Medicine LIST OF ABBREVIATIONS CNS, central nervous system; DASS, Depression Anxiety Stress Scales; FSS, functional somatic syndrome; GIT, gastrointestinal track; GRAS, generally regarded as safe; IBD, inflammatory bowel disease; IBS, irritable bowel syndrome; IBS-C, constipation-predominant subtype of irritable bowel syndrome; IBS-D, diarrhea-predominant subtype of irritable bowel syndrome; IBS-PI, post-infectious subtype of irritable bowel syndrome; IBS-M, mixed subtype of irritable bowel syndrome; IBS-U, unclassified subtype of irritable bowel syndrome; QIDS, Quick Inventory of Depressive Symptomatology; SDH, sulfite dehydrogenase; SRB, sulfate-reducing bacteria; UL, Tolerable Upper Intake Level; VSC, volatile sulfur compound

TERMINOLOGY

To improve the readability of this article, the following new terms will be introduced and used in this article:

The term "sulfur-handling disorder" will refer to a disorder with the metabolism of sulfur in humans. For example, a hypersensitivity to sulfites.

The term “Agent” will refer to the administration of a medical treatment. The medical treatment could be: a drug, a supplement, a diet modification or a routine.

The term "SDH-lacking bacteria" will refer to intestinal symbiotic bacteria in which the activity of sulfite dehydrogenase enzymes is low.

The term "SDH-containing bacteria" will refer to intestinal symbiotic bacteria in which the activity of sulfite dehydrogenase enzymes is high.

The term “component” will refer to something that adversely changes a body system, e.g. lactose can trigger diarrhea, in a lactose intolerance individual. 2 2016101768 06 Oct 2016

BACKGROUND ART

[0001] Irritable bowel syndrome (IBS) is a functional somatic syndromes (FSSs). FSSs are health disorders that lack a medical explanation. They are, therefore, explained by a pattern of symptoms observed in affected individuals. Chronic fatigue syndrome (CFS), functional dyspepsia and fibromyalgia are other examples of the different types of FSSs (Wessely et al., 1999; Talley, et al., 2005). A substantial overlap of symptoms exists between the different types and the similarities between them outweigh the differences. As a consequence, a medical treatment for one type of the FSSs could be used to treat another (Warren and Clauw, 2012; Wessely et al., 1999).

[0002] IBS is typically a disorder affecting the function of the nerves and muscles of the small and large intestines (Owyang, 2009). The syndrome was first recognized in 1892 and given the name "mucous colitis” (Osier, 1892). Since then various names have been applied to the syndrome, including “spastic colon (Ryle, 1928), “irritable colon” (Jordan and Kiefer, 1929) and “irritable colon syndrome” (Chaudhary and Truelove, 1962). The current name, irritable bowel syndrome, was applied in 1966 (DeLor, 1967).

[0003] The syndrome can be divided into four main subtypes under the Rome III criteria. They are: the constipation-predominant subtype of IBS (IBS-C); the diarrhea-predominant subtype of IBS (IBS-D); the mixed subtype of IBS (IBS-M), i.e. alternating between diarrhea and constipation; and the unclassified subtype of IBS (IBS-U) (Owyang, 2009). There are also subtypes of IBS based on different sets of criteria. For example, if IBS developing after an episode of acute gastroenteritis, the term post-infectious subtype of IBS (IBS-PI) can be used (Beatty et al., 2014).

[0004] In the last 15 years, there has been increasing focus on a subtle inflammatory change in the gut immune system in IBS-PI. Although the evidence is accumulating in favor of such a change, identifying the underlining immune mechanism is challenging current scientific techniques (Camilleri et al., 2012).

[0005] To date there is evidence of increased activity in mast cells, T lymphocytes and enteroendocrine cells in the intestinal mucosa of individuals with post-infectious subtype.

The inflammatory mediators released by these immune cells could be at the origin of the pathological changes seen in this subtype. There is a breakdown of the intestinal epithelial barrier, visceral hypersensitivity and heightened enteric nervous system signaling in the post-infectious subtype. These changes are probably adversely stimulating the gut neurons, 3 2016101768 06 Oct 2016 causing spastic smooth muscle contractions. Studies have found increased numbers of activated mast cells throughout the intestinal tract of this subtype, which show, slight, but statistically significant correlation to visceral hypersensitivity (Camilleri et al., 2012).

[0006] Tryptase, a specific protease released by mast cells, seems particularly relevant to the intestinal epithelial barrier. Tryptase can activate Protease-activated Receptor 2 on epithelial cells resulting in modulation of tight junction macromolecular assemblies. These junctions join the membranes of adjacent cells that line the bowel walls and form a virtually impermeable barrier to fluids. The breakdown of these junctions allows an abnormal flow of fluids between the cells that line the bowel walls. This results in translocation of macromolecules across the cell barrier, which in turn, up-regulates the intestinal immune system (Camilleri et al., 2012). It has been suggested that IBS-PI and the subtype it normally manifests itself as, IBS-D, may be a low-grade inflammatory bowel disease (IBD; Bercik et al., 2005).

[0007] Symptoms and the severity of the symptoms on the sufferer can vary considerably between affected individuals. The typical symptoms are: abdominal pain; abdominal discomfort and/or bloating; and irregular bowel movements. Additional symptoms may include: flatulence, an urgency to pass stools, difficulty passing stools, a sensation of incomplete bowel evacuation, agitation and psychological symptoms (Owyang, 2009). The syndrome can manifest itself in a mild form, i.e. the sufferer may describe it as an “inconvenience”, to as debilitating as IBD or bowel cancer. Symptoms are generally exacerbated by: foods, beverages and stress; and in women during the premenstrual and menstrual phases of her menstrual cycle (Owyang, 2009).

[0008] Many sufferers have psychological symptoms. A cognition dysfunction like a clouding of consciousness ("brain fog") is commonly seen in IBS (Kennedy et al., 2012). High levels of depression, anxieties and phobias are also seen in this group (Owyang, 2009). There is evidence of excessive biochemical signaling taking place between the neurons in the gut and the central nervous system (CNS). The term "gut-brain axis" is used to described this phenomenon (Kennedy et al., 2012).

[0009] Sufferers tend to have a certain psychological profile and may suffer from psychological distresses. People with IBS tend to be perfectionists, tend to want to please others. There is a high incidence of physical and emotional abuse in this group (Creed, 2007). 4 2016101768 06 Oct 2016

TECHNICAL PROBLEM

[0010] IBS is the most gastrointestinal disorder encountered in clinics. Depending on the criteria used, the prevalence of IBS in a given population, range from 9% to 22%. It accounts for 12% of the visits to non-specialized clinics and 28% of the referrals to gastroenterologists. As stated in the Background Art section of this article, the syndrome can be extremely debilitating. The pathogenesis and pathophysiology of the syndrome is complex and is poorly understood. Due to its high prevalence, for some sufferers its debilitating nature and its complexity, the cost of the syndrome to society is substantial (Owyang, 2009).

[0011] Scientists believe that a complex combination of biological factors, are responsible for the symptoms seen in IBS. The factors include: environmental, genetics, hormones, gut microbiota, past acute gut infections, psychological profile and psychological distress (Owyang, 2009). This complexity may be why the syndrome is difficult to treat. Current treatments for IBS have limited efficacy and tolerability and, consequently, there is a great need to find new medical treatments for sufferers (Owyang, 2009).

SOLUTION TO PROBLEM

[0012] The new medical treatment paradigm or model described in this article, addresses three new and probably important components in the pathophysiology of IBS or an IBS subgroup. For this article the term “component” will refer to something that adversely changes a body system, e.g. lactose can trigger diarrhea in lactose intolerant individuals. These three components have not been dealt with by current treatments for IBS. The components are: (1) a new type of disorder with the metabolism of sulfur in humans, (2) a pronounced startle reflex response to auditory or tactile stimuli and (3) gut microbes placing stress on the defective biological system involved in the sulfur-handling disorder or directly causing the disorder. The term "sulfur-handling disorder" will be use from here on to describe this new type of disorder with the metabolism of sulfur.

DESCRIPTION OF EMBODIMENTS

[0013] The invention described herein, is a new medical treatment paradigm for IBS. The treatment consists of: (1) diet changes, (2) dietary supplementations, (3) drugs and (4) other routines. For this article the term “agent” will refer to the administration of a medical treatment. The medical treatment could be: a drug, a supplement, a diet modification or a 5 2016101768 06 Oct 2016 routine. These agents aim to address the three components described in the Technical Problem section of this article.

[0014] The treatment may consist of the following steps and agents:

Step 1:

Agent 1: Benzodiazepine drug

Protocol: Administer continually or intermittently; e.g. administer intermittently by administering for four consecutive days and then stop for the other three consecutive days of each week.

Note: The patient should not take stimulate drugs, e.g. caffeine Agent 2: Removal of sulfites from diet; i.e. remove foods, beverages, supplements, probiotics containing yeast (e.g. Saccharomyces boulardiicontaining probiotics) and if possible, medications, from diet that contain or could contain a concentration of sulfites > 10 ppm

Step 2:

Agent 3: Relaxation routine (e.g. mindfulness)

Protocol: Daily and when stressed

Notes: The benzodiazepine drug and the relaxation routine aims to treat the pronounced startle reflex response to auditory or tactile stimuli. The 4 d on and 3 d off protocol for administrating benzodiazepines aims at reducing the likelihood of the development of a tolerance to benzodiazepine.

Agent 4: Oral cobalamin (vitamin B12) supplement Dose: 1000 < 5000 μg

Protocol: Administer to manage visual, psychological and bowel symptoms Notes: Monitor vision for improvements. Improvements may be an early sign that the cobalamin supplementation is helpful.

Step 3:

Agent 5: Oral molybdenum (Mo) supplement Dose: 1000 ^ 3000 μg/d for adults

Protocol: Administer to manage visual, psychological and bowel symptoms. Also administer after episodes of diarrhea. Divide the dose into two or more smaller doses throughout the day. Do not take Mo with copper (Cu) or sulfate compounds. 2016101768 06 Oct 2016 6

Notes: Monitor vision for improvements. Improvements may be an early sign that the cobalamin supplementation is helpful. If the patient has not responded after implementing this step, the treatment probably will not help the patient.

Agent 6: Oral Cu supplement

Dose: Recommended daily intake (RDI), i.e. 1.7 mg/d for men (National Health and Medical Research Council, 2006)

Protocol: Administer if Mo intake is greater than the tolerable upper intake levels (UL) i.e. > 2000 μg/d for adults (National Health and Medical Research Council, 2006). Do not take with Mo.

Notes: Taken to prevent a Cu deficiency induced by intakes of Mo > UL. Mo and Cu compete for the same ligands (Barceloux, 1999) and, therefore, should not be taken together.

Step 4:

Agent 7: Oral sulfate supplement

Dose: Not yet determined

Protocol: Intermittently administer to manage psychological symptoms. Do not take with Mo.

Agent 8: Oral protein isolate supplement

Doses: Administer to produce flatus with the characteristic odor attributed to foul flatus.

Protocol: Intermittently administer to manage psychological and bowel symptoms. If the patient is taking a digestive aid, e.g. pancrelipase, stop the aid when the patients starts the protein isolate.

Notes: The Mo, cobalamin, sulfate and protein isolate supplements; and the low sulfite diet; aims to treat the sulfur-handling disorder. Mo and cobalamin also seem to be very effective at stopping episodes of diarrhea. The patient may note relief within 15 min after taking 1000 μο of Mo or 1000 to 5000 μο of cobalamin supplements. Mo is also taken as well as the protein isolate, to stimulate the growth of gut microbes that may compete with the gut microbes that are placing stress on the body system involved in the sulfur handling disorder. Some malabsorption of nutrients seems to be important in the managing bowel symptoms. 7 2016101768 06 Oct 2016

The key agents for the pronounced startle reflex response are probably: (1) a benzodiazepine and (2) a relaxation routine. The key agents for IBS are: (1) a low sulfite diet, (2) cobalamin, (3) Mo and (4) a protein isolate.

[0015] The patient will need to self-evaluate his or her symptoms during the treatment and probably make changes to the dose and the timing of the doses. For example, belching and a full gut feeling could be a symptom that transit time has slowed and the patient should take a laxative, e.g. for adults > 5.00 mmol of sulfate (Christl et al., 1992). Another example may be what is best described as a “burning sensation” while passing stools. In this case the patient may need to increase intake of Mo and protein.

[0016] Symptoms used to monitor treatment can be subtle and can even manipulate the mind of the patient. A subtle type of symptom indicating an episode of diarrhea is about to begin, may be a lack of energy when trying to walk up a flight of stairs. A mind manipulating symptom, again indicating an episode of diarrhea, can be a sudden improvement in mood and confidence. This symptom can have devastating consequences, as it can lead to: confusion, frustration and depression; and the patient concluding that the treatment will never work. The patient may need counselling if he or she has this mind manipulating symptom.

EXAMPLE

1. DETAILS, SYMPTOMS, CLINICAL DIAGNOSES AND TREATMENTS

1.1 DETAILS AND SYMPTOMS

[0017] The subject of this example is a middle aged Caucasian man. His clinical symptoms fit with the Rome III Diagnostic Criteria and are best described by the diarrhea-predominant subtype. The subject tends to be a perfectionist and tends to want to please others. This man developed this syndrome after an episode of acute gastroenteritis. He notes that the gastroenteritis was not severe, i.e. he was not hospitalized and it resolved in what would be considered a normal duration for an acute gut infection. The only abnormality the subject can recall is that his urine was dark-colored for a few days. The pathogenesis described by this subjects fits with the post-infectious subtype.

[0018] The subject notes that he finds that the syndrome can be extremely debilitating. His symptoms include: 8 2016101768 06 Oct 2016 • Flatulence, • Abdominal pain associated with loose stools, • Anal rash and a burning sensation around the anus when passing stools, • Fatigue, • Sensitivities to multiple chemical, • Muscle pain, • Joint pain, • Agitation, • Dizziness, • Brain fog, • Fleadaches, • Pronounced startle reflex response to auditory or tactile stimuli; • Depression and • Anxieties.

The most troubling symptom the subject noted, was diarrhea. The diarrhea occurs almost daily when he is not taking medication to control it. When off medications he rated his stools on the Bristol Stool Form Scale (Lewis and Heaton, 1997) from four to seven. The diarrhea tends to be triggered by foods, beverages or psychological stress.

[0019] An IBS family cluster exists in his family. His father has IBS-D and he is from a large family with eight other siblings and most of them have IBS. Although, some have the constipation-predominant subtype. His sister developed CFS around the same time that the subject developed IBS.

1.2 ABNORMAL CLINICAL TEST RESULTS

[0020] The subject has had a battery of biochemical tests. The tests identified "borderline deficiencies" for: • Serum vitamins A, D and E; • Serum Ca; and • Plasma essential fatty acids.

The tests also identified that his: • Serum cholesterol was abnormally low, • Serum free testosterone could be abnormally low and • Urinary free cortisol was mildly elevated. 9 2016101768 06 Oct 2016 [0021] A Three-day Fecal Fat Test on fecal specimens from the subject, identified that his fecal fat level can be elevated. Fecal fat levels were elevated in the initial Three-day Fecal Fat Test. A subsequent test was normal. Despite finding that his fecal fat levels can be normal at times, the subject was diagnosed with steatorrhea.

[0022] Food elimination and challenge testing identified that dairy, wheat and salicylates triggered symptoms. Dairy and salicylates triggered diarrhea. Dairy also induced brain fog; and nasal and throat mucus. Wheat induced agitation and a general feeling of being unwell. Consequently, the subject was put on a low dairy and wheat diet; and a "moderate approach" diet for salicylates (Swain et al., 2011).

[0023] The subject has also had a battery of radiography tests. Barium (Ba) Meal Testing found that transit of Ba through his gastrointestinal track (GIT) ranged from normal to rapid. The Ba could be well through the large bowel in only 25 min. The x-ray film showed that the small bowel was dilated with a lumen diameter of 30 mm. Flence, the subject was diagnosed with malabsorption of nutrients.

[0024] Macroscopic and microscopic examinations of the GIT of the subject were unable to identify an abnormality that could be responsible for his IBS symptoms. Macroscopic examinations of his upper GIT using gastroscopic methods found an esophageal and a duodenal ulcer. Both ulcers resolved after medical intervention. Macroscopic examination of his lower GIT using colonoscopic methods did not find an abnormality. Histological examinations and biochemical testing of the biopsy specimens taken from the GIT were also normal.

[0025] The subject has also been tested for psychological disorders. His Depression Anxiety Stress Scales (DASS) ratings showed that he had sporadic periods of mild depression, moderate anxieties and severe stress. His Quick Inventory of Depressive Symptomatology (QIDS) rating showed that he had mild depression.

1.3 MEDICATIONS AND SUPPLEMENTATIONS

[0026] The subject was taking a battery of drugs to manage his symptoms. Fie was taking at least 4 mg of loperamide almost every week. The loperamide was used to stop the episodes of diarrhea. Fie has been taking this drug for about 15 y to manage this symptom. The subject also routinely took at least 2 g of cholestyramine daily, for the diarrhea; and 100 or 150 mg of amitriptyline hydrochloride and 5 to 20 mg of dexamphetamine sulfate daily for the 2016101768 06 Oct 2016 10 psychological symptoms. He also took 5 mg of diazepam daily or for four consecutive days of each week, for the psychological symptoms.

[0027] The subject was also taking a battery of supplements, again to manage his symptoms. He took a multivitamin and mineral supplement ("Blackmores Sustained Release Multi + Antioxidants", Blackmores Ltd., Warriewood, New South Wales, Australia) and 14 mL of medium chain triglycerides oil (Nutricia Australia Pty. Ltd., Talavera Corporate Centre, Level 4, Building D, 12-24 Talavera Rd., Macquarie Park, New South Wales, Australia) each day, because of malabsorption. The subject also took about 24 g of psyllium husk daily, because he found that soluble fiber helps to form a more consistent stool.

1.4 MOLYBDENUM SUPPLEMENTATION

[0028] In May 2015, the subject added a Mo supplement to his diet and reported a sudden overall improvement in his health a few days later. The most substantial beneficial effect was that he no longer needed to take loperamide to stop the episodes of diarrhea. After Mo supplementation he rated his stools on the Bristol Stool Form Scale as four. He also reported that he had: • Diminished need to go again shortly after a completed evacuation; • More energy; • Better cognition; • Better mood and confidence; • Less agitation; • Clearer hearing and voice; • Better ability to focus on near objects; • Less episodes of headaches and dizziness; • Better color vision at night; and • Better perception of the depth of distant objects.

His DASS rating may had improved (Table 1) but his QIDS rating was unchanged. 2016101768 06 Oct 2016 11 TABLE 1. DASS Ratings Before and After Mo Supplementation

Scale Rating Before Mo After Mo January 2015 April 2015 July 2015 October 2015 Depression Mild Normal Mild Normal Anxiety Moderate Moderate Mild Mild Stress Severe Severe Moderate Moderate [0029] The Mo supplement used was Mo(ll) glycinate chelate (Thorne Research, Inc., Drover, Idaho, U.S.A.) and the doses were 500 to 3000 μg/d. He also notes that a 1000 or 2000 μg dose of Mo taken when he feels the need to go to the toilet again soon after a completed evacuation, diminishes the need to go again. His blood Mo was also measured after supplementation and Mo could not be detected in the specimen, i.e. < 0.02 μΓηοΙ/L and was in the normal range (QML Pathology, 11 Riverview Place, Metroplex on Gateway, Murarrie, Brisbane, Australia). Finally, the Blackmores Sustained Release Multi + Antioxidants, vitamin and mineral supplement does not list Mo as an ingredient.

[0030] The subject did, however, note a puzzling symptom that coincided with the improvements in his health, after Mo supplementation. He says that a few days after adding Mo to his diet, he produced large volumes of flatus; and his flatulence and stools had a foul odor. He describes the odor as being similar to the odor emitted from raw sewage. This man says that he had periods with and without the foul odor. He says that during the periods when his flatulence and stools had a foul odor, his overall health was better. This includes bowel symptoms. 12 2016101768 06 Oct 2016

2 PILOT TRIALS

2.1 SULFITE ELIMINATION AND CHALLENGE PILOT TRIAL

[0031] The element Mo supported by an organic molecule, is the coenzyme of biological metalloenzymes involved in carbon, sulfur and nitrogen metabolism. Of the five Mo-containing enzymes in humans, only the sulfite oxidase enzyme is crucial for health (Sardesai, 1993; Wahl et al., 2010). Sulfite oxidase is a very efficient enzyme, that oxidizes sulfite anions (SO32') to sulfate anions (SO42"; Feng, Tollin et al., 2007). Defects with this enzyme in humans result in systemic sulfite toxicity (Sardesai, 1993).

[0032] Sulfites include the gas sulfur dioxide (S02), sulfurous acid (H2SO3) and inorganic sulfite salts. Examples of inorganic sulfite salts are: sodium sulfite, sodium bisulfite and sodium metabisulfite (Taylor et al., 2014). Sulfite anions adversely reacts with biological molecules and can cause metabolic disorders in organisms. The mechanism by which sulfites react with biological molecules is not fully understood, but it is thought it interact with disulfide bonds of macromolecular molecules (Gunnison and Jacobsen, 1987).

[0033] Human exposure to sulfites can be the result of environmental or endogenous agents. Sulfites can be added to foods, beverages and medications to extend the shelf life of these products. Sulfites can also occur naturally in some foods and beverages as a result of fermentation (Taylor et al., 2014). A sulfite anion is an intermediate anion in the catabolism of sulfur-containing amino acids (i.e. L-cysteine, L-methionine and taurine (Stipanuk and Ueki, 2011)), sulfolipids and other "sulfur-containing metabolic waste" (Blachier et al., 2010). Sulfites can also be produce by microbes that inhabit humans in a symbiotic relationship. Almost all yeast and some bacteria that inhabits humans rely on its host to detoxify its sulfite waste (Zhang and Gladyshev, 2008). Therefore, the subject may have a metabolic problem with the handling of sulfites.

[0034] A sulfite elimination and challenge pilot trial was carried out with the aim of confirming the hypothesis that the subject has a metabolic problem with the handling of sulfites. The trial design used was the “Elimination Diet - Food Challenge Protocols” published by the Royal Prince Alfred Hospital, Allergy Unit (9-11 Layton St., Camperdown, Sydney, N.S.W., Australia). The subject removed foods and beverages from his diet that could contain a concentration of sulfites > 10 ppm (Taylor et al., 2014). The foods and beverages he removed were: 13 2016101768 06 Oct 2016 • Dried fruit and vegetables; • Processed lemon juice; • Canned fish; and • Fresh crustaceans.

The subject continued to take Mo throughout this trial.

[0035] The subject noted that after removing these foods and beverages, that his overall health improved. He noticed that he had: • More energy; • Better cognition; • Better mood and confidence; • Less agitation; • Less episodes of headaches and dizziness; and the • Odor of his flatulence and stools improved.

He did not notice a change in the form of his stools.

[0036] The challenge was performed after 3 wk on the elimination diet and after five consecutive days with milder symptoms. The food chosen for the challenge was preserved dried pears without skins (“Fruit Salad”, Angas Park Fruit Company, Sunbeam Ave., Irymple, Victoria, Australia). The product listed sulfur dioxide (220) and potassium sulfite (224) as preservatives. The challenge dose was 30 g. This is about 1/3 of the dose recommended by Elimination Diet - Food Challenge Protocols.

[0037] The subject suffered from gut rumbling, dizziness, headaches and numbness after the challenge. About 5 min after ingesting the dried pears, he notes gut rumbling. About 25 min into the challenge he noted dizziness, headaches and numbness of the left-hand side of his face. The following day his stool was slightly looser with a rating of five on the Bristol Stool Scale. The trial was stopped at this stage, because, it was decided that the subject should have medical supervision if challenged with the higher 100 g dose.

2.2 SULFATE COMPOUND SUPPLEMENTATION ABAB PILOT TRIAL

[0038] Christl and coworkers (1992) have shown that the ingestion of a sodium sulfate (Na2S04) supplement can increase the production of hydrogen sulfide (H2S) gas in the large intestine of humans. They then concluded that Na2S04 stimulates the growth of sulfate-reducing bacteria (SRB) in the large intestine. H2S gas has a foul "rotten eggs" odor (Suarez 14 2016101768 06 Oct 2016 et al., 1998; Tangerman, 2009) and is the main gas that gives raw sewage its characteristic foul odor attributed to it.

[0039] Given these findings by Christl and coworkers (1992) an ABAB pilot trial was carried-out on the subject, using an oral Na2S04 supplement as the "treatment". The trial aim was to identify if the foul odor after Mo supplementation could be reproduced by ingesting Na2S04.

[0040] The trial reported a worsening of flatus odor during the treatment phases. There were similarities between the treatment phases of this trial and the symptoms observed a few days after Mo supplementation (Table 2). Despite this, the subject notes that the amount of flatus was less and the odor of his flatus and stools were not as strong, when compared to that observed a few days after Mo supplementation. He says that he did not have ratings to choose from to report these observations. TABLE 2. Results of the Sodium Sulfate Supplementation ABAB Pilot Trial

Criteria Rating or Description Baseline Treatment Baseline Treatment Day 1 - 3 Day 4 - 6 Day 7 -9 Day 10-12 Flatus frequency Very high Very high Very high Very high Flatus odor Sweet Foul Nil Foul Stool frequencya 0 1 1 1 Stool odor N.A. Foul Fecal odor Foul Stool form b N.A. 4 4 4 a Number of bowel movements/d b Grade on the Bristol Stool Form Scale [0041] The subject noted an overall improvement in this health during the treatment phases. He recorded improvements in energy, brain fog and agitation; and an extreme improvement in mood and confidence. He had sporadic periods during the treatment phases that lasted for about 2 h, when his mood and confidence were the best that it has been since developing IBS. The flatus frequency and odor recordings for the B phases were done during these periods; and selecting another time to record results would have resulted in different ratings. 2016101768 06 Oct 2016 15

2.3 SULFUR-CONTAINING AMINO ACID SUPPLEMENTATION ABAB PILOT TRIAL

[0042] Magee and colleagues (2000) have shown that fecal sulfide concentration correlates (r2 = 0.56, P < 0.001) with protein intake in humans. When a large amount of protein is ingested, volatile sulfur compounds (VSCs) can be produced in the ileum and the colon. VSCs are produced, because there is a chance that proteins may only be partially digested in the duodenum, not absorbed in the duodenum or jejunum and may reach the ileum or colon where it can be digested and fermented by anaerobic bacteria. Species from the Bacteroides, Propionibacterium, Streptococcus, Clostridium, Bacillus and Staphylococcus genera ferment amino acids in the ileum or colon (Macfarlane et al., 1986). To ferment amino acids, species from these genera, must first secrete proteases to fully digest the proteins or polypeptides into amino acids. Once digested the fermentation of sulfur-containing amino acids; L-cysteine and L-methionine by these species; produce H2S and methanethiol (CH3SH) gases; respectively (Magee et al., 2000). Anaerobic bacteria in the ileum and colon of the subject may be fermenting sulfur-containing amino acids and the foul gases may be predominately a mixture of H2S and CH3SH gases.

[0043] To test the hypothesis that anaerobic bacteria are producing the foul gases, an ABAB pilot trial was carried-out on the subject, using a soy protein isolate supplement as the treatment. The aim of the trial was to reproduce the foul odor observed a few days after Mo supplementation, during the B phases.

[0044] The trial recorded that the bowel symptoms during the B phases of the trial, were similar to the odor observed a few days after Mo supplementation. The frequency and the flatus odor worsened during the B phases (Table 3). As with the Sodium Sulfate Supplementation ABAB Pilot Trial, the periods when the odor of his flatus worsened were sporadic and lasted for about 2 h. He also noted similar improvements in his health, to that observed on the Sodium Sulfate Supplementation ABAB Pilot Trial. That is, he says his energy, brain fog and agitation improved; and that he had an extreme improvement in mood and confidence. 16 TABLE 3. Results of the Sulfur-Containing Amino Acid Supplementation ABAB Pilot Trial

Criteria Rating or Description Baseline Treatment Baseline Treatment Day 1 - 3 Day 4 - 6 Day 7 -9 Day 10-12 Flatus frequency Normal Very high High Very high Flatus odor Nil Foul Nil Foul Stool frequencya 0 1 0 0 Stool odor N.A. Foul N.A. N.A. Stool form b N.A. 4 N.A. N.A. 2016101768 06 Oct 2016 a Number of bowel movements/d b Grade on the Bristol Stool Form Scale

2.4 COBALAMIN SUPPLEMENTATION PILOT TRIAL

[0045] Ji (1993) has shown that aquocobalamin can oxidize sulfite anions to sulfate anions, in vitro. Furthermore, Anibarro and colleagues (1992) showed that abnormally high oral intakes of cyanocobalamin can prevent asthma in sulfite hypersensitive asthmatics, when challenge with an oral sulfite compound. Aquocobalamin and cyanocobalamin are different form of cobalamin (Vitamin B12).

[0046] The absorption of cyanocobalamin in the small intestine decreases as oral intake increases. This is probably why effects have not been associated with excess intake of cyanocobalamin (National Health and Medical Research Council, 2006). Therefore, if sulfites were accumulating in the human bowels, an abnormally high intake of cyanocobalamin could be used as a treatment. Consequently, it was hypothesized that abnormally high intake of cyanocobalamin would decrease the concentration of sulfite anions in stools.

[0047] To test the hypothesis that a physiologically abnormally high intake of cyanocobalamin would decrease the concentration of sulfite anions in his stools, a cyanocobalamin supplement was added to his diet and his stools tested for sulfites. The supplement used was "High Strength Vitamin B12" (Sanofi Consumer Healthcare, 87 Yarraman Place, Virginia, Queensland, Australia). He took single oral doses of cyanocobalamin ranging from 1000 to 5000 pg and a total daily dose of 6000 to 15000 pg. 17 2016101768 06 Oct 2016

The concentration of sulfite anions in stool slurries made from stools of the subject were measured before and while taking cyanocobalamin. His blood vitamin B12 concentration was also measured (Mater Pathology, Level 6, Mater Hospital Brisbane, Raymond Terrace,

South Brisbane, Australia). The subject continued to take Mo and consume a low sulfite diet throughout this pilot trial.

[0048] The subject noted improvements in his overall health while on cyanocobalamin supplementation. His stools were firmer with a rating from two to four on the Bristol Stool Form Scale. This man noted that his flatus and stools did not have a foul odor during the trial. The subject was able to stop cholestyramine after supplementation. Before the cobalamin supplementation, attempts to stop cholestyramine resulted in loose stools a few days later, with a rating of five or six on the Bristol Stool Form Scale. The subject also notes a gradual improvement in vision over the 1 mo treatment period of the trial. There were marked improvements in depth perception of distant objects and in color vision at night. His blood vitamin B12 concentration was > 1476 pmol/L. The normal range is 120 to 600 pmol/L (Mater Pathology, Level 6, Mater Hospital Brisbane, Raymond Terrace, South Brisbane, Australia).

[0049] The stool slurry experiment found that the concentration of sulfite anions decreased after cyanocobalamin supplementation. Before cobalamin supplementation, the concentration of sulfite anions in stool slurries made from stools of the subject were ^ 10 and < 50 ppm. After supplementation, sulfite anions could not be detected in a stool slurry. That is the concentration of sulfite anions was < 10 ppm.

2.5 BENZODIAZEPINE ABAB PILOT TRIAL

[0050] As stated in the Details and Symptoms subsection of this article, the subject has a pronounced startle reflex response to auditory or tactile stimuli. He describes the worse cases of this response, is like having an electric shock, in a mild form, that is felt in every skeletal muscle of his body. This symptom has not responded to any of the agents used so far in this case report. Benzodiazepines have been successfully used to treat this symptom (Lenz and Ptacek, 2004). Therefore, a pilot trial was performed on the subject again with a ABAB design, using diazepam as the treatment, to determine if the benzodiazepines improved this symptom. The trial reported pronounced responses during the A phases which did not occur during the B phases. 18 2016101768 06 Oct 2016

3 DISCUSSION

[0051] In this case report a pattern has been found between sulfur and the health of the subject. For example, he notes that his health improved during the elimination phase and he had symptoms during the challenge phase, of a sulfite elimination and challenge trial. The element sulfur links what seems to be unrelated observations after medical treatments in this man. The observations are: (1) beneficial effects and symptoms after Mo glycinate chelate supplementation; (2) beneficial effects on a low sulfite diet and symptoms after a sulfite challenge; (3) beneficial effects and symptoms after Na2S04 supplementation; (4) beneficial effects and symptoms after protein isolate supplementation; (5) beneficial effects of cyanocobalamin supplementation; and (6) a decrease in the concentration of sulfites in a stool slurry after cyanocobalamin supplementation.

[0052] The pilot data generated in this case report, has led to the discovery of a new type of metabolic disorder with the handling of sulfur in humans. This sulfur-handling disorder may be a major trigger of diarrhea and the other symptoms seen in this man.

[0053] Cyanocobalamin may be oxidizing a toxic sulfur compound in this subject. The toxic sulfur compound being oxidized is probably sulfite anions and the oxidized compound is probably sulfate anions. As stated earlier, the human body decreases the absorption of cobalamin when intake is high. Therefore; detecting sulfite anions in a stool slurry before cobalamin supplementation and being unable to detect it after cobalamin supplementation; supports the hypothesis that physiologically abnormally high intakes of cyanocobalamin is oxidizing sulfites in his bowels.

[0054] Although the vision improvements do not fit with the hypothesis, the finding does seem plausible. This is because, systemic sulfite toxicity is known to cause vision impairments (Abumrad et al., 1981; Tan et al., 2005) and his blood vitamin Bi2 was high.

This indicates that a considerable amount of the ingested cobalamin was absorbed from his bowels. These finding suggests that the subject probably suffered from a systemic accumulation of sulfites in this tissues.

[0055] The subject of this case report may suffer from a low activity of sulfite oxidase enzymes and this could be causing the accumulation of sulfites in his tissues. Given that most of the foods and beverages that could contain a concentration of sulfite anions of > 10 ppm have been removed from the diet of this man, the sulfite anions are probably being produced endogenously. As stated earlier in this example, a sulfite anion is an intermediate 19 2016101768 06 Oct 2016 anion in the catabolism of sulfur-containing metabolic waste (Blachier et al., 2010; Stipanuk and Ueki, 2011). This catabolic process generates large quantities of sulfite anions (Stipanuk and Ueki, 2011). If the activity of human sulfite oxidases is low, the catabolism of sulfur-containing metabolic waste would reach a "bottleneck" at the sulfite to sulfate anion oxidation step. This causes an accumulation of sulfites in the tissues and systemic sulfite toxicity (Abumrad et al., 1981). Therefore, systemic sulfite toxicity could be yet another component in the pathogenesis of IBS.

[0056] The coenzyme of the sulfite oxidase enzyme consists of a backbone molecule, molybdopterin and a cofactor, a Mo atom (Feng et al., 2007). In most eukaryotes molybdopterin is synthesized in the mitochondria and cytosol of cells. Organisms take up Mo in its oxyanion form, molybdate (Mo042~). Molybdate is transported by membrane transporter proteins into the cytosol of cells and an enzyme inserts Mo into molybdopterin.

[0057] Four proteins have been identified that handle molybdate in eukaryotic cells. There are three membrane transporter proteins; i.e. Mot1, Mot2 and SFIST1 and one enzyme; Mo-insertase. It is possible that Mo may regulate eukaryotic Mo-membrane transporter genes. This because, prokaryotic operons containing Mo-membrane transporter genes can be regulated by the availability of Mo (McNicholas et al., 1997). Despite this, there is considerable amount of evidence that genetic errors in Mo-insertase genes in eukaryotes, can be restored or at least partially by physiologically abnormally high concentrations of molybdate (Mendel et al., 1981; Falciani et al., 1994; Fleck et al., 2002).

[0058] Research on Mo-insertase enzymes has found that defects in Mo-insertases are restored or at least partially, by physiologically abnormally high amounts of Mo (Mendel et al., 1981; Falciani et al., 1994;Fleck et al., 2002). These findings support the conclusion that a genetic error in the Mo-insertase gene, resulting in low activity sulfite oxidases in humans, can be restored or at least partially, by ingesting abnormally high amounts of Mo. Consequently, there may be individuals that respond positively to the ingestion of abnormally high amounts of Mo.

[0059] When this man ingested Mo, this may have increased the rate of insertion of Mo into molybdopterin. Thus increasing the concentration of the metallo-organic coenzyme in cells and possibly increasing the activity of sulfite oxidases. This change may have somewhat overcome the bottleneck described earlier. It is unlikely that this phenomenon has occurred in this man in the past, this is because this man probably has not ingested such a high dose of Mo in his life. This is because, Blackmores Sustained Release Multi + Antioxidants, 20 2016101768 06 Oct 2016 vitamin and mineral supplement does not list Mo as an ingredient and many of the vitamin and mineral supplements that are currently on the market also do not list Mo. Therefore, he may have a problem with the insertion of Mo into molybdopterin.

[0060] The subject may have suffered from a sulfur deficiency. The tentative low activity of sulfite oxidases in this individual could result in a considerable amount of sulfur-containing metabolic waste not being recycled. If sulfur is lacking in his diet, this fault may cause a sulfur deficiency. The improvements during the Sodium Sulfate Supplementation ABAB Pilot Trial and the Sulfur-containing Amino Acid Supplementation ABAB Pilot Trial may be, because more sulfur is available for the synthesis of sulfur containing macromolecules.

[0061] Mo-insertase enzyme is coded by GPHN, in humans and GPH/V also codes for the structure protein gephyrin. Macromolecular assemblies of gephyrin proteins clusters glycine and GABA type A (GABAa) receptors in inhibitory post-synapses. This function is completely unrelated to the Mo insertion function (Mendel and Kruse, 2012).

[0062] Defective gephyrin proteins can result in poor clustering of neurotransmitter receptors. Poor clustering of glycine receptors causes a condition called hyperekplexia. Hyperekplexia is characterized by a pronounced startle reflex response to auditory or tactile stimuli (Lenz and Ptacek, 2004). Poor GABAa receptor clustering may be indirectly involved in the pathogenesis of depression and anxieties (Luscher et al., 2011). Depending on the mutation, gephyrin malfunction may be involved in age-associated disabilities (personal correspondence; Guenter Schwarz, Institute of Biochemistry, Department of Chemistry and Center for Molecular Medicine, Cologne University, Zuelpicher St. 47, Koeln, Germany). Reiss et al. (2001) hypothesized that there could be a genetic disease in humans with what seems to be unrelated symptoms, that could be linked by a genetic error in GPHN that responds to Mo supplementation.

[0063] The subject of this report may suffer from mild hyperekplexia. Hyperekplexia is a medical condition characterized by a pronounced startle reflex response to auditory or tactile stimuli. Severe cases of this response results in an abnormal increase in muscle tone, a condition called hypertonia. Hyperekplexia can be a relatively mild neurological disorder (Lenz and Ptacek, 2004). This may be because, over time the CNS can adapt to this defect. The pronounced reflex response during the treatment phases of the Benzodiazepine ABAB Trial, supports the diagnoses that the subject sufferers from hyperekplexia. This possible finding indicates that this man should continue to take the diazepam and not take stimulates, i.e. caffeine and dexamphetamine drugs. 21 2016101768 06 Oct 2016 [0064] Poor clustering of GABAa receptors is particularly relevant to this man. This is because, low activity of sulfites oxidases and other bowel abnormalities seen in this man, poorly explains his depression and anxieties. Individuals suffering with depression, which can be successfully treated with antidepressants, have been shown to have alterations in the numbers of GABAa receptors (Luscher et al., 2011). Poor clustering of GABAa receptors may have contributed to the depression and the anxieties seen in this man and in other IBS sufferers.

[0065] A GPHNgenetic error may be the underlining cause of IBS in this subject. This hypothesis was made because: (1) it explains the sulfur-handling disorder in this man; (2) he has a pronounced startle reflex response to stimuli; and (3) an IBS family cluster exists in his family. This family cluster suggests a genetic error. Therefore, poor glycine and GABAa receptor clustering in post-synapses could be causing the neuropathy in this man and may yet be another component in IBS. Therefore, benzodiazepines may be useful in treating IBS sufferers with a pronounced startle reflex response to auditory or tactile stimuli.

[0066] Most yeast and some bacteria that inhabit the bowels of humans in a symbiotic relationship, rely on or somewhat rely on its host to detoxify sulfite waste. Most yeasts do not have sulfite oxidase enzymes and some species from the Bacteroides and Clostridium genera, have a low occurrence or the absence of sulfite dehydrogenase enzymes (Zhang and Gladyshev, 2008; Bojanova and Bordenstein, 2016). Sulfite dehydrogenase is the equivalent enzyme in prokaryotes to the eukaryotic sulfite oxidase enzyme (Kappler, 2011). Sulfites have antimicrobial properties and the cells lining the gut must detoxify sulfites produce by these microbes, for them to grow. Therefore, a certain microbiome may result in the cells lining the bowels of humans being exposed to a relatively high concentration of sulfites.

[0067] The subject may have an imbalance of intestinal symbiotic bacteria in his bowels. The imbalance may be between bacteria with low and high activity of sulfite dehydrogenase enzymes. It is unlikely that yeast have had an impact on this imbalance. This is because, it is estimated that yeast make up only 0.03% of the microbes in feces, in healthy humans (Bojanova and Bordenstein, 2016) and, therefore, probably do not contribute to the imbalance. This may have occurred during the acute GIT infection which resulted in the development of IBS. This change to the intestinal microbiome may have led to an accumulation of sulfites in the lumen of his bowels. This accumulation may have been the component that “tipped the scales” of a system already under stress, because of a genetic 22 2016101768 06 Oct 2016 error in GPHN. For this article, intestinal symbiotic bacteria in which the activity of sulfite dehydrogenase enzymes is low will be referred to as, "SDH-lacking bacteria". The term "SDH-containing bacteria" will refer to intestinal symbiotic bacteria in which the activity of sulfite dehydrogenase enzymes is high.

[0068] A species or species from one or more of the following genera: Bacteroides, Propionibacterium, Streptococcus, Clostridium, Bacillus and Staphylococcus·, may be responsible for the foul gases. SRB probably play only a minor role in the bowel symptoms observed in this individual. This is because, the Sodium Sulfate Supplementation ABAB Pilot Trial poorly reproduced the bowel symptoms, observed a few days after Mo supplementation. SRB can also utilize sulfites almost as well as sulfates (Cypionka, 1987). Given that there may be an accumulation of sulfites in his bowels, there should not have been a marked increase in H2S gas production when Na2S04 was ingested. This conclusion is somewhat supported by the trial.

[0069] The hypothetical low activity of sulfite oxidase enzymes should have resulted in the health of the subject deteriorating during the B phases of the Sulfur-containing Amino Acid Supplementation ABAB Pilot Trial. This is because, highly active sulfite oxidases are needed to breakdown the sulfur-containing amino acids that are not needed. However, the overall health of the subject seemed to improve during the B phases. The foul odor of his flatus and stools when he consumed the soy protein isolate, suggests the reduction of sulfur-containing amino acids to sulfides. This tentative finding suggests that sulfites is being detoxified in the bowels of this man.

[0070] H2S gas is as toxic as cyanide and CH3SH gas is "highly toxic", however, there are very efficient biochemical pathways in the human colonic epithelia to detoxify these gases. The pathways have not been completely elucidated, however, tentative pathways have been proposed. CH3SH is oxidized to H2S and formaldehyde (CH20) by an unknown process in the colonic epithelia (Levitt et al., 1999; Fume et al., 2001). CH20 is also highly toxic and is oxidized to formic acid (CH202) by formaldehyde dehydrogenase in the colonic epithelium. CH202 is non-enzymatically oxidized to tetrathionate (S4062'; Hedberg et al., 2000; Boeniger, 1987). H2S is probably oxidized to S032' by colonic sulfide quinone oxidoreductase and sulfur dioxygenase (Blachier et al., 2010). Colonic sulfur transferase probably reduces S032' to thiosulfate (S2032 ). Colonic sulfite oxidase probably oxidizes S032' to S042" (Fig. 1). The somewhat toxic thiosulfate is either non-enzymatically oxidized to S4062'or enter the colon venous blood and is oxidized to sulfate in the liver (Koj et al., 1967). For this report, this 23 2016101768 06 Oct 2016 biochemical pathway (in the colonic epithelia) will be called the "colonic sulfide detoxification pathway".

[0071] When the subject ingested abnormally high amounts of protein and Mo; these substances may have had a prebiotic effect on the bacteria in his ileum and colon. The substances may have stimulated the growth of SDH-containing bacteria. This growth may have suppressed the growth of SDH-lacking bacteria. The overall effect of this, may have decrease the hypothetical accumulation of sulfites in his intestinal lumen.

[0072] The SDH-containing bacteria may be fermenting the unabsorbed protein and its genes involved in Mo metabolism may have been upregulated. These bacteria may be secreting proteases, digesting the unabsorbed proteins into amino acids. L-cysteine and L-methionine amino acids may then be fermented by these bacteria. The proteases secreted by these bacteria may also be digesting the Mo glycinate chelate in his bowels (Fig. 2) The Mo in his large bowels may be activating prokaryotic operons with Mo-containing enzyme genes and Mo-membrane transporter genes. As stated earlier in this article, Mo can up-regulated operons containing Mo-membrane transporter genes (McNicholas et al., 1997).

[0073] As stated earlier, the fermentation of L-cysteine and L- methionine produce H2S and CH3SH gases, respectively. H2S and CH3SH gases in the lumen and the tissues of his ileum and colon may have induced the colonic sulfide detoxification pathway. Since a sulfite anion is an intermittent compound in this pathway, this pathway may also detoxify the hypothetical accumulation of sulfites in his intestinal lumen (Fig. 2).

4 MATERIALS AND METHODS

4.1 STOOL SLURRY SULFITE ANION ASSAY

[0074] The concentration of sulfite anions in stool slurries made from the subject, were measured using sulfite test strips (Precision Laboratories, 415 S Airpark Road, Cottonwood, Arizona, U. S. A). Measurements were taken within 30 min after the subject passed the fecal specimen. About a glass marble size sample was taken from each fecal specimen and added to about 20 mL sodium hydroxide (NaOH) solution. The sample was mixed until it was suspended in the solution, i.e. it formed a slurry. NaOH salt was then added to the suspension to increase the pH to about 12. The concentration of sulfite anions was measured as per the manufactures protocol. The only exception was that the test strip was 24 2016101768 06 Oct 2016 rinsed, by dipping it into a second NaOH solution at a pH of about 12. The negative control was a NaOH solution at about a pH of 12.

4.2 SUPPLEMENTATION ABAB PILOT TRIALS

[0075] The "baseline" (Phase A) and the treatment (Phase B) phases of each trial had a period of 3 d. The dose of the oral Na2S04 anhydrous supplement ("Blackmores Professional Celloids S.S. 69", Blackmores Ltd., 20 Jubilee Avenue, Warriewood, New South Wales, Australia) and the soy protein isolate supplement during the B Phases: were 16.5 mmol/d, in three 5.5 mmol doses and about 105 g/d in at least three doses per day, respectively. The soy protein isolate supplement increased the total amount of protein in his diet from about 70 to about 175 g/d. The subject continued to take Mo and consume a low sulfite diet throughout these trials. The beneficial effects and the symptoms were assessed and recorded by the subject, on the third day of each phase. The criteria assessed were his: flatus and stool frequencies; flatus and stool odor; and stool form.

[0076] Possible ratings for each criterion (within quotation marks) were: • Flatus frequency: “low”, frequency < 10/d; “normal’, frequency ^ 10/d and < 20/d; “high” frequency ^ 20/d and < 50/d; and “very high” frequency ^ 50/d. Flatus frequency was measured for 2 h and extrapolated to 24 h. • Flatus and stool odor: “nil”; “sweet”, unpleasant sweet odor; “vegetable”, decomposing vegetable odor; and “foul”, rotten eggs odor. • Stool frequency was the number of bowel movements on the third day of each phase. • Stool form was graded using the Bristol Stool Form Scale.

4.3 BENZODIAZEPINE PILOT TRIAL

[0077] The baseline (Phase A) phases of the trial were 1 wk. The treatment (Phase B) phases were until a pronounce startled reflex response were felt by the subject or 1 wk. The dose of diazepam during the A phases was 5 mg/d. Pronounced startled reflex responses were recorded by the subject. The subject recorded a pronounce startled reflex response, if he felt a mild electric shock in his skeletal muscles, in response to an event that startled him. The subject continued to take Mo, cyanocobalamin, Na2S04, soy protein isolate and consume a low sulfite diet throughout this pilot trial. 25 2016101768 06 Oct 2016

SUMMARY OF INVENTION

[0078] The invention is a new medical treatment paradigm for IBS. The treatment paradigm consists of: dietary changes, dietary supplementations, drugs and other routines. The treatment aims to address what is thought be three important components, in the pathophysiology of IBS or an IBS subgroup. The components are: (1) a sulfur handling disorder; (2) a pronounced startle reflex response to auditory or tactile stimuli; and (3) gut microbes placing stress on the defective biological system that leads to development of the sulfur-handling disorder or produces the disorder directly. The key agents for the pronounced startle reflex response are probably: (1) a benzodiazepine and (2) a relaxation routine. The key agents for IBS are: (1) a low sulfite diet, (2) cobalamin, (3) Mo and (4) a protein isolate.

INDUSTRIAL APPLICABILITY

[0079] The IBS symptoms described in this example are seen in other IBS sufferers and it is expected that at least a subgroup of sufferers will respond positively to this medical treatment paradigm. The tentative: (1) mild hyperekplexia, (2) low activity of sulfite oxidase enzymes, (3) sulfur deficiency and (4) overgrowth of gut microbes; probably can be well managed by this treatment, or an improved version of the treatment. Therefore, sufferers should be able to live a relatively normal life. In addition to this, this treatment may also help with treating depression, anxieties and phobias.

[0080] The medical treatment paradigm used in this article would be relatively easily applied in clinical settings. The supplements, diet modifications and drugs are relatively easy to obtain, are inexpensive and are generally regarded as safe (GRAS). The foul gases are unlikely to cause bowel disease and the subject of this case report describes the odor as an “inconvenience”. The data generated in this case report, could be used as a diagnostic test for patients to those described in this article. For example, the oral administration of Mo could be used to identify sufferers. Finally, Mo has been used to successfully treat functional dyspepsia (personal correspondence; Gary Deed, 314 Old Cleveland Rd., Coorparoo, Brisbane, Australia). Therefore, this treatment may be an effective treatment for other FSSs (Wessely et al., 1999; Warren and Clauw, 2012) and even inflammation of the bowels.

[0081] There is considerable scope for improvements to this medical treatment paradigm. These improvements may eliminate the current symptoms caused by the current treatment, i.e. the foul gases. Thus making the treatment more appealing to patients. Drugs that oxidize 26 2016101768 06 Oct 2016 sulfites may reduce the need to administer agent that are likely to produce foul gases in patients. Some candidate drugs are: sodium cromoglycate, doxepin and antileukotriene (Taylor et al., 2014). An antifungal (e.g. nystatin) may also help, if the patient is suspected of suffering from an overgrowth of yeast in the bowels of the patient. Mo metabolism is related to Cu and sulfate metabolism (Vyskocil and Viau, 1999) and a better understanding of the interactions between these two elements and sulfate in humans, may improve the effectiveness of the treatment. A bacterial probiotic that can overgrow the gut microbes that may be placing stress on the body system involved in the sulfur-handling disorder, or producing it directly, may provide substantial relieve from symptoms. Finally, given that an underlining cause for IBS may have been identified, i.e. a genetic error in GPHN leading to biochemical abnormalities and neuropathies, substances targeted to rectify the defect may eliminate symptoms.

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Brian James Grech, BAppSc(MedSc) BAppSc(Hons) PhD 4 Corrie Ct., Capalaba, Brisbane, Australia Email: bjgrech@ozonline.com.au Tel: 61 (0)404 857 631 6 October 2016

Claims (4)

1. The use of diazepam, 1000 to 3000 pg/d doses of a molybdenum(ll) glycinate chelate supplement, a low sulfite diet and a soy protein isolate supplement; to successfully treat a pronounced startle reflex response to auditory or tactile stimuli in IBS-D; or a pronounced startle reflex response to auditory or tactile stimuli in an embodiment of symptoms similar to those observed in IBS-D.

2. The use of 1000 to 3000 pg/d doses of a molybdenum(ll) glycinate chelate supplement, a low sulfite diet a soy protein isolate supplement; to successfully treat the diarrhea-predominant subtype of irritable bowel syndrome (IBS-D); or an embodiment of symptoms similar to those observed in IBS-D.

3. The use of 6000 to 15000 pg/d oral doses of a cyanocobalamin supplement, 1000 to 3000 pg/d doses of a molybdenum(ll) glycinate chelate supplement, a low sulfite diet, a soy protein isolate supplement; to successfully treat IBS-D; or an embodiment of symptoms similar to those observed in IBS-D.

4. The use of a sodium sulfate supplement, 1000 to 3000 pg/d doses of a molybdenum(ll) glycinate chelate supplement, a low sulfite diet, a soy protein isolate supplement; to successfully treat IBS-D; or an embodiment of symptoms similar to those observed in IBS-D.